A review of information on nornicotine

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Title:
A review of information on nornicotine
Physical Description:
24 p. : ill. ; 27 cm.
Language:
English
Creator:
Markwood, L. N
United States -- Bureau of Entomology and Plant Quarantine
Publisher:
U.S. Dept. of Agriculture
Place of Publication:
Washington, D.C.
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Subjects

Subjects / Keywords:
Alkaloids   ( lcsh )
Tobacco   ( lcsh )
Genre:
bibliography   ( marcgt )
federal government publication   ( marcgt )
non-fiction   ( marcgt )

Notes

Bibliography:
Includes bibliographical references (p. 20-24).
General Note:
Caption title.
General Note:
At head of title: United States Department of Agriculture, Bureau of Entomology and Plant Quarantine.
General Note:
"E-561."
General Note:
"March 1942."
Statement of Responsibility:
by L.N. Markwood.

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University of Florida
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All applicable rights reserved by the source institution and holding location.
Resource Identifier:
aleph - 030272381
oclc - 17864802
System ID:
AA00023032:00001

Full Text
- - - -.. -. '.. ". .
".. .". . . . .



-..S
March 1942





UNITED STATES I PARENT OF AGRICULTURE

BUREAU OF ENTOMOLOGY AND PLANT QUARANTINE


E-561


'. :- ~ A REVIEW OF INFORMATION ON NORNICOTINE


'y L. N. Markwood, Division of Insecticide Investigations



Contents
....;. .. ;" ..a g
Page
Introduction ...................................... 2
.. : :-. .* *
Physical properties ...................... 3

Occurrence .................. ........................................ 4
. ; ." *. :
:- < Synthesis .. . .......... ........... ......... 10
S"'.. .
..-Analytical determination .......................... 14


S.Pharmacology .................................. 15
,.4 6 :. ..V:.., '5'


Patents . . .................................... 19
! ' <.',.. .



Reviews and popular accounts ...................... 19


'Summary ..... . ................. ................ 19
^ ::.:.:: :. ,"









';Literature cited ............. .................... 20
%'. ":!"













A' A
4,..
A ., k












4K44
... .. .

4:-
..gh: T.



41
It... ...* .....


0H.
I,
.... ,,
p.i""
::.E"S ::






Introduction K. :.:.
rornicotine is a liquid alkaloid, closely related to tniaolX
in chemical structure and in physiological behavior. It ocpur 0'n.."i
tobacco, Nicotiana tabacum L., in other species of Nicotiana,':'-'"3W4
one other plant as well, viz, Daboisia hovwoodii ?. l .- 'el.: ..:
Nornicotine is an alkaloid of relatively recent Jhaistory. Th;aw
nornicotine, which means the normal or parent form of nicotine, ap,.-....
first occurred in chemical literature in 1879 when Andreoni (3a) at i
to make it from nicotine by the action of hydrochloric and hydriodi6e( $I
Pictet and Rotschy (30) in 1901 also referred to nornicotine. The'P t4
actual claim for its existence appeared -in '1927 on the part of'M. and E
Polonovski (32,33), who described a method for preparing various nor-
alkaloids, of which nornicotine was one. In 1928 3hrenstiin (10-; 11-)"ti;
ported finding it in tobacco. In 1930 Von Braun and Weissbach (5) obtau
it by demethylating nicotine. Since then a number of papers dealing wit4i1
nornicotine have appeared. The most important contributions have been
by SpWth and coworkers (3d to 46S), who have greatly elaborated the entire
field of tobacco alkaloids. i "
The relationship bctwircen nicotine, CloHlN2, and nornicotine, CgH12Nia'
is seen from the structural formulas. ":
H 1 H--2 I__ 2 .


HC~ T 13 CNNJ 11
"^, '^
I^J (, >,,j


Nicotine


Nornicotine


Where nicotine has a methl group joined to the pyrrolidino-nitroge Fii
atom, nornicotine has a hydrogen atom;- The latter alkaloid- is therefore a. ::...M
I ,! K=,,, ,
secondary-tertiary base, in distinction from nicotine, which is ditortiary. :.:
Because of its secondary character nornicotine enters into a number of re-...'.
actions which are not possible for nicotine. A more descriptive name for :
nornicotino is beta-pyridyl-aliha-pyrrolidine, or 2-(3'-pyridyl)pyrrold.iie. a .i
Nornicotine, like nicotine, is optical'y7 active. All three forms ,;:
d-, 1.-, and dl- occur in nature rnd have also boen synthesized. ....
The most prominent source of nornicotino, from the standpoint of : -i
present available supply, is ordinary tobacco, Ticotiana tabacum. The pro- 'i,.
portion of nornicotino in the total alka.loids is variable; in most strains ;J4|
the percentage is low, being of the order of a fe.' percent, but it can alsQ. .'4;
occur in nuantities up to 95 pqrc nt of the total alkaloids. The latter ."..t
case is apparently connected with low-nicotine tobaccos. The 1-form le the. ,'
principal isomer occurring in tobacco, but the dl-form also occurs. :.
14. silvestris Spog. and Comes is a plant also containing a notable.: ".;
quantity of th, 1-form of nornicotine, which is usually the predomiuati4...
alkaloid therein. Other a-ecies of iicotiana no doubti.ontai.:.tg ler.
!.O0d, but for the most part their alk loidal nature not:S.A.: 4PqK
investigated. :- Jl^ ..."







W;, ',, 0 ..,i . . .. .. N .=w :. :.:r. ....:. .r o...
kO :id h e' l...ot;..-",>; a,':
V 4-W. U.to"an& dl-r.aornicotine occur in. the Matralian plant
:..... ,., (.., : ,', .: -... .. .
1 7. ... AN .. .. .. ..

K., ,, .* Pbysical Pronert ies
24 M..:: ,, 47 :, F.... .. .
n. . ..4 .
; ; ..: %on o tiIn.. Isa colorless, hygroscopic, somewhat viscous liquid
..... t q..t atifp oeot which is definitely less pungent than the odor
* tl6i:{i'ti I- aIppnaur to be more stable than nicotine, as it colors
, L Meat-ey..(yllo in a clear-glass bottle exposed to light, and
r 4 't',vs Vl air. It is miscible with water and orwanrn solvents in
Ip4: lotons. Strong alkali causes it to separate from aqueous solu-
['. . E :.: . :" .'* y'.; :.." & "l< * 1

7", ^ : ,
... Lt ';.| is .only slightly vrclatile with stmun, in which res-oect it differs
m -nctine, which is r,'adily volatile. This difference has been
*ryt"t/-^'dtai~i. in" .separating the two bases.
' :'# A 'I;:" : i.' "'"/ ,* ..: -
::',, ..The oicrate of nornicotine is more solub1.e than that of nicotine, a
:*4t: frr.ence which has also serv&t in the r.eirnatinn -f the;:e basse.
;,: >-.0 The most trisptworthv va-ues of the ')hysicaI yrooerties )f nor-
A...4 '..'otine h?.ve been Felected from the literature and are recorded below.
' il'A though such.properties do r-ot usually, very "ith ortica:. isoners, the
-Ui; rt"iture.,of the isomer is noted with the value cited. ThjQ values for
ki Mp:"icbtine are also recorded, for ccpparison.

i' sstfj^.ravity

'.01Q737 .(EhrLnstein, 13). 1 L.nd d.l.
= 1.07 (Smith, 37). 1.

LL.^, = 1.07 (Spaith and Zajic, 6). 1.

1"||.A..".. i. = 1.70 ( Marion, an6 Z;i1c
71 d" ..-.d i...
*'rr% 5).' ..
f'.",."~H ... c s s. .T ~~h ai nuri.or,]:,L5
,.l;i': i:=:;" :"i'7..= 1.0757- (Hick-s ;'.nc. TLe14,,8..:ra ort 18) (1 c..rd ,41.
'46
L ':; " 20 on
1.:.::. 20. = 1.072 (Spah, Hicks, r.d Z.jic, 39) d.
1"(,,: "h 39).. ", .






PS ,^i^l3O. si at .12 mm. (ronsten, 13)'. Wtn1 and1 d .
.tJ., 339l at -2mm. (Craig, 7)dl
rr-4t9-140 at. .12 mm. (von Braun a-nd Weisabach, 5)13 and dl.
^ft: fl&7frti at atm. pes'. (von Braun and Weissbach, 5). 1 and dl.
.;:: ~~~ ~ P e s ",'.., .., '




;r .:.::.s. ,r s. (Smith, 3
:.......-. Hicks :and LeMessurier, 16). d and dl.
BB ^..,i.c.,! 0 iJ.lan. -:i ae sle -a3). d and d l. 1 .
'. 59.'Eo.ckt,.21.aztaagi)., 1


.~~~~ 41!
4:- i d and "d

A .i
4: .WWr ) a








.4 .. .:: :.
NY1. 1.537 Cl.breastein. i 3. I .1.. ,.
18. 6
.6 = 1.5490-1.5518 (Hick. an &dK..xrier. i). a aI,: A ;A
-.""-,. '"['.ff"::".tH B
^.3 =1.5490 (spth, xicka anM z&e.,3s). aeata. A)dt.. :

Nicotine, n= 1.5300. 1 '

icll rotation .''

