the thiamine content of herbs and medicinal plants
TRANSCRIPT
SCIENTIFIC EDITION
The Thiamine Content of Herbs and Medicinal Plants*
By ALEXANDER S. CHAIKELISt
By means of a photofluorometric assay tech- nique, eliminating the elution phase, devised to investigate the thiamine content of plant material ranging from rhizomes and roots to barks, leaves, and flower tops, the thiamine concentration in herbs and medicinal plants was determined. A spot check analysis of 41 genera belonging to 20 distinct and separate botanical families showed a thiamine range from 125 to 2880 micrograms per 100 Gm. of dried substance. This vitamin B1 content was postulated as a possible modus operandi of the many infusions and de- coctions of medicinal herbs prescribed i n cases of anorexia, dyspepsia, general debility,
and nervousness.
OR MANY CENTURIES the.use of plants in the alleviation and cure of bodily
ills received wide attention but little official sanction until in the reign of King Henry VIII Parliament enacted what is now popularly known in England, “The Herba- lists’ Charter” (1).
Healers, like Nicholas Culpepper (1616- 1654) in England and Samuel Thompson (1769-1843) in America, did much to ad- vance the study and practice of herbalism. Apart from the claims made by the herba- lists, the use of herbs as sources of drugs, like those described in articles and books on pharmacognosy and/or materia medica, has done much to justify their usefulness in medicine (2-7).
With our present knowledge of the various sources of vitamins, can i t be that the cumu- lative beneficial results produced by the use of herbs, exclusive of their medicinal proper- ties, might be the effects of increased vita- min intake? To obtain information along this line of thought, it was decided to make a spot check on some herbs and other plants,
* Received July 9 , 1946, from the Biology Department, The City College, and the Department of Industrial Hygiene, School of Public Health, Columbia University, New York City. t The author expresses his gratitude to Dr. Frederick B. Flinn, Director of the Department of Industrial Hygiene, School of Public Health, Columbia University, for his en- couragement, and to Hoffman-LaRoche, Inc., Nutley, N. J., for donations of thiamine.
343
emphasis to be on those which were and are, a t present, described as possessing tonic, stomachic, and stimulant actions in cases of dyspepsia, constipation, general debility, and nervousness. Since all higher plants are able to not only synthesize thiamine but to excrete i t through the roots, it seemed logical to investigate the existence and quantity of thiamine in these and other parts of the dried plants that had recognized medicinal value.
Samples of the herbs and medicinal plants were purchased in the dried state from a reliable distributor of such material. The plants so bought represented 41 genera belonging to 20 separafe botanical families. This number of specimens was considered to be an adequate sampling of the entire group of medicinals.
EXPERIMENTAL
Based on the idea of procedure for thiochrome determination described by Hennessy (8) and the digestion method of plant material to release the thiamine in the free state as well as the phosphory- lated form (cocarboxylase) suggested by Platt and Block (9), the thiamine content of the plant material was analyzed by the following method devised to fit the demands of the material under investigation and the requirement of a quick, reliable technique. A. Reagents Required.-The many reagents
needed to effectively carry out the thiochrome determinations included those listed in the U. S. P., those enumerated by Platt and Block (9), and those, such as the needed standard solutions, described in (10).
The digesting enzymes papain and clarase before use were treated in the following manner: A 5-Gm. batch of each enzyme is suspended in 50 cc. distilled water and shaken thoroughly. To this suspension is added 50 cc. isobutanol, and the mixture again shaken in a separatory funnel. Discard the super- natant isobutanol fraction, repeating the process until the isobutanol portion when oxidized with NaOH and KIFe(CN)B shows no evidence of thio- chrome when examined in a photofluorometer.
The aqueous fraction is then treated with ethanol to precipitate the enzymes, filtered, and the residue dried in vucuo. The dry amorphous powder is then ready for use in the digestion phase of the treatment section of the procedure.
TA
BL
E C
OM
POSI
TE
SUM
MARY
OF T
HIA
MIN
E
CO
NTE
NT IN
V
ARI
OU
S DR
IED
HE
RB
S AN
D OT
HER M
ED
ICIN
AL
PLANTS
Av.
