Citation
Volatile flavor components of celery stalks (Apium graveolens VAR. dulce) as related to temperature and time in storage - with further investigations on component distribution within the stalk

Material Information

Title:
Volatile flavor components of celery stalks (Apium graveolens VAR. dulce) as related to temperature and time in storage - with further investigations on component distribution within the stalk
Creator:
Ezell, Danny Odell, 1941-
Publication Date:
Language:
English
Physical Description:
95 leaves : ill. ; 28 cm.

Subjects

Subjects / Keywords:
Boiling ( jstor )
Celery ( jstor )
Flavors ( jstor )
Hydrocarbons ( jstor )
Juices ( jstor )
Odors ( jstor )
Petioles ( jstor )
Sensation ( jstor )
Simulations ( jstor )
Solvents ( jstor )
Celery ( lcsh )
Dissertations, Academic -- Vegetable Crops -- UF
Vegetable Crops thesis Ph. D
Genre:
bibliography ( marcgt )
non-fiction ( marcgt )

Notes

Thesis:
Thesis--University of Florida, 1968.
Bibliography:
Includes bibliographical references (leaves 91-94).
General Note:
Typescript.
General Note:
Vita.
Statement of Responsibility:
by Danny Odell Ezell.

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University of Florida
Holding Location:
University of Florida
Rights Management:
Copyright [name of dissertation author]. Permission granted to the University of Florida to digitize, archive and distribute this item for non-profit research and educational purposes. Any reuse of this item in excess of fair use or other copyright exemptions requires permission of the copyright holder.
Resource Identifier:
029982704 ( ALEPH )
37526033 ( OCLC )
ACG1417 ( NOTIS )
AA00004940_00001 ( sobekcm )

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Full Text


VOLATILE FLAVOR COMPONENTS OF
CELERY STALKS (Apium graveolens VAR.
dulce) AS RELATED TO
TEMPERATURE AND TIME IN STORAGE-
WITH FURTHER INVESTIGATIONS ON
COMPONENT DISTRIBUTION WITHIN THE
STALK
By
DANNY ODELL EZELL
A DISSERTATION PRESENTED TO THE GRADUATE COUNCIL OF
THE UNIVERSITY OF FLORIDA
IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE
DEGREE OF DOCTOR OF PHILOSOPHY
UNIVERSITY OF FLORIDA
1968


ACKNOWLEDGEMENTS
The author wishes to express sincere appreciation to
the Chairman of the Supervisory Committee, Dr. D. D. Gull,
for his guiding assistance in these experiments. Appreci
ation is expressed to Dr. R. C. Smith, Dr. B. D. Thompson,
and Mr. R. K. Showalter, who have contributed to his academic
training, direction of the research, and preparation of this
dissertation.
Special recognition is given to his wife, Wanda, who
has given understanding, encouragement, and long hours as a
dedicated typist to make this graduate study possible.
ii


TABLE OP CONTENTS
Page
ACKNOWLEDGEMENTS li
LIST OP TABLES v
LIST OP FIGURES vii
INTRODUCTION 1
REVIEW OP LITERATURE 3
MATERIALS AND METHODS 9
Storage 9
Phase 1 9
Phase II 10
Phase III 10
Component Distribution 10
Analytical Procedures 10
Organoleptic 10
Extraction 12
Chromatographic 13
RESULTS AND DISCUSSION 16
Organoleptic Evaluation 25
Storage 26
Phase I 26
Phase II 35
Phase III 46
iii


Page
Component Distribution 8l
SUMMARY AND CONCLUSIONS 88
BIBLIOGRAPHY 91
\
iv


LIST OP TABLES
Table Page
1 Storage treatments used in Phase III 11
2 Oven temperature and time relationships
for the matrix sequence used in all
chromatographic measurements 15
3 Retention times on Apiezon L column and
average peak heights of peaks with re
tention time greater than 90 minutes 20
4 Odors detected at the exhaust port of an
Apiezon L column after injection of
1 ul of celery solvent extract residue. ... 21
5 The effect of low temperature (38P)
storage on the ratio of high/low
boiling fraction in celery flavor
extracts 27
6 The effect of storage at 38P for 2 or 1|
weeks upon the relative per cent of 9
major chromatogram components 28
7 The effect of storage at 70F for 5 days
upon the relative per cent of 9 major
chromatogram components 3&
8 The effect of storage under market sim
ulated conditions upon the relative
per cent of 9 major chromatogram
components 4.4.
9 Observod retention times of volatile
components of celery head-space
samples, using 2 column stationary
phases 50
10Relative per cent of total peak area
for the 9 peaks measured in head-
space analyses. $1
v


Table
Page
11 Mean peak area (fresh weight basis)
for each of the 9 peaks analyzed
in head-space measurements as re
lated to storage temperature and
duration 52
12 Per cent dry weight of celery as
related to storage temperature
and duration 65
13 Mean peak area (dry weight basis)
for each of the 9 peaks analyzed
in head-space measurements as re
lated to storage temperature and
duration 66
llj. Ratios of high/low boiling fractions
from chromatograms of extracts pre
pared from various parts of the
celery plant 8l
vi


LIST OP FIGURES
Page
Figure
1 Chromatograms prepared from (A) dry
ice-acetone and (B) liquid nitrogen
extracts 17
2 Chromatogram prepared from aqueous
extract as described in extraction
procedures 18
3 Infrared spectrum of compound
believed to be n-butyl phthalide 23
4 Chromatograms from celery extracts
prepared (A) after 10 weeks' storage
at 38F and (B) at harvest 30
5 Mean ratio of high/low boiling com
ponents as affected by storage for
5 days at 70F (difference signif
icant at 0.05 level) 32
6 Mean peak area of chromatograms pre
pared from celery extracts taken at
harvest and after 5 days' storage at
70F (difference significant at 0.01
level) 33
7 Chromatograms from celery prepared (A)
after 5 days'storage at 70F and (B)
at harvest 34
8 Mean ratio of high/low boiling com
ponents of the chromatograms from
the control and market simulation
treatment (difference significant
at 0.05 level) 38
9 Mean peak area of low and high boiling
fractions of the chromatograms of
the control and market simulation
treatment 39
vii


Page
Pigura
10 Mean peak area of d-limonene and
n-butyl phthalide for the control
and market simulation treatment
("""'change in peak area significant
at 0.01 level) l0
11 Mean peak area of all peaks in the
lou and high boiling range ex
cluding limonene and butyl
phthalide, respectively 42
12 Chromatograms of extracts prepared
from (A) celery stored under
market simulated conditions and
(B) freshly harvested celery 45
13 Chromatogram from head-space mea
surements of celery volatiles with
peak numbers according to reten
tion time (Apiezon L) 47
14 Chromatogram obtained when celery
head-space volatiles were chro
matographed on Carbowax 20 M 49
15 Trends in the change of peak area
(fresh weight basis) as a rgsult
of duration in storage at 38 F 57
16 Peak area (fresh weight basis) of
celery samples after 2 and 4 weeks *
storage at 38? and subsequent
storage after each for 1 day at
70F and 8 days at 50? (including
all peaks) 59
17 Peak area (fresh weight basis) of
celery samples at harvest, after
storage at 38? for 2 and 4 weeks,
and after storage at 45P1 for 2 and
4 weeks for each of the 9 peaks
measured in head-space analyses 62
18 General trends in total peak area
(fresh weight basis) changes
according to temperature and dura
tion of storage at 45F> 38F, and
subsequent storage at 50P and 70F
after storage at 3$? 63
viii


Figure Page
19 Trends in the change of peak area
(dry weight basis) as a result of
duration in storage at 38F 70
20 Peak area (dry weight basis) of
celery samples after 2 and 4
weeks' storage at 38F and sub
sequent storage after each for
1 day at ?0F and 8 days at 50F
(including all peaks) 73
21 Peak area (dry weight basis) of
celery samples at harvest, after
storage at 38F for 2 and 4 weeks,
and after storage at 45? for 2
and 4 weeks for each of the 9 peaks
measured in head-space analyses 75
22 General trends in total peak area
(dry weight basis) according to
temperature and duration of storage
at 45P, 38P> and subsequent storage
at 50F and 70F after storage at
38F 77
23 Retention time of 5> 8, and 7 carbon
straight chain alcohols for polar
and non-polar chromatographic phases. ... 83
24 Chromatogram prepared from an extract
of the top (leafy) portion of the
celery plant 84
25 Chromatograms prepared from celery
extracts of (A) the inner and (B)
outer portion of the stalk 85
ix


INTRODUCTION
With the increasing sophistication in analytical
techniques and instrumentation has come a greater knowledge
on the volatile constituents of foods. Although the flavor
of celery (Anlum graveolens var. dulce) is quite distinctive,
it has received relatively little attention as compared to
other fruit and vegetable flavors.
Vegetables are in many cases quite difficult to
examine chemically. Flavor substances frequently occur in
concentrations of a few parts per million, while accompanied
by other organic materials and large amounts of water.
Several extraction (38) and analytical (39) techniques have
allowed increased resolution of these flavor substances and
a decrease in interference from foreign materials.
Celery is a perishable agricultural commodity; how
ever, with proper handling it can be maintained in a market
able condition for moderate periods of time. The long
storage durations involved in shipping Florida celery to
Europe have imposed more stringent demands on the keeping
quality of fresh celery. Quality reduction of celery due to
storage duration is usually manifest by water loss, pithiness,
loss of green color, decrease in sucrose and increasing
toughness. While flavor changes do exist as a result of
1


2
storage, it has not been determined if these flavor changes
are caused by changes in the volatile flavor constituents
or if these changes result in a detriment to quality.
This research was initiated to: i. select a proce
dure for extraction and measurement of the volatile flavor
components in celery stalks; ii. observe whether differences
occur in the chromatograms of celery extracts as a result of
storage temperature and duration; iii. observe differences
in the chromatograms of head-space volatiles (vapor pressures
sufficient to permit analysis in the vapor phase without ex
traction) obtained from celery as a result of storage temper
ature and duration, iv. investigate the flavor profile of the
various portions of the stalk of celery.


REVIEW OP LITERATURE
The maintenance of quality of fresh celery in storage
involves several aspects of applied physiology. Corbett (8)
indicated that several physical and chemical changes were
apparent during the storage of celery. Chemical analyses
of different parts of the plant showed marked changes in
reducing and total sugars and also both soluble and insoluble
nitrogen. Reducing sugars decreased in the leaves from har
vest to the end of storage. Young (45) and White-Stephens
(43) also concluded there was an increase in dry weight in
the crown portion of the celery plant with a corresponding
decrease in the outer leaf portion, and the outer leaves of
the pascal celery plant were of definite value in growth of
the heart petioles during storage. White-Stephens also
observed a marked increase in sucrose and polysaccharides in
the inner petioles coincident with a similar decrease in the
outer petioles. Corbett (8) concluded that soluble nitrogen
increased in both leaves and stalks from harvest to near the
end of the storage period,at which time there was a very
marked decrease in the leaves. This decrease of soluble
nitrogen in the leaves resulted in an increase in the inner
petioles.
Hall, et_ al. (24) cautioned against making quality
3


4
comparisons between different varieties of celery at differ
ent maturities. Single varieties of celery seemed to follow
the same pattern of change in chlorophyll content, alcohol
insoluble solids, total sugars, crude fiber, and dry weight
with regard to increasing maturity. Three varieties followed
the same general trend, but varied in the time the lowest and
highest points were reached.
Organoleptic (taste) comparisons have been made be
tween celery (Utah 52-70 variety) grown in Florida and the
same type celery produced in California. When considering
celery from an April harvest, the taste panel indicated the
Florida celery had a superior flavor, but was tougher and
had more fiber. However, when comparisons were made with
celery from a May harvest, the rankings indicated that the
California celery had a more desirable flavor and was less
bitter, but was tougher and more fibrous than the Florida-
grown celery. In these experiments it was noted that a low
potassium-sodium ratio was associated with a bland flavor.
The celery from the two areas was grown by commerical pro
ducers, was the same age, and was handled under similar
conditions.
The price of Florida celery has frequently been below
that of celery produced in other areas. While these opinions
were not unanimous, the reasons given for the reduced price
included poor appearance, bitter flavor and toughness (22).
While making observations on the flavor of celery,
Kali (23) described a bitter fraction and a burning-numbing


5
sensation from celery. The bulk of the bitter flavor was
associated with the dark green outer layer of the petiole
and did not appear to be associated with the burning-numbing
sensation. Description of the various flavors were: salty
flavor, radish-like flavor, and a hydrocarbon-type flavor
described as kerosene-or turpentine-like. The panel occa
sionally noted a sweet flavor. Kali also found a considerable
difference in the composition and organoleptic ratings of
outer, inner, and heart petioles of celery (21). The com
position of these petioles was influenced by the temperature
at which the celery was stored. In addition to the dif
ferences found between petioles according to position, there
were differences in flavor and composition between middle
and upper portions of the outer, inner, and heart petioles.
Using chromatographic analyses, Gold and Wilson (II4.)
showed that not all flavor constituents were in the juice
of celery stalks. The chromatograms prepared from juice and
puree showed common peaks, although the chromatograms were
not identical. An organoleptic study was performed on the
juice of celery from the top (leafy) and basal portion of
the celery stalk. The taste panel was able to differentiate
between the juice from the top (leafy) and basal portions
of stalks at the 0.01 level of significance. However, Gold
and Wilson were not able to differentiate between the chro
matograms prepared from the two juices. There was no dif
ference between chromatograms prepared from fresh and
frozen juice samples.


6
Pan (36) characterized the burning-numbing taste of
celery using gas-liquid chromatography. This chemical had
no unsaturated bonds or nitro groups. He found the presence
of aldehydes, carbonyl, phenolic, hydroxy, and aromatic
groups but the fraction was not a lactone. In further re
search (37), Pan described a new technique in the isolation
of a bitter principle from celery. This principle was
cationic at pH greater than 7, soluble in polar solvents,
and fluorescent under ultraviolet light. The bitter
principle could not be steam distilled from celery and was
located in the dark green portion of the petiole.
The complex mixture which constitutes the flavor and
odor of celery received considerable attention as early as
l897 Ciamician and Silber (7) reported the following
terpenes contributing to the flavor of celery: limonene,
myrcene, and an isomer of apiol. Of the lactone fractions
identified, sedanolide and sedanonic anhydride were pro
posed as being of primary importance in the odor of celery
seed. Following the identification of sedanolide and
sedanonic anhydride by Ciamician and Silber, Berlingozzi and
Cione (i|) undertook a study of the chemistry and odor
characteristics of alkyl and alkylidene phthalides. Working
2 6 A
with A -dihydrophthalide, A-tetrahydrophthalides, and
hexahydrophthalides, they found when one of the 9^-carbon
hydrogens was replaced by an alkyl group, a celery odor
was noted. When both were replaced by alkyl groups, the
odor was less intense. Celery odor was most intense when


7
the 7^-carbon hydrogen was replaced by the alkylidene group.
Intensity increased as carbons increased from 1 to 4*
Barton and DeVaries (2) analyzed celery oil and re
ported the isolation of butyl phthalide instead of sedano-
lide when using Ciamicians method (7)* They assumed that
an unstable sedanolide might be changed to butyl phthalide,
and proposed the structure of an /-unsaturated lactone,
neocnidilide. Mitsuhashi and Muramatsu (33) further pro
posed that sedanolide is at least a mixture of neocnidilide
and butyl phthalide.
Recent investigations concerning the flavor and odor
of celery were conducted by Gold and Wilson (14> 15, l) and
Wilson (i|4) In 1963 (l), they listed 38 compounds which
were identified as volatile components of celery. While
most of the compounds listed probably make some contribution
to the composite flavor and aroma of celery, 6 are of pri
mary importance: 3_iso butylidene-and 3~iso validene-3a,
4-dihydrophthalide; 3~isobutylidene-and 3isovalidene-
phthalide; cis-3-hexen-l-yl pyruvate; and diacetyl. The
4 phthalide derivitives were found in a ratio of 6:3:1:1*
The difficulty of separating the elements of a
chemical mixture into their respective flavor potencies has
been pointed out (17 40)- Guadagni, e_t al_. (l8) found
an additive effect of various chemicals even when the
chemicals were present in sub-threshold concentrations.
This difficulty was further emphasized in attempts to mea
sure sensory responses for food products by direct injection


8
of aqueous vapors into a gas chromatograph (6, 29, 35)*
Newar (35) noted that the concentration of a given compound
in a vapor phase at a given temperature is affected by:
vapor pressure of the compound, type of media in which it is
distributed, degree of solubility in the media, concentration
of compound in liquid phase, and its miscibility with other
organic compounds. Therefore, a decrease in liquid con
centration does not necessarily mean a decrease in vapor
concentration.
While there has been much research concerned with
the quality of celery in storage and with the volatile flavor
components of celery, little research has been conducted to
investigate changes in these volatile components during
storage, and it is to this question that the following work
is addressed.


MATERIALS AND METHODS
Florimart and Utah 52-70-2-13 (commonly known as
Florida-2-13) cultivars of celery were obtained from commer
cial growers. Size Number 3 stalks (36/crate) were harvested
90 days after transplanting, packed, hydrocooled, and trans
ported immediately to the laboratory. Preliminary investi
gations were conducted to determine the most satisfactory
method for extraction of celery volatiles and measurement of
the various parameters.
Storage
Phase I
The initial study was established to determine the
effect of low and high temperature storage on the composition
of flavor extracts prepared from celery stalks. Low tempera
ture effects were determined by comparing extracts from
freshly harvested Florimart celery with those from celery
Oo
stored 2 or 4 weeks at 3 F. High temperature effects were
determined by comparing extracts from freshly harvested
o
celery with those from celery stored 5 days at 70 F. A
randomized block design was used with 3 harvest dates con
stituting blocks. A non-replicated study involved weekly
extracts of celery stored at 3SF for 10 weeks.
9


10
Phase II
Floriraart cultivar was also used in a storage treat
ment to simulate market conditions. Treatment involved
placing the stalks at 45F for 2 weeks and transferring
o
samples to 50 F for an additional week. The experiment was
a randomized block with 2 harvest dates as blocks and a
duplicate analysis of each sample.
Phase III
Florida 2-13 cultivar was used for aroma (head-space)
measurements. Treatments employed for these analyses are
listed in Table 1. Statistical design of the experiment was
a randomized block with 3 replications and triplicate
analyses of each sample; harvest dates constituted blocks.
Dry weights were determined according to conventional pro
cedures .
Component Distribution
Celery stalks of the Florimart cultivar were divided
vertically into top (leafy), middle, and bottom and, hori
zontally into outer and inner portions. Extracts were pre
pared of each sample to derive further information on the
distribution of the volatile components of the vapor profile
within the plant. Three replications were used with a
completely randomized design.
Analytical Procedures
Organoleptic
A triangular test was used for all organoleptic


11
Table 1.
Storage treatments
used in
Phase III.
Treatment
Storage
Temperature
Duration
Subsequent
Storage
0
(P)
At harvest
(Weeks)
- -
1
38
1
--
2
38
2
--
3
38
3
--
1+
38
4
--
5
38
2
1
day at 70
6
38
4
1
day at 70
7
38
2
8
days at 5>0
8
38
4
8
days at 50
9
45
2
--
10
45
4



12
measurements (38)- Organoleptic comparisons were made be
tween freshly harvested celery and celery stored 2 weeks at
}8F. All organoleptic tests were conducted with a con
sumer-type panel composed of staff personnel. Panel scores
were obtained in a taste panel room with subdued lighting.
Samples were taken by dicing the center one-third of each
inner petiole, excluding all heart petioles, from 20 stalks.
Petioles with great visual differences were excluded.
Extraction
Celery petioles were washed and stripped of leaves.
All petioles were ground in a Waring blendor and the juice
expressed by hand. Each extraction sample consisted of
ij.,000 ml of juice which was taken from approximately 12 lb.
of petioles. The juice was introduced in 4-00 rol portions
into the evaporation chamber of a Nester-Faust model 500
rotary spray evaporator. The residue was removed from the
evaporation chamber before each new aliquot of juice was
introduced. The evaporation chamber was immersed in a water
bath at 70C and maintained at a pressure of 30 mm of mercury,
while the condensor and collection chamber were maintained
at 0C and 30 mm of mercury. Preliminary procedures differed
in that traps for collection of the flavor components were
at temperatures of dry ice-acetone and liquid nitrogen and
were placed after the cold water condensor (14, lb).
The clear aqueous condensate containing the volatile
flavor constituents was retained for solvent extraction.