[alpha]|3 -8S.8. (Spith ant Zejic, 16). 1.
[alpha]0 = +86.3 (Spth, Hicks, an SJic, 39). :
NhaJ 0 16...."3:
Nicotine, [al-ha] .320 '.'


Ultra-violet absorption ;'J
. ...'
14Maximum at 2600 A.U.; log e 3.15 (Hicks and LeMessurier, IS). d and 4.1 .
;,icotine, maximum at 2604 A.U.; log e = 3.37. '


Occurrence ,:

Ehrenstein (10iO, ii) in 1928 presented preliminary results of an '!
investigation of the secondary alkaloids of tobacco. A further statement :I
(12) appeared in 1930, and in 1931 (13) a complete account of his work .:'
appeared. He reported the finding of two new alkaloids in tobacco, one I
of which was 1-nornicotine. .R. :i'

The existence in tobacco of alkaloids other than nicotinq- goes 3Lback
to Pictet and Rotschy (30), who in 1901 reported finding three new alka- .. ;
loids. One of these was called nicotinene, b. p. 266-7; the formula'
C H N and structure were assigned to it. 4

CA~
3..
Noga (25) in 1914 confirmed the presence of this body. Ehrenstoin, .
however, doubted the purity of nicotinen" and reinvestigated the matter. .
He readily obtained a fraction of alkaloids, b. p. 269-70, whi* ,"
corresponded well with nicotine. A methylimino (N-CHO) determination nva "
negative, however, and hence the structure assigned abuve was incorrect. .
He found that both from the steam-volatile and the nonvolatile portions .......,
of the original tobacco extract this fraction of alkaloids oould 'be ,.":,
isolated. After several fractionations of the crude base by distilatioai;
the picrate was precipitated and then fractionally crystalliec. from .,
different solvents. In this way two different pirates were obtained, ..',. :.!..
higher-melting one corresponding to a higher-boiling alkaloid ad a :loar lp
molting one corresponding to a lower-oiling alkaloid. The. ft:mW" .':::.o.......
these is not considered here, except to note that he coacludi 'eit.^i *
anabasine, which was an error, as 4pith-and Keastlee ) .. 431y4,4
was anatabine. . .


.' 1. s...-..i.i i .. ".
,;, :,: ,@ R
.: ..'.~ ~ "1 .: .


Opt









SThe lower-boiling alkalQid was found to be 1-nornicotine, C H N.
Sb. p. 130.5-131.3 (11 imm.) or 134-50 (14 mm.), d49.5 = 1.0737, [aipha]d =
S-17.700, .1' 1.5378, It was a nearly colorless liquid of a slight
amine odor, which hardly colored after 1 year in a sealed tube. The H01
salt was dextrorotator?. Oxidation with MIO gave nicotinic acid; the
H* NO-CH determination showed absence of alkyla ed nitrogen. The methiodide
derivative made from it was identical in properties with the corresponding
S derivative made from nicotine, as shown by the following scheme:


CH3--I *
(HI formed in the case
of nornicotine)


Nornicotine


Other compounds made to characterize the base were: picrate, m. p.
191-2; picrolonato, m. p. 250-2, urea, m. p. 167-70; phenylthiourea,
m. p. 176-7*. The natural 1-nornicotine isolated here agreed well in
properties with that already ?eportad by von Braun and Weissbach (5),
which was made by demethylating nicotine. Ehrenstein quantitatively de-
hydrogenated nornicotino ov.r ?t-&asbestos at 320-30 to 2-[bota-pyridyl]-
pyrrolo, or nornicotyrine,



i^,-J In m. -D. 100-102,

picrate m. p. 202-3. By this investigation Ehrenstoin proved for the
first time the presence of norr.nicotine in tobacco.

Shmuck (35) in 1934 published results on the investigation of
several species and hybrids of Nicctiana. The chief feature was the
attempt to characterize the several types by division into steam-volatile
and nonvolatile alkaloids. The following tablo' illustrates the character
of the data in tho paper.


Nicotiana alata Link and Otto
N. lon.iflora Cav.
: Petunia violacea Lindl.
N.&- glauca Graham
N : silvestris Speg. and Oomes
:,.. gl. tino sa L.
% ..tabac.Nu'L.
j.; -.-, .gt .. . .. .
SI :.' tab.cu'ma Xa NzS., ilvestrls.
.. j.. j", u
."7"


Total
alkaloid
content
Percent
0.1130
.0766
.1201
.4774
.5577
.3342
1.94
.91
.19


Alkaloids


Volatile
Percent
0.0
0.0
0.0
23.8
11.8
0o.6
100.0
24.6
80.0


Nonvolatile
Percent
100.0
100.0
100.0
76.2
S8.2
99.4
0.0
75.4
20.0


.*:' ~






The division of alkaloids into volatile and nonvolai. .e" 4-
a bearing on their nature in that nicotine Is readily volatile' .t
..while the other, tobacco alkaloids are much less volatile, .llhwugh
volatile enough on prolonged distillation to- be detected in th'4
The above table shows complete volatility of tje alkaloid b
which is indicative of nicotine only. The first three plants,
interest here, have completely nonvolatile alkaloids; thftirbt4..
interesting in other connections. Three species, IT. Ulutino s
vestris, and N. glauca, are of great interest because of their -h* ib
alkaloid content and their'high proportion of nonvolatile alaloi 'a

Shnnimuck did not positively identify'.any of the alkaloids eXo.. .ti%.
nicotine. However, in the following 'tablo, which gives hte t .iUtt N"t jAji.'a.,i
of the pirates of the volatile alkaloids, some insight is:gainOn in Co;,..... :...
their character. . .. **..
...... .., .....
* : ..* .,: ; .r "" *
Volatile alkaloid of-- Picrate, m.p. .

Nicotiana glauca Graham 207-80 I
N. silvestris Speg. and Cones 176-8
N. glutinosa L. '208-160
N. tabacum L. 21S-9
N. tabacum x N. glauca 198-200
N. tabacum x N. silvestris 178-80 :

The lowest-melting picrate of the group nicotine, nornicotina, and
anabasine is that of nornicotine (m. p. 191-2); honce N. silvestriu ..s
(Smith, 37), as well as the hybrid N. tabacum x F. silvestris, contains
nornicotine as the chief volatile alkaloid constituent. The alkaloids of :
N. glauca and of N. glutinosa, forming pirates of closely agreeing .
melting points, are obviously alike. Smith (36) later showed that the .
alkaloid of N. glauca is largely anabasine, and this alkaloid has since
been reported in N. glutinosa by H. H. Smith and 0. R. Smith (unpublished). A4

Spdth and Zajic (46) in 1935, in the third of a series of papers
on tobacco alkaloids, reported on the isolation of 1-nornicotino'therein. ,
Starting with a mixture of Kentucky and Virginia tobacco. .:.cy prepared "
a concentrated extract, which "ras made strongly alkaliui zn.i exhaustively .,
extracted with ether. The ether extract was fractionally distilled,
giving a main fraction distilling at 94-5 (i mm.) and a higher-boiling
fraction. The ether solution of the main fraction was fractionally ex-
tracted with a saturated NaOl solution containing a small quantity of HOl.
Each fraction was made alkaline and steam-distilled under vacuum at low '
temperature until all the nicotine was expelled. Tho alkaloid recovered
*frnm +t.ai rsntdiminf ,n rnnnwnr'tsdt int.n *n inflrft Cn.n ni- i. 1R48- hri4d the a i bs..:.."


still net pure, was recovered and distilled (1 uan.). There was thus
obtained an almost colorless liquid having a specific rotation
[alpha)2=- -39.7. This crude nornicotine was purified by crystafllization
as the perchlorate from methanol-ether solution in the form of white .
crystals. The free base recovered from the perchlorate hadb after dis-",
tlllation at I mn., a specific rotation of [alpha.]3- -_88. (ths density
oaldulated therefrom is d43 1.070). Analysis for carbon aind h,.rog'b1.
agreed with the composition CqH1g14. The picrate melted atkl9,M1n a dk
the melting point of a mixture of hhis picrate with t p Ipt f t j
Daboimia base (35)', which was a mixtureof d- an l-.,,
'A~