T
hiam
ine
Thi
amin
e T
hiam
ine
Com
mon
Nam
e A
grim
ony
Ang
ostu
ra
Bal
mon
y B
arbe
rry
Bay
Lau
rel
Bla
ck A
lder
B
less
ed T
hist
le
Bon
eset
B
uchu
B
ugle
wee
d C
alam
us
Car
away
Cas
cari
lla
Cat
nip
Cel
erv
Ciy
mom
ile
Che
rry
Tre
e C
ondu
rang
o D
ande
lion
D
ogw
ood
(Jam
aica
) Fe
verf
ew
Gar
den
Sag
e G
enti
an
Gho
st R
oot
Gro
und
Ivy
H
orse
radi
sh
Moo
nsee
d M
usk
Roo
t P
arsl
ey
Par
sley
P
rick
ly A
sh
Qua
ssia
S
cull
cap
Sna
kero
ot (
Can
.)
Sna
kero
ot (
Va.
) S
outh
ernw
ood
Str
awbe
rry
(wild
) T
ansy
T
hym
e (c
omm
on)
Whi
te A
sh
Whi
te C
inna
mon
Y
ello
w Dock
Gen
us a
nd S
peci
es
Agr
imon
y eu
pafo
rium
Lin
n.
Gal
ipea
o$i
cina
lis
Che
lone
gla
bra
Lin
n.
Ber
beri
s A
puif
oliu
m (
Pur
sh.)
L
auru
s no
bilis
Lin
n.
Ilcx
wer
fici
llala
Lin
n.
Cni
cus
bcna
dicf
us (G
oetn
.)
Eup
afor
ium
per
folia
lum
Lin
n.
Bar
osm
a be
lulin
a B
artl
ing
and
Wen
dl.
Lyc
opus
uni
joru
s Linn.
Aco
rus
Cal
amus
Lin
n.
Car
um C
arvi
Lm
a.
Cro
fon
Elu
lcli
a B
enne
tt
Nep
efa
cafa
ria
Lin
n.
Api
um
gra
veol
cns
Lin
n.
Maf
rica
ria
Cha
mom
illa
Lin
n.
Pru
nus
sero
fina
Lin
n.
Mar
sdcn
ia C
ondu
rang
o R
eich
enha
ch f.
T
arax
acum
ofl
cina
lc W
eber
P
isci
dia
eryf
hrin
a (J
acqu
es)
Chr
ysan
fkem
um P
arfk
cniu
m B
ernh
. Sa
lwia
ofil
cina
lis L
inn.
G
enlia
na lu
fca
Lin
n.
Ang
elic
a ar
chan
gelic
a L
inn.
N
epcf
a hc
dcra
cca
Lin
n.
Coc
hlea
ria
arm
orac
ia L
inn.
M
enis
perm
um c
anad
cnse
Lin
n.
Fer
ula
Sum
bul
Hoo
ker
fili
us
Car
um p
cfro
selin
um (B
enth
.)
Car
um p
elro
selin
um (
Ben
th.)
X
anfh
oxyl
um am
eric
anum
Mil
ler
Pic
rasm
a ex
cels
o P
anch
on
Scut
ella
ria
1afc
riR
ora
Lin
n.
Asa
rum
can
aden
se L
inn.
A
risl
oloc
hia
scrp
enfa
ria
Lin
n.
Arf
emis
ia a
brof
anum
Lin
n.
Fra
gari
a ni
rgin
iana
Lin
n.
Tan
acet
um ou
lgar
c Lin
n.
Thy
mus
vul
gari
s L
ina.
F
raxi
nus
amer
ican
a L
inn.
C
inna
mom
um C
assi
a (B
lum
e)
Rum
ex c
risp
us L
inn.
Fam
ily
Ros
acea
e R
utac
eae
Scr
ophu
lari
acea
e B
erbe
rida
ceae
L
aura
ceae
A
quif
olia
ceae
C
mpo
sita
e 8
ompo
sita
e R
utac
eae
Lab
iata
e A
race
ae
Um
bell
ifer
ae
Eup
horb
iace
ae
Lab
iata
e U
mbe
llif
erae
C
ompo
sita
e R
osa c
e a e
Asc
lepi
adac
eae
Com
posi
tae
Leg
umin
osae
C
ompo
sita
e L
abia
tae
Gen
tian
acea
e U
mbe
llif
erae
L
abia
tae
Cru
cife
rae
Men
ispe
rmac
eae
Um
bell
ifer
ae
Um
bell
ifer
ae
Um
belli
fera
e R
utac
eae
Sim
arub
acea
e L
abia
tae
Ari
stol
ochi
acea
e A
rist
oloc
hiac
eae
Com
posi
tae
Ros
acea
e C
ompo
sita
e L
ahia
tae
Ole
acea
e .