13
Dichloromathane, amounting to approximately 10 per cent of
the volume of essence, was shaken with the essence of 2
minutes. The samples were washed 2 times and dried with
sodium sulfate according to conventional methods. Solvent
was partially removed in a rotary evaporator at 10ij.P with
a reduced pressure of 660 mm of mercury. The residue was
removed from the rotary evaporator when approximately 10 ml
remained in the flask. Further removal of the solvent was
accomplished at room temperature and atmospheric pressure.
Head-space samples were prepared by placing 500 grams
of diced celery into a 1,000 ml erlenmeyer flask. The
flasks were sealed and placed in a water bath at 55C for 2
hours. At the end of this period, the head-space gas was
taken by syringe for direct injection into the chromatograph.
Standards were prepared by placing known quantities of the
various chemicals on tissue paper and positioning in the
center of the flask's contents.
Chromatographic
All chromatograms were prepared on an Aerograph Model
600 D chromatograph equipped with a flame ionization detec
tor. Injection sample size and instrument parameters were
maintained constant for each experiment. The sample volumes
were 0.5 ul and 2.5 ml for solvent residue and head-space
samples, respectively. The injection port was maintained
at 235C. Helium was used as a carrier gas with an inlet
pressure of 1^0 psi and flow rate of 2 0 ml/min at room


14
temperature. A manual matrix programmed sequence was used
from 100C to 240C at 3/min as shown in Table 2. Hydrogen
flow was 20 ml/min while the air was maintained at 250 ml/rain.
Chart speed was ^ in/rain.
Treatment comparisons were made on 12 x l/8 copper
columns packed with 5 per cent W/W Apiezon L and Carbowax
20 M, both supported on 80/l00 mesh Chromasorb G.
Subtractive chromatography for alcohols and aldehydes
was performed by procedures described by Ikeda, e_t al_. (27)
and Allen (1).
Infrared analyses of certain compounds were performed
on a Perkin-Elmer Model 237 spectrophotometer equipped with
beam condensor. Spectrophotometer cells were used with .005 in
spacers and methylene chloride as a solvent.
Statistical analyses of paired samples were computed
by employing students' distribution (4D Treatment means
were compared by using Duncan's multiple range test (11)
following analysis of variance.


15
Table 2.
matrix
Oven temperature and time relationships for the
sequence used in all chromatographic measurements.
Time
Cumulative Interval
Temperature
Program power*"'
(Minui
;es)
(C)
0-10
10
100
50
10-25
15
120
50
25-35
10
11*0
50
35-50
15
160
50
50-58
8
180
60
58-73
15
200
60
73-77
4
220
60
77
--
240
60
Power indicator for programming rate of Aerograph Model
600 D oven.


RESULTS AND DISCUSSION
In 1966, preliminary extractions were made from
celery stalks of cultivar Florimart according to the pro
cedures previously discussed. Figure 1 shows chromatograms
obtained from the extracts of the liquid nitrogen and dry
ice-acetone traps. All peaks are measured at range 1, at
tenuation 8, unless otherwise designated. Little difference
was observed between the component distribution as an
effect of the two trapping procedures (liquid nitrogen vs.
dry ice-acetone). A larger quantity of extract was obtained
from the dry ice-acetone trap than from the liquid nitrogen
trap.
High boiling components are designated as those
which have retention times greater than $0 minutes: the
time at which the oven temperature is set at l80C. Low
boiling fractions are those components which emerge from
the column in less than 50 minutes. Figure 2 shows a chro
matogram prepared from an extract sample of the aqueous
condensate procedure described earlier. Reference to
Figure 1 shows that there is a proportionately smaller
quantity of high boiling components in the chromatograms
from the dry ice-acetone and liquid nitrogen traps than in
the chromatogram from the aqueous condensate shown in
16


32
Figure 1. Chromatograms prepared from (A) dry ice-acetone and (B) liquid nitrogen
extracts. ^
M


*12
rW
90
Figure 2. Chromatogram prepared from aqueous extract as described in extraction
procedures.
CD


19
Figure 2. Peaks with retention times of 72 minutes or more
made up more than l+O per cent of the total peak area of the
chromatograms prepared from these aqueous extracts, while
they made up less than 5 per cent of those from the liquid
nitrogen and dry ice-acetone traps. While both methods
yielded many low boiling compounds, trapping with liquid
nitrogen and dry ice-acetone gave a larger proportion.
The data obtained from the chromatograms prepared
from the aqueous collections revealed several additional
peaks which had retention times greater than 90 minutes, on
the Apiezon column, when the instrument was isothermally
controlled at 24GC subsequent to the sequence listed in
Table 2. The retention times and average peak heights of
these peaks are presented in Table 3* These peaks are not
considered in treatment comparisons.
In order to establish qualitative meaning of the chro
matogram in Figure 2, an odor characterization was estab
lished according to the odors noted at the exit port of the
column during the programmed sequence. These odors are
presented in Table 4 with their estimated strengths. Few
celery-like odors were observed in the low boiling range,
while a large proportion of the odors in the high boiling
range were characteristic of celery. It should be noted
that additional odors may have been present but were not
detected due to concentration or sensitivity effects.
Since many of the phthalide compounds which impart
the typical aroma of celery are sterioisoraers (33)> it is


20
Table 3.
average
greater
Retention times
peak heights of
than 90 minutes
on Apiezon L column and
peaks with retention time
Retention time
Average peak height
(Minutes)
(Millimeters)
105
11
108
52
120
31
130
42
11|2
42
147
17
156
455
170
63
214
42
272
42


21
Table 4* Odors detected at the exhaust port of an Apiezon L
column after injection of 1 ul of celery solvent extract
residue.
Time
Odor
Time
Odor
(Minute
s)
(Minutes)
11
Turpentine (f)
40
Fishy (f)
12
Woody (f)
41
Undescribed (f)
IS
Orange (m)
44
Diesel exaust (m)
20
Spinach-sx-ieet (m)
45
Celery (f)
22
Bitter weed (f)
55
Sweet (f)
24
Sour milk (f)
56
Orange peel (m)
25
Plastic glue (m)
58
Cotton (s)
26
Bananas (s)
59
Cotton (f)
27
Green bananas (m)
68
Apple (s)
28
Milk weed (f)
72
Celery (f)
29
Celery (f)
73
Celery (s)
30
Undescribed (f)
74
Apple (f)
31
Undescribed (f)
75
Celery (m)
32
Undescribed (f)
78
Cooked celery (m)
33
Undescribed (f)
79
Celery (m)
36
Undescribed (f)
80
Celery (m)
37
Mint (m)
81
Celery-carrot-like (m)
38
Terpinyl acetate (s)
82
Celery-like (s)
39
Undescribed (f)
85
Cooked celery (f)
Odor descriptive terms selected by author.
(f), (m), and (s) designate relative strength of odors as
faint, medium,and strong, respectively.


22
difficult to determine which compound is of most importance.
Host of these isomers readily convert to butyl phthalide, re
suiting in butyl phthalide contamination in any isolation.
A sample of n-butyl phthalide which strongly yielded the
characteristic aroma of celery was obtained.^ This compound
had a retention time of 77 minutes on Apiezon L and 69 min
utes, 30 seconds on Carbowax 20 M. In order to substanti
ate further the true structure of this compound, an infrared
spectrum was prepared. The trace of the infrared spectrum
(Figure 3) very closely resembles that presented by Mitsu-
hashi, et_ al_. (31) for Ligustilide, a sterioisomer of butyl
phthalide. However, the spectrum of Figure 3 shows absorp
tion at 3,030 cm ^ with 1,600 and 1,535 cm~^ indicating the
presence of all three double bonds around the benzene ring
(3)- These data suggest that the compound is n-butyl phtha
lide rather than Ligustilide or a mixture of neocnidilide
and butyl phthalide (33)*
On the basis of these data it is assumed that the
peak appearing at 77 minutes retention time from solvent
front of Apiezon L is n-butyl phthalide. Collection of the
actual peak would have been desirable; however, needed fcil
ities were not available.
When using a fractional collection apparatus, Gold
and Wilson (llj) found 25 compounds present in the dry ice
^United States Department of Agriculture Fruit and Vegetable
Laboratory, Winter Haven, Florida.


absorbance
23
Figure 3. Infrared spectrum of compound believed to be
n-butyl phthalide.


trap. Among these compounds they discovered no acids,
aldehydes or phenols. They also indicated only ¡4 compounds
were present in the liquid nitrogen condensate, and the
principal odor constituent of the dry ice trap was cis-3-
hexen-l-yl pyruvate, and that of the liquid nitrogen trap
was diacetyl (16). The presence of more compounds in these
extracts seems to indicate the fractionating system of Gold
and Wilson was more efficient in reducing the vapor tempera
ture than the system used in these experiments.
The importance of phthalides and their derivitives
as flavor contributors in celery have been pointed out by
several authors (2, 7> li|> 15 16, 19, 31). It therefore
seems essential that a representative celery extract contain
a portion of these high boiling phthalide compounds. On the
basis of previous investigations (16, 26), the low boiling
compounds shown in the previous chromatograms were believed
to be primarily C H-^ hydrocarbons and related compounds.
The data obtained from the aqueous essence sample are
in agreement with those of Gold and Wilson (16). By separa
ting the high boiling and relatively low boiling fractions,
they found the phthalides primarily responsible for the
celery odor located in the column bottom. They did, however,
indicate that the addition of this material to tomato juice
did not reproduce the tomato-celery juice blends unless
material from the dry ice or liquid nitrogen traps was
included. Since the solvent extract prepared from the
aqueous essence contains both fractions, it is believed


25
that the profile shown in Figure 2 most adequately repre
sents the flavor profile of celery.
Organoleptic Evaluation
Triangular taste test comparisons between freshly
harvested celery and celery stored 2 weeks at 38F indicated
there was a significant (0.05 level) difference between
treatments. Celery used for this first test was harvested
on April 2I4. (stored 2 weeks) and May 9 (fresh). However,
when comparison was made between celery harvested May 9
(stored 2 weeks) and May 21+ (fresh), no significant differ
ence was observed. In the first test 61 per cent of the
judges paired the samples correctly,while in the second test
only 39 per cent were able to pair them correctly. From
these data it is difficult to determine if differences did
exist as a result of storage or if the differences were
inherent within each harvest. Hall (21) found that the
flavor of celery did change while in storage at I|DF, and
that there was an interaction between storage temperature
and petiole position on the organoleptic measurement. When
comparing Florida-grown with California-grown celery,
variations were found between April and May harvests (22).
These differences were attributed to a change in sucrose
content and not to any large change in the volatile flavor
constituents. Hotijever, Hall (23) indicated that a taste
panel could easily be influenced by the presence of the
bitter substance which is associated with the outer petioles.


26
It is assumed that if differences do exist in the volatile
flavor constituents between celery stored 2 weeks at 33F
and freshly harvested celery, storage at higher temperatures
and for longer durations should serve to compound these
differences. Texture changes in celery stored at higher
temperatures make triangular taste tests quite difficult.
Storage
Phase I
Chromatograms used for computation of these data are
similar in appearance to the chromatogram shown in Figure 2
and were prepared using the aqueous trapping procedure.
There seemed to be little change in the chromatograms from
the celery stored 2 or 4 weeks at 38F when compared to their
respective controls.
The importance of the odor characteristics of the
high boiling phthalides has already been discussed. How
ever, any true flavor change will be a result of the inter
actions of all compounds present, whether in threshold or
sub-threshold concentrations (18). Changes in the ratio
of high/low boiling components will be used as an indicator
of change for all chromatograms. Changes in this ratio do
not necessarily indicate flavor changes since no research
has established a correlation.
The average ratio of high boiling/low boiling com
ponents for all chromatographed samples was 1.32. Ratios
for high boiling/low boiling fractions for the low temperature


27
storage treatments are presented in Table 5* The ratio for
the freshly harvested sample (1.33) was very near the aver
age for all treatments. While no significant change was
observed between freshly harvested samples and those stored
2 weeks at 38F, there was a slight increase in the mean
ratio after 4 weeks' storage.
Table 5* The effect of low temperature (38 p) storage on
the ratio of high/low boiling fractions in celery flavor
extracts.
Treatment
Harvest
2 weeks' 38P
4 weeks' 38P
(Fraction ratio-high/low)
1.33
1.30
1.65
While a large number of peaks were resolved for each
chromatogram, only a few contribute substantially to the total
peak area of these chromatograms. The 9 peaks which contrib
ute most (more than 1.0 per cent of total area) to the total
peak area are listed in Table 6. Also presented in this
table is the mean per cent each contributed to the chromato
gram total for the respective treatments. The only signif
icant change observed was an increase in the peak at 17
minutes (tentatively identified as d-limonene) when celery
was stored 2 weeks at 38P. This is difficult to explain
since the mean for this treatment is higher than the control
and the sample taken at 4 weeks'storage. It is noted that
there was a large increase in the mean per cent of total area


28
Table . The effect of storage
upon the relative per cent of
components.
at 38P for 2 or 4 weeks
9 major chromatogram
Retention
Time
Treatment
At harvest
2 weeks
4 weeks
(Minutes)
(Per cent)
17
20.3 a*
23.4b
20.2a
24
6.9
5.6
6.7
36
3-5
2.5
1.7
56
1.0
1.1
1.0
62
1.8
2.7
2.7
72
11.3
7.7
8.4
78
23.6
31.5
29.9
80
9.9
4.4
7.5
81
3.1
3-4
3.6
8O.4
(Total per cent)
82.3
81.7
Values with differing letters in horizontal rows are
significantly different at the 0.05 level according
to Duncan's multiple range test.


29
for the peak at ?8 minutes when the celery was stored 2 or
4 weeks as compared with the control. This increase was
reflected in an increase in the ratio of high/low boiling
components for the 4 weeks'storage treatment; however,
increases in the low boiling fraction negated this effect
at 2 weeks' storage. No significant difference was observed
between the total per cent represented by these 9 peaks.
Chromatograms were prepared from celery extracts at
weekly intervals during storage for 10 weeks at 38?* After
celery had been stored 8 weeks at 38o? it was of no market
value, and approximately 25 per cent of the petioles had to
be removed before the extracts were prepared.
Figure 4 shows a chromatogram prepared from celery
stored 10 weeks at 38F versus freshly harvested celery.
Peaks in areas of highest concentration (12 to 22 minutes
and 72 to 90 minutes) seemed to have been maintained while
peaks in areas of low concentration (24 to 70 minutes) have
been reduced or are missing from the chromatogram. Note
worthy is the fact that the low boiling components are pre
sent in abundance after storage for long periods of time and
regardless of the condition of the celery.
Observation of chromatograms prepared from freshly
harvested celery show no peaks of large area between 85 and
95 minutes* retention time. Also, no build-up in concentra
tion was experienced in this range when celery was stored 2
or 4 weeks at 38F. However, when celery extracts from
celery stalks stored 8 weeks or more at 38F were


Figure 1;. Chromatograms from celery extracts prepared (a) after 10 weeks' storage at
38F and (B) at harvest.
VjJ
o


31
chromatographed, 2 large (peak height greater than 300 ram)
peaks were present at 8? and 91 minutes' retention time.
Celery stored 5 days at 70P was quite dehydrated
and of poor market quality. The freshly harvested control
for this study had a high/low component ratio of 0.98, which
was lower than the average. This ratio increased to 2.i|5
when the celery was stored 5 days at 70P (Figure 5)*
This increase in component ratio was accompanied by
a corresponding highly significant decrease in total peak
area when compared to the freshly harvested control. The
mean peak area for the control chromatograms was 1+8,162.3
p
mm while the mean peak area from the chromatograms of the
70P storage treatment was only 23,296.5 mm^ (Figures 6 and
7). Such a large change in peak area would not normally be
expected because the extracts were prepared from a constant
volume of juice. Since the moisture content of the celery
decreased during storage, it is possible that under these
dehydrated conditions a larger proportion of the volatile
components remained in the pulp and were not extracted.
However, Matthews (30) found a reduction in total volatiles
of beans after storage for 5 days at 70P. A conversion of
the volatile flavor components to.non-volatile forms through
enzymatic action would result in such a change. However,
no data have been collected to substantiate this hypothesis.
The changes that occurred in the extracts prepared
from celery stored 5 days at 70P can be seen in Figure 7*
There was a decrease in the peak area of all peaks,


Ratio (High/Lovj)
32
\
Figure 5. Mean ratio of high/low boiling components
as affected by storage for 5 days at 70F (differ
ence significant at 0.05 level).


33
Figure 6. Mean peak area of chromatograms prepared
from celery extracts taken at harvest and after 5
days' storage at 70F (difference significant at
0.01 level).


Figure 7 Chromatograms from celery extracts prepared (A) after 5> days'storage at
70 F and (B) at harvest.
Vjo


particularly those in the low boiling range which appeared
before d-limonene (17 minutes). The high boiling components
were moderately stable and the peak area decreased only
slightly.
Distribution data of the 9 large peaks from the chro
matograms prepared at harvest and after 5 days' storage at
70F are presented in Table 7- There was a significant
decrease in the proportion of the total peak area repre
sented by the peak at 17 minutes. Noteworthy is the fact
that this change is opposite that observed when celery was
stored at 3$?. However, there was a corresponding increase
in the per cent represented by the large peak at 78 minutes
when the celery was stored 5 days at 70F. From these data
it seems likely that the components of high vapor pressure
were metabolized or lost through evaporation as the celery
was dehydrated. Table 7 also presents the per cent of the
total peak area represented by the sum of the peak areas of
the 9 major peaks. The sum of the areas increased when
celery was stored 5 days at 70F. Reference to Table 6
shows that there was a similar increase when celery was
stored 2 and 4 weeks at 38F. This would suggest the pos
sible elimination or decrease of many of the small peaks
not considered in these data.
Phase II
Celery used for the market simulation study had a
very high peak ratio of high/low boiling components at


Table 7- The effect of storage at 70P for 5 days upon the
relative per cent of 9 major chromatogram components.
Retention
Time
Treatment
At harvest
70P for 5 days
(Minutes)
(Per
cent)
17
19.5
9.0*
24
5-7
7-5
36
3-3
1.4
56
1.0
1.6
62
2.1
4-1
72
13.6
8.0
78
22.0
36.9
80
9.3
7.2
81
3-3
9.9
(Total
per cent)
79.8
85.6
''Difference significant at the 0.05 level.