H,






dt a.P.. n The. lspornicotine was. methylated with for.aldew de and
B formtt.I acid. into a Sailwhleh agreed in properties with those of 1-
> niootino., As. shown by optical rotation and by the melting points of the
:..pic.rat, ttin itro-m-cesolate, and picrolonate (223-, 207 219,
reap.). The diimethifide of this base also agreed in optical rotation
7: ..and r. p. with.tbat of 1-nicotine. Spath'and Zajic were accordingly the
fvrst to isolate ,l-nornicotine in pure forn, having the specific rota-
J ..tion'o$ 8S -f 0
Hicks.and .LeMqssurier (18) in 1035 investigated the alkaloid of
J"ae Australian plant 'Thboisia horwoodii, a plant used b;r natives to
catch eus,. b poisoning waterholes, and also chewed on occasion by the
nattvps where "piturin was unobtainable. "Pituri', which is plant
material chew6d by the natives, was found to be a variable product;
normally it consists of the leaves of at least two species of Nicotiana,
vizI N. excelsior (Black) Black and N.'gossei Domin ?but not N. suaveolens
'Lehm.T. .Bbcause of the variability of. the product, the lack of botanical.
I 'identification, and incorrect chemical deduction, confusion existed in
I;.: ,th..6hemica., literature as to the nature of the alkaloid of pitulri; some
S. wor rsreported the presence of nicotine, others denied this. The
S, present -work wasoon authentic 2. horwoodii and indicated the presence of
ll: lnortdoot ine.
'".:..An '' :s alcoholic extract of the leaves led to an ilka loid fraction. of
'. b h.At, p.. 1170 (4.35 mml.), or 266-S (atm. preos.). It was a colorless oil,
...'gradeually, browning on exposure to air ad -light, with a so-nehat pungent
Sodor which was definitely sweeter than that of nicotine. It was soluble
3" i .n water, alcohol, and ether. Various salts were prepared, of which-the
AN.'. picrate, picrolonate, and chbloroplatinate were obtained crystalline, the
NO:. Latter in orange and yellow forms. The refractive index was e1.490 to
2; ;551S9- (at 1-.6), compared s ith.1.529 for nicotine at 16.6'. The
specific rotation, [alpha]J-1.5% +3..59 (d = 1.0757). Molecul?.r
'b..y 'eent, worast's (camphor) method, ws 149.5; by analysis of the orange
| chlorop3atinate, of probable fornula B-2Z03-?tCi4, the combining weight
;- could not be accurately determined, but a-reared to have the value
130.1; by analysis of the silicotvngstate, 232'S21320.SiO212W0, the
value 149,9 was obtained. Carbon apnd hydrogen determinations on the
free base and on the picrolonate (m. p. 252-30) agreed more closely
5 vith nornicotine (m. w. 145) than with nicotine (m. w. 162). KMn104
WEV' :, .: " "







I'T, oxidation (in collaboration with Splth) gave nicotinic acid, hence the
compound was a beta-pyridine derivative, probably d-nornicotine. The
!i:,, ultra-violet absorption agreed with this view; the absorption coefficient
M4 (log e = 3.15) was slightly lower than for nicotine (log e 3.37), but
the maximum (2600 A.) was at the same point as for nicotine (2604 A.).
F -or toxicological tests see under Pharmacology (p. 16).

l :7 0 Spath, Hicks, and Zajic (3h) in 1935 reported on the examination
O.f,-the Anstralian plant Duboisia hopwoodii. The leaves of this plant
are used by the-natives of Australia as a chewing material and also as
M &nial poison. Some chemical work had already been done by earlier
^:';, b s. n the chewing material, known as "pituri," but with contra-
il-4o4rpixlta The reason for the.confusion in regard to the alkaloidh
.rz $ iJta punted out by Hicks, Briicke, and Hueber (17),
B^^ coul"Ant'be accu-r a deterin, wso appearibled Hitkes ,t alue
:1t.A y anatyismat&rial was of a variable nature, not always
'1W fe ate(.W WC-.4 5*aglreed wih varcinnsey
MH. i,.
M!ate
::: oareun a bta-yriinederiatie, rcbbly -nonictin Th




.-.- .; . ..


Nicotiana species and with another species of Duboisia. The present
investigation on an authentic sample of D. hopwoocli established th'e.o
presence of d-nornicotine.


From an alcoholic extract of the ,plant was obtained a sain froa.-
tion of alkaloid distilling at 117 (3.6 mm.), which proved t6 be a
mixture of d-nornicotir.e and dl-nornicotine, Analysis indicated the
formula C9H12N ; the molecular weight, found by the camphor method. was
150.2, compared with 4i8.11 calculated. Other values recorded for this
alkaloid are: Density, d = 1.0757; refractive index, nlS-3 = 1.5490;
specific rotation, [alpha]24 = +35.3. The ultra-violet absorption was
a maximum at 2600 A. U., log e (epsilon) ='3.15. For nicotine the maxi-
mum absorption is at 2604 A. U., log e = 3-37. The dipicrate of this
alkaloid melted at 191-2 (without bubble formation); the dipicrolonate
melted at 252-3. The base was methylated with formaldehyde and formic
acid to nicotine, from which the picrate of m. p. 224 was obtained.
The nicotine regenerated fron,.the picrate had in aqueous solution a
specific rotation of [alpha]2 = +48.3, hence was a mixture of the d
and dl forms. Pure 1-nicotine when tested under similar conditions (in
KOH at low concentration) gave [alpha]J4 = -77.78. By dividing 77.78
into '43.3 there results 62 percent as the relative amount of d-nornicot.ine
in the alkaloid and 3S percent as the amount of dl-nornicotine. The
approximate specific rotation of pure d-nornicotine was then calculated:
+38.3 divided by 0.62 = +61.7. It was realized, however, that this
value requires confirmation with the pure d-base. Examination of the
methylated base, with respect tq the methiodide, trinitro-m-cresolate,
and picrolonate, gave results consistent for a mixture of d- and dl-
nicotine. A further test made on the Daboisia alkaloid (via bensoylation)
proved the absence of nicotine and other tertiary bases.

Hicks (15) in 1936 further investigated the chemistry of the
alkaloid of Diboisia hopwoodii. Its methiodide (m. p. 208-9) was in
agreement with the dimethiodide of nicotine (m. p. 208.5-209.5).
Identity had previously been shemwn by Ehrenstein for the methiodides of
1-nornicotine and nicotine (10).

The Duboisia alkaloid was dehydrogenated (Pt-asbestos catalyst)
according to procedures used by Wibaut and Overhoff (50) and by
Shrenstein (13). The product, b. p. 194.2 (30 mm.), after re-
crystallization from a mixture of ether and petroleum ether (large
needles), had the m. p. 100-2; picrate, m. p. 202-3. These values
agreed with those obtained by Ehrenstein for nornicotyrine, 2-[beta-
pyrielyl] pyrrole. Hence the alkaloid in Duboisia hopwoodii has the
nornicotine structure.

Spnth, Hicks, and Zajic (39) reported in 1936 that they were able
to separate d-norr.icotine in practically pure form from the mixed
Duboisia base, as the perchlorate. This method had proved successful in
isolating pure 1-nornicotine from the tobacco bases (46), whereby the
value for specific rotation, [alpha])3 = -8.8 0 was obtained. For d-
nornicotine from Duboisia was now found [alpha]) = +86.30, which
agreed closely in absolute value with that for 1-nornicotine. The
melting points of the picrate, trinitro-m-cresolate," nd picrolonate,
of both forms agreed (190-1, 200, 2520, resp.) %. .


I L " JhL: A.
....it; bit.1h.4


'1'

H~"
"pH


I
: ." ".j
- *'i.
f=


. *=


ii

















J .,$aua .auu"v' ..h..t r..tJ wSA. ,&.. ~J&- L,. &.IJJU *,i550
fTh:.e was a possibility that the dl-nornicotine so isolated might ..
.* .b~r 60$oe dby racemi-szation of 1-nornicotine luring chemical in-
vMK&#,iOt. experiments were made by heating 1-nornicotine for 4t
W' t.100c-with 10 percent HI01 and with 10 percent KOH. The decrease
aimOounted to only about 2 percent. Since mild conditions
^,|$ emplboye, 'throughout the course of treatment of these alkaloids,.it
i t'se assumed that hardly any racemization occurred, and therefore
tca-Aorricotlne exists .asq such in tobacco. In a test of a certain
."'tobacco the..'alkaloid found therein was practically pure 1-
'Q'S4$.tinehavng alpha)]2
0NUOt "aha~vLn =-o9.7' '
^^P,..LiT !,;,' .. i'.! .. *. ..j *
I1: 1j more. drastic treatment of-.1-nornicotine considerable racemiza-
tloai..* u-dizce4. vThtas, by hating with the calculated quantity of H2S0 .s.'
..... 3 O% the specific rotation fell after 24 hours to -36.7, and after ':
-4 i T,,. o 2.6. o
.|| l| ;3a -,." .. .. '..;. ,.,
M ; :N.R 1 "..... ,
r"..,Kovalenko (21) in 1937 reported a study of the alkaloids of : .
u.$-'A '.`Britt. and Rusby and N. silvestris Speg. and Comes. These 4 .
Ael|ed ..h alkaloid, or r.lkaloids, the picrate of which had the
.'..'.-S '." To alkaloid, other than nicotine, was definitely .
Qi'.' tt'i it was established that another steam-volatile alkaloid ..
i vp eMifwhich foruied an insoluble'picrate. It had the character of "
MrU $iiK. amine.' 7(The chief alkaloid in N. silvestris was shown by
.3 T`) t be nornicotitie.)
Smith(f)l -n 1937 reported finding 1-nornicotine in Nicotiana
M Ontris, grown in Virginia, in an amount equal to about 95 percent
i the .-totai alklobids, which latter amounted to about 1 percent of the
aetlal. examined'. Associated with the nornicotine was a small quantity
ST-ii! tcotine1,. whibh was separated by steam-distillation through a frac-
tin ng-bcolumin. The nicotine was identified by its picrate (m. p.
iKl -4uE.r:)and 'by :pt.cal rotation. The base recovered from the
..iether traction had the following properties, whidh identi-
ii 4 t iirily pur0'-1-nornicotine: Sp. gr. 1.07; [alpha]g =-80.0; ....,
,ii '0 :'COt.); b, p, 27DZ1. For comparison, some nor-
o : nlcotiae, by KnO)4 -oxidation (45) The pirates
'.i".the..norA.cotie fromN:. silvestris was "
nd-"fori aid (WI) to nicotine, the
p.." ,2b.0. Smith also
o ''. _$ Akti Y of anbasine,
E *, : : .. , r t' :~.
".,',: K y',, '"i ..;.',,.,*:-- : "2 4"i'.;!.. 4T. ; tj
J' .. .. ., . .
..:. ... :jq.': ..! .. ...: ,J.." =, -'.e.l.." k :. '
li i !~~iw ;;i .: :,: F .. .. ... :