Lau
race
ae
Ari
stol
ochi
acea
e
Par
ts U
sed
Ent
ire
herb
B
ark
~ Lea
ves
Roo
t ba
rk
Lea
ves
Bar
k E
ntir
e he
rb
Lea
ves
Lea
ves
Ent
ire
herb
R
hizo
me
and
root
s S
eeds
Bar
k L
eave
s S
eeds
F
low
er t
ops
Lea
ves
Bar
k R
hizo
me
and
root
s R
oot
bark
E
ntir
e he
rb
Lea
ves
Rhi
zom
e an
d ro
ots
Rhi
zom
e an
d ro
ots
Ent
ire
herb
R
hizo
me
and
root
s R
hizo
me
and
root
s R
hizo
me
and
root
s L
eave
s R
hizo
me
and
root
s B
ark
Hea
rtw
ood
Ent
ire
herb
R
hizo
me
and
root
s R
hizo
me
and
root
s E
ntir
e he
rb
Lea
ves
Ent
ire
herb
L
eave
s B
ark
Bar
k R
hizo
me
and
root
s
Dos
e in
Gm
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m.
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26.1
Act
ion
and
Met
hod
of A
dmin
istr
atio
n T
onic
. as
an
infu
sion
: 1
oz. t
o 1
pt.
wat
er
Bitt
e;
toni
c;
as a
bove
, 2 f
l. 0
2.
t. i.
d.
Ton
ic, f
or c
onst
ipat
ion;
as
abov
e, 2
fl. o
z. t
. i. d
. B
itte
r to
nic;
po
wde
red
bark
, 20
gr.
t.
i. d
. F
lavo
ring
: sp
ice
for m
eats
, pou
ltry
, m
eat s
auce
s T
onic
; as
infu
sion
, 2 fl
. 01. q. i.
d.
Ton
ic.
as a
bove
T
onic
! as
abo
ve, 2
fl.
02
. q.
i.
d.
Sti
mui
ant;
as
abov
e, 2
fl. 0
2. q. i
. d.
C
alm
ativ
e;
as a
bove
A
rom
atic
hit
ter;
as
abov
e S
tim
ulan
t, c
arm
inat
ive:
2
fl. o
z. t
. i.
d. a
s an
Bit
ter;
as
infu
sion
, a 2
-8. o
z. d
ose
t. i.
d.
Ton
ic;
as in
fusi
on, 2
fl. 0
2.
q. i
. d.
S
tim
ulan
t;. a
s in
fusi
on, 2
8.
oz. t.
i.
d.
Bit
ter;
as
infu
sion
, 2 8
. 02
. q.
i.
d.
Bit
ter
toni
c; a
s in
fusi
on, 2
8. o
z. t
. i.
d.
Bit
ter.
as
infu
sion
, as
abov
e S
impl
e bi
tter
; as
infu
sion
, 2 fl
. oz
. q
. i.
d.
Bit
ter t
onic
; as
infu
sion
. 2 A
. oz.
t. i
. d.
Cal
mat
ive
in h
yste
ria;
as i
nfus
ion,
2 fl
. oz.
t. i.
d.
Car
min
ativ
e; a
s inf
usio
n fo
r dys
peps
ia. a
s abo
ve
Sim
ple
bitt
er;
as in
fusi
on, 2
fl.
oz.
q. i.
d.
Bit
ter;
as
abov
e T
onic
. as
inf
usio
n in
dys
peps
ia,
as a
bove
S
tom
akhi
c; a
s in
fusi
on, 2
fl.
02
. t.
i. d
. S
impl
e bi
tter
; as
abo
ve, 2
fl.
oz. t.
i.
d.
Cal
mat
ive;
as
an e
xtra
ct i
n 8-gr. d
oses
F
lavo
ring
; us
ed a
d li
bitu
m
Cal
mat
ive;
as
infu
sion
, 2 fl
. oz
. t.
i. d
. S
tim
ulan
t; a
s in
fusi
on.
2 fl.
oz.
q.
i. d
. B
itte
r to
nic;
0.5
cc.
as
flui
dext
ract
T
onic
; as
infu
sion
, 4 fl
. oz
. q.
i.d
. A
rom
atic
bit
ter;
as
infu
sion
, 2 fl
, oz.
t. i
. d.
