Journal of Eihno~ha~Qeoiogy, 7 (1983) 239-242 239

Elsevier Scientific Publishers Ireland Ltd.

Short Co~~ication

PANGAMIC ACID, A STAMINA BUILDING, ANTISTRESS AND ANTI-

HYPERLIPIDEMIC PRINCIPLE FROM CZCER ARZETZNUM L.

J. SINGH, G. HANDA, P.R. RAO and C.K. ATAL

Regional Research Laboratory, Jammu-Tawi (India)
(Accepted June 30,1982)

summary
The stamina building, antistress and antibyperllpidemic principle of
Cicer arietinum has been isolated and identified for the first time to be
pangamic acid (n-~ucon~~e~yl~ino~etic acid) by chemical and spec-
tral methods.

Introduction

Cicer ~~e~~#u~ (Bengal gram) is a very important staple food in India

due to its highly nutritive constituents i.e. proteins, carbohydrates, fats,
vitamins and minerals, etc. Exceptionally nutritive and body building proper-
ties are ascribed to it. Seeds soaked overnight in water and the sprouted
grain are both considered to be extremely nourishing and constitute a
regular item‘ of diet for athletes and professional wrestlers in India. Testa
of Bengal gram is used as a food for horses which gives them an untiring
stamina (Siddiqui, 1945; Wealth of India, 1950).
Here, we wish to report on the isolation of this stamina building principle
identified as pangamic acid (n-gluconodimethylaminoacetic acid, probable
Vitamin Bts) from Cicer wietinum. It is also useful in the treatment of
ischemic heart disease, disorders of lipid metabolism, liver dysfunction
and various types of dermatoses (Dutta and Gupta, 1978; Atal et al., 1980;
Singh et al., 1980). Pangamic acid has previously been reported to be present
in apricot kernels, rice bran, brewer’s yeast, ox blood, horse liver, barley,
maize, rice and oats, and was assigned the structure of n-gluconodimethyl-
aminoacetic acid, having the molecular formula C10H19N08, mol. wt. 281
(Krebs et al., 1951; Telegdy et al., 1969). This product has been listed in
the 8th (1968) and 9th (1976) edition of the Merck Index and the 9th

0378.8741/83/020239-004/$01.50
o 1983 Elsevier Scientific Publishers Ireland Ltd. - Published and printed in Ireland
240

edition (1976) also lists its structure as D-ghCOniC (6-bis(l-methyl-ethyl))-

amino acetate which actually is a synthetic higher homologue of pangamic
acid (n-gh.rCOnOdimethylaminOaCetiC acid) (Yurkevich et al., 1970) not
present in nature. This has also been clarified by us in a recent paper (Singh
et al., 1981) and in Singh et al. (1982) where we have identified and es-
timated pangamic acid to be present in Anacardium occidentale, Arachis
hypogea, Cajanus cajan, Glycine soja, Phaseolus aconitifolius, P. aureus,
P. mungo, P. vulgaris, Pinusgerardeana, Pisum sativum and Vigna sinensis.
In both these reports (Singh et al., 1981, 1982) we have stressed that the
continued designation of pangamic acid as a “B” vitamin is based on its
presence in common foods and on the basis of its broad spectrum of physio-
logical activities, though no deficiency disease has been attributed to it.
Therefore, its status as a true vitamin still remains unsettled.

Isolation and identification of paugamic acid from Cicer arietinum (Bengal

gram)

Bengal gram (1.2 kg) were extracted with hot water and the strained
water extract concentrated to about 5 9, adjusted to pH 4.2 with CCl,COOH,
concentrated to about 500 ml and centrifuged to remove the precipitated
proteins. The mother liquor was made to a syrupy consistency, chromato-
graphed over silica gel using ethanol/water (70 : 30) as an eluant and collect-
ing 50-ml fractions. Fractions 3-6 on evaporation and drying yielded
crude pangamic acid (n-gh_rconodimethyhWninoacetic acid) as a light brown
semisolid (9.7 g) some of which was further purified by preparative TLC
for spectral analysis; IR (KBr): 3500-3100, 1760, 1730, 1625-1600,
1380, 1370 cm-‘; NMR (D,O): S 3.8 (s, N (CH,),), 6 4.14.35 (m, COCH,);
MS: M’ at 281 (major peaks at m/z 281, 267, 265, 250, 237, 236, 235,
228,223,208,203,179,163,159,145,142,116,115,103,102,87,86,
85,73, 58, 57,44,43 (b.p.). The fragments in the high resolution mass at
exact masses at m/e 228.0225 (CIOH14NOI),203.0217 (CsHIJNOS), 159.0584
(C,HIJNOJ), 142.0267 (CbH604), 117.0530 (C5H903), 115.0303 (CSH9N02),
103.0327 (C4H,03) further confirmed the molecular formula C,,,H19NOs for
the structure of pangamic acid as n-ghrconodimethylaminoacetic acid.
The compound gave a triacetate derivative (dark brown semisolid) in
acetic anhydride and pyridine; IR (KBr): 1790-1770,1380-1370,1240-
1210, 1070, 1050, 1020 cm-‘; NMR (CDCI,): 6 1.8-2.2 (d, N-(CH&
and 3 X OAc), 6 3.4-4.4 (m, COCH,), 6 4.8-5.6 (m, 6H); M’ 389 and the
major peaks appearing at m/e, 359, 346, 333, 318, 303, 289, 271, 257,
215,201,145,126,116,103,102,87,73,58,57,44,43.
Acetylation using ZnCl? and acetic anhydride followed by treatment
with diazomethane gave a tetra acetyl methyl ester; IR (KBr): 1760,1740,
1735, 1730,1440-1420,1385, 1370, 1230, 1210 cm-‘; NMR (CDC13):
S 2.20 (18H, s, (NCH& and 4 X OAc), 6 3.8 (s, COOCHJ), 6 4.2-4.7 and
5.4-6.3 (8H, m, COCHz and sugar residues).
 241