37
harvest as compared to the average for all treatments. The
ratio was 2.38 for the freshly harvested celery and decreased
significantly (0.05 level) during storage to 1.09 (Figure
8). The peak areas for the treatment moans used in com
puting the high/low ratio are shown in Figure 9. The dif
ference between the areas of each treatment are 21, 39, and
19 per cent for low, high, and total fraction areas, respec
tively. While there was change in both the low and high
component areas, these data indicate a greater change in the
high fraction.
A comparison between the mean peak areas of two com
pounds, d-limonene and n-butyl phthalide (identification
based on retention times of known compounds on polar and
non-polar chromatographic columns), extracted from the con
trol and market simulated lots of celery,is presented in
Figure 10. There was a highly significant increase in the
peak area of d-limonene in the chromatograms of the market
simulation treatment when compared to those of the control.
Considerable variation was observed in the areas of the peak
representing butyl phthalide; however, there was a substan
tial decrease in the mean peak area after storage under
market simulated conditions. The changes observed for these
two components were in opposite directions.
Any comparison between limonene and butyl phthalide
is quite critical from an organoleptic point of view. The
importance of the high boiling phthalides has been stressed
by the odors noted in Table Ij. and by the research of other


Ratio-High/Lovj
38
Figure 8. Mean ratio of high/low boiling components
of the chromatograms from the control and market
simulation treatment (difference significant at
0.05 level).


Area (mm x 1,000)
39
Figure 9. Mean peak area of low and high boiling frac
tions of the chromatograms of the control and market
simulation treatment.


Figure 10. Mean peak area of d-limonene and n-butyl
phthalide for the control and market simulation
treatment (^change in peak area significant at
0.01 level).


4i
investigators (I4, 16). Terpanes, however, seen to be less
dominant in determining vegetable flavor than in many of the
fruit flavors. It is difficult to determine whether a
decrease in the high boiling fraction during storage would
result in a decrease in the actual potency of the celery
flavor. Any discussion of the decrease in proportion of
these elements should take into c'onsideration the additive
effects of the various sub-threshold concentrations which
are affecting the flavor profile (18). This additive effect
of the various components further complicates the discussion
of the actual organoleptic effect of a proportionate change
in high or low boiling components.
C-rouping other peaks of the chromatograms seems to
substantiate the change in limonene in the low boiling range.
When peaks in the low boiling range (excluding limonene) were
considered as a whole, little change was observed between the
control and the market simulated study (Figure 11). There
was a considerable decrease in the peak area total for those
peaks which occurred in the high boiling range (excluding
butyl phthalide) as a result of storage under market condi
tions. It is quite possible that an imbalance of these com
ponents at harvest stimulated a change to a more balanced
situation. It has not been determined if there could be a
conversion of high boiling components to low boiling com
ponents during storage, but enzymatic processes are active
in the formation of volatile flavor components (25).
In addition to a slight decrease in total peak area


Low boiling components
30
\ High boiling components
25
o
\
o
o
20 -
X
l1
N
X
\
C2
V
B
15
\
\
N
N

<
10
K
N
\
\
\
5 '
\
\
\
\
\
\
Control
Market simulation
Figure 11. Mean peak area of all peaks in the
low and high boiling range excluding limonene
and butyl phthalide, respectively.


43
as a result of storage under market conditions there was a
highly significant decrease in the proportionate amount of
this total represented by the 9 previously mentioned peaks
(Table 8). Also shown is a decrease in the proportionate
amount contributed by each of the 9 peaks except peaks at
1? minutes (d-limonene) and at 8l minutes. However, indi
vidual peak changes of substantial importance are those at
17 72, and 78 minutes. Others in combination might also
contribute substantially as noted above. These data also
indicate a net decrease in those high boiling flavor com
ponents most responsible for the aroma of celery.
Chromatograms prepared from the celery extracts of
the control and the market simulation treatments are con
tained in Figure 12. As was pointed out previously, there
was an increase in the low boiling fraction, particularly
limonene, and a proportionate decrease in the high boiling
fraction between 72 and 36 minutes. When celery was stored
at 45? for 2 weeks and 50? for an additional week, 2
large peaks appeared in the chromatograms at 87 and 91
minutes. The 2 peaks shown in the market simulated treatment
chromatogram at 87 and 91 minutes have peak areas of 1,305
2 2 p 2
mm and 2,004 mm as opposed to 72.3 mm and 176 mm for the
chromatogram prepared at harvest. These data are in agree
ment with those received from chromatograms of celery ex
tracts prepared from celery stored 8 to 10 weeks at 38o?
(Figure 4)*
Considerable variation occurred in peaks on these


44
Table 8. The effect of storage under market simulated condi
tions upon the relative per cent of 9 major chromatogram
components.
Retention
Time
Control
Market
Simulation
Change
(Minutes)
(Per cent)
Low boiling
17
16.6
28.7
+72.9*
24
3.6
3.6

36
2.8
2.4
- 7.1
High boiling
56
3-4
2.8
-17-6
62
5.8
4.8
-I7.2
72
11.9
9.6
-19.3
78
33-5
7-7
-77.0
80
9.8
8.9
- 9.1
81
1-5
2.0
+330
(Total per cent)
94-4
80.1**
'"'Significant at 0.05 level
'"'"'Significant at 0.01 level


Figuro 12. Chromatograms of extracts prepared from (A) celery stored under market
simulated conditions and (B) freshly harvested celery.
-p-
vn


46
chromatograms between 37 and 45 minutes'retention time.
Peaks in this area were always present in measurable con
centrations at range 1, attenuation 8; however, the relative
peak areas were not constant within replications.
While the celery used in Phase I and Phase II was of
the same chronological age, it is doubtful that the physio
logical maturity was the same. An indication of this vari
ation could be observed differences between the ratio of
high/low boiling components for the freshly harvested
o
samples used for the JO P storage treatment and for the
market simulation. Celery used for the market simulation
was harvested in July while that used for the J0F treatment
was harvested in May. If the physiological maturity was
not the same for all treatments, generalized comparisons
cannot be rendered between treatments (9, 12, 13, 24).
Phase III
Nine separate peaks were resolved from Florida 2-13
celery using the head-space analysis technique. Identifi
cation of 4 of these compounds was accomplished by compari
sons of their retention times on polar (Carbowax 20 M) and
non-polar (Apiezon L) columns and by spiking these chemicals
Into flasks containing the celery for head-space measurement.
Since all comparisons are made with chromatograms from the
Apiezon L columns, numbers have been assigned to the re
solved compounds on the basis of retention time on this
phase (Figure 13). The chromatographic profile obtained


47
J
8
6
Figure 13. Chromatogram from head-space measurements of
celery volatiles with peak numbers according to reten
tion time (Apiezon L).


48
when these samples were chromatographed on Carbowax 20 M is
shown in Figure 14* Comparison data were prepared on the
basis of an instrument sensitivity of range 0.1 and attenu
ation 16, while peaks 6 and 8 were measured at attenuation
32 and 64> respectively, for both columns.
Observed retention times of the 9 volatile constit
uents are presented in Table 9. These are measured from
injection and are somewhat different from those expected
when pure compounds are chromatographed. This deviation in
time is an effect of the large amount of water vapor present
in the holding flask (34)- However, the relative retention
times are not greatly affected by the presence of water.
The retention data presented in Table 9 are in agreement
with those of Hunter and Brogden (34)* These investigators
found c*-terpinene to be present in volatile extracts and
the location of this compound in their chromatograms sub
stantiates the idea that it is responsible for peak 9.
However, no retention data have been collected to support
this hypothesis.
The relative per cent of total peak area of the 9
measured peaks is presented in Table 10. Approximately 68
per cent of the total peak area of these chromatograms is
accounted for by the four C-, qH^ hydrocarbons identified,
while d-limonane and ^-pinene resulted in approximately 83
per cent of the total peak area.
The mean peak areas (fresh weight basis) of the 9
peaks are listed in Table 11 according to treatment. Total


49
o
Figure II4..
volatiles
Chromatogram obtained when celery head-space
were chromatographed on Carbowax 20 M.


Table 9. Observed retention times of volatile components of celery head-space
samples, using 2 column stationary phases.
Peak
Number
Compound
Apiezon
Unknown
L
Kn own
Carbowax
Unknown
20 M
Known
1
..
7:00
(Minutes)
1:10
_ ,
2

8:40
--
1:55
--
3
--
10:48
--
1:35

4
<^-Pinene
11:35
11:35
2:56
2:55
5
/^-Myrcene
12:40
12:38
6:00
6:00
6
y^-Pinene
13:50
13:50
4:26
4:26
7

16:05

2:20

8
d-Limonene
17:50
17:52
7:15
7:15
9
-
20:20
- -
9:23
- ~
vn
o


51
Table 10. Relative per cent of total peak area for
the 9 peaks measured in head-space analyses.
Peak number
Per cent of total area
1
0.30
2
2.30
3
0.18
4
1-46
5
2.84
6
11.43
7
3-90
8
72.23
9
5.30
/


52
Table 11. Mean peak area (fresh weight basis) for each of
the 9 peaks analyzed in head-space measurements as related
to storage temperature and duration.
Storage
Treatment
1
Peak number
2 3
4
p
(Area mm )
Control
16.5a*
277.4a
6.3a
122.2a
1 wk @ 38F
19.5b
310.4ab
12.3a
145.9ab
2 wk @ 38F
43-3e
407.5ab
20.4bc
263.5cd
+**1 day @ 70F
37.6c
296.5c
16.labe
177.3ab
+ 8 days @ 50F
52.4g
310.Oab
25.3c
227-lbcd
2 wk @ 45F
47- 6f
441.1b
27.8c
251.7cd
3 wk @ 38F
44* 6e
389.8ab
25.8c
228.0bcd
4 wk @ 38F
40.2d
258.7a
22.6bc
199.Oabcd
+ 1 day @ 70F
39.3cd
309.lab
24.3bc
l8l.4abc
+ 8 days @ 50F
58.3h
411.Oab
21.8bc
281.3d
4 wk @ 45f
47.4f
389.lab
25.7c
269.8d
"Those means in vertical columns not followed by
letter are significantly different at the 0.05
the same
level.
designates
at 38F.
subsequent storage treatment after storage


53
Table 11. Continued
Storage
Treatment
5
Peak number
6
7
(Area mm^)
Control
220.7a*
1,187.7a
304.9ab
1 wk § 38P
30.0ab
1,430.2ab
46l.2ab
2 wk @ 38P
497.0c
1,814.5c
875.9b
+""'1 day @ 70F
412.5bc
1,532.6abc
593.4ab
o
+ 8 days @ 50 f
464.9bc
1,803.Obc
642.5ab
2 wk @ i|5F
521.1c
2,141.2c
732.lab
3 wk @ 38P
500.5c
1,930.0c
660.8ab
4 wk @ 38P
406.7abc
l,5l4-2ab
255.8a
+ 1 day @ 70P
388.9abc
1,649Oabc
3o9.0ab
+ 8 days @ $0F
494.5c
2,137.5c
766.3ab
4 wk @ 45p
500.8c
1,973.8c
708.3ab
Those means in vertical columns not followed by the same
letter are significantly different at the 0.05 level.
designates subsequent storage treatment after storage
at 38P.


54
Table 11. Continued
Storage
Treatment
8
Peak number
9
Total(l-9)
. 2,
(Area ram )
Control
5,454-9a*
552.9a
8,144*la
1 wk @ 38F
7,002.3a
5716abc
10,260.8ab
2 wk @ 38F
11,650.8a
897.labcd
16,456.4^8
+**1 day @ 70F
11,362.2a
688.Oabc
15,117.lab
+ 8 days @ 50F
12,149.8a
804.labcd
16,481.5ab
2 wk @ 45F
14,232.3a
1,069.2d
19,475.4b
3 wk @ 38F
11,440.3a
826.6abcd
16,030.lab
4 wk @ 38F
11,986.7a
760.7abcd
I5,389.5ab
+ 1 day @ 70F
10,501.9a
666.labe
14,150.2ab
+ 8 days @ 50 F
12,118.4a
l,459.2e
17,348.0ab
4 wk @ 45p
14,064.7a
899.led
19,381.4b
"Those means in vertical columns not followed by the same
letter are significantly different at the 0.05 level.
+ designates subsequent storage treatment after storage
at 38bF.


55
area for these peaks as a result of treatment is also pre
sented in this table. While all values were not signifi
cantly different, the mean peak area for the freshly har
vested control was lower than any other treatment for all
peaks except peaks 2 and 7 The lowest peak area for these
peaks occurred when celery was stored weeks at 38F.
Statistical differences were observed between treatments for
all peaks except peak number 8 (limonene).
Without exception, the mean peak area of all peaks
progressively increased as a result of storage at 38P for
1 and 2 weeks as compared with the control (Figure 15).
This increase in peak area continued for some compounds
until the third week of storage; however, all peaks except
peak 8 showed a decrease in peak area after 3 weeks' storage
at 38F.
Figure lb shows a comparison of the volatile changes
o
(FWB) resulting from storage of celery for 1 day at 70 F
and 8 days at 50F after storage for 2 and i| weeks at 38 F.
Considering only the treatments subsequent to 2 weeks' storage
at 38F, without exception, all compounds from the 70F
treatment had a lower peak area than those from the 50F
treatment. There was only one exception to this trend (peak
o 0
3) when 70 F and 50 F treatments were prepared subsequent to
[|. weeks' storage at 38?* Also, storage of celery for 1 day
at 70F after 2 weeks at 38F resulted in a decrease in peak
area of all peaks as compared to the freshly harvested
sample. The results of the same treatment after I4. weeks at


Figure 15.
basis) as
Trends in the change of peak area (fr
a result of duration in storage at 38
sh weight
F.


WEEKS
Vn.
PEAK


Figure l6
storage
at 50dF
Peak area (fresh weight basis) of celery samples after 2 and
at 38? and subsequent storage after each for 1 day at 70F and
(including all peaks).
weeks*
8 days


WEEKS WEEKS WEEKS "" WEEKS
(X1000)
o
5
K
CD
(X 100)
(X100)
65


60
38P are moro variable, peaks 4> 5> 8, and 9 showing the
decrease after 1 day at 70P. In some treatments, storage
for 8 days at 50F resulted in an increase in peak area
when compared to the corresponding 3? treatment. These
effects were, however, more prevalent after 4 weeks at 38F
(all peaks except peak 2 resulted in the increase).
Peak areas for the control and storage treatments of
2 and 4 weeks at 38F are presented in Figure 17. There
was no significant difference between the two storage treat
ments at 45F. A comparison of the 45? treatments at 2
and 4 weeks reveals that the peak areas resulting from the
45F treatment tended to be higher than those at 38F. For
both 2 and 4 weeks'storage, the only exceptions were peaks
4, 6, and 7 at 2 weeks, no exceptions being noted at 4 weeks'
storage. Noteworthy is the fact that total peak area of all
9 compounds for the 45F treatments, both 2 and 4 weeks,
were the only total values significantly different from the
control.
The general trends for these treatments are summa
rized in Figure 18. The points in this graph are based on
the total peak area of all peaks. Storage at 38F for 1
and 2 weeks resulted in a linear increase in peak area,
while further storage at 38F caused a reduction in peak
o Oo
area. Storage at 70 F for 1 day after storage at 30 F
. o
resulted in a decrease in peak area, while storage at 50 F
for 8 days after 38 F holding resulted in an increase in
peak area. The degree of increase at 50F was greatly


Figuro 17* Peak area (fresh weight basis) of celery samples at harvest,
at 30F for 2 and 4 weeks, and after storage at 1|5P for 2 and 4 weeks
the 9 peaks measured in head-space analyses.
after storage
for each of


LEGEND
EZ3 -AT HARVEST
-38*F STORAGE
IOI -45F STORAGE
(X1000) (X10)
O
rv)


Area (rom x 1,000)
63
Figure 18. General trends in total peak area (fresh weight
basis) changes according to temperature and duration of
storage at 45? 38P, and subsequent storage at $0F
and 70F after storage at 38ip.


affected by the increase and large value of peak area for
peak 8. The greatest total peak areas were observed from
celery stored 2 or 4 weeks at 45F* Interpolation of the
change between harvest and 2 weeks' storage at 45F Is not
possible for these data.
After one experimental block had been analyzed, there
appeared to be differences associated with apparent moisture
loss. In attempts to define these effects, dry weight
measurements were prepared from celery in the second and
third block and these data are presented in Table 12. Pew
significant differences were observed as a result of storage
treatment. However, the per cent dry weight of celery
stored 2 and 4 weeks at 45? was significantly higher than
most other storage treatments. Also, the control was signif
icantly different from only 2 treatments: 2 weeks' storage
o
at 45 P and 3 weeks' storage at F*
Since these differences in dry weight were observed,
the peak areas were calculated on a dry weight basis (DWB)
and are presented in Table 13. When calculated on the basis
of dry weight, there seemed to be more variation in the peak
areas than when calculated on a fresh weight basis. The
peak area (DWB) for the control was not lowest; instead,
there was a general decrease in 4 the compounds to 1 week
storage at 38P (Figure 19). After this storage duration,
the peak areas of the various peaks increased as when the
data were calculated on a fresh weight basis. All peaks
increased in peak area from 1 week to 2 weeks' storage at


65
Table 12. Per cent dry weight of celery as
related to storage temperature and duration.
Storage
Treatment
Dry weight
Control
(Per cent)
4. llbcd'*
1 wk @ 38P
3 99abc
2 wk @ 38F
3-95abc
+**1 day @ 70P
4.19bcd
+ 8 days @ 50F
3.94abc
2 wk @ 45F
4.82e
3 wk @ 38F
3.65a
i; wk @ 38F
+ 1 day @ 70F
3.78abc
3.67ab
+ 8 days @ %0F
4.17bcd
4 wk @ 45F
4 60de
Those means in vertical
by the same letter are
ent at the 0.05 level.
"'r+ designates subsequent
storage at 38F.
columns not followed
significantly differ-
storage treatment after


66
Table 13. Mean peak area (dry weight basis) for each of
the 9 peaks analyzed in head-space measurements as re
lated to storage temperature and duration.
Storage
Treatment
1
Peak
2
number
3
4
Control
0.4a*
(Area
177ab
ram^)
0.3a
6. Oabc
1 wk @ 38F
0.5a
17.lab
0.4a
4* 9a
2 wk @ 38F
1.2ab
19Oab
0.8bc
9.2d
+**1 day @ 70F
0.9ab
l6.Oab
0.7ab
5.6ab
+ 8 days @ 50?
1.3ab
21.Oab
1.2c
8.7cd
2 wk @ 45F
l.Oab
21.2ab
1.2c
8.0bcd
3 wk @ 38F
l.Oab
24.1b
0.8bc
8.6cd
4 wk @ 38?
0.9ab
14.5a
0.7ab
8.Obcd
+ 1 day @ 70F
1.3ab
l8.9ab
l.Obc
8.4cd
+ 8 days @ 50F
1.6b
22.4ab
1. Obc
10.6d
4 wk @ 45F
0.8ab
l8.5ab
1. Obc
8.4cd
Those means in vertical columns not followed by the same
letter are significantly different at the 0.05 level.
+ designates subsequent storage treatment after storage
at 38F.