Aro
mat
ic b
itte
r; a
s in
fusi
on, a
s ab
ove
Em
men
agog
ue;
as in
fusi
on, 2
fl. 02. q
. i.
d.
Bit
ter
toni
c; a
s in
fusi
on, 2
fl. 0
2.
t. i
. d.
S
tim
ulan
t, a
nhpa
smod
ic;
as in
fusi
on, a
s ab
ove
Sti
mul
ant,
ant
ispa
smod
ic; as a
bove
B
itte
r to
nic;
as
infu
sion
, 2 fl
. 02
. t.
i.
d.
Aro
mat
ic b
itte
r; a
s ab
ove
Sto
mac
hic,
ton
ic;
as a
bove
Ser
ved
cold
infu
sion
SCIENTIFIC EDITION 345 B. Treatment of the Substance to Be Ana-
lyzed.-l. Place 10 Gm. of the substance, dried and finely ground, in a 250-cc. flask.
2. Add 90 cc. N/lO HzSO~, mix thoroughly, and then add 0.2 Gm.-batches each of the purified papain and clarase.
Now add 10 cc. of the M/10 sodium acetate- acetic acid buffer solution. Mix well (check PH with bromcresol green so that i t is 4.0; correct with either 10% HCl or 10% NaOH).
4. Stopper flask and heat the mixture a t 40-45' for from sixteen to eighteen hours.
5. Remove flask, add 0.2 cc. glacial acetic acid, and heat mixture to 80" to coagulate the proteins.
6. Cool flask to room temperature; centrifuge a 20-cc. portion, and save the clear supernatant aqueous layer for the analysis.
Oxidation (by means of alkaline ferricyanide) of thiamine t o thiochrome, which is extracted by isobutanol, is followed by measurement of the fluorescence in ultraviolet light of the thiochrome formed. The thiamine pyrophosphate also forms a fluorescent thiochrome, which is not, however, soluble in isobutanol-hence one reason for the enzyme treatment to hydrolyze the thiamine pyro- phosphate to thiamine.
With a Coleman Universal Spectrophotometer #11 adapted with an ultraviolet illuminator and the requisite filters for thiamine study, determine the degree of fluorescence using the substitute working standard of quinine sulfate (3 gamma per cc.). See details in the direction booklet (11).
3.
7.
8.
RESULTS
The values for the vitamin B1 content of the 41 genera of herbaceous medicinal plants are repre- sented in Table I. The thiamine, in micrograms per gram of dried substance, as listed in the table represents the mean value of three distinct and separate samples of, each plant studied.
Of the total number of plants studied in this spot check, it can be seen by reference to Table I that a group of seven medicinal plants contain thiamine in excess of 1500 ?/lo0 Gm. of substance. These seven plants arranged in order of the highest thia- mine content are: Prickly Ash (Xanthoxylum americanum Miller), Black Alder (Ilex verticilluta Linn.), White Ash (Fraxinus americana Linn.). Buchu (Barosm betulina Bartling & Wendland), Cherry (Prunus serotina Linn.), Quassia (Picrasma excelsa Panchon), and Dandelion (Taraxacum oficinale Weber) .
On the other hand, five genera show a thiamine content of less than 300 ~/100 Gm. of dried sub- stance. Arranged in order from the lowest to the highest concentration these five herbs are: Parsley (root) (Petroselinum sativum Hoffman), Southern- wood (Artemisia abrotanum Linn.), Agrimony (Agrimony eupatorium Linn.) , Strawberry (wild) (Fragaria virginiana Linn .) , and Bugleweed (Lycopus uni$orus Linn.). The remaining herbs
in the series range from 365-1154 y/100 e m . of substance.
DISCUSSION
Many factors influence the effectiveness with which a plant synthesizes vitamins. Therefore the values reported in this paper cannot be considered as constant but merely as indicative of the concentration of thia- mine contrary to the claim made that leaves, for instance, belonging to higher plants regardless of the botanical family source contain a constant thiamine concentration equal to 75 micrograms per 100 Gm.
It is an accepted fact that animals (in- cluding man) require a daily intake of thiamine because this vitamin is not stored in the organism although, as Westerbrink (12) reported, relatively higher concentra- tions are found in the liver, kidneys, heart, muscle, and brain than in the blood stream. The animal organism absorbs only as much thiamine as is needed for the time being, all excess is excreted and to a small extent destroyed in the body.