N .M..
in the nicotine-free portion of the '. slrveetria alkaloioA
*, .M . *. :.
Kostoff and Sara-a (20) in 1939 dewribed the result QCi4. jAj!.
breeding experiments of tobacco. Under 'the direction of ,A:A.A' 9
the nature of the plants was determined Wi part by the diffeW. AAi: ... 0,
4.J. 4 .....
tween total. and volatile alkaloids, and by'the m. p. of the 'on,-TE i .
The percentage of total alkaloid was determined by Keller', ste.tho Oa",k.
the volatile alkaloid'(which was chiefly nicotine) by Bertrad'J RO's4oA N,
The nonvolatile alkaloid group could be determined by differenceP. T .. .;
differences for most cases of N. tabacum were small, :indicating aatii J.
pure nicotine types, whereas in N. ailvestris (omnb of the anicestb rs" .-.'.- .
N. tabacum) they were considerable.' Hence N. silvestris, according to.';;:
Shmuck, contains a large percentage of nornicotine. (Smith (37) in I9?i-;":'
showed that the alkaloid in this species is almost entirely 1-norniCooitA.4 .):.
.' I


Hicks lIb) in 1940 discussed reasons for the confusion existing
over the nature of tha alkaloid of Duboisia ho.woodii. He pointed out
that earlier workers had probably unknowingly worked on mixture of
D. hopwoodii and Ilicotiana excelsior and had therefore found nicotine,
whereas in fact the alkaloid of D. hopwoodii is nornicotine.


*- '* .*


";' =.
...! :i


A :.. .


Markwood (26, 27) in 194O discovered that a certain strain of"
ordinary tobacco, N. tabacum, groom in Maryland, contained a high per- ..ii
centage of nornicotine. It wras estimated that approximately 95 percent "_A
of the total alkaloid (the latter equal to about 0.73 percent of the 4."
leaf) was nornicotine and the remaining 5 percent was nicotine. The .J:.
melting point of the picrate of the unseparated alkaloid fraction war .
197-9, which was indicative of nearly pure nornicotine. After rbioval I
of nicotine, the picrate melted at 1S8.5-190. The base was methylated r
with formaldehyde and formic acid, forming nicotine, which was identified .,*,'
by the melting point of its picrate. *
S .:..'% i: .. ^ :?:
The tobacco material examined here was a low-nicotine type. 8bt :..
sequent unpublished work indicates an association of nornicotine and tow-i:.'1
nicotine types. This is in agreement with the opinion of Koenig, cited. .
by Wenusch (Q7). It is likely that other strains or types of tobacco will
also be found to harbor appreciable proportions of nornicotine. A method :
for readily recognizing a nornicotine type of tobacco has been pro- .,
liminarily worked out by the writer and awaits further confirmation before
publication. ii

n rt _. *v-.l.
Synthesis

Pictit andi Crepieux (29) in 1895 were the first to synthesize noric- "
otyrine, which is a dehydrogenated nornicotine. By distilling a mixture '
of beta-aminopyridine and mucic acid the compound N-(beta-pyridyl)pyrrole -.. I.:
was formed. This compound underwent rearrangement when heated to a low: : .
red heat, forming a low-melting compound (m. p. 72), a C-(bet&-pyrids1)Y; : .:
pyrrole, which was regarded as. alpha-(beta-pyridyl)pyrrole(nornicotyvine) :;!
The monopicrate, m. p. 1620; chloroplatinate (C0H N .HOl) pPtCl 4H2 O,d- \d
composing at 150; a mercury compound, m. p. 17.6; and the. 1ethiodiAe, V -
m. p. 170-1, were described. 4 .. "
*. j ". ... .' .:* / '*,:.* ' "'." iiB..


...,* .**:' '.. .-j 'ilfi *"-,i=g


^ ^ .. .


. **.. ^ .na.S1; R


" j





11 -

pyrrole, starting with alpha-aminopyridine and mucic acid, which were
reacted In the presence of Al 0 as a contact substance for splitting
out H20. By passing the N-(alp a-pyridyl)pyrrole through a slightly
glowing tube they effected rearrangement into an isomeric C-(alpha-
pyrid yl)pyrrole, which they identified as alpha-(alpha'-pyridyl)pyrrole
(alpha-nornicotyrine), m. p. 87-8.

Wibaut and Dingemanse (49) in 1923 followed along the line des-
cribed by Pictet and Crepieux (29) but started with alpha-aminopyridine.
The product obtained by reaction with mucic acid, and subsequent thermal
.rearrangement of the N-(alpha-pyridyl)pyrrole, was a mixture of two.
isomeric C-(alpha-pyridyl)pyrroles; the main product had the m. p. 90,
while the other isomer had the m. p. 132. One of these compounds was
alpha-nornicotyrine, but which one was not then determined. The compound
of 900 m. p. was identical with the pyrrole (m. p. 87-8) reported by
Chichibabin and Bylinkin (6).

Wibaut (48) in 1926 established the structure of the two isomeric
C-(alpha-pyridyl)pyrroles prev': ousiy prepared by Wibaut and Dingemanse
(49). By the reaction of ethyl piwlylacotate, chloroacetaldehyde, and
NH- was obtained a C-(alpha-n 'riC.yl)pyrrole which could have only the
alpha-alpha structure. This pyrrolo, m. p. 37.5-U9.2, was identical
with the pyrrole of 90 m. p. of Wibaut and Dingemanse, and hence the
latter compound was alpha-nornicotyrine (alpha-(alpha'-pyridyl)pyrrole).
SThe other pyrrole (m. p. 1320) was therefore beta-(alpha'-pyridyl)pyrrole.

M. and M. Polonovski (31) in 1927 described a general method for
dealkylating tertiary amines to secondary amines, chief among such being
the tropine alkaloids. The method was then extended to include nicotine,
whereby nornicotine was claimed to be formed (32, 33). The method con-
sisted in treating nicotine oxide (the "oxynicotine" of Pinner and
Volffenstein, Ber. 24, 61-7, 1891) with either acetic or benzoic an-
hydrides to. form the corresponding acyl derivative, which was then
saponified, preferably with alcoholic KOH, to nornicotine. The inter-
mediates, acetyl-nornicotine and benzoyl-nornicotine, were thick oils.
Nornicotine, best purified as the chloroaurate, w.ras an oil, very soluble
in water, from which it wvras separated by strong alkali, and was readily
extracted by ether. The freshly distilled base, b. p. 150-155 at 30 mm.,
was colorless, but rapidly darkened and resinified in air; it was
difficultly volatile with steam, of an odor recalling nicotine. It had
the specific rotation, alpha] = -20 (C = 3.8, in methanol). They
prepared the dipicrate, m. p. 1350; chloroaurate, CHI12IN2.2HCl-2AuCI3,
m. p. 210-2; nitroso derivative, an oil extractable with ether; and an
oily phenyl isocyanate derivative. From later, more trustworthy work,
it is seen that the nornicotine obtained here was at best very impure.

Von Braun and Weissbach (5) in 1930 confirmed the inadequacy of
the Polonovski procedure, and proceeded along a similar line with better
results.

S They reacted nicotine with benzoic or with hydrocinnamic acids
(the latter was preferable), thus obtaining the acyl derivative, which
was then Maponified to nornicotine. A secondary reaction in the acyla-
.-tion resulted i the opening of the pyrrolidine ring with the formation
; bf a metaico e erivative. By treatment of the crude reaction product
O.l "
:,L :: ... j. : .:::, .. .., ... q .
.... ..