67
Table 13. Continued
Storage
Treatment
Peak number
5 6
7
2
(Area ram )
Control
9.6ab*
68.9ab
9.8ab
1 wk 38F
8.3a
60.6a
5-3a
2 wk @ 3oF
l4-9cd
72.2ab
10.Sab
+*wl day 70F
10.9abc
64.. 2ab
8.2ab
+ 8 days @ 50F
13.6bcd
74* 8ab
9.9ab
2 wk U5F
15-Ocd
82.Oab
12.0b
3 wk 38F
15.7d
92.5b
11.3ab
4 wk 38F
11.7abcd
67lab
8.2ab
+ 1 day 70F
13* 6bcd
72.7ab
9.7ab
+ 8 days 50F
15.led
91.6b
13.3b
4 wk i;50F
13-9cd
752ab
11.6ab
"""Those means in vertical columns not followed by the same
letter are significantly different at the 0.05 level.
+ designates subsequent storage treatment after storage
at 38F.


68
Table 13. Continued
Storage
Treatment
Peak
8
number
9
Total(1-9)
(Area
mm^)
Control
233.2ab:
30.4abc
376.4ab
1 wk @ 38F
l64 3a
23.4a
284.8a
2 wk @ 38F
232.4ab
37-5cd
397-9ab
+**1 day @ 70F
251.7ab
27.8ab
386.Oab
+ 8 days @ 50F
279.8ab
34.1bc
444.5b
2 wk @ U5F
318.5b
45.2d
505.2b
3 wk @ 38F
301.6b
36.Obcd
491.6b
4 wk @ 38F
293.7b
31.labe
431.6ab
+ 1 day @ 70F
286.2b
37-Obcd
450.6b
+ 8 days @ 50F
315.4b
35* 6bcd
506.7b
4 wk @ 45f
321.4b
37.6cd
488.6b
'"Those means in vertical columns not followed by the same
letter are significantly different at the 0.05 level.
+ designates subsequent storage treatment after storage
at 38bF.


Figure 19.
basis) as
Trends in the change of peak area (dry weight
a result of duration in storage at 38F.


AREA(M M
70
0
1
2
WEEKS
3
4
kvim ro


71
38p. Also, all peaks showed a decrease in peak area from
3 to 4 weeks when storage was at 38F.
Figure 20 shows a comparison of the volatile changes
(DWB) resulting from storage of celery for 1 day at 70P
and 8 days at 50F after storage for 2 or 4 weeks at 38P.
Considering only the treatments subsequent to 2 weeks' storage
at 38P, without exception, all peaks from the 70P treat
ment had a lower peak area than those from the 50F treat
ment. This trend was not true for peaks 3 and 9 when celery
was sampled after 4 weeks at 38F. Also, storage of celery
for 1 day at 70P after 2 weeks at 38P resulted in a de
crease in peak area for all compounds. The results of the
same treatment subsequent to 4 weeks at 38P do not agree
with those after 2 weeks at 38P when compared with the 38P
check at 4 weeks' storage. All peaks except peaks 3> 8, and
9 showed a progressive increase in peak area for 70P and
50P storage, respectively. This trend was noted in peaks
2, 6, and 7 for the data calculated on a fresh weight basis.
Peak areas (DWB) for the control and storage treat
ments of 2 and 1; weeks at 38P and 45F are presented in
Figure 21. All peaks except 1, 4 and 5 showed an increase
in peak area when stored at 45>F (2 and 4 weeks) as compared
to 38P storage. The differences between these treatments
and the control are not as great as when the data were
calculated on a fresh weight basis.
When considering the peaks on a dry weight basis, the
celery stored 2 and 4 weeks at 45F did not result in the


Figure 20. Peak area (dry weight basis) of celery samples gfter 2 and 4 weeks* storage
at 38F and subsequent storage after each for 1 day at 70 F and 8 days at 50F (in
cluding all peaks).


AREA (MM2) AREA (MM
WEEKS
WEEKS
WEEKS
PEAK 7
WEEKS
OCK-2WKjt 3&F
IC11+- 1 DAYat 7cTf
!5S3 4- 8 DAYS at SO'F
CK-4 WK at 38*F
mi + 1 DAY at 7CfF
^2+8DAYSat5CTF
(OOIX)
WEEKS WEEKS
WEEKS
->1
U)


Figure 21. Peak area (dry weight basis) of celery samples at harvest, after storage
at 38F for 2 and 4 weeks, and after storage at 45P for 2 and 4 weeks for each of
the 9 peaks measured in head-space analyses.


AREA (MM2) AREA (MM
LE££N£
ELU -AT HARVEST
"38*F STORAGE
IC3I -45*F STORAGE
350
250
200
PEAK 8
n
n
mm
|T
|
n r] I1
IB 1 B
0 2 4
WEEKS
-J
Vn.


76
highest area for all peaks; however, the total peak area
for these treatments in each case was higher than the corre
sponding treatment at 38F.
A summary of the net effects of these variations is
presented in Figure 22. This graph varies from that pre
sented in Figure 18 in two ways. First, there was not a
progressive increase in the total peak area from harvest
through 2 weeks' storage at 38F when calculated on a dry
weight basis. There was, however, an increase in the total
peak area from 1 through 3 weeks' storage at 38? with a
decrease after storage for 4 weeks. Second, when calculated
on a dry weight basis, the trend of lowering peak area with
storage of celery at 70F was not consistent. The same
decrease was noted in these data as in those calculated on
a fresh weight basis when celery was stored 1 day at 70F
after 2 weeks'storage at 38F; however, when subsequent
storage at 70F was performed after storage for 4 weeks at
38F, the peak area (DWB) increased slightly. This reflects
the inconsistency of peak 8 (Figure 20) which was the only
peak showing a decrease after 2 weeks' storage at 38F and
o o
1 day at 70 F. Those treatment effects experienced at 45 F
and at 50F for fresh weight determinations were also true
when those data were calculated on a dry weight basis.
Peak areas for 45F treatments remained higher than any
38F treatment and treatment at 5>0F for 8 days after 38F
storage resulted in increases in peak area.
Considering the peak area on a fresh weight basis,


77
Figure 22. General trends in total peak area (dry weight
basis) according to temperature and duration of storage
at l45F> 38F, and subsequent storage at 50F and 70F
after storage at 38F.


78
there was an increase in volatile content from harvest to
2 weeks' storage whereas there was little change through 4
weeks' storage. In fact, all storage treatments except
those at 70F resulted in increases in measured volatile
content at some time during storage. From these data it
appears that there is a synthesis of the hydrocarbons
in storage at temperatures of 38F, 45F and 50F. However,
a loss of these hydrocarbons during storage at 70F indicates
they are metabolized or otherwise partially lost through a
physical system. It is not possible to determine from these
data what limiting factors caused a decrease in the buildup
of these components after 2 weeks' storage of celery at 38F
and 45F.
The reactions taking place in the holding flask have
not been studied in these experiments ; however, the temper
ature of the holding flask (55C) is near the destructive
point of most enzymes. If the enzymes controlling the pro
duction of hydrocarbons were functional at this
temperature, the amount of volatile components present after
2 hours' holding time would relate directly to the amount
of substrate present in the tissue.
The biogenesis of terpenes is not fully understood;
however, it does involve the condensation of isoprene units
through mevalonic acid and squalene; mevalonic acid being
formed from acetic acid in plant tissue (5)* The mechanism
involves condensation of acetyl CoA and acetoacetyl CoA to
beta-hydroxy-beta-methyglutary1 CoA. The latter then is


79
reduced to form the mevalonic acid. Since the precursors
of the terpenes are common metabolic agents, the synthesis
or degradation of these products should be temperature
dependent and subject to the same limitations of other
metabolic activities. A buildup of the terpenes could only
be explained as a result of the degradation of other meta
bolites (such as beta oxidation of fats) to form available
acetyl CoA and acetoacotyl CoA. The amounts of available
acetyl CoA and acetoacetyl CoA required to influence volatile
changes are not known.
It is a known phenomenon that cutting of tissue re
sults in an increase in metabolic activity. The injury to
the tissue in these experiments was maintained constant and
should not have affected the results.
The changes in moisture content did not greatly effect
measured peak area in the head-space experiment. There was
no significant correlation between celery dry weight and
measured volatile content as based on correlation coeffi
cients. The increase in dry weight of celery stored 2 or
4 weeks at 45? was a result of inadequate control of
humidity, even though celery was placed in plastic bags to
minimize water loss. A larger buildup of the volatile
constituents with these treatments could have been indi
rectly affected by the reduced moisture content.
The 9 volatile components measured in the head-space
analyses correspond roughly to the chromatographic fraction
of the solvent extracts with retention times of 8 to 20


80
minutes. There was a considerable decrease (Figure 7) in
the quantity of these components being measured when celery
was stored at 70F for 5 days. Also, storage of celery for
2 weeks at 45? and 1 additional week at 50F resulted in
an increase in these components and a highly significant
increase in the amount of limonene (Figure 10). The changes
in the C]_o^l6 hydrocarbons in the head-space analyses
corresponded to those observed when celery solvent extracts
were prepared from celery stored under market conditions
and at 70F. Storage of celery for 2 weeks at 38F resulted
in a significant increase in the peak area of limonene
(solvent extraction) while after I). weeks storage at 38F
the content of limonene (solvent extraction) was not differ
ent from the control (Table 6). While no significant differ
ence was observed between the limonene content (head-space)
of the control celery and samples stored 2 weeks at 38F\
the treatment at 38? did result in a larger peak area and
all 4 ^10^16 hydrocarbons analyzed followed the same general
trend.
The data substantiate the idea that storage of celery
at 70F results in a high ratio of high/low boiling com
ponents and presumably in a stronger (but not necessarily
more desirable) celery flavor, while storage at temperatures
between 38F and 50F result in lower ratios of high/low
boiling components and a possible reduction in strong celery
flavor. These assumptions should be confirmed through
rigorous experiments using organoleptic procedures.


81
Component Distribution
Considerable quantitative difference was observed
between the high and low boiling components in the chromato
grams prepared from the various parts of the stalk. Table
II4. presents the data for the ratio of high/low boiling com
ponents. The mean ratio for the top was significantly
different from the middle and outer portions of the stalk,
while the ratio for the outer portion was significantly
higher than any other portion.
Table I4.. Ratios of high/low boiling fractions from chro
matograms of extracts prepared from various parts of the
celery plant.
Top
Inner
Bottom
Middle
Outer
0.66*
0.88
1.09
1.23
1.80
Those values not connected by a continuous line are
significantly different (0.05 level) according to
Duncan's multiple range test.
A qualitative difference was observed between the
chromatograms from the top (leafy) portion and all other
portions of the celery stalk. The chromatograms prepared
from the top portion had a large peak at 5 minutes, 25
seconds when chromatographed on a column of Apiezon L, and
5 minutes, 15 seconds when chromatographed on Carbowax 20 M.
This component occupied as much as 4 per cent of the total
peak area of these chromatograms and was absorbed by a


82
column containing boric acid which indicated it was an
alcohol (27). The odor of this compound was very ether-like
when detected at the exhaust port of the gas chromatograph.
Standard alcohols were chromatographed to aid in prediction
of carbon number and are plotted against retention times
in Figure 23.
The chromatogram from the top (leafy) extract is
presented in Figure 2i[. In addition to the unknown alcohol
previously mentioned, other qualitative variations in this
sample occurred near the retention time of 72 minutes. In
all other chromatograms the peak at 72 minutes appeared as
a major peak with a small shoulder. However, observation
of the chromatogram from the top (leafy) extract shows that
two peaks are completely resolved and present in large
quantities.
Chromatograms from celery extracts of inner and outer
portions are presented in Figure 25. While both fractions
showed a reduced amount of limonene as compared to other
fractions, the samples extracted from the inner portion of
the stalk had a larger quantity of the other components
with retention times of 10 to i|0 minutes, yielding a lower
ratio of high/low boiling components. In addition to the
comparatively low amount of low boiling fraction in the
extracts of the outer portion, there was a larger amount
of those components with retention times of 60 to 90 minutes
thus there was a higher ratio of high/low boiling components
While few qualitative differences can be seen between these


83
Figure 23. Retention time of 5, 6, and 7 carbon
straight chain alcohols for polar and non-polar
chromatographic phases.


Figure 24. Chromatogram prepared from an extract of the top (leafy) portion of the
celery plant.
CD


Figure 25. Chromatograms prepared from celery extracts of (A) the Inner and (B) outer
portion of the stalk.
CD
vn


86
chromatograms, the fact that the inner portion of the stalk
resulted in a low, high/low ratio and the outer portion a
high, high/low ratio substantiates the differences in ratio
of high/low fractions previously mentioned.
Chromatograms from the middle and bottom portions of
the stalk are comparable to those of the storage treatment
extracts and little quantitative or qualitative difference
was observed. These results should be expected since the
celery used in the preparation of the storage treatment
extracts did not contain leafy portions and was primarily
composed of middle and bottom portions of the stalk.
While Pan (38, 37) and Hall (29) did not measure
differences in the volatile constituents of celery, their
research seems to substantiate these data. It was noted
that the strong bitter celery flavor was associated with the
outer petioles and with the more green portions of the stalk.
Prom these data, it is not possible to determine if the high
ratio of high/low boiling components is associated with a
strong celery flavor. However, according to the odor char
acteristics presented in Table 3 this would appear true.
Also, in accordance with these data, Hall (21) found organ
oleptic differences between middle and upper portions of
the outer and inner petioles of the same stalk.
Taste panel comparisons have been made between juice
expressed from the top (leafy) section of the celery plant
and that expressed from the bottom portion below the leaves
(ll|). The panel was able to differentiate between the two


87
samples at the 0.01 level of statistical significance.
However, these investigators indicated it was not possible
to differentiate between the two juices on the basis of the
gas chromatograms of their distillates.
Leaf alcohol (hex-cis-3 enol) is widely distributed
in green plants and is reported to be primarily responsible
for the odor of foliage plants (40 42). Since the alcohol
in the celery extracts had a retention time similar to the
6 carbon group (Figure 23), it is possible that it is
closely related to the above-mentioned leaf alcohol and not
a specific product of celery. Noteworthy is the fact that
no peak corresponding to this retention time could be de
tected when leafy portions of celery were analyzed using
head-space procedures.


SUMMARY AND CONCLUSIONS
Odor detection of the various components in the
celery extracts revealed that those components with reten
tion times of 72 to 85 minutes possessed the characteristic
odor of celery. Analysis of the various chromatograms
revealed that two areas of high concentration were present
and were classified into high and low boiling components.
Further investigation of these areas revealed that net area
changes in the chromatograms could be detected by use of
the ratio of high/low boiling components.
Storage-induced flavor changes in celery stalks
appear to be quite temperature dependent when analyzed by
methods used in this research. Storage of celery at 70F,
whether fresh or previously stored at 38F, resulted in a
decrease in the amount of hydrocarbons. Also,
complete flavor profile analysis showed a highly significant
decrease in total peak area of the chromatograms from
celery stored 5 days at 70F. While all components decreased
when celery was stored under these conditions, the ratio
of high to low boiling components increased significantly.
When celery was stored under market simulated con
ditions (2 weeks at 45P and 1 subsequent week at $0F)
the ratio of high/low boiling components decreased
88


89
significantly. This decrease was probably due to increases
in the peak area of the hydrocarbons. When celery
was stored at 38P or 45F, and analyzed using head-space
techniques, the hydrocarbons content was higher
after 2 weeks' storage than for freshly harvested celery
(PWB); no further increase was noted with longer storage
at these temperatures. Storage at £0? subsequent to
storage at 38P also resulted in increases in these com
ponents. Previous investigations (5) have established that
the basic mechanism of synthesis for these components is
the condensation of acetyl CoA and acetoacetyl CoA; however,
it is not known what limitation might cause a reversal in
the trend of component increase at 70F storage.
Since other investigators have discussed the impor
tance of the high boiling phthalides in celery flavor (10, 14,
15, 16, 20, 28, 32), the changes in the ratio of high/low
boiling fractions seerr, to have importance. In view of
these changes, it would appear that storage of celery at
70P would result in a stronger celery flavor, while storage
at 38P, 45P, or combinations of these temperatures with $0F
storage would result in celery with less intense celery
flavor as compared to the same celery at harvest.
The same theory applies to the ratio of these com
ponents in various portions of the stalk and is supported
by several investigators (14, 21, 22, 23). In general, the
strong portion of the celery flavor is associated with the
greener portions of the stalk, in particular the outer


90
petioles. This is further supported by the fact that the
inner (heart) petioles had a ratio of high/low boiling
fraction which was significantly lower than either the
middle or outer portions.
Further research should establish the organoleptic
association with these data. This would require rigorously
controlled conditions since celery stalks have considerable
changes in texture and appearance with long time storage.


