Department of Animal Science Florida Agricultural
Mimeograph Series No. 62-8 Experiment Station
December, 1961 Gainesville, Florida
THE EFFECT OF ANTE-MORTEM INJECTION OF SODIUM CHLORIDE, PAPAIN
AND PAPAIN DERIVATIVES ON THE TENDERNESS OF BEEF CATTLE. ./
A. Z. Palmer, D. L. Huffman, J. W. Carpenter,
R. L. Shirley and J. F. Hentges, Jr. 2-
The tenderization of beef cattle by ante-mortem vascular injection of
papain has been reported by Beuk et al. (1959), Goeser (1961) and Huffman
et al. (1961b). Huffman et al. (1961a, 1961b), further reported significant
(P< 0.05) tenderization from the,injection of saline solution, crude papain
ash, and heat denatured crude papain. When crude papain ash was combined
with crystalline papain, a complimentary effect was noted by Huffman et al.
(1961a, 1961b) indicating a possible enzyme activation by a component of
the ashed crude papain.
The objective of this study was to .determine the effect of ante-mortem
injection of salt (NaCI) on beef tenderness. A further objective.was to
develop additional data or confirm.previous findings of the complimentary
:effect of crude papain ash and crystalline papain.
A.preliminary study.indicated that young.rabbits could tolerate one
ml. of a saturated salt (NaCI) solution per pound of live weight without
Trial 1. Twenty-two steers of mixed breeding averaging 606 Ibs...dressed
carcass weight.and grading U..S. Good were used in this study. Eleven steers
were.used as non-injected controls' eleven steers were injected 30 minutes
ante-mortem with 500 ml. of'saturated salt (NaCI) solution. The cattle
were restrained in a squeeze chute and the head held to the side exposing the
jugular vein. The injection was made with a- 16 guage, two inch needle
and a glass syringe. Veterinarian inspectors performed ante-mortem and post-
1/ The cooperation of Central Packing Co., Center Hill, Florida, in supplying
the cattle used in this study is acknowledged. Ante-mortem and post-
mortem inspection was provided by the Meat Inspection Section of the
Florida Livestock Board, Florida Department of Agriculture..
2/ Palmer, Associate Meat Scientist; Huffman, Research Assistant) Carpenter,
Assistant Meat Scientist; Shirley, Animal Nutritionist; Hentges, -
Associate Animal Nutritionist; Department of Animal Science, Universi'ty':.-,
of Florida, Gainesville, Florida. Mrs. Norma Bowles and Mr. Howg'd,.
Povey, Laboratory Assistant and Meats Laboratory Manager respectively,
Animal Science Department, University of Florida, Gainesville, assisted
in this study. .
Short loin steaks, one inch thick, were removed from each carcass 72
hours after slaughter. The steaks were broiled medium well done (1600F.).
Warner-8ratzler shear values were obtained on three one-half inch cores taken
from each broiled steak. A four-member trained panel rated tenderness on a
one to nine scale with- I being inedible, 5 average tenderness, 7 very tender,
8 extremely tender-grainy, and 9 mushy over tenderized. Color of the
rib-eye was visually determined by panel with I being bright cherry red,
2 slightly dark and 3 objectionably dark.
Trial 2. Eighty'steers were used in this trial, forty were Hereford
steers and forty were Brahman crossbred steers. The animals were grouped
according to previous feeding and breeding. The average slaughter weight
was 100 Ibs.; carcass grades ranged from high Standard to low Choice with
an average slightly above average Good.
Treatment I steers served as non-injected controls. Treatment 2
steers were injected with 0.9% NaCI at the level of .07 mi. per lb, live
weight. Treatment 3 steers received ash of crude papain at the level
of 0.2 mg. per Ib. live weight; the ash was in 0.9% NaCI solution which was
injected at the rate of .07 ml. per lb. live weight. Treatment 4 steers were
injected with crystalline papain at the level of 0.2 mg. per lb. live weight;
the papain was in 0.9% NaCI solution which was injected at the rate of .07 ml.
per lb. live weight. Treatment 5 steers were injected with ashed crude
papain (0.2 mg. per lb. live weight) plus crystalline papain (0.2 mg.
per lb. live weight). The crude papain ash and crystalline papain were in
0.9% NaCI solution which was injected at the rate of .07 ml. per lb. live
The steers were injected 5 to 52 minutes ante-mortem using the technique
previously described; the average time interval was 25 minutes from time of
injection to time of slaughter. A veterinarian provided ante-mortem and post-
A one inch steak was cut from the short loin of each carcass 48 hours
after slaughter, wrapped and frozen for later testing. The steaks were
defrosted 24 hours at 340 F., broiled and evaluated for tenderness by taste
panel and by shear technique as previously described.
Three-quarter inch thick liver slices were removed from the livers
after a 24 hour chill, and cooked in covered petri dishes in a 3500 F.
oven for twenty minutes. The cooked samples were tested for tenderness by
a four member panel.
,. .. : :
RESULTS AND DISCUSSION
Trial I data are presented in Table I. It may be noted that the steers
which were injected with the salt solution provided steaks which were on
the average more tender by taste panel and by Warner-Bratzler shear evaluations
than steaks from non-injected steers; differences in tenderness between the
injected and non-injected steers were not statistically significant, however.