... .. : r': :. ::': i
with HBr a separation wars WfM4ei, &ndat the smaN ti t S
cinnamoyl-nornicotine was hydilrted to nornicotia&,*rbtehvh&( )91^
moved in pure condition with ethelr the yield was abou, 20. p
based on hydrocinnamic acid, but less on the nicotine, hia'hnCBiCt
in excess. ..:. ....
The nornicotine obtained had the following properties: 3as, ''
266-7 at atmospheric pressure; 139-40 at 12 ann. It was colorless; 464 .
stable in air (no darkening or resinifying as with the Polonoc i,::. *;.:'..
product), miscible with water, strongly basic, of pxperidine-lik.' Odb.I.4.
Density, d9 = 1.o044. Specific rotation, [alpba] -5.5, ,but t .is"" -"..l
value was believed to be lower than the 'maximam rotation because O. ..:::' ::'. .
partial racemization. The picrate was obtained fairly, pure'by .re-. '.. ,,.+..
crystallization from alcohol; m. p. 18S-90. Purer than the picrat..o'..." .
was the finely crystalline picrolonate, from alcohol; m. p. 239-4. O'. ,7 .'i
(sharp). The hydrochloride oould be obtained only as an oil The .: ..,
chloroplatinate was obtained as small, dark-red crystals, darkening 1 :".2
at 270 and decomposing at 295. The chloroaurate was obtained as ^
yellow crystals, m. p. 217. : k'
; . : -y
Nitroso-nornicotine was readily formed. by reaction of the base .r
with NaNO2 in HOl1 solution; it was a yellow, viscous oil of b.p. ,
190-2 at 0.5 mm., miscible with water but separated therefrom by ,
saturation with K2O0 and extracting with ether, or better, methylene ,
chloride. The picrate, HCol salt, and Au salt were oily. The Pt salt ..
formed fine crystals of m. p. 190. The monomethiodide was well A"
crystallized, m. p. 144. The nitroso derivative was converted into ,
nornicotine by treatment with excess of strong HC1. By nitriting a .
mixture of nicotine and nornicotine a separation of these tNo bases., :
can be effected, thus: The product is nitrite, then fractionated, .
whereby the lower-boiling unchanged nicotine is separated, and the ;
nitroso-nornicotine is converted as above. [Such conversion and regeniea-
tion of nornicotine, however, probably results in partial racemizatioa. )i

Nornicotine readily formed an acetyl derivative with acetic an". ,
hydride. This derivative was a viscous oil; b. p. 212-40 at 12 m. I: t
specific rotation, [alpha]20 = -3.24'0..(in benzene). It formed .i- '
picrate, m. p. 151; an oily Au salt; a Pt salt which decomposed at
245; and a methiodide, colorless needles, m. p. 201. i
*' : : i: ]
Nornicotine formed a colorless, crystalline carbamide with cyVni6 'b
acid, which was nornicotinyl carbamide, C H ON m. p. 164-6. Its :
Pt salt, small red crystals, darkened atl256 and decomposed at 270Q i
The pirate was oily. Nornicotine also formed a colorless-, crstallie s
derivative with phenyl isothiocyanate, of composition C0IH17N3S and m. p :
1710.
:A
By the action of ethyl iodide on nornicotine, Nethyl-nornictia ..
was made, of b. p. 127-8 at 12 mm. The HC1 salt was a hygroscopic T'V
solid; the Au salt, yellow crystals which decomposed at 203f, the ..:'
picrate, m. p. 174-6'. By sitlar treatment of nornicotine with allY! ,.. .
bromide was formed N-allyl-nornicotine, a colorless liquid, b. p. :
136-7 at 12mm. It formed a picrate, s. p..l80-20;.A AU.lt, deowO r .'."
ing at 145 -; a Pt salt, orange crystals, darken at 23Q a -d doi. !|3
Scomposing at 2550. ,

.. ...
.... "* ..... ,. ...... : : ";, "" _':. .:::::.c' ...... -;: .L.... L "2 ': :-" "' .:-'': '" = ",







.#p...i: '.path, Marion:, and ZajiQc (45) in 1936 effected the demethylation.
S Of nicotine into noaricotirie ii two ways. A. By .,treatment of nicotine
*i:t: .... itiM a 0in aqueeus, solution & partial demethylation occurred..
fthe cr46 mixed basip product was dissolved in ether and fractionally
.',extracted with 0.1 1 0 saturated with NaC1, whereby the.nornicotine
ya ws removed with only a small admixture of nicotine. The nicotine was
,. ',then expelled by vacuum distillation with water. The crude product was
S purified by repeated recrystallization of the picrate, which was finally
;. obtained pure with a melting point of 191. The pirate was decomposed
with HCi, the free base recovered and distilled. There was thus obtained,
: from 6.6 g. of nicotine, 0.41 g. of nornicotine. Specific rotation,
:' ;[alpha)0 s-76.1; by crystallization as the perchlorate the rotation was
Increased t0o Calpha = 832L. [Calculates to a density, d20 = 1.070.]
T? T he picrolonate, m- p. 252, and the trinitro-m-cresolate, m. p. 200 were
i: made, S. The second method employed by these workers for demethylating
nicotine consisted in treatment with silver oxide. This reaction dates
back to Blau (Ber. 27, 2535-9, 194), who thereby prepared nicotyrine, a
S 4ehydrogenation product of nicotine, but did not report the formation of
S nornicotine. The crude reaction product was fractionated as before.
P From 40 g. of nicotine there was obtained 3.9 g. of nornicotine di-
l picrate, m. p. 190-191. Before conversion to picrate the base had a
f, specific rotation of '40; after purification via the picrate it showed
Saipa] =-Sg.8 (djC = 1.070).
by Craig (7) in 1933 synthesized nornicotine, and from it nicotine,
', by a series of.aggregating steps starting with pyridine. Hence this
Ic method, while lengthy, represents more truly a synthesis than does the
demethylation of nicotine. Pyridine v'as sulfonated to sodium beta-
j:." pyridine sulfonate and the latter converted with NaCN to nicotinic acid
Snitrile. The nitrile was then reacted with the Grignard reagent made
; from gamma-bromo-propyl ethyl ether, whereby was formed beta-pyridyl-
gamma-ethoxypropyl ketone, which was a new compou-na. From this ketone
S was formed the oxime with hydroxylamine. The oxime was reduced to the
:; amine, l-[beta-pyridylj-l-amino- 4-ethoxybutane, which was dealkylated
| with HBr to the hydroxy compound. Ring closure to make the pyrrolidine
tV, ring followed, and by addition of KOH a basic oil separated from which
Swas obtained an oil distilling at 139-l40 (12 mm.). The properties of
This oil agreed. throughout with those reported by von Braun and Weissbach
S' '(5) for nernicotine, and similarly the picrate and phenylthiourea com-
', pounds were in agreement. Methylation with CH3I to racemic nicotine
followed. The author noted that nornicotine is a stronger base than
: .nicotine. The foregoing process' represented a complete new synthesis
S from.pyridine of the two bases in racemic form.
*..,, '.. Craig (s) in 1934 by a similar procedure prepared so-called alpha-
S nornicotine and alpha-nicotine, in which linkage to the pyridine ring is
S in,the alpha.position instead of the beta position. The alpha-nornicotine
l.;,,i'..WVas described as a rather pleasant-smelling oil, soluble in water and in
O|;.rgadie solvents in all proportions. The picrate melted at 1660. A
l. .phepylthiourea ,.derivative was made but could not be obtained pure.

;::.. .Spath. and Kesztler (41) in 1936 resolved synthetic dl-nornicotine
0.. d.i, by the method of Craig (9) into the two optical antipodes.
.i.ettCoh (.. 37, 1225-35,y 1904) were able to resolve ,,;
A ~ :.;'"=,' i ,". 7 ..,.,,., .. ,
:a.41 S.. " "" " ":
a"s.t" : ..:: ,;::: .. ,..:: ., .::, .. : .e,:. ., ,. .?.. :,:.. .....,. : ,: .. .. .. :.: .. ., ., @ .,. :,''. ,' ,. : .t :. -









IS resolution into aana I-rormlibrouqfqj, U.quf leSa4i p
cussed. The dll-nornicatine was first treMted in metbanol. soA`U4 ......
l-6,6'-dinitro-2,2'-diphenio aqid; 'i ylgw. of l-norniZomi!nk p"'.
d.initrod.iphenate wqre obtained. The. base recoveredd ftO the
decomposition with HOI1 first had a rotation1. C alph ;' .-a'=.-4,
purification through the peroblorate it. heA. "taphabLl 7-
mother liquor, on treatment with the d-diphden ea.IR, a %ve:&zt .3# ^Sp.
d-nornicoqine-d-dinitrodiphenate, .the recoteredt base of whic- : f .f,
a rotation, Ealpha]7 +4s.o, and after, purification as the pehrcASt4
had [salphah8 = +86.08'. A ..-..*'A.
S .. .. ... ..
. . ...., ; .. . . .
For 1-nornicotine diperchlorate eo prepared, 0H91O820O12, aJ""'... '"
1853-6, [alpha]j8 = +13.6 (in water;. c = 5.28) For the' diper9hlot of natural 1-nornicotine, [alpha])8 +13.4 (in water, c 5.23) Z "':t;A
d-nornicotine diperchlbrate, [alpha = -12.98 (in water, .- 5.a .''.