Full Text

VOLATILE FLAVOR COMPONENTS OF
CELERY STALKS (Apium graveolens VAR.
dulce) AS RELATED TO
TEMPERATURE AND TIME IN STORAGE-
WITH FURTHER INVESTIGATIONS ON
COMPONENT DISTRIBUTION WITHIN THE
STALK
By
DANNY ODELL EZELL
A DISSERTATION PRESENTED TO THE GRADUATE COUNCIL OF
THE UNIVERSITY OF FLORIDA
IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE
DEGREE OF DOCTOR OF PHILOSOPHY
UNIVERSITY OF FLORIDA
1968

ACKNOWLEDGEMENTS
The author wishes to express sincere appreciation to
the Chairman of the Supervisory Committee, Dr. D. D. Gull,
for his guiding assistance in these experiments. Appreci¬
ation is expressed to Dr. R. C. Smith, Dr. B. D. Thompson,
and Mr. R. K. Showalter, who have contributed to his academic
training, direction of the research, and preparation of this
dissertation.
Special recognition is given to his wife, Wanda, who
has given understanding, encouragement, and long hours as a
dedicated typist to make this graduate study possible.
ii

TABLE OP CONTENTS
Page
ACKNOWLEDGEMENTS li
LIST OF TABLES v
LIST OP FIGURES vii
INTRODUCTION 1
REVIEW OP LITERATURE 3
MATERIALS AND METHODS 9
Storage 9
Phase 1 9
Phase II 10
Phase III 10
Component Distribution 10
Analytical Procedures 10
Organoleptic 10
Extraction 12
Chromatographic 13
RESULTS AND DISCUSSION 16
Organoleptic Evaluation 25
Storage 26
Phase I 26
Phase II 35
Phase III 46
iii

Page
Component Distribution 8l
SUMMARY AND CONCLUSIONS 88
BIBLIOGRAPHY 91
\
iv

LIST OP TABLES
Table Page
1 Storage treatments used in Phase III 11
2 Oven temperature and time relationships
for the matrix sequence used in all
chromatographic measurements 15
3 Retention times on Apiezon L column and
average peak heights of peaks with re¬
tention time greater than 90 minutes 20
4 Odors detected at the exhaust port of an
Apiezon L column after injection of
1 ul of celery solvent extract residue. ... 21
5 The effect of low temperature (38°P)
storage on the ratio of high/low
boiling fraction in celery flavor
extracts 27
6 The effect of storage at 38°P for 2 or 1|
weeks upon the relative per cent of 9
major chromatogram components 28
7 The effect of storage at 70°F for 5 days
upon the relative per cent of 9 major
chromatogram components 3&
8 The effect of storage under market sim¬
ulated conditions upon the relative
per cent of 9 major chromatogram
components Í4.Í4.
9 Observod retention times of volatile
components of celery head-space
samples, using 2 column stationary
phases 50
10Relative per cent of total peak area
for the 9 peaks measured in head-
space analyses. $1
v

Table
Page
11 Mean peak area (fresh weight basis)
for each of the 9 peaks analyzed
in head-space measurements as re¬
lated to storage temperature and
duration 52
12 Per cent dry weight of celery as
related to storage temperature
and duration 65
13 Mean peak area (dry weight basis)
for each of the 9 peaks analyzed
in head-space measurements as re¬
lated to storage temperature and
duration 66
llj. Ratios of high/low boiling fractions
from chromatograms of extracts pre¬
pared from various parts of the
celery plant 8l
vi

LIST OP FIGURES
Page
Figure
1 Chromatograms prepared from (A) dry
ice-acetone and (B) liquid nitrogen
extracts 17
2 Chromatogram prepared from aqueous
extract as described in extraction
procedures 18
3 Infrared spectrum of compound
believed to be n-butyl phthalide 23
4 Chromatograms from celery extracts
prepared (A) after 10 weeks' storage
at 38°F and (B) at harvest 30
5 Mean ratio of high/low boiling com¬
ponents as affected by storage for
5 days at 70°F (difference signif¬
icant at 0.05 level) 32
6 Mean peak area of chromatograms pre¬
pared from celery extracts taken at
harvest and after 5 days' storage at
70°F (difference significant at 0.01
level) 33
7 Chromatograms from celery prepared (A)
after 5 days'storage at 70°F and (B)
at harvest 34
8 Mean ratio of high/low boiling com¬
ponents of the chromatograms from
the control and market simulation
treatment (difference significant
at 0.05 level) 38
9 Mean peak area of low and high boiling
fractions of the chromatograms of
the control and market simulation
treatment 39
vii

Page
Pigura
10 Mean peak area of d-limonene and
n-butyl phthalide for the control
and market simulation treatment
("""'change in peak area significant
at 0.01 level) l±0
11 Mean peak area of all peaks in the
lou and high boiling range ex¬
cluding limonene and butyl
phthalide, respectively 42
12 Chromatograms of extracts prepared
from (A) celery stored under
market simulated conditions and
(B) freshly harvested celery 45
13 Chromatogram from head-space mea¬
surements of celery volatiles with
peak numbers according to reten¬
tion time (Apiezon L) 47
14 Chromatogram obtained when celery
head-space volatiles were chro¬
matographed on Carbowax 20 M 49
15 Trends in the change of peak area
(fresh weight basis) as a rgsult
of duration in storage at 38 F 57
16 Peak area (fresh weight basis) of
celery samples after 2 and 4 weeks '
storage at 38°?’ and subsequent
storage after each for 1 day at
70°F and 8 days at 50°? (including
all peaks) 59
17 Peak area (fresh weight basis) of
celery samples at harvest, after
storage at 38°?’ for 2 and 4 weeks,
and after storage at 45°? for 2 and
4 weeks for each of the 9 peaks
measured in head-space analyses 62
18 General trends in total peak area
(fresh weight basis) changes
according to temperature and dura¬
tion of storage at 45°P’> 38°P, and
subsequent storage at $0°F and 70°F
after storage at 38°? 63
viii

Figure Page
19 Trends in the change of peak area
(dry weight basis) as a result of
duration in storage at 38°F 70
20 Peak area (dry weight basis) of
celery samples after 2 and 4
weeks' storage at 38°F and sub¬
sequent storage after each for
1 day at ?0°F and 8 days at 50°F
(including all peaks) 73
21 Peak area (dry weight basis) of
celery samples at harvest, after
storage at 38°F for 2 and 4 weeks,
and after storage at 45°? for 2
and 4 weeks for each of the 9 peaks
measured in head-space analyses 75
22 General trends in total peak area
(dry weight basis) according to
temperature and duration of storage
at 45°P, 38°P> and subsequent storage
at 50°F and 70°F after storage at
38°F 77
23 Retention time of 5> 8, and 7 carbon
straight chain alcohols for polar
and non-polar chromatographic phases. ... 83
24 Chromatogram prepared from an extract
of the top (leafy) portion of the
celery plant 84
25 Chromatograms prepared from celery
extracts of (A) the inner and (B)
outer portion of the stalk 85
ix

INTRODUCTION
With the increasing sophistication in analytical
techniques and instrumentation has come a greater knowledge
on the volatile constituents of foods. Although the flavor
of celery (Anlum ,-?raveolens var. dulce) is quite distinctive,
it has received relatively little attention as compared to
other fruit and vegetable flavors.
Vegetables are in many cases quite difficult to
examine chemically. Flavor substances frequently occur in
concentrations of a few parts per million, while accompanied
by other organic materials and large amounts of water.
Several extraction (38) and analytical (39) techniques have
allowed increased resolution of these flavor substances and
a decrease in interference from foreign materials.
Celery is a perishable agricultural commodity; how¬
ever, with proper handling it can be maintained in a market¬
able condition for moderate periods of time. The long
storage durations involved in shipping Florida celery to
Europe have imposed more stringent demands on the keeping
quality of fresh celery. Quality reduction of celery due to
storage duration is usually manifest by water loss, pithiness,
loss of green color, decrease in sucrose and increasing
toughness. While flavor changes do exist as a result of
1

2
storage, it has not been determined if these flavor changes
are caused by changes in the volatile flavor constituents
or if these changes result in a detriment to quality.
This research was initiated to: i. select a proce¬
dure for extraction and measurement of the volatile flavor
components in celery stalks; ii. observe whether differences
occur in the chromatograms of celery extracts as a result of
storage temperature and duration; iii. observe differences
in the chromatograms of head-space volatiles (vapor pressures
sufficient to permit analysis in the vapor phase without ex¬
traction) obtained from celery as a result of storage temper¬
ature and duration, iv. investigate the flavor profile of the
various portions of the stalk of celery.

REVIEW OP LITERATURE
The maintenance of quality of fresh celery in storage
involves several aspects of applied physiology. Corbett (8)
indicated that several physical and chemical changes were
apparent during the storage of celery. Chemical analyses
of different parts of the plant showed marked changes in
reducing and total sugars and also both soluble and insoluble
nitrogen. Reducing sugars decreased in the leaves from har¬
vest to the end of storage. Young (45) and White-Stephens
(43) also concluded there was an increase in dry weight in
the crown portion of the celery plant with a corresponding
decrease in the outer leaf portion, and the outer leaves of
the pascal celery plant were of definite value in growth of
the heart petioles during storage. White-Stephens also
observed a marked increase in sucrose and polysaccharides in
the inner petioles coincident with a similar decrease in the
outer petioles. Corbett (8) concluded that soluble nitrogen
increased in both leaves and stalks from harvest to near the
end of the storage period,at which time there was a very
marked decrease in the leaves. This decrease of soluble
nitrogen in the leaves resulted in an increase in the inner
petioles.
Hall, et_ al. (24) cautioned against making quality
3

4
comparisons between different varieties of celery at differ¬
ent maturities. Single varieties of celery seemed to follow
the same pattern of change in chlorophyll content, alcohol
insoluble solids, total sugars, crude fiber, and dry weight
with regard to increasing maturity. Three varieties followed
the same general trend, but varied in the time the lowest and
highest points were reached.
Organoleptic (taste) comparisons have been made be¬
tween celery (Utah 52-70 variety) grown in Florida and the
same type celery produced in California. When considering
celery from an April harvest, the taste panel indicated the
Florida celery had a superior flavor, but was tougher and
had more fiber. However, when comparisons were made with
celery from a May harvest, the rankings indicated that the
California celery had a more desirable flavor and was less
bitter, but was tougher and more fibrous than the Florida-
grown celery. In these experiments it was noted that a low
potassium-sodium ratio was associated with a bland flavor.
The celery from the two areas was grown by commerical pro¬
ducers, was the same age, and was handled under similar
conditions.
The price of Florida celery has frequently been below
that of celery produced in other areas. While these opinions
were not unanimous, the reasons given for the reduced price
included poor appearance, bitter flavor and toughness (22).
While making observations on the flavor of celery,
Hall (23) described a bitter fraction and a burning-numbing

5
sensation from celery. The bulk of the bitter flavor was
associated with the dark green outer layer of the petiole
and did not appear to be associated with the burning-numbing
sensation. Description of the various flavors were: salty
flavor, radish-like flavor, and a hydrocarbon-type flavor
described as kerosene-or turpentine-like. The panel occa¬
sionally noted a sweet flavor. Kali also found a considerable
difference in the composition and organoleptic ratings of
outer, inner, and heart petioles of celery (21). The com¬
position of these petioles was influenced by the temperature
at which the celery was stored. In addition to the dif¬
ferences found between petioles according to position, there
were differences in flavor and composition between middle
and upper portions of the outer, inner, and heart petioles.
Using chromatographic analyses, Gold and Wilson (II4.)
showed that not all flavor constituents were in the juice
of celery stalks. The chromatograms prepared from juice and
puree showed common peaks, although the chromatograms were
not identical. An organoleptic study was performed on the
juice of celery from the top (leafy) and basal portion of
the celery stalk. The taste panel was able to differentiate
between the juice from the top (leafy) and basal portions
of stalks at the 0.01 level of significance. However, Gold
and Wilson were not able to differentiate between the chro¬
matograms prepared from the two juices. There was no dif¬
ference between chromatograms prepared from fresh and
frozen juice samples.

6
Pan (36) characterized the burning-numbing taste of
celery using gas-liquid chromatography. This chemical had
no unsaturated bonds or nitro groups. He found the presence
of aldehydes, carbonyl, phenolic, hydroxy, and aromatic
groups but the fraction was not a lactone. In further re¬
search (37), Pan described a new technique in the isolation
of a bitter principle from celery. This principle was
cationic at pH greater than 7, soluble in polar solvents,
and fluorescent under ultraviolet light. The bitter
principle could not be steam distilled from celery and was
located in the dark green portion of the petiole.
The complex mixture which constitutes the flavor and
odor of celery received considerable attention as early as
l897« Ciamician and Silber (7) reported the following
terpenes contributing to the flavor of celery: limonene,
myrcene, and an isomer of apiol. Of the lactone fractions
identified, sedanolide and sedanonic anhydride were pro¬
posed as being of primary importance in the odor of celery
seed. Following the identification of sedanolide and
sedanonic anhydride by Ciamician and Silber, Berlingozzi and
Cione (i|) undertook a study of the chemistry and odor
characteristics of alkyl and alkylidene phthalides. Working
2 6 A
with A ' -dihydrophthalide, A-tetrahydrophthalides, and
hexahydrophthalides, they found when one of the ^-carbon
hydrogens was replaced by an alkyl group, a celery odor
was noted. When both were replaced by alkyl groups, the
odor was less intense. Celery odor was most intense when

7
the 7^-carbon hydrogen was replaced by the alkylidene group.
Intensity increased as carbons increased from 1 to 4*
Barton and DeVaries (2) analyzed celery oil and re¬
ported the isolation of butyl phthalide instead of sedano-
lide when using Ciamician’s method (7)* They assumed that
an unstable sedanolide might be changed to butyl phthalide,
and proposed the structure of an /á’-unsaturated lactone,
neocnidilide. Mitsuhashi and Muramatsu (33) further pro¬
posed that sedanolide is at least a mixture of neocnidilide
and butyl phthalide.
Recent investigations concerning the flavor and odor
of celery were conducted by Gold and Wilson (14> 15, ló) and
Wilson (44)• In 1963 (ló), they listed 38 compounds which
were identified as volatile components of celery. While
most of the compounds listed probably make some contribution
to the composite flavor and aroma of celery, 6 are of pri¬
mary importance: 3~iso butylidene-and 3~iso validene-3a,
4-dihydrophthalide; 3~isobutylidene-and 3“isovalidene-
phthalide; cis-3-hexen-l-yl pyruvate; and diacetyl. The
4 phthalide derivitives were found in a ratio of 6:3:1:1*
The difficulty of separating the elements of a
chemical mixture into their respective flavor potencies has
been pointed out (17» 40)- Guadagni, e_t al_. (l8) found
an additive effect of various chemicals even when the
chemicals were present in sub-threshold concentrations.
This difficulty was further emphasized in attempts to mea¬
sure sensory responses for food products by direct injection

8
of aqueous vapors into a gas chromatograph (6, 29, 35)*
Newar (35) noted that the concentration of a given compound
in a vapor phase at a given temperature is affected by:
vapor pressure of the compound, type of media in which it is
distributed, degree of solubility in the media, concentration
of compound in liquid phase, and its miscibility with other
organic compounds. Therefore, a decrease in liquid con¬
centration does not necessarily mean a decrease in vapor
concentration.
While there has been much research concerned with
the quality of celery in storage and with the volatile flavor
components of celery, little research has been conducted to
investigate changes in these volatile components during
storage, and it is to this question that the following work
is addressed.

MATERIALS AND METHODS
Florimart and Utah 52-70-2-13 (commonly known as
Florida-2-13) cultivars of celery were obtained from commer¬
cial growers. Size Number 3 stalks (36/crate) were harvested
90 days after transplanting, packed, hydrocooled, and trans¬
ported immediately to the laboratory. Preliminary investi¬
gations were conducted to determine the most satisfactory
method for extraction of celery volatiles and measurement of
the various parameters.
Storage
Phase I
The initial study was established to determine the
effect of low and high temperature storage on the composition
of flavor extracts prepared from celery stalks. Low tempera¬
ture effects were determined by comparing extracts from
freshly harvested Florimart celery with those from celery
o
stored 2 or 4 weeks at 3° F. High temperature effects were
determined by comparing extracts from freshly harvested
o
celery with those from celery stored 5 days at 70 F. A
randomized block design was used with 3 harvest dates con¬
stituting blocks. A non-replicated study involved weekly
extracts of celery stored at 3S°F for 10 weeks.
9

10
Phase II
Floriraart cultivar was also used in a storage treat¬
ment to simulate market conditions. Treatment involved
placing the stalks at 45°F for 2 weeks and transferring
o
samples to 50 F for an additional week. The experiment was
a randomized block with 2 harvest dates as blocks and a
duplicate analysis of each sample.
Phase III
Florida 2-13 cultivar was used for aroma (head-space)
measurements. Treatments employed for these analyses are
listed in Table 1. Statistical design of the experiment was
a randomized block with 3 replications and triplicate
analyses of each sample; harvest dates constituted blocks.
Dry weights were determined according to conventional pro¬
cedures .
Component Distribution
Celery stalks of the Florimart cultivar were divided
vertically into top (leafy), middle, and bottom and, hori¬
zontally into outer and inner portions. Extracts were pre¬
pared of each sample to derive further information on the
distribution of the volatile components of the vapor profile
within the plant. Three replications were used with a
completely randomized design.
Analytical Procedures
Organoleptic
A triangular test was used for all organoleptic

11
Table 1.
Storage treatments
used in
Phase III.
Treatment
Storage
Temperature
Duration
Subsequent
Storage
0
(°P)
At harvest
(Weeks)
- —
1
38
1
--
2
38
2
--
*3
38
3
--
4
38
4
--
5
38
2
1
day at 70°
6
38
4
1
day at 70°
7
38
2
8
days at 5>0°
8
38
4
8
days at 50°
9
45
2
--
10
45
4
—

12
measurements (38)- Organoleptic comparisons were made be¬
tween freshly harvested celery and celery stored 2 weeks at
}8°F. All organoleptic tests were conducted with a con¬
sumer-type panel composed of staff personnel. Panel scores
were obtained in a taste panel room with subdued lighting.
Samples were taken by dicing the center one-third of each
inner petiole, excluding all heart petioles, from 20 stalks.
Petioles with great visual differences were excluded.
Extraction
Celery petioles were washed and stripped of leaves.
All petioles were ground in a Waring blendor and the juice
expressed by hand. Each extraction sample consisted of
ij.,000 ml of juice which was taken from approximately 12 lb.
of petioles. The juice was introduced in 4-00 rol portions
into the evaporation chamber of a Nester-Faust model 500
rotary spray evaporator. The residue was removed from the
evaporation chamber before each new aliquot of juice was
introduced. The evaporation chamber was immersed in a water
bath at 70°C and maintained at a pressure of 30 mm of mercury,
while the condensor and collection chamber were maintained
at 0°C and 30 mm of mercury. Preliminary procedures differed
in that traps for collection of the flavor components were
at temperatures of dry ice-acetone and liquid nitrogen and
were placed after the cold water condensor (14, lb).
The clear aqueous condensate containing the volatile
flavor constituents was retained for solvent extraction.

13
Dichloromathane, amounting to approximately 10 per cent of
the volume of essence, was shaken with the essence of 2
minutes. The samples were washed 2 times and dried with
sodium sulfate according to conventional methods. Solvent
was partially removed in a rotary evaporator at 10ij.°P with
a reduced pressure of 660 mm of mercury. The residue was
removed from the rotary evaporator when approximately 10 ml
remained in the flask. Further removal of the solvent was
accomplished at room temperature and atmospheric pressure.
Head-space samples were prepared by placing 500 grams
of diced celery into a 1,000 ml erlenmeyer flask. The
flasks were sealed and placed in a water bath at 55°C for 2
hours. At the end of this period, the head-space gas was
taken by syringe for direct injection into the chromatograph.
Standards were prepared by placing known quantities of the
various chemicals on tissue paper and positioning in the
center of the flask's contents.
Chromatographic
All chromatograms were prepared on an Aerograph Model
600 D chromatograph equipped with a flame ionization detec¬
tor. Injection sample size and instrument parameters were
maintained constant for each experiment. The sample volumes
were 0.5 ul and 2.5 ml for solvent residue and head-space
samples, respectively. The injection port was maintained
at 235°C. Helium was used as a carrier gas with an inlet
pressure of 1^0 psi and flow rate of 2 0 ml/min at room

14
temperature. A manual matrix programmed sequence was used
from 100°C to 240°C at 3°/min as shown in Table 2. Hydrogen
flow was 20 ml/min while the air was maintained at 250 ml/rain.
Chart speed was ^ in/rain.
Treatment comparisons were made on 12’ x 1/8” copper
columns packed with 5 per cent W/W Apiezon L and Carbowax
20 M, both supported on 80/l00 mesh Chromasorb G.
Subtractive chromatography for alcohols and aldehydes
was performed by procedures described by Ikeda, e_t al_. (27)
and Allen (1).
Infrared analyses of certain compounds were performed
on a Perkin-Elmer Model 237 spectrophotometer equipped with
beam condensor. Spectrophotometer cells were used with .005 in
spacers and methylene chloride as a solvent.
Statistical analyses of paired samples were computed
by employing students’ distribution (4D« Treatment means
were compared by using Duncan's multiple range test (11)
following analysis of variance.