These data do not confirm previous findings reported by Huffman et al. (1961a,
1961b) that the ante-mortem injection of salt improves tenderness. Previous
effects, however, were obtained using 0.9% NaCI solution whereas this trial
involved the use of saturated NaCI solutions; this difference in salt treat-
ment might account in part for differences in results.
The rib eyes of the salt injected steers were darker in color, as
determined subjectively, than the rib-eyes of the control non-injected
steers; differences in color between treatments were highly significant
Trial 2 data are presented in Table 2. None of the ante-mortem injection
treatments had a significant effect on steak tenderness as evaluated by taste
panel and by the Warner-Bratzler shear. It should be noted that most steaks
were above average in tenderness thus allowing less opportunity for tenderizing
effect's due to injection to be obtained.
The Hereford steers were more tender than the Brahman crossbred steers;
differences in tenderness between the Hereford and the Brahman crossbred
steers were highly significant (PO.01O) according to taste panel data and
significant (P< 0.05) according to Warner-Bratzler shear data.
The livers from injection treatments 4 and 5, those treatments using
active papain, were more tender than livers from treatments I, 2 and 3. In
general treatment 4 and 5 livers were considered over-tenderized. Liver
tenderness was not influenced appreciably by breed group; interestingly,
however, on a within treatment basis, the Hereford livers appear to be
more tender than the Brahman crossbred livers.
The ante-moretm injection of a saturated salt solution (NaCI) in
beef steers brought about only a slight but statistically non-significant
tenderization of broiled steak whereas the injection had a highly significant
(P 0.01) darkening effect on the color of lean in the rib eyes.
In Trial 2 steak tenderness was not influenced by the ante-mortem
injection treatments although tenderness differences were highly significant
(P<0.01) between breed groups. -Treatments containing active papain
over-tenderized the livers in this study.
TABLE I. THE EFFECT OF ANTE-MORTEM INJECTION OF SODIUM CHLORIDE
ON THE COLOR AND TENDERNESS OF BEEF. TRIAL I.
Treatment No. of Av. color Broiled Steak Tenderness
TSteers .Score I/ Panel Av.-/ Shear Av-/
Non-injected control II 1.38 4.34 11.76
Salt injected II 1.86 4.77 11.41
./ Difference between color scores were highly significant (P<0.01).
2/ Differences were not significant.
TABLE 2. THE EFFECT OF ANTE-MORTEM INJECTION OF SALINE, PAPAIN
AND PAPAIN DERIVATIVES ON THE TENDERNESS OF BEEF. TRIAL 2
Steak Tenderness Liver
No. of Panel Shear Tenderness
Treatment / Breed Group Steers Av. 2/ Av. 3/ Panel Av. ./
1. Non-injected control Hereford 8 5.66 7.82 6.69
Bra. Crossbred 8 5.69 9.55 6.04
2. 0.9% NaCd..07 ml./Ib., Hereford 8 6.03 8.94 6.31
live wt. Bra. Crossbred 8 5.28 9.45 5.63
3. Crude.~apain washed Hereford. 8 5.91 8.10 6.03
0.2 mg./Ib.. Iivewt. Bra. Crossbred 8 5.66 8.95 5.68
4. Crystalline Papain Hereford 8 5.97 9.35 8.91
0.2 mg./lb. live wt. Bra. Crossbred 8 5.28 10.46 8.05
5. Crude Papain ash 0.2 Hereford 8 6.22 9.31 8.61
mg./lb. live wt. plus Bra. Crossbred 8 4.84 9.97 7.90
crystalline Papain 0.2
mg./lb.: ive wt.
I/ Treatments 3, 4 and 5 materials were held in a 0.9% NaCI solution which was
injected at the rate of .07 ml. per Ib. live weight.
2/ Differences due to treatment were not significant; differences between breed
groups were highly significant (P :.0l).
3/ Differences due to treatment were pot significant; differences between breed
groups were significant (P<0.05).
4/ Treatments 4 and 5 were more tender than treatments I, 2 and 3 (P<0.01).
Beuk, J. F., A. L. Savich, P. A. Goeser and J. M. Hogan. 1959. Method
of tendering meat. U. S. Patent 2,903,362. Issued Sept. 8, 1959.
Goeser, P. A. 1961. Tendered meat through ante-mortem vascular in-
jection of proteolytic enzymes. Proc. Res. Conf., Amer. Meat
Institute Foundation, 13:55.
Huffman, D. L.,
A. Z. Palmer, J. W. Carpenter and R. L. Shirley. 1961a.
of ante-mortem injection of papain on tenderness of
J. Poultry Sci. (In press).
Huffman, D. L., A. Z. Palmer, J. W. Carpenter and R. L. Shirley. 1961b.
The effect of ante-mortem injection of papain on the tenderness
of beef cattle. Fla. Agr. Exp. Sta., An. Husb. and Nutr. Mimeo
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Snedecor, G. W. 1956. Statistical Methods (5th edition). The Iowa
College Press, Ames, Iowa.