Spath and Kainrath (40) in 1935, in their work. on the pimplifla..10
tion of the classical Pictet nicotine synthesis, prepared the coqipout :I i
1-(3'-pyridyl)pyrrole from 3-aminopyridine and music acid, Thispyrroile*,'...
by thermal treatment at 700 rearranged into two isomeric pyrroles,, of (
which one was 2-(3'-pyridyl)pyrrole (nornicotyrine), m. p. 97 (piirato,.i: .':.
m. p. 200-2); the dther isomer was 3-(3'-pyridyl)pyrrole. Separatio -:.;..
was effected by repeated crystallization of the free bases and their ;.i
pirates. NornicotSTine was reduced witt hydrogen'and Pd-sponge as ,
catalyst to dl-nornicotine (dipicrate, m. p. 194). The crude hydct- '"": ;...
genation product was also methylated with formaldehyde and formic aci :. .
dl-nicotine. '".i'i
.- ' "/ ; ".'" 4 :' p
Lions and Ritchie (22) in 1940 published a new synthesis of nor" ..
nicottrine, 2-(3'-pyridyl)pyrrole, in order definitely to confirm its 2..*3|
constitution. It was synthesized from ethyl nicotinylacetat "e, : "::
NCO5HICOCH20C02C2H, ethoxy ethylenedichloride, 0H20C1RClO2Hr, and Gtriql"
NH3. The purified product, nornicotyrine, was isolated in the form af..":.,
needles, m. p. 98-9, which gave a blue fluorescence in a mi:tlre :i.,
benzene and petroleum ether (picrate, m. p. 203-4 (d.)). Whis type o*tf.
synthesis, involving NH5, represents a departure from most former .mo.e. '.ik
3~ ~~~ rereensa eprtr
but is similar to the synthesis of alpha-nornicotyrine, employed by .,4
Vibsat (4s) in 1926. '
' .'" ..(
-.:..R ILI:

Analytical Determination .-1. T"":
Qualitative tests for nornicotine are. based upon iuch factor asi.,..-,V.M
the appearance and melting point of salts, ea. g., picrate 191-2, e4.'.& '..
trinitro-m-cresolate 200, picrolonate 252, urea 167-70, phenrWlt4iota4
176-7, and Qhloroaurate 217, :':A.:
; i "' "'* :'' i B
Kethylation to nicotine, .with identification of the.iattor .jf.. ;A
valuableaevidence of notnicottne, .. ,.:.;.,..,

e J,.1.A. : :" A : .
.,. .. 3 .. ..
.. .A'.'>:. f l.:t.: A A.:d a :'.:


A:


ii
"::..: ::' '


i l;'"." :


".:, .%






- 15 -


The optical rotation and other physical properties are also
valuable means of identification.

The quantitative determination of nornicotine has not been fully
iwestigated. It may be said, however, that any methods applicable to
nicotine (with due regard to the difference in volatility with steam)
are also applicable to nornicotine. The chief problem most likely to
be encountered is the quantitative separation of nornicotine and nicotine
as they occur in tobacco.

Von Braun and Weissbach (5) in 1930 pointed out that a separation
of nornicotine from nicotine could be effected by forming the nitroso
derivative of nornicotine with KN02 in HC1 solution; nicotine was not
changed thereby. They then fractionally distilled the isolated bases
and obtained nicotine as the lower-boiling fraction. [This method may
be satisfactory for a quantitative determination of nornicotine, but it
is open to objection in the preparation of 1-nornicotine because sub-
sequent regeneration of nornicotine from the nitroso compound by means
of strong HC1 may result in racemization.]

P. Koenig (19) in 1934 observed that the distillates of certain
tobaccos gave pirates of melting point departing considerably from that
for pure nicotine (218-222.5). These pirates melted at 214, 205,
193, and even 173; the depression in melting point was attributed to
foreign bodies, of which the chief one was nornicotine. He described a
method for aquantitatively separating these bases, by virtue of their
difference in basicity and in volatility with steam. A distillation
from MgO solution yielded only the nicotine in the distillate, while the
NaOH distillation yielded both alkaloids. This was explained by the
fact that nornicotine, being the stronger base, was less readLily
liberated from its salts, and also that it was less volatile with steam.
The bases in the respective distillates were precipitated with -icric
acid, and the picric acid content of the pirates was determined by
titration with NaOH. [Note: This separation is only approximate, and
it favors a high value for nicotine, as nornicotine is volatile enough
to distil over partially from MgO solution.]


Pharmacology

Dingemanse, Laqueur, and Wibaut (9) in 1926 reported on the
pharmacology of certain pyridine and pyrrole derivatives. An-ong them
was alpha-nornicotyrine (alpha-(alpha'-pyridyl)pyrrole), m. o. 900. A
frog injected with 1 cc. of a 1-percent solution of alpha-nornicoty'rine
showed symptoms of paralysis. As an anesthetic this compound was about
twice as active as novocaine. The general conclusion reached from this
and other tests was that the alpha-pyridine compounds have about the
same degree of physiological activity as the beta-pyridine compounds.
(This conclusion, however, is not borne out by the tests of other in-
vestigators.)

The first information concerning the toxicity of nornicotine
comes from-A. Bergwall, whose report of pharmacological tests was cited
by Ehrenstein () 1931, 'It was the latter's nornicotine, isolated
,from tobacco, Ich fWas tested, and this it should be noted was not
~~A ... ..;. .




~. ..:: ......


pure 1-nornicotine but a-mixture of-the 1- and dl-forms. (approxz3int4 '
20 percent 1- and 80 percent dl-nornicotine). .....:....

Tests on the contraction of frog muscle showed nornicotine top-.l;?
have only one-tenth the activity of nicotine. Blood pressure 'tests .onf
cats (injection in the Vena jugularia) showed that nornicot-ine,. Z.--.
nicotine, causes a rise in arterial pressure which is accoarpnie& :1W .,.::...
irregular heart action, but the effect was weaker than that produced. ."
by nicotine. It was difficult to give numerical evaluation to tou t-s::.-.':: :A.
of this kind, as repeated injections diminished the effects. Nor-. ..:il
nicotyrine (2-[beta-pyridyl]-pyrrole), .on the other hand, caused a "'..."..
lowering of blood pressure, but the action was relatively weak. :

Macht and Davis (23) in 1934 reported toxicity tests on the ......
following compounds, which were prepared by L. 0. Craig: "'*'

1-beta-nicotine ;.i
dl-beta-nicotine :...
dl-beta-nornicotine :J:,
dl-alpha-nicotine ,!
dl-alpha-nornicotine ; 3
(l-beta-Nornicotine, the form naturally occurring in'tobacco, ....
was not included.) =

The tests embraced growth studies on seedlings of Lupinus i
albus L., and mortality studies on tadpoles, goldfish,'water turtles, .1
land turtles, white mice, white rats, guinea pigs, and cats.

All five compounds were toxic, but the order of toxicity varied ::
with the different test objects. In nearly all cases 1-beta-nicotine *.^
(natural nicotine) was the most toxic. The beta compounds without ex- :
ception were more toxic than the alpha. In general dl-beta-nornicotlne.
was more toxic than dl-beta-nicotine, but there were exceptions. As +"'j
between dl-alpha-nicotine and dl-alpha-nornicotine, the order of toxicoity ...
was about equally divided. ;

A typical picture of toxicity, showing the effect of intravenous -
injection of cats, in terms of mg. of compound per kg, of cat for lethal ',
dose, is given in the following set of values (the order of compounds is
that given above): 1.3, 2.0, 0.9, 6.1, 13.1. .

Macht and Davis (24) in 1935 made a special report on the toxicity' :
of the several compounds tested previously (23). The test here was con- .i
fined to inhibition of root growth of seedlings of Lupinus albus. The I |
data and conclusions of the previous article were substantiated. In *"
addition, new data were given on the effect of combinations of these ,*"i"
bases. The results showed that certain combinations gave simple additivet .'
effects, agreeing with the calculated summation, while others produced a ',L
synergistic (greater than the calculated) effect, and still other corn- .i
binations were antagonistic.

Hicks and LeMessurier (18) in 1935 investigated the toxioological, =1
behavior of the alkaloid .of Duboisia hopwoodil, whih was nornlcotine, 'Y.J
roughly in 1:1 ratio of the d- and dl-formps. The l. 8. for rats ic"'i
(peritoneal injection) was 0.0017 gg; the .. 1. n ie wa :


>.:.,, ,, ,5ii g'i: .S .,, .









[' s o31cotine. The action on the isolated perfused. mammalian heart was
u ". imiiar to.'that of nicotine. Intravenous injection into the intact
il' i. uai al" (rabbit) caused a rise of blood pressure similar to that given
t'h"reInaline, followed by paralysis of the vague. This effect was
sfmiiar td that .prodfue& by stimulation of the splanchnic ganglia by
Saiipotine. The actior'on isolated frog's gastrocnemius preparation
m mwa similar to that of nicotine. It was noted that the symptoms of
poisoning in the rat were distinct from those produced by nicotine,
I:' "'although death in'both cases was due to respiratory failure, The'
- mmm' Dubom ia plant is also. known to be very texic to camels, as one mouth-
S;- till of the bush is fatal to these animals.

SHicks, lrdacke, and Hueber (17) in 1935 reported more completely
,'1. on the pharmacology of d-nornicotine, which was isolated- from Duboisia
m holpwooii. libood-pressure experiments on cats were conducted, and the
.' action on-'eripheral ganglia was studied, both with reference to heart
i.' q-anglia (isolated frog heart) and to the lion cervicale supremum
c''' (oat)'. The action on Isolated frog muscle. musc. rectus abdominis)
SI/ and. on the posterior extremities of a dog were also studied. The con-
cl., .,.usions reached were that the action of d-nornicotine on blood press
respiration, and vasomotor centers, .as well as on peripheral vagal and
sympathetic ganglia and on the heart, is not different from that of
i nicotine. The isolated frog muscle was somewhat more sensitive to d-
... nomnicotinethan to nicotine, but the difference was only slight.