15
Table 2.
matrix
Oven temperature and time relationships for the
sequence used in all chromatographic measurements.
Time
Cumulative Interval
Temperature
Program power'"'
(Minui
;es)
(°c)
0-10
10
100
50
10-25
15
120
50
25-35
10
140
50
35-50
15
160
50
50-58
8
180
60
58-73
15
200
60
73-77
4
220
60
77 —
--
240
60
Power indicator for programming rate of Aerograph Model
600 D oven.

RESULTS AND DISCUSSION
In 1966, preliminary extractions were made from
celery stalks of cultivar Florimart according to the pro¬
cedures previously discussed. Figure 1 shows chromatograms
obtained from the extracts of the liquid nitrogen and dry
ice-acetone traps. All peaks are measured at range 1, at¬
tenuation 8, unless otherwise designated. Little difference
was observed between the component distribution as an
effect of the two trapping procedures (liquid nitrogen vs.
dry ice-acetone). A larger quantity of extract was obtained
from the dry ice-acetone trap than from the liquid nitrogen
trap.
High boiling components are designated as those
which have retention times greater than $0 minutes: the
time at which the oven temperature is set at l80°C. Low
boiling fractions are those components which emerge from
the column in less than 50 minutes. Figure 2 shows a chro¬
matogram prepared from an extract sample of the aqueous
condensate procedure described earlier. Reference to
Figure 1 shows that there is a proportionately smaller
quantity of high boiling components in the chromatograms
from the dry ice-acetone and liquid nitrogen traps than in
the chromatogram from the aqueous condensate shown in
16

«12
Figure 1. Chromatograms prepared from (A) dry ice-acetone and (B) liquid nitrogen
extracts. ^

■í*
Figure 2. Chromatogram prepared from aqueous extract as described in extraction
procedures.
CD

19
Figure 2. Peaks with retention times of 72 minutes or more
made up more than 1+0 per cent of the total peak area of the
chromatograms prepared from these aqueous extracts, while
they made up less than 5 per cent of those from the liquid
nitrogen and dry ice-acetone traps. While both methods
yielded many low boiling compounds, trapping with liquid
nitrogen and dry ice-acetone gave a larger proportion.
The data obtained from the chromatograms prepared
from the aqueous collections revealed several additional
peaks which had retention times greater than 90 minutes, on
the Apiezon column, when the instrument was isothermally
controlled at 24G°C subsequent to the sequence listed in
Table 2. The retention times and average peak heights of
these peaks are presented in Table 3* These peaks are not
considered in treatment comparisons.
In order to establish qualitative meaning of the chro¬
matogram in Figure 2, an odor characterization was estab¬
lished according to the odors noted at the exit port of the
column during the programmed sequence. These odors are
presented in Table 4 with their estimated strengths. Few
celery-like odors were observed in the low boiling range,
while a large proportion of the odors in the high boiling
range were characteristic of celery. It should be noted
that additional odors may have been present but were not
detected due to concentration or sensitivity effects.
Since many of the phthalide compounds which impart
the typical aroma of celery are sterioisoraers (33)> it is

20
Table 3.
average
greater
Retention times
peak heights of
than 90 minutes
on Apiezon L column and
peaks with retention time
Retention time
Average peak height
(Minutes)
(Millimeters)
105
11
108
52
120
31
130
42
11|2
42
147
17
156
455
170
63
214
42
272
42

21
Table 4* Odors detected at the exhaust port of an Apiezon L
column after injection of 1 ul of celery solvent extract
residue.
Time
Odor’"
Time
Odor
(Minute
s)
(Minutes)
11
Turpentine (f)
40
Fishy (f)
12
Woody (f)
41
Undescribed (f)
IS
Orange (m)
44
Diesel exaust (m)
20
Spinach-sx-ieet (m)
45
Celery (f)
22
Bitter weed (f)
55
Sweet (f)
24
Sour milk (f)
56
Orange peel (m)
25
Plastic glue (m)
58
Cotton (s)
26
Bananas (s)
59
Cotton (f)
27
Green bananas (m)
68
Apple (s)
28
Milk weed (f)
72
Celery (f)
29
Celery (f)
73
Celery (s)
30
Undescribed (f)
74
Apple (f)
31
Undescribed (f)
75
Celery (m)
32
Undescribed (f)
78
Cooked celery (m)
33
Undescribed (f)
79
Celery (m)
36
Undescribed (f)
80
Celery (m)
37
Mint (m)
81
Celery-carrot-like (m)
38
Terpinyl acetate (s)
82
Celery-like (s)
39
Undescribed (f)
85
Cooked celery (f)
Odor descriptive terms selected by author.
(f), (m), and (s) designate relative strength of odors as
faint, medium,and strong, respectively.

22
difficult to determine which compound is of most importance.
Most of these isomers readily convert to butyl phthalide, re
suiting in butyl phthalide contamination in any isolation.
A sample of n-butyl phthalide which strongly yielded the
characteristic aroma of celery was obtained. This compound
had a retention time of 77 minutes on Apiezon L and 69 min¬
utes, 30 seconds on Carbowax 20 M. In order to substanti¬
ate further the true structure of this compound, an infrared
spectrum was prepared. The trace of the infrared spectrum
(Figure 3) very closely resembles that presented by Mitsu-
hashi, et_ al_. (31) for Ligustilide, a sterioisomer of butyl
phthalide. However, the spectrum of Figure 3 shows absorp¬
tion at 3,030 cm ^ with 1,600 and 1,535 cm”^ indicating the
presence of all three double bonds around the benzene ring
(3)- These data suggest that the compound is n-butyl phtha¬
lide rather than Ligustilide or a mixture of neocnidilide
and butyl phthalide (33)*
On the basis of these data it is assumed that the
peak appearing at 77 minutes’ retention time from solvent
front of Apiezon L is n-butyl phthalide. Collection of the
actual peak would have been desirable; however, needed fácil
ities were not available.
When using a fractional collection apparatus, Gold
and Wilson (li|) found 25 compounds present in the dry ice
^United States Department of Agriculture Fruit and Vegetable
Laboratory, Winter Haven, Florida.

absorbance
23
Figure 3. Infrared spectrum of compound believed to be
n-butyl phthalide.

trap. Among these compounds they discovered no acids,
aldehydes or phenols. They also indicated only 1+ compounds
were present in the liquid nitrogen condensate, and the
principal odor constituent of the dry ice trap was cis-3-
hexen-l-yl pyruvate, and that of the liquid nitrogen trap
was diacetyl (16). The presence of more compounds in these
extracts seems to indicate the fractionating system of Gold
and Wilson was more efficient in reducing the vapor tempera¬
ture than the system used in these experiments.
The importance of phthalides and their derivitives
as flavor contributors in celery have been pointed out by
several authors (2, 7> ll|, 15» 16, 19, 31). It therefore
seems essential that a representative celery extract contain
a portion of these high boiling phthalide compounds. On the
basis of previous investigations (16, 26), the low boiling
compounds shown in the previous chromatograms were believed
to be primarily H-^ hydrocarbons and related compounds.
The data obtained from the aqueous essence sample are
in agreement with those of Gold and Wilson (16). By separa¬
ting the high boiling and relatively low boiling fractions,
they found the phthalides primarily responsible for the
celery odor located in the column bottom. They did, however,
indicate that the addition of this material to tomato juice
did not reproduce the tomato-celery juice blends unless
material from the dry ice or liquid nitrogen traps was
included. Since the solvent extract prepared from the
aqueous essence contains both fractions, it is believed

25
that the profile shown in Figure 2 most adequately repre¬
sents the flavor profile of celery.
Organoleptic Evaluation
Triangular taste test comparisons between freshly
harvested celery and celery stored 2 weeks at 38°F indicated
there was a significant (0.05 level) difference between
treatments. Celery used for this first test was harvested
on April 21± (stored 2 weeks) and May 9 (fresh). However,
when comparison was made between celery harvested May 9
(stored 2 weeks) and May 2\ (fresh), no significant differ¬
ence was observed. In the first test 61 per cent of the
judges paired the samples correctly,while in the second test
only 39 per cent were able to pair them correctly. From
these data it is difficult to determine if differences did
exist as a result of storage or if the differences were
inherent within each harvest. Hall (21) found that the
flavor of celery did change while in storage at I|D°F, and
that there was an interaction between storage temperature
and petiole position on the organoleptic measurement. When
comparing Florida-grown with California-grown celery,
variations were found between April and May harvests (22).
These differences were attributed to a change in sucrose
content and not to any large change in the volatile flavor
constituents. Kouever, Hall (23) indicated that a taste
panel could easily be influenced by the presence of the
bitter substance which is associated with the outer petioles.

26
It is assumed that if differences do exist in the volatile
flavor constituents between celery stored 2 weeks at 33°F
and freshly harvested celery, storage at higher temperatures
and for longer durations should serve to compound these
differences. Texture changes in celery stored at higher
temperatures make triangular taste tests quite difficult.
Storage
Phase I
Chromatograms used for computation of these data are
similar in appearance to the chromatogram shown in Figure 2
and were prepared using the aqueous trapping procedure.
There seemed to be little change in the chromatograms from
the celery stored 2 or 4 weeks at 36°F when compared to their
respective controls.
The importance of the odor characteristics of the
high boiling phthalides has already been discussed. How¬
ever, any true flavor change will be a result of the inter¬
actions of all compounds present, whether in threshold or
sub-threshold concentrations (lS). Changes in the ratio
of high/low boiling components will be used as an indicator
of change for all chromatograms. Changes in this ratio do
not necessarily indicate flavor changes since no research
has established a correlation.
The average ratio of high boiling/low boiling com¬
ponents for all chromatographed samples was 1.32. Ratios
for high boiling/low boiling fractions for the low temperature

27
storage treatments are presented in Table 5* The ratio for
the freshly harvested sample (1.33) was very near the aver¬
age for all treatments. While no significant change was
observed between freshly harvested samples and those stored
2 weeks at 38 F, there was a slight increase in the mean
ratio after 4 weeks' storage.
Table 5* The effect of low temperature (38 ?) storage on
the ratio of high/low boiling fractions in celery flavor
extracts.
Treatment
Harvest
2 weeks' 38°P
4 weeks' 38°?
(Fraction ratio-high/low)
1.33
1.30
1.65
While a large number of peaks were resolved for each
chromatogram, only a few contribute substantially to the total
peak area of these chromatograms. The 9 peaks which contrib¬
ute most (more than 1.0 per cent of total area) to the total
peak area are listed in Table 6. Also presented in this
table is the mean per cent each contributed to the chromato¬
gram total for the respective treatments. The only signif¬
icant change observed was an increase in the peak at 17
minutes (tentatively identified as d-limonene) when celery
was stored 2 weeks at 38°P. This is difficult to explain
since the mean for this treatment is higher than the control
and the sample taken at 4 weeks'storage. It is noted that
there was a large increase in the mean per cent of total area

28
Table ó. The effect of storage
upon the relative per cent of
components.
at 38°P for 2 or 4 weeks
9 major chromatogram
Retention
Time
Treatment
At harvest
2 weeks
4 weeks
(Minutes)
(Per cent)
17
20.3 a*
23.4b
20.2a
24
6.9
5.6
6.7
36
3-5
2.5
1.7
56
1.0
1.1
1.0
62
1.8
2.7
2.7
72
11.3
7.7
8.4
78
23.6
31.5
29.9
80
9.9
4.4
7.5
81
3.1
3-4
3.6
8O.4
(Total per cent)
82.3
81.7
Values with differing letters in horizontal rows are
significantly different at the 0.05 level according
to Duncan's multiple range test.

29
for the peak at ?8 minutes when the celery was stored 2 or
I4. weeks as compared with the control. This increase was
reflected in an increase in the ratio of high/low boiling
components for the Í4 weeks'storage treatment; however,
increases in the low boiling fraction negated this effect
at 2 weeks' storage. No significant difference was observed
between the total per cent represented by these 9 peaks.
Chromatograms were prepared from celery extracts at
weekly intervals during storage for 10 weeks at 38°?* After
celery had been stored 8 weeks at 38o? it was of no market
value, and approximately 25 per cent of the petioles had to
be removed before the extracts were prepared.
Figure ]+ shows a chromatogram prepared from celery
stored 10 weeks at 38°F versus freshly harvested celery.
Peaks in areas of highest concentration (12 to 22 minutes
and 72 to 90 minutes) seemed to have been maintained while
peaks in areas of low concentration (2i| to 70 minutes) have
been reduced or are missing from the chromatogram. Note¬
worthy is the fact that the low boiling components are pre¬
sent in abundance after storage for long periods of time and
regardless of the condition of the celery.
Observation of chromatograms prepared from freshly
harvested celery show no peaks of large area between 85 and
95 minutes* retention time. Also, no build-up in concentra¬
tion was experienced in this range when celery was stored 2
or 1+ weeks at 38°F. However, when celery extracts from
celery stalks stored 8 weeks or more at 38°F were

Figure 1;. Chromatograms from celery extracts prepared (a) after 10 weeks' storage at
38°F and (B) at harvest.
VjJ
O

31
chromatographed, 2 large (peak height greater than 300 mm)
peaks were present at 8? and 91 minutes' retention time.
Celery stored 5 days at 70°P was quite dehydrated
and of poor market quality. The freshly harvested control
for this study had a high/low component ratio of 0.98, which
was lower than the average. This ratio increased to 2.i|5
when the celery was stored 5 days at 70°P (Figure 5)*
This increase in component ratio was accompanied by
a corresponding highly significant decrease in total peak
area when compared to the freshly harvested control. The
mean peak area for the control chromatograms was 1+8,162.3
p
mm while the mean peak area from the chromatograms of the
70°P storage treatment was only 23,296.5 mm^ (Figures 6 and
7). Such a large change in peak area would not normally be
expected because the extracts were prepared from a constant
volume of juice. Since the moisture content of the celery
decreased during storage, it is possible that under these
dehydrated conditions a larger proportion of the volatile
components remained in the pulp and were not extracted.
However, Matthews (30) found a reduction in total volatiles
of beans after storage for 5 days at 70°P. A conversion of
the volatile flavor components to.non-volatile forms through
enzymatic action would result in such a change. However,
no data have been collected to substantiate this hypothesis.
The changes that occurred in the extracts prepared
from celery stored 5 days at 70°P can be seen in Figure 7*
There was a decrease in the peak area of all peaks,

Ratio (High/Low)
32
\
Figure 5. Mean ratio of high/low boiling components
as affected by storage for 5 days at 70°F (differ¬
ence significant at 0.05 level).

33
Figure 6. Mean peak area of chromatograms prepared
from celery extracts taken at harvest and after 5
days' storage at 70°F (difference significant at
0.01 level).

Figure 7» Chromatograms from celery extracts prepared (A) after 5> days'storage at
70 F and (B) at harvest.

35>
particularly those in the low boiling range which appeared
before d-limonene (17 minutes). The high boiling components
were moderately stable and the peak area decreased only
slightly.
Distribution data of the 9 large peaks from the chro¬
matograms prepared at harvest and after 5 days' storage at
70°F are presented in Table 7- There was a significant
decrease in the proportion of the total peak area repre¬
sented by the peak at 17 minutes. Noteworthy is the fact
that this change is opposite that observed when celery was
stored at 3$°?. However, there was a corresponding increase
in the per cent represented by the large peak at 78 minutes
when the celery was stored 5 days at 70°F. From these data
it seems likely that the components of high vapor pressure
were metabolized or lost through evaporation as the celery
was dehydrated. Table 7 also presents the per cent of the
total peak area represented by the sum of the peak areas of
the 9 major peaks. The sum of the areas increased when
celery was stored 5 days at 70°F. Reference to Table 6
shows that there was a similar increase when celery was
stored 2 and 4 weeks at 38°F. This would suggest the pos¬
sible elimination or decrease of many of the small peaks
not considered in these data.
Phase II
Celery used for the market simulation study had a
very high peak ratio of high/low boiling components at

Table 7. The effect of storage at 70°P for $ days upon the
relative per cent of 9 major chromatogram components.
Retention
Time
Treatment
At harvest
70°P for 5 days
(Minutes)
(Per
cent)
17
19.5
9.0*
24
5-7
7-5
36
3-3
1.4
56
1.0
1.6
62
2.1
4-1
72
13.6
8.0
78
22.0
36.9
80
9.3
7.2
81
3-3
9.9
(Total
per cent)
79.8
85.6
’''Difference significant at the 0.0$ level.

37
harvest as compared to the average for all treatments. The
ratio was 2.38 for the freshly harvested celery and decreased
significantly (0.05 level) during storage to 1.09 (Figure
8). The peak areas for the treatment moans used in com¬
puting the high/low ratio are shown in Figure 9. The dif¬
ference between the areas of each treatment are 21, 39, and
19 per cent for low, high, and total fraction areas, respec¬
tively. While there was change in both the low and high
component areas, these data indicate a greater change in the
high fraction.
A comparison between the mean peak areas of two com¬
pounds, d-limonene and n-butyl phthalide (identification
based on retention times of known compounds on polar and
non-polar chromatographic columns), extracted from the con¬
trol and market simulated lots of celery,is presented in
Figure 10. There was a highly significant increase in the
peak area of d-limonene in the chromatograms of the market
simulation treatment when compared to those of the control.
Considerable variation was observed in the areas of the peak
representing butyl phthalide; however, there was a substan¬
tial decrease in the mean peak area after storage under
market simulated conditions. The changes observed for these
two components were in opposite directions.
Any comparison between limonene and butyl phthalide
is quite critical from an organoleptic point of view. The
importance of the high boiling phthalides has been stressed
by the odors noted in Table Ij. and by the research of other

Ratio-High/Lovj
38
Figure 8. Mean ratio of high/low boiling components
of the chromatograms from the control and market
simulation treatment (difference significant at
0.05 level).

Area (mm x 1,000)
39
Figure 9. Mean peak area of low and high boiling frac¬
tions of the chromatograms of the control and market
simulation treatment.

Figure 10. Mean peak area of d-limonene and n-butyl
phthalide for the control and market simulation
treatment (-"-“Change in peak area significant at
0.01 level).

4i
investigators (IÍ4, 16). Tarpanes, however, seem to be less
dominant in determining vegetable flavor than in many of the
fruit flavors. It is difficult to determine whether a
decrease in the high boiling fraction during storage would
result in a decrease in the actual potency of the celery
flavor. Any discussion of the decrease in proportion of
these elements should take into c'onsideration the additive
effects of the various sub-threshold concentrations which
are affecting the flavor profile (18). This additive effect
of the various components further complicates the discussion
of the actual organoleptic effect of a proportionate change
in high or low boiling components.
C-rouping other peaks of the chromatograms seems to
substantiate the change in limonene in the low boiling range.
When peaks in the low boiling range (excluding limonene) were
considered as a whole, little change was observed between the
control and the market simulated study (Figure 11). There
was a considerable decrease in the peak area total for those
peaks which occurred in the high boiling range (excluding
butyl phthalide) as a result of storage under market condi¬
tions. It is quite possible that an imbalance of these com¬
ponents at harvest stimulated a change to a more balanced
situation. It has not been determined if there could be a
conversion of high boiling components to low boiling com¬
ponents during storage, but enzymatic processes are active
in the formation of volatile flavor components (25).
In addition to a slight decrease in total peak area

o
o
o
x
(V
30
25
20
A 15
tí
O
< 10
Low boiling components
\ High boiling components
Control
Market simulation
Figure 11. Mean peak area of all peaks in the
low and high boiling range excluding limonene
and butyl phthalide, respectively.