Efforts to make a permethylated derivative with CHJ and Ag,O in DMF

resulted in its decomposition giving dimethylglycine (m.p. 155-157’C)
identified by JR and NMR and gluconic acid identified by TLC.
Hydrolysis with aqueous 2 N NaOH for 1 h yielded dimethylglycine
and gluconic acid as identified by TLC.
Chromatography of the isolated compound on a Pye Unicam series 204
gas chromatograph: column 1.5 m X 0.4 mm (i.d.), SE -30; Nz flow rate
50 ml; temp. programming, 120-240% at 8”C/min gave one major peak
(Rt 5 min) and two minor peaks (Rt 2.0 min and 11.6 min); its triacetate
derivative gave two peaks; one major (Rt 6.5 min) and one minor (Rt 14.8
min). The standards used for comparison were synthetic pangamic acid
(n-gluconodimethylaminoacetic acid; C1,-,H19N08,mol. wt. 281) as reported
previously (Singh et al., 1981) and its triacetate derivative, respectively.
High performance liquid chromatography gave one major (Rt 60 s, 93.1%)
and two minor peaks (Rt 72 s, 6.3% and 120 s, 0.6%, respectively) when
chromatographed on a Water Associates 51 cm X 2 mm (i.d.) stainless
steel column packed with CX/Corasil microparticulate (37-50 pm) con-
nected to a Water Associates ALC/ GPC Model 244 absorbance detector
with a fixed wave length of 254 nm; mobile phase, phosphate buffer (pH
5.0); flow rate, 1.0 ml/min; sample injected, 10 ~1; synthetic pangamic
acid was used as standard (Singh et al., 1981) (n-gluconodimethylamino-
acetic acid, Rt 60 s, 93%; 72 s, 7.0%).,
The compound isolated from Cicer arietinum, therefore, agreed in TLC,
IR, NMR, MS, GLC, HPLC behaviour with synthetic pangamic acid (Singh
et al., 1981) and in the IR, NMR and GLC systems with the acetate deriv-
ative, and IR, NMR of the tetra-acetyl methyl ester.

Isolation and identification from Cicev arietinum testa

Bengal gram testa when processed as detailed above gave identical results
for the presence of pangamic acid.

Results and discussion

Pangamic acid (n-gluconodimethylaminoacetic acid, trade name Vitamin

BIL) has been isolated and identified for the first time in Cicer arietirzum by
chemical and spectroscopic methods. The triacetate derivative (semisolid)
of the natural pangamic acid agreed on TLC, GLC, IR, NMR and MS with
the triacetate of the synthetic sample and similarly the tetra-acetyl methyl
ester in TLC, IR and NMR with the tetra-ace@1 methyl ester of the authen-
tic sample. Hydrolysis of natural pangamic acid with alkali yielded dimethyl
glycine and gluconic acid as identified by TLC. The GLC pattern of natural
and synthetic pangamic acid were identical indicating some decomposition.
Appearance of two peaks in GLC for their triacetate derivatives may be due
to their lactonised and carboxylic forms, respectively. This is further sup-
242

ported by the appearance of two identical peaks in HPLC for the synthetic
as well as the natural pangamic acid.
The natural pangamic acid (Cicer urietinum) and the synthetic one (D-
glucono~e~yl~ino~etic acid) (Singh et al., 1981) have been found to
possess marked antihyperlipidemic, antistress and increase of physical
performance activities which support the nutritional and stamina building
properties of Bengal gram (AM et al., 1980; Singh et al., 1980). Therefore,
pangamic acid can safely be recommended as a food additive or as a thera-
peutic agent which is contrary to the campaign of Herbert (1979) against
the use of pangamic acid (trade name Vitamin B,,) and its formulations.

References

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