Smm Riclhardson, Craig, and Hansberry (34) in 1936 summarized the
-'' toxic action of a group of N-heterocyclic compounds upon the bean aphi
Aphis rumicis L. The compounds were applied in 0.25 percent sodium
I oleate solution. The concentrations required for 50 and 100 percent
; mortalities of the insect were:
? ' ..
m 50 percent 100 percent
Compound net mortality net mortal
S~u~m' g. per 100 cc. mg. per 100 cc


re,






d,




t
ty


.V 1-beta-nicrotine 49 1,185
di' -beta-nornicot ine 145 490
dl-alpha-nicotine 1,496 10,960
Sdl-alpha-nornicotine l,514 10,3470
i-. dl-beta-nicotine 96 1,259
S' anabasine 5 166

-, m m The results show the marked superiority of the beta compounds.
m Anabasine stands out as the most toxic aphicide. There was no great
difference between the comparable methyl- and nor-forms. Natural
19m m i .,l nicotine Was about twice as toxic as its racemic isomer. Although
Ii :l-beta-.nornicotine, the alkaloid occurring in tobacco, was not in-
S eluded in this study, the racemic form of beta-nornicotine was shown
... to, ben somewhat more toxic than natural nicotine, and hence 1-beta-
llIn^ otine quite likely would have a toxicity of the same order. It
hbuici m be morne .in mind that there results ported here apply only to
PeO aphidr and that the degree and. order of toxicity can be different .
e Useetmmm . Thi$"ork -Q eprepresenZte the. first insecticide investi- ... im
,...... ... ..y i .. JAi.:i.,, ... .*..,. .
,," ...W e,..e.:J ;",,:, '..':,"% %" . " r : "
ilk m~ m~m' I,%: .. ."r'. .m








Wenusch (47) In 193b prtblishedkAv intereogtia-catawvtp
toxicity of 1-nornic6tine as it rblateB t. the. Mekins'
Cigarettes made of a nornicotize' tobacco were smoke4 'btSr
puffs, and the smoke was caught in &i.ute alfuc ai6d.. J
alkaline liquid was exhaustively steam-distilledlM ah&d t h. i
the alkaloid was formed in the distillate. Since noraicotcen0&4
is more soluble in water and in alcohol tha&'is nicotine. p1 i
was possible to effect a'separation of the's two bases by
crystallization of the pirates. Compared with the -.eamomuat: e
:in the original tobacco, the amount in the main tmoke stream ni'
small; about 25 percent of nicotine passes into the samoke.by
mittent smoking but the corresponding percentage of nornicoti Bi:m. VXAPX
much less. :
t In the soluble picrate fraction, beside one of a p 2,
there was obtained a picrate that blackened at 150, sublimed: t
melted at 192-4, and when made alkaline smelled strongly 1ike m' s
(mousy. odor) but was not myosmine4 A picrate of the same propt
; *was prepared from nornicotine by EMn04'oxidation.7 Since thfl .e.-iA
could not be obtained directly from tobacco, it must bwe its orpgi4 4
air-oxidation of nornic6tine on smoking. It appeared to be micii 4ItdN:Vl
toxic than nicotine. ''':...'.
:. ...:B
Sl-Nornicotine was physiologically less active than l-nico ,\"' ..
as revealed by tests on the contraction of leech muscle. A sol1ti*^,:l
of 1:300,000 of 1-nlcotine produced approximately the same derre'ett
contraction as a 1:60,000 solution of lr-nornic6tt'le, hence the ,
was roughly one-fifth. k.0 .
~. 17 :.4:,:
SThe conclusion reached was that the presence of 1-nernlot.
.., tobacco is toxicologically of minor importance since it is lessti
'.. than nicotine and passes over into the smoke to a much less d.egree:l..
<.: nicotine. .. -.,
Hansberry and Norton (14) in 19O40 reported on aphicidal (i
rumicis) tests with 1- and d-nicotines and 1-, d-, and dl-norniQotiit
: ... The 1-nicotine was obtained pure by the method of Batz (Monath, 2i&ib :"
1241-52, 1905); [alpha] =-16S.90. The d-nicotine was cryst4al4 iuf lg
the 1-tartrate from racemized nicotine, prepared by heating naturst.v"ii
nicotine with H2;04. The 1-nornicotine was isolated from..' Bilv-, ato4iS
grown at Ithaca, N. Y.; the plant contained approximately 1.4 pero .....
crude alkaloids, of which 57 percent was nicotine and 43 parpent A.rvii.
,;. ~ nicotine. The purified 1-nornicotine had a specific rotatios'n,'..'!
S[alpha] = -82. The d-nornicotine was the base prepared from AAub i'4
'hopwoodii and was furnished by C. S. Hicks of Australia. The dl- M'..
nornicotlne was prepared from nicotine by the method of "ln Braun 3ia
WVelssbach (5); the rotation was not determined; the pirate melt "'mti.A
191-191.50. '
4. .. 0
'The compounds were applied in 0.25 percent scrdtd 0 ,
tim. The remslts are fhown in the following table .

710r, .: .' r i t: : .L:':'. :Jr
.. 4. ,~ ... :*..":..". / ,. :, : -,r' v!: i :. .. "..I: : .
... ... 2. .:. r~A.... *1: .... .









.l *..?."Material


.1 nicotine
d ..:-.nicotine
0 '.... n.1-ornicotine
*, -' / 'd.- nornicotine
i ; : d-nornicotlne
I ;;"-' : *


Concentrat ions used,
0.005 0.02 0.05


percent
0.2

.87
145
100
100
100


6.4_ 2.0


100
90


100


It is apparent that d-nicotine was substantially less toxic'
than any.of the other alkaloids. All the nornicotines were more
S.toxic than natural (I-) nicotine. 1-Nornicotine was doubtfully more
toxic than the d--or dl-forms. No pronounced difference existed be-
tween the d- and 1-forms of nornicotine. The data indicate that ex-
tracts of Nicotiana silvestris and Duboisia hopwoodii may be better
insecticides than nicotine, toward some insects.

The results of the tests with aphids indicate that nornicotine
should be tested as an insecticide against other species. It may
also have other uses, such as a vermifuge for poultry and other
Animals, especially if the nornicotine, when used for this purpose,
proves to be less toxic to the host than is nicotine.


Patents

U. S. patent 2,219,287, dated October 29, 1940, to Robert B.
Arnold (4), assignor to Tobacco By-Products and Chemical Corporation,
specifies the group anabasine,. nornicotine, and nicotine in combina-
tion with a base-exchange polysilicate, as a parasiticidal spray com-
position.


Reviews and Popular Accounts

Marion (25) in 1938 presented a review of the tobacco alka-
loids, which included a discussion of nornicotine.


Spath and Kuffner (44) in 1939 presented a comprehensive review
on the entire subject of tobacco alkaloids, of which nornicotine is one.

Popular accounts of Markwood's discovery of nornicotine in Mary-
land tobacco appeared in Science News Letter (3), Science Digest (2),
Modern Medicine (i), and in newspapers, such as the New York Times
(Sept. 8, 1940) and Buffalo Evening News (Oct. 16, 1940).


Summary


*NorAicotine is a colorless liquid alkaloid which occurs in the.
2a1 -speies tabacum L. and silvestris Speg. and Comes, and in
1:hoapwodii F. Muell. It probably also occurs in other incom- ;.
t ..j ivws-tigatteL :spOcies of Niootiana. In chemical composition it '.

.. .. ......
.. -,' .:. 0i . .. . .


iT


" ,:. *


r..
:'. .... ..
i1 '. .


*-1"
if. *.'
*"* i,, .
: *' *


4"
V

-
*


,44
1*









** .....* ..y ,j
Nornicotine appears to be more stable than nic'ot ME.ie '
higher boiling point, and is less volatile with steam. >:*

Pharmacological tests indicate that in general it Ai4' :.:-
than nicotine toward warm-blooded animals, ":it that to.wardin:7
judged by behavior with A lhis ruaicis L.) it ie equally tozip f
even superior. This conclusion is only provisional, however, :; ..
few insecticidal tests have been made. 1'

The prospects for a supply of nornicotine are excellent t"4
a fairly rich source of it has been found ih. a domestic tobacco'. 1i
the insecticidal point of view it deserves thorough testing by bto
mologists. It may alsb have other applications, such as..a vermaiftig
for poultry and animals. "


Literature Cited


1. ANONYMOUS
1941.


../ k
'~~~~ : ***^
.. : .* ,, :y
...: .. . :4



It~
," b 4.
.41:2461~ :
r :.'. .,.. :1 ,


Modern Medicine. January 1941. Page' 84.


1940. Science Digest. Nov. 1940. Page 66.


1940. Science News Letter. Aug. 30, 1940. Page 204.


3a. ANISRONI,
1879.