43
as a result of storage under market conditions there was a
highly significant decrease in the proportionate amount of
this total represented by the 9 previously mentioned peaks
(Table 8). Also sho’wn is a decrease in the proportionate
amount contributed by each of the 9 peaks except peaks at
1? minutes (d-limonene) and at 8l minutes. However, indi¬
vidual peak changes of substantial importance are those at
17» 72, and 78 minutes. Others in combination might also
contribute substantially as noted above. These data also
indicate a net decrease in those high boiling flavor com¬
ponents most responsible for the aroma of celery.
Chromatograms prepared from the celery extracts of
the control and the market simulation treatments are con¬
tained in Figure 12. As was pointed out previously, there
was an increase in the low boiling fraction, particularly
limonene, and a proportionate decrease in the high boiling
fraction between 72 and 36 minutes. When celery was stored
at 45°? for 2 weeks and 50°? for an additional week, 2
large peaks appeared in the chromatograms at 87 and 91
minutes. The 2 peaks shown in the market simulated treatment
chromatogram at 87 and 91 minutes have peak areas of 1,3C5
2 2 2 2
mm and 2,00i| mm as opposed to 72.3 mm and 176 mm for the
chromatogram prepared at harvest. These data are in agree¬
ment with those received from chromatograms of celery ex¬
tracts prepared from celery stored 8 to 10 weeks at 38°?
(Figure 4)*
Considerable variation occurred in peaks on these

U4
Table 8. The effect of storage under market simulated condi¬
tions upon the relative per cent of 9 major chromatogram
components.
Retention
Time
Control
Market
Simulation
Change
(Minutes)
(Per cent)
Low boiling
17
16.6
28.7
+72.9*
24
3.6
3.6
—
36
2.8
2.4
- 7.1
High boiling
56
3-4
2.8
-17.6
62
5.8
4.8
-I7.2
72
11.9
9.6
-19.3
78
33-5
7.7
-77.0
80
9.8
8.9
- 9.1
81
1.5
2.0
+330
(Total per cent)
94-4
80.1**
'"'Significant at 0.05 level
'"'"'Significant at 0.01 level

Figuro 12. Chromatograms of extracts prepared from (A) celery stored under market
simulated conditions and (B) freshly harvested celery.
-p-
vn

46
chromatograms between 37 and 45 minutes'retention time.
Peaks in this area were always present in measurable con¬
centrations at range 1, attenuation 8; however, the relative
peak areas were not constant within replications.
While the celery used in Phase I and Phase II was of
the same chronological age, it is doubtful that the physio¬
logical maturity was the same. An indication of this vari¬
ation could be observed differences between the ratio of
high/low boiling components for the freshly harvested
o
samples used for the JO F storage treatment and for the
market simulation. Celery used for the market simulation
was harvested in July while that used for the J0°F treatment
was harvested in May. If the physiological maturity was
not the same for all treatments, generalized comparisons
cannot be rendered between treatments (9, 12, 13, 24).
Phase III
Nine separate peaks were resolved from Florida 2-13
celery using the head-space analysis technique. Identifi¬
cation of 4 of these compounds was accomplished by compari¬
sons of their retention times on polar (Carbowax 20 M) and
non-polar (Apiezon L) columns and by spiking these chemicals
Into flasks containing the celery for head-space measurement.
Since all comparisons are made with chromatograms from the
Apiezon L columns, numbers have been assigned to the re¬
solved compounds on the basis of retention time on this
phase (Figure 13). The chromatographic profile obtained

47
J
8
6
Figure 13. Chromatogram from head-space measurements of
celery volatiles with peak numbers according to reten¬
tion time (Apiezon L).

48
when these samples were chromatographed on Carbowax 20 M is
shown in Figure 14* Comparison data were prepared on the
basis of an instrument sensitivity of range 0.1 and attenu¬
ation 16, while peaks 6 and 8 were measured at attenuation
32 and 64> respectively, for both columns.
Observed retention times of the 9 volatile constit¬
uents are presented in Table 9. These are measured from
injection and are somewhat different from those expected
when pure compounds are chromatographed. This deviation in
time is an effect of the large amount of water vapor present
in the holding flask (34)- However, the relative retention
times are not greatly affected by the presence of water.
The retention data presented in Table 9 are in agreement
with those of Hunter and Brogden (34)* These investigators
found c*-terpinene to be present in volatile extracts and
the location of this compound in their chromatograms sub¬
stantiates the idea that it is responsible for peak 9.
However, no retention data have been collected to support
this hypothesis.
The relative per cent of total peak area of the 9
measured peaks is presented in Table 10. Approximately 68
per cent of the total peak area of these chromatograms is
accounted for by the four C-, qH^¿ hydrocarbons identified,
while d-limonane and ^-pinene resulted in approximately 83
per cent of the total peak area.
The mean peak areas (fresh weight basis) of the 9
peaks are listed in Table 11 according to treatment. Total

49
o
Figure II4..
volatiles
Chromatogram obtained when celery head-space
were chromatographed on Carbowax 20 M.

Table 9. Observed retention times of volatile components of celery head-space
samples, using 2 column stationary phases.
Peak
Number
Compound
Apiezon
Unknown
L
Kn own
Carbowax
Unknown
20 M
Known
1
..
7:00
(Minutes)
1:10
_ —,
2
—
8:40
--
1:55
--
3
--
10:48
--
1:35
—
4
<^-Pinene
11:35
11:35
2:56
2:55
5
/^-Myrcene
12:40
12:38
6:00
6:00
6
y^-Pinene
13:50
13:50
4:26
4:26
7
--
16:05
—
2:20
—
8
d-Limonene
17:50
17:52
7:15
7:15
9
--
20:20
--
9:23
--
vn
o

51
Table 10. Relative per cent of total peak area for
the 9 peaks measured in head-space analyses.
Peak number
Per cent of total area
1
0.30
2
2.30
3
0.18
4
1-46
5
2.84
6
11.43
7
3-90
8
72.23
9
5.30
/

52
Table 11. Mean peak area (fresh weight basis) for each of
the 9 peaks analyzed in head-space measurements as related
to storage temperature and duration.
Storage
Treatment
1
Peak number
2 3
4
p
(Area mm )
Control
16.5a*
277.4a
6.3a
122.2a
1 wk @ 38°F
19.5b
310.4ab
12.3a
145.9ab
2 wk @ 38°F
43-3®
407.5ab
20.4bc
263.5cd
+**1 day @ 70°F
37.6c
296.5c
16.labe
177.3ab
+ 8 days @ 50°F
52.4g
310.Oab
25.3c
227-lbcd
2 wk @ 45°F
47.6f
441.1b
27.8c
251.7cd
3 wk @ 38°F
44* 6e
389.8ab
25.8c
228.0bcd
4 wk @ 38°F
40.2d
258.7a
22.6bc
199.Oabcd
+ 1 day @ 70°F
39.3cd
309.lab
24.3bc
l8l.4abc
+ 8 days @ 50°F
58.3h
411.Oab
21.8bc
281.3d
4 wk @ 45°f
47.4f
389.lab
25.7c
269.8d
"'Those means in vertical columns not followed by
letter are significantly different at the 0.05
the same
level.
designates
at 38°F.
subsequent storage treatment after storage

53
Table 11. Continued
Storage
Treatment
5
Peak number
6
7
(Area mm^)
Control
220.7a*
1,187.7a
304.9ab
1 wk § 38°P
30ó.0ab
1,430.2ab
46l.2ab
2 wk @ 38°P
497.0c
1,814.5c
875.9b
+‘""“'1 day @ 70°F
412.5bc
1,532.6abc
593.4ab
o
+ 8 days @ 50 f
464.9bc
1,803.Obc
642.5ab
2 wk @ i|5°P
521.1c
2,141.2c
732.lab
3 wk @ 38°P
500.5c
1,930.0c
660.8ab
4 wk @ 38°P
406.7abc
l,5l4*2ab
255.8a
+ 1 day @ ?0°P
388.9abc
1,649•Oabc
3o9.0ab
+ 8 days @ $0°F
494.5c
2,137.5c
766.3ab
4 wk @ 45°p
500.8c
1,973.8c
708.3ab
Those means in vertical columns not followed by the same
letter are significantly different at the 0.05 level.
"+ designates subsequent storage treatment after storage
at 38°P.

54
Table 11. Continued
Storage
Treatment
8
Peak number
9
Total(1-9)
, 2,
(Area ram )
Control
5,454-9a*
552.9a
8,144*la
1 wk @ 38°F
7,002.3a
571•6abc
10,260.8ab
2 wk @ 38°F
11,650.8a
897.labcd
16,456.4^8
+**1 day @ 70°F
11,362.2a
688.Oabc
15,117.lab
+ 8 days @ 50°F
12,149.8a
804.labcd
16,481.5ab
2 wk @ 45°F
14,232.3a
1,069.2d
19,475.4b
3 wk @ 38°F
11,440.3a
826.6abcd
16,030.lab
4 wk @ 38°F
11,986.7a
760.7abcd
I5,389.5ab
+ 1 day @ 70°F
10,501.9a
666.labe
14,150.2ab
+ 8 days @ 50 F
12,118.4a
l,459.2e
17,348.0ab
4 wk @ 45°p
14,064.7a
899.led
19,381.4b
"Those means in vertical columns not followed by the same
letter are significantly different at the 0.05 level.
"ft
+ designates subsequent storage treatment after storage
at 38bF.

55
area for these peaks as a result of treatment is also pre¬
sented in this table. While all values were not signifi¬
cantly different, the mean peak area for the freshly har¬
vested control was lower than any other treatment for all
peaks except peaks 2 and 7« The lowest peak area for these
peaks occurred when celery was stored weeks at 38°F.
Statistical differences were observed between treatments for
all peaks except peak number 8 (limonene).
Without exception, the mean peak area of all peaks
progressively increased as a result of storage at 38°P for
1 and 2 weeks as compared with the control (Figure 15).
This increase in peak area continued for some compounds
until the third week of storage; however, all peaks except
peak 8 showed a decrease in peak area after 3 weeks' storage
at 38°F.
Figure lb shows a comparison of the volatile changes
o
(FWB) resulting from storage of celery for 1 day at 70 F
and 8 days at 50°F after storage for 2 and i| weeks at 38 F.
Considering only the treatments subsequent to 2 weeks' storage
at 38°F, without exception, all compounds from the 70°F
treatment had a lower peak area than those from the 50°F
treatment. There was only one exception to this trend (peak
o 0
3) when 70 F and 50 F treatments were prepared subsequent to
[|. weeks' storage at 38°?* Also, storage of celery for 1 day
at 70°F after 2 weeks at 38°F resulted in a decrease in peak
area of all peaks as compared to the freshly harvested
sample. The results of the same treatment after I4. weeks at

Figure 15.
basis) as
Trends in the change of peak area (fr
a result of duration in storage at 38
sh weight
F.

WEEKS
vjt.
PEAK

Figure l6
storage
at 50dF
Peak area (fresh weight basis) of celery samples after 2 and
at 38°F and subsequent storage after each for 1 day at 70°F and
(including all peaks).
weeks*
8 days

WEEKS WEEKS WEEKS "" WEEKS
AREA IMM2)
(X100)
(X1000)
(X 100)
(X100)
65

60
38°P are moro variable, peaks 4> 5> 8, and 9 showing the
decrease after 1 day at 70°P. In some treatments, storage
for 8 days at 50°F resulted in an increase in peak area
when compared to the corresponding 3®°? treatment. These
effects were, however, more prevalent after 4 weeks at 38°F
(all peaks except peak 2 resulted in the increase).
Peak areas for the control and storage treatments of
2 and 4 weeks at 38°F are presented in Figure 17. There
was no significant difference between the two storage treat¬
ments at 45°F. A comparison of the 45°? treatments at 2
and 4 weeks reveals that the peak areas resulting from the
45°F treatment tended to be higher than those at 38°F. For
both 2 and 4 weeks'storage, the only exceptions were peaks
4, 6, and 7 at 2 weeks, no exceptions being noted at 4 weeks'
storage. Noteworthy is the fact that total peak area of all
9 compounds for the 45°F treatments, both 2 and 4 weeks,
were the only total values significantly different from the
control.
The general trends for these treatments are summa¬
rized in Figure 18. The points in this graph are based on
the total peak area of all peaks. Storage at 38°? for 1
and 2 weeks resulted in a linear increase in peak area,
while further storage at 38°F caused a reduction in peak
o n o
area. Storage at 70 F for 1 day after storage at 38 F
. o
resulted in a decrease in peak area, while storage at 50 F
for 8 days after 38 F holding resulted in an increase in
peak area. The degree of increase at 50°F was greatly

Figuro 17* Peak area (fresh weight basis) of celery samples at harvest,
at 30°P for 2 and 4 weeks, and after storage at 1|5°P for 2 and 4 weeks
the 9 peaks measured in head-space analyses.
after storage
for each of

LEGEND
EZ3 -AT HARVEST
-38*F STORAGE
ini -45’F STORAGE
(X1000) (X10)
O
rv)

Area (rom x 1,000)
63
Figure 18. General trends in total peak area (fresh weight
basis) changes according to temperature and duration of
storage at 45°?» 38°P, and subsequent storage at $0°F
and 70°? after storage at 38°ip.

affected by the increase and large value of peak area for
peak 8. The greatest total peak areas were observed from
celery stored 2 or 4 weeks at 45°F* Interpolation of the
change between harvest and 2 weeks' storage at 45°F Is not
possible for these data.
After one experimental block had been analyzed, there
appeared to be differences associated with apparent moisture
loss. In attempts to define these effects, dry weight
measurements were prepared from celery in the second and
third block and these data are presented in Table 12. Pew
significant differences were observed as a result of storage
treatment. However, the per cent dry weight of celery
stored 2 and 4 weeks at 45°? was significantly higher than
most other storage treatments. Also, the control was signif¬
icantly different from only 2 treatments: 2 weeks' storage
o °
at 45 P and 3 weeks' storage at F*
Since these differences in dry weight were observed,
the peak areas were calculated on a dry weight basis (DWB)
and are presented in Table 13. When calculated on the basis
of dry weight, there seemed to be more variation in the peak
areas than when calculated on a fresh weight basis. The
peak area (DWB) for the control was not lowest; instead,
there was a general decrease in 4 the compounds to 1 week
storage at 38°P (Figure 19). After this storage duration,
the peak areas of the various peaks increased as when the
data were calculated on a fresh weight basis. All peaks
increased in peak area from 1 week to 2 weeks' storage at

65
Table 12. Per cent dry weight of celery as
related to storage temperature and duration.
Storage
Treatment
Dry weight
Control
(Per cent)
4. llbcd"*
1 wk @ 38°P
3« 99abc
2 wk @ 38°F
3-95abc
+**1 day @ 70°P
4.19bcd
+ 8 days @ 50°F
3.94abc
2 wk @ 45°F
4.82e
3 wk @ 38°F
3.65a
i; wk @ 38°F
+ 1 day @ 70°P
3.78abc
3.67ab
+ 8 days @ 50°F
4.17bcd
4 wk @ 45°P
4» 60de
Those means in vertical
by the same letter are
ent at the 0.05 level.
"'r+ designates subsequent
storage at 38°F.
columns not followed
significantly differ-
storage treatment after

66
Table 13. Mean peak area (dry weight basis) for each of
the 9 peaks analyzed in head-space measurements as re¬
lated to storage temperature and duration.
Storage
Treatment
1
Peak
2
number
3
4
Control
0.4a*
(Area
17•7ab
ram^)
0.3a
6. Oabc
1 wk @ 38°F
0.5a
17.lab
0.4a
4* 9a
2 wk @ 38°F
1.2ab
19•Oab
0.8bc
9.2d
+**1 day @ 70°F
0.9ab
l6.Oab
0.7ab
5.6ab
+ 8 days @ 50°?
1.3ab
21.Oab
1.2c
8.7cd
2 wk @ 45°F
l.Oab
21.2ab
1.2c
8.0bcd
3 wk @ 38°F
l.Oab
24.1b
0.8bc
8.6cd
4 wk @ 38°?
0.9ab
14.5a
0.7ab
8.Obcd
+ 1 day @ 70°F
1.3ab
l8.9ab
l.Obc
8.4cd
+ 8 days @ 50°F
1.6b
22.4ab
1. Obc
10.6d
4 wk @ 45°F
0.8ab
l8.5ab
1. Obc
8.4cd
Those means in vertical columns not followed by the same
letter are significantly different at the 0.05 level.
+ designates subsequent storage treatment after storage
at 38°F.

67
Table 13. Continued
Storage
Treatment
Peak number
5 6
7
2
(Area ram )
Control
9.6ab*
68.9ab
9.8ab
1 wk © 38°F
8.3a
60.6a
5-3a
2 wk @ 3o°F
l4-9cd
72.2ab
10.Sab
+*'"'l day © 70°F
10.9abc
6Í4.. 2ab
8.2ab
+ 6 days @ 50°F
13.6bcd
74•8ab
9.9ab
2 wk © U5°F
15-Ocd
82.Oab
12.0b
3 wk © 38°F
15.7d
92.5b
11.3ab
4 wk © 38°F
11.7abcd
67•lab
8.2ab
+ 1 day © 70°F
13* 6bcd
72.7ab
9.7ab
+ 8 days © 50°F
15.led
91.6b
13.3b
4 wk © i;50F
13-9cd
75•2ab
11.6ab
"""Those means in vertical columns not followed by the same
letter are significantly different at the 0.05 level.
+ designates subsequent storage treatment after storage
at 38°F.

68
Table 13. Continued
Storage
Treatment
Peak
8
number
9
Total(1-9)
(Area
mm^)
Control
233-2ab*
30.4abc
376.4ab
1 wk @ 38°F
164« 3a
23.4a
284.8a
2 wk @ 38°F
232.4ab
37-5cd
397-9ab
+**1 day @ 70°F
251.7ab
27.8ab
386.Oab
+ 8 days @ 50°F
279.8ab
34.1bc
444.5b
2 wk @ U5°F
318.5b
45.2d
505.2b
3 wk @ 38°F
301.6b
36.Obcd
491.6b
4 wk @ 38°F
293.7b
31.labe
431.6ab
+1 day @ 70°F
286.2b
37-Obcd
450.6b
+ 8 days @ 50°F
315.4b
35* 6bcd
506.7b
4 wk @ 45°f
321.4b
37.bed
488.6b
'"Those means in vertical columns not followed by the same
letter are significantly different at the 0.05 level.
-ft#
+ designates subsequent storage treatment after storage
at 38bF.