T.. I:. ;


'~~ el^
*.. .l: : ,:,,-

,* .. .:: .; :


G.
Nicotine. Gazi. Chim. Ital. 9: 169-73.


4, ARNOLD, Rt. 3. :* LI
1940. Insecticide and process of making the same. United Stai*
Patent 2,219,87 (Octi 29, 1940; appl..Jan, 22, 2.193J),A::-';

5. BEAUN, J. von, and WZISSBAOH, K. *,':..
1930. Dealkylation of tertiary amines by organic acids,. 11-:.j,'
Nicotine. Ber. Deut. Chem. Gesell. 63B; 2018-26. f


a.
It
N'





U;


r .f
1.' *.


6. CHICHIBABIN, A.. E., and BYLIINKIN, J. G. ;
1923. Alpha-pyridylpyrroles. Ber. Dent. Chem. GeaeL1' ;563.
1745-9.
,.."f . .; 1 ...
7. CRAIG, L. C. ,:
1933. A new synthesis of nornicotine and nicotine. Jotu '41
Chem. Soc. 55' 2854-7.
r~~ :"". *^:

1934. Synthesis of alpha-nicotine and alpha-norn'ctt .if =U
Amer. Chem. Soc. 56; 1144-7.' ,,,.

J:.. .
".:..).. .. ..I' 'a.;: 3 a


I


, ..:




4 4-
4)
I.


- 21 -


9. DINGEMANSE, Z., LAQUE UR, E., and WIBAUT, J. P.
1926. The pharmacological properties of alpha-aminopyridine
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10. EHRENSTEIN, M.
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11.


1928.


Two new alkaloids from tobacco. Naturwissenschaften 16
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12.
1930. The newer development of the chemistry and biochemistry
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V.
13.- -- -
W 1931. Alkaloids of tobacco. Arch. Pharm. 269: 627-59.
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A 1940. Toxicities of optically active nicotines and nornicotines
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15. HICKS, C. S.
;; 1936. Further observations on the chemistry of d-nornicotine.
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16. HICKS C. S.
1940. The chemistry of the alkaloid of Duboisia hopwoodii.
t Australian Jour. Sci. 11: 110-12.

,: 17. HICKS, C. S., BRUCKE, F. T., and HUEBER, E. F.
1935. The pharmacology of Duboisia hopwoodii (d-nornicotine).
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S 18. -------- and LeMESSURIER, H.
S1935. Preliminary observations on the' chemistry and pharmacology
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19. KOEMIG, P.
1934. Tobacco. In Handbuch der Lebensmittelchemie, v. 6, pp.
296-8. Berlin.

20. KOSTOFF, D., and SARANA, M.
S1939. Heritable variations in Nicotiana tabacum L. induced by
abnormal temperatures and their evolutionary sig-
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ic.: 21. KOVALEbKO, E. I.
".1. :, ': .1937. Alkaloids of Nicotiana rusbyi and N. silvestris. Vsesoyuz.
..:;... T
Inst. Tabach. i Makhoroch. Prom. No. 133: 39-46.
.' .






-22- : 'WI "
22. LIONS, F., and RITCHIE, E. ,
1940. A new synthesis of nornicotyrine,. an' of itso g 'oa.:.
analog. Roy. Soc. N. S. -Wales, Jour. and Prod qt'.a.....
74: 110-16. '
........................ ."......."..."..... ",
.. . ,; :, .' ,
23. MACHT, D. I., and DAVIS, M. E. '' "
1934. Toxicity of alpha- and beta-nicotines and nornicotin9;m. <:. 0
Jour. Pharmacol. and Sxper. Therap. 50: 93-9 .: 1i
24. MACHT, D. I., and. DAVIS, M. E. i
1935. Toxicity of alpha- and beta-nicotines and nornicotines: i
for Lupinus albus. Amer. Jour. Botany 22: 329-32.... L.

25. MARION, L. ..
1938. Alkaloids of tobacco. Rev. Trimestr. Canad. 24: 70-81 4.
26. MALKWOOD, L. N. .
1940. Nornicotine as the predominating alkaloid in certain 'Pt
tobaccos. Science 92: 204-5. "
27. --- -
1940. Determination of nicotine in fresh tobacco leaf. Jour. : il
Assoc. Official Agr. Chem.: 23: 80O4-10.

28. NOGA, E. :
1914. The alkaloids in tobacco extract. Fachl. Mitt. Osterr. A,
Tabakregie, Nos. 1 and 2. .i:
29. PIOTET, A., and CREPIBtX, P.
1895. Phenyl- and pyridyl-pyrroles and the constitution of .:.i.
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30 -------- and ROTSCHY, A.
1901. New alkaloids of tobacco. Ber. Deut. Ohem. Gesell. 34): ..P .
696-708.

31. POLONOVSKI, M. and M...
1927. A new method for converting tertiary heterocyclic bases .'i
into secondary (dealkylated) bases. Acad. des Sci. -
Compt. Rend. 184: 331-3. ...
32. POLONOVSKI, M. and M. 0- '.
1927. Beta-pyridyl-alpha-pyrrolidine (nornicotine). Acad. den, :'y: R|f
Sci. Compt. Rend. 184: 1333-5. -

33 -.---...-
1927. Amine oxides of the alkaloids. III. Action of anhydriteu-.!.j
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34. RICHARDSON, C. H., CRAIG, L. C., and HAITSBERY, T. R. ..'*'
1936. Toxic action of nicotine, nornicotines and anabaslne S ;{,
upon Aphis rumicis L. Jour. Scoa..Snt. 219, ( 50 Ogi5
S :"i
..*.. ,






:I ;: ,. -"
,,F. .:iiI.,:
-23-

35. SHMUCK, A.
1934. Researches on the chemistry of tobacco. 7.- The alka-
..... loids contained in some iT.ic'tJana species. Vsesoyuz
Inst. Tabach. i Makhoro-h. Prom. No. 109: 24-39.
36. SMITH, 0. R.
1935. Occurrence of anabasine in Nicotiana glauca R. Grab.
(Solanaceae). Jour. Amer. Chem. Soc. 57: 959.-6o.

37. --
1937. Occurrence of 1-nornicotine in Nicotiana silvestris.
Jour. Econ. Ent. 30: 724-7.
38. SPATH, E., HICKS, C. S., and ZAJIC, E.
1935. d-NTornicotine, an alkaloid of Duboisia hopwoodii F.
:: v. Muell. Ber. Deut. Chem. Gesell. 68B: 1388-93.
.i. .
S39. SPITH, E., HICKS, C. S., and ZAJIC, E.
: 1936. d-Nornicotine. 3er. Deut. Chem. Gesell. 693: 250-1.
40. -------- and KAINRATH, P.
..'i 1938. Tobacco alkaloids. XV. The Pictet nicotine synthesis.
,Ber. Deut. Chem. Gesell. 713: 1276-81.
-- -------- and KESZTLER, F.
1936. Tobacco alkaloids. IX. Synthesis of 1-nornicotine and
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I;' 42 ------- and KESZTLER, F.
1937. Tobacco bases. XI. 1-Anatabine, a new tobacco alkaloid.
|;".:Ber. Deut. Chem. Gesell. 70B: 239-43.
143. ------- and KESZTLER, F.
p:" 1937. Tobacco alkaloids. XII. Occurrence of dl-nornicotine,
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I. 44.- ---- and KUFFNER, F.
1:- .1939. Tobacco alkaloids. Fortschr. Chem. Org. Naturstoffe
2: 248-300.

|. 45. MARION, L., and ZAJIC, E.
l," 1936. Tobacco bases. IV. Synthesis of 1-nornicotine. Ber.
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46. SPATH, E., and ZAJIC, E.
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HGesell. 68B: 1667-70.
Jk y47. WENMUSCH, A.
1936. Nornicotine. Pharm. Zentralhalle 77: 141-3.



B"A..:k .... ,
i@ ': ..%







-ti&-.
4.


4g. WIBAIT,
1926.


09.



50.


UNIERIT OF s ULRD


.-.,, ,. . I 11111111111 i ll I III IIII III Ilii I11 1111I liiil11111I1111 III I
3 1262 09224 7724

A ".M:: .
*' ** " a*" WB
.**. '**a a


J P. ...
Synthesis of C-(alpha-pyr4idyl) -alpha-pyrrole .o.... .
structure of the isomeric O-Calpha-pyridflypyf410
and. the corresponding alpharnicotyrines. l ea. kV
Ohim. 45: 657-70. "
:.* K
"; .. I


---- and DINGDMAITSE, E.. .
1923. Synthesis of alpha-pyridylpyrroles and two isoOe 4' W
nicotyrine. oec. Tray. Ohimi. 42: 1033-49,....

-- and OVfERHOFIT, tT. *
1928. The catalytic dehydrogenation of nicotine. A suitaNA-t
method for the preparation of N-methyl-(3-pylrJdy:1Y.
2'-pyrrole (3,2'-nicotyrine). Rec. Trav. 0hilm.
477: 935-9. 4* ';:


.. .. ....... ....... .......... .
.... . '; '. .. ". ,. .." ;
^- - ..... ......... . ................... ....... .. ....::^ g. ..:,;:^. .. *....u,. L: 5s,