Figure 19.
basis) as
Trends in the change of peak area (dry weight
a result of duration in storage at 38°F.

AREA(M M
70
0
1
2
WEEKS
3
4
â– kvim ro

71
38°p. Also, all peaks showed a decrease in peak area from
3 to 4 weeks when storage was at 38°F.
Figure 20 shows a comparison of the volatile changes
(BWB) resulting from storage of celery for 1 day at 70°P
and 8 days at 50°F after storage for 2 or 4 weeks at 38°P.
Considering only the treatments subsequent to 2 weeks' storage
at 38°P, without exception, all peaks from the 70°P treat¬
ment had a lower peak area than those from the 50°F treat¬
ment. This trend was not true for peaks 3 and 9 when celery
was sampled after 4 weeks at 38°F. Also, storage of celery
for 1 day at 70°P after 2 weeks at 38°P resulted in a de¬
crease in peak area for all compounds. The results of the
same treatment subsequent to 4 weeks at 38°P do not agree
with those after 2 weeks at 38°P when compared with the 38°P
check at 4 weeks' storage. All peaks except peaks 3> 8, and
9 showed a progressive increase in peak area for 70°P and
50°P storage, respectively. This trend was noted in peaks
2, 6, and 7 for the data calculated on a fresh weight basis.
Peak areas (DWB) for the control and storage treat¬
ments of 2 and 1; weeks at 38°P and 45°F are presented in
Figure 21. All peaks except 1, 4» and 5 showed an increase
in peak area when stored at I).5>0F (2 and 4 weeks) as compared
to 38°F storage. The differences between these treatments
and the control are not as great as when the data were
calculated on a fresh weight basis.
When considering the peaks on a dry weight basis, the
celery stored 2 and 4 weeks at 45°F did not result in the

Figure 20. Peak area (dry weight basis) of celery samples gfter 2 and 4 weeks* storage
at 38°F and subsequent storage after each for 1 day at 70 F and 8 days at 50°F (in¬
cluding all peaks).

AREA (MM2) _ AREA (MM
PEAK 7
WEEKS
L£££H£
â–¡ CK-2WKjt 33^
IC11+- 1 DAYat 7cTf
!Jg§ 4- 8 DAYS at SO'F
OCK-4 WK at 30*F
mi + 1 DAY at 7CfF
^2+SDAYSat5CTF
WEEKS
(OOIX)
WEEKS WEEKS
U)

Figure 21. Peak area (dry weight basis) of celery samples at harvest, after storage
at 38°F for 2 and 4 weeks, and after storage at 45°F for 2 and 4 weeks for each of
the 9 peaks measured in head-space analyses.

AREA (MM2) AREA (MM
LEGEND
EZ3 -AT HARVEST
C55 "38*E STORAGE
IDS I -45*F storage
HI'S
Vn.

76
highest area for all peaks; however, the total peak area
for these treatments in each case was higher than the corre¬
sponding treatment at 38°F.
A summary of the net effects of these variations is
presented in Figure 22. This graph varies from that pre¬
sented in Figure 18 in two ways. First, there was not a
progressive increase in the total peak area from harvest
through 2 weeks' storage at 38°F when calculated on a dry
weight basis. There was, however, an increase in the total
peak area from 1 through 3 weeks' storage at 38°F with a
decrease after storage for 4 weeks. Second, when calculated
on a dry weight basis, the trend of lowering peak area with
storage of celery at 70°F was not consistent. The same
decrease was noted in these data as in those calculated on
a fresh weight basis when celery was stored 1 day at 70°F
after 2 weeks'storage at 38°F; however, when subsequent
storage at 70°F was performed after storage for 4 weeks at
38°F, the peak area (DWB) increased slightly. This reflects
the inconsistency of peak 8 (Figure 20) which was the only
peak showing a decrease after 2 weeks' storage at 38°F and
o . , o
1 day at 70 F. Those treatment effects experienced at 45 F
and at 50°F for fresh weight determinations were also true
when those data were calculated on a dry weight basis.
Peak areas for 45°F treatments remained higher than any
38°F treatment and treatment at 5>0°F for 8 days after 38°F
storage resulted in increases in peak area.
Considering the peak area on a fresh weight basis,

77
Figure 22. General trends in total peak area (dry weight
basis) according to temperature and duration of storage
at l45°F> 38°F, and subsequent storage at 50°F and 70°F
after storage at 38°F.

78
there was an increase in volatile content from harvest to
2 weeks' storage whereas there was little change through 4
weeks' storage. In fact, all storage treatments except
those at 70°F resulted in increases in measured volatile
content at some time during storage. From these data it
appears that there is a synthesis of the hydrocarbons
in storage at temperatures of 38°F, 45°F and 50°F. However,
a loss of these hydrocarbons during storage at 70°F indicates
they are metabolized or otherwise partially lost through a
physical system. It is not possible to determine from these
data what limiting factors caused a decrease in the buildup
of these components after 2 weeks' storage of celery at 38°F
and 45°F.
The reactions taking place in the holding flask have
not been studied in these experiments ; however, the temper¬
ature of the holding flask (55°C) is near the destructive
point of most enzymes. If the enzymes controlling the pro¬
duction of C]_oHib hydrocarbons were functional at this
temperature, the amount of volatile components present after
2 hours' holding time would relate directly to the amount
of substrate present in the tissue.
The biogenesis of terpenes is not fully understood;
however, it does involve the condensation of isoprene units
through mevalonic acid and squalene; mevalonic acid being
formed from acetic acid in plant tissue (5)* The mechanism
involves condensation of acetyl CoA and acetoacetyl CoA to
beta-hydroxy-beta-methyglutary1 CoA. The latter then is

79
reduced to form the mevalonic acid. Since the precursors
of the terpenes are common metabolic agents, the synthesis
or degradation of these products should be temperature
dependent and subject to the same limitations of other
metabolic activities. A buildup of the terpenes could only
be explained as a result of the degradation of other meta¬
bolites (such as beta oxidation of fats) to form available
acetyl CoA and acetoacetyl CoA. The amounts of available
acetyl CoA and acetoacetyl CoA required to influence volatile
changes are not known.
It is a known phenomenon that cutting of tissue re¬
sults in an increase in metabolic activity. The injury to
the tissue in these experiments was maintained constant and
should not have affected the results.
The changes in moisture content did not greatly effect
measured peak area in the head-space experiment. There was
no significant correlation between celery dry weight and
measured volatile content as based on correlation coeffi¬
cients. The increase in dry weight of celery stored 2 or
4 weeks at 45°F was a result of inadequate control of
humidity, even though celery was placed in plastic bags to
minimize water loss. A larger buildup of the volatile
constituents with these treatments could have been indi¬
rectly affected by the reduced moisture content.
The 9 volatile components measured in the head-space
analyses correspond roughly to the chromatographic fraction
of the solvent extracts with retention times of 8 to 20

80
minutes. There was a considerable decrease (Figure 7) in
the quantity of these components being measured when celery
was stored at 70°F for 5 days. Also, storage of celery for
2 weeks at 45°? and 1 additional week at 50°F resulted in
an increase in these components and a highly significant
increase in the amount of limonene (Figure 10). The changes
in the Cio^l6 hydrocarbons in the head-space analyses
corresponded to those observed when celery solvent extracts
were prepared from celery stored under market conditions
and at 70°F. Storage of celery for 2 weeks at 38°F resulted
in a significant increase in the peak area of limonene
(solvent extraction) while after I). weeks’ storage at 38°F
the content of limonene (solvent extraction) was not differ¬
ent from the control (Table 6). While no significant differ¬
ence was observed between the limonene content (head-space)
of the control celery and samples stored 2 weeks at 38°?»
the treatment at 38°? did result in a larger peak area and
all 4 ^10^16 hydrocarbons analyzed followed the same general
trend.
The data substantiate the idea that storage of celery
at 70°F results in a high ratio of high/low boiling com¬
ponents and presumably in a stronger (but not necessarily
more desirable) celery flavor, while storage at temperatures
between 38°F and 50°F result in lower ratios of high/low
boiling components and a possible reduction in strong celery
flavor. These assumptions should be confirmed through
rigorous experiments using organoleptic procedures.

81
Component Distribution
Considerable quantitative difference was observed
between the high and low boiling components in the chromato¬
grams prepared from the various parts of the stalk. Table
II4. presents the data for the ratio of high/low boiling com¬
ponents. The mean ratio for the top was significantly
different from the middle and outer portions of the stalk,
while the ratio for the outer portion was significantly
higher than any other portion.
Table IÍ4.. Ratios of high/low boiling fractions from chro¬
matograms of extracts prepared from various parts of the
celery plant.
Top
Inner
Bottom
Middle
Outer
0.66*
0.88
1.09
1.23
1.80
S
Those values not connected by a continuous line are
significantly different (0.05 level) according to
Duncan's multiple range test.
A qualitative difference was observed between the
chromatograms from the top (leafy) portion and all other
portions of the celery stalk. The chromatograms prepared
from the top portion had a large peak at 5 minutes, 25
seconds when chromatographed on a column of Apiezon L, and
5 minutes, 15 seconds when chromatographed on Carbowax 20 M.
This component occupied as much as 4° per cent of the total
peak area of these chromatograms and was absorbed by a

82
column containing boric acid which indicated it was an
alcohol (27). The odor of this compound was very ether-like
when detected at the exhaust port of the gas chromatograph.
Standard alcohols were chromatographed to aid in prediction
of carbon number and are plotted against retention times
in Figure 23.
The chromatogram from the top (leafy) extract is
presented in Figure 2i[. In addition to the unknown alcohol
previously mentioned, other qualitative variations in this
sample occurred near the retention time of 72 minutes. In
all other chromatograms the peak at 72 minutes appeared as
a major peak with a small shoulder. However, observation
of the chromatogram from the top (leafy) extract shows that
two peaks are completely resolved and present in large
quantities.
Chromatograms from celery extracts of inner and outer
portions are presented in Figure 25. While both fractions
showed a reduced amount of limonene as compared to other
fractions, the samples extracted from the inner portion of
the stalk had a larger quantity of the other components
with retention times of 10 to 1+0 minutes, yielding a lower
ratio of high/low boiling components. In addition to the
comparatively low amount of low boiling fraction in the
extracts of the outer portion, there was a larger amount
of those components with retention times of 60 to 90 minutes
thus there was a higher ratio of high/low boiling components
While few qualitative differences can be seen between these

83
Figure 23. Retention time of 5, 6, and 7 carbon
straight chain alcohols for polar and non-polar
chromatographic phases.

Figure 24. Chromatogram prepared from an extract of the top (leafy) portion of the
celery plant.
CD
-p-

Figure 2S>« Chromatograms prepared from celery extracts of (A) the Inner and (B) outer
portion of the stalk.
CD
vn

86
chromatograms, the fact that the inner portion of the stalk
resulted in a low, high/low ratio and the outer portion a
high, high/low ratio substantiates the differences in ratio
of high/low fractions previously mentioned.
Chromatograms from the middle and bottom portions of
the stalk are comparable to those of the storage treatment
extracts and little quantitative or qualitative difference
was observed. These results should be expected since the
celery used in the preparation of the storage treatment
extracts did not contain leafy portions and was primarily
composed of middle and bottom portions of the stalk.
While Pan (38, 37) and Hall (29) did not measure
differences in the volatile constituents of celery, their
research seems to substantiate these data. It was noted
that the strong bitter celery flavor was associated with the
outer petioles and with the more green portions of the stalk.
Prom these data, it is not possible to determine if the high
ratio of high/low boiling components is associated with a
strong celery flavor. However, according to the odor char¬
acteristics presented in Table 3» this would appear true.
Also, in accordance with these data, Hall (21) found organ¬
oleptic differences between middle and upper portions of
the outer and inner petioles of the same stalk.
Taste panel comparisons have been made between juice
expressed from the top (leafy) section of the celery plant
and that expressed from the bottom portion below the leaves
(li|). The panel was able to differentiate between the two

87
samples at the 0.01 level of statistical significance.
However, these investigators indicated it was not possible
to differentiate between the two juices on the basis of the
gas chromatograms of their distillates.
Leaf alcohol (hex-cis-3 enol) is widely distributed
in green plants and is reported to be primarily responsible
for the odor of foliage plants (40» 42). Since the alcohol
in the celery extracts had a retention time similar to the
6 carbon group (Figure 23), it is possible that it is
closely related to the above-mentioned leaf alcohol and not
a specific product of celery. Noteworthy is the fact that
no peak corresponding to this retention time could be de¬
tected when leafy portions of celery were analyzed using
head-space procedures.

SUMMARY AND CONCLUSIONS
Odor detection of the various components in the
celery extracts revealed that those components with reten¬
tion times of 72 to 85 minutes possessed the characteristic
odor of celery. Analysis of the various chromatograms
revealed that two areas of high concentration were present
and were classified into high and low boiling components.
Further investigation of these areas revealed that net area
changes in the chromatograms could be detected by use of
the ratio of high/low boiling components.
Storage-induced flavor changes in celery stalks
appear to be quite temperature dependent when analyzed by
methods used in this research. Storage of celery at 70°F,
whether fresh or previously stored at 38°F, resulted in a
decrease in the amount of hydrocarbons. Also,
complete flavor profile analysis showed a highly significant
decrease in total peak area of the chromatograms from
celery stored 5 days at 70°F. While all components decreased
when celery was stored under these conditions, the ratio
of high to low boiling components increased significantly.
When celery was stored under market simulated con¬
ditions (2 weeks at 45°P and 1 subsequent week at $0°F)
the ratio of high/low boiling components decreased
88

89
significantly. This decrease was probably due to increases
in the peak area of the hydrocarbons. When celery
was stored at 38°P or 45°F, and analyzed using head-space
techniques, the hydrocarbons content was higher
after 2 weeks' storage than for freshly harvested celery
(PWB); no further increase was noted with longer storage
at these temperatures. Storage at 5>0°F subsequent to
storage at 38°P also resulted in increases in these com¬
ponents. Previous investigations (5) have established that
the basic mechanism of synthesis for these components is
the condensation of acetyl CoA and acetoacetyl CoA; however,
it is not known what limitation might cause a reversal in
the trend of component increase at 70°F storage.
Since other investigators have discussed the impor¬
tance of the high boiling phthalides in celery flavor (10, 14,
15, 16, 20, 28, 32), the changes in the ratio of high/low
boiling fractions seerr, to have importance. In view of
these changes, it would appear that storage of celery at
70°P would result in a stronger celery flavor, while storage
at 38°P, 45°F, or combinations of these temperatures with $0°F
storage would result in celery with less intense celery
flavor as compared to the same celery at harvest.
The same theory applies to the ratio of these com¬
ponents in various portions of the stalk and is supported
by several investigators (14, 21, 22, 23). In general, the
strong portion of the celery flavor is associated with the
greener portions of the stalk, in particular the outer

90
petioles. This is further supported by the fact that the
inner (heart) petioles had a ratio of high/low boiling
fraction which was significantly lower than either the
middle or outer portions.
Further research should establish the organoleptic
association with these data. This would require rigorously
controlled conditions since celery stalks have considerable
changes in texture and appearance with long time storage.

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91

92
12. Pink, P. ¥. 1963* Rate of growth and nutrient absorp¬
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13* Godwin, R. Marshall and Billie S. Lloyd. 1961. Com¬
petition between Florida and California celery in
the Chicago market. Pla. Agr. Exp. Bui. 636.
14. Gold, Harvey J. and Charles W. Wilson. 1961. Tech¬
niques in isolation of volatile materials from celery
and identification of some compounds with acedic
properties. Pla. Sta. Hort. Soc. Proc. 74*291.
15* and . 1963. Alkylidene phthalides
and dihydrophthalides from celery. Jour. Org. Chem.
28:985.
16. and . 1963. Volatile flavor
substances of celery. Jour. Pood. Sci. 28(4)*484"
488.
17. Gould, R. P. 1966. Flavor chemistry. Adv. in Chem.
Series. American Chemical Society. pp. I45“l52.
18. Guadagni, G. G. , Ron G. Buttery, S. Okano, and H. K.
Burr. 1963. Additive effects of subthreshold con¬
centrations of some organic compounds associated
with food flavours. Nature 200:1288.
19. Guenther, E. 1948* The essential oils. 1:1-427.
Van Nostrand. New York.
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4:591-602. Van Nostrand. New York.
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evaluation of portions of celery stalks. Pla. Sta.
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differences between celery grown in Florida and Cali¬
fornia. Pla. Sta. Hort. Soc. Proc. 72:142-145*
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24. , H. W. Burdine, and V. L. Guzman. 1961.
The composition of three celery( varieties at several
stages of maturity. Proc. Amer. Soc. Hort. Sci.
78:361-366.

93
25* Hewitt, E. J., D. A. M. Mackay, K. Kanigsbacker, and
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of the terpene and sesquiterpene hydrocarbons in
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of volatile flavor constituents of several vegetables.
Doctoral Dissertation, Cornell University.
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6:2i;3.
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stitution of umbilliferae plants. IX. Structure of
Cnidilide and Neocnidilide. Tetrahedron 20:1971-
1982.
3U. Newar, W. W. and I. S. Fagerson. 1962. Direct gas
chromatographic analysis as an objective method of
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35* • 1966. Some considerations in interpretation
of direct head-space gas chromatographic analyses of
food volatiles. Food Tech. 20(2) :115-H7.
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. 1961. Bitterness in celery. Jour. Food
Sci. 26:337*
37.

94
38. Roessler, E. B., J. Warren, and J. P. Guymon. 1948*
Significance in triangular taste tests. Food Res.
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to the determination of flavor component changes in
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Master of Science Thesis, University of Florida.
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The chemistry and physiology of flavors. Symposium
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necticut. pp. 465-509.
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methods. Iowa State University Press. Ames, Iowa.
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Primary products of photosynthesis in leaves of
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Proc. Amer. Soc. Hort. Sci-. 35:697-698.

BIOGRAPHICAL SKETCH
Danny Odell Ezell, the son of Odell W. and Madge G.
Ezell, was born June 3, I9J4.I, at Spartanburg, South Carolin
In June, 1958» be was graduated from Chesnee High School,
Chesnee, South Carolina. In June, 1962, he received the
degree of Bachelor of Science with a major in Agricultural
Education from Clemson University. After graduation, he
enrolled in the Graduate School of Clemson University. He
worked as a graduate assistant in the Department of Horti¬
culture until June, 1964, when he received the degree of
Master of Science. Prom September, 1964» until the present
time he has pursued his work toward the degree of Doctor
of Philosophy.
Danny Odell Ezell is married to the former Elwanda
Dayle Henderson. He is a member of Alpha Tau Alpha, Alpha
Zeta, Gamma Sigma Delta, and the American Society for Horti
cultural Sciences.

This dissertation was prepared under the direction
of the chairman of the candidate's supervisory committee
and has been approved by all members of that committee. It
was submitted to the Dean of the College of Agriculture and
to the Graduate Council, and was approved as partial ful¬
fillment of the requirements for the degree of Doctor of
Philosophy.
June 1968
Dean, Graduate School
Supervisory Committee:

AGIN»
CULTURAL
library
UNIVERSITY OF FLORIDA
3 1262 08556 7393


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