The physiological action of thiamine in the animal organism in the form of its pyrophosphoric acid ester is intimately concerned with carbohydrate, fat, and water metabolism. Thus, glycogen, the specific animal carbohydrate, requires the presence of this vitamin to be properly metabolized. The fat metabolism is in- fluenced by vitamin B1 only in so far as the synthesis of fat from carbohydrate is con- cerned. The water metabolism in the animal organism seems connected with the action of this vitamin, probably through the carbohydrate metabolism. Edema and water imbibitions in the heart and in other organs are symptoms of thiamine inade- quacy. Vitamin B1 deficiency affects, first of all, the emotions and the tonus of the nervous system, for this substance takes part in the conduction of nerve impulses as shown by Minz (13) that acetylcholine and vitamin B1 are liberated during impulse conduction.
Further symptoms of thiamine deficiency are loss of appetite (anorexia), lowered physical endurance associated with unusual susceptibility to fatigue, gastrointestinal
346 JOURNAL OF THE AMERICAN PHARMACEUTICAL ASSOCIATION
disturbances (dyspepsia, constipation), mus- cular weakness, and a decrease of the blood pressure.
Attention is called to the fact that it is precisely for such discomforts that many herb infusions and decoctions, known as “tonics,” have been prescribed. The indica- tions are that these prescriptions materially benefited the ailing individual, a t least in a small measure, by providing him with readily absorbable thiamine and its phos- phorylated form a t a time when his system was in great need of these items.
SUMMARY
1. The thiamine content of a spot check analysis of 41 genera of herbs and medicinal plants was determined by means of the photofluorometric technique.
2. A photofluorometric assay technique was devised employing the suggestions of Hennessy (8) and those of Platt and Block
These 41 genera of herbs and medici- (9) -
3.
nal plants belonging to 20 separate botan- ical families showed a thiamine content ranging from 125-2880 micrograms per 100 Gm. of dried substance.
The rational basis for the use of herb infusions and decoctions in the form of “tonics” is suggested on the basis of the physiological action of thiamine in the animal organism.
4.
REFERENCES
(1) Warcl. 11.. “Herbal Manpl,” London. 1936. L. I < . . “A Modern Herbal.” ork, 1931. Wirth. E. H., “Pharmacog-
iiaaeiphia, 1936. , “Textbook of Pbarmacognosy.” adelphia, 1543 rhe Spice Handbook.” Chemical w York. 1945. A Textbook of Pharmacognosy.” ,ondon, 1945. ull. Torrey Bolan. Club. 73, 60-72
(8 ) Hennessy, D. J . , Ind. Eng . Chem., Anal . Ed . , 13,
(9) Platt. B. S.. and Block, C. F., Biochem. J . , 37, 436
\ - - I ” , .
2 16-17 ( 1941 ) .
f 19431. ~ ~ ~ ~ . , . (10) Hoffman-LaRoche. Inc., Nut!ey. N. J.. “Determina-
tion of Vitamin BI in Food Products (11 ) “Zirections for Model 11 ’Universal Spectropho-
tometer, D-142, Coleman Electric Co., Maywood. 111..
1944.
1945, p. 11.
560(1932). (12) Westerbrink, H. C. K . , Arch. nlcrland. physiol . , 17,
(13) Minz, B., C o m p f . r e n d . sot. b i d . , 127, 1251(1938).
Persistence of Penicillin in Saliva After Local Oral Application*
By MILTON GJELHAUG LEVINE and ROBERT E. HOYT
Penicillin for local application to the mouth area was administered in a chicle base chew- ing gum, in flavored parafKn, and in a mouth- wash. The retention time of penicillin in the saliva for each of these dosage forms with varying concentrations of the drug is recorded
in this report.
HE USE of penicillin in the buccal cavity Thas been suggested in hemolytic strepto- coccus infections, tonsillitis, Vincent’s disease (acute ulcerative gingivostomatitis), Ludwig’s angina, for lesions of the oral cavity involving loss of surface, and for prophylactic use in operative procedures of the mouth. Of all these, the most con- vincing therapeutic evidence has been ad-
*Received Aug. 12, 1946, from the Institute of Experi- mental Medicine. College of Medical Evangelists, 1.0s Angeles, Calif.
duced for the use of local preparations in Vincent’s angina.
Our own experiences and those reported by other investigators (1, 2, 9, 10) indicate that in most cases Vincent’s disease may be cured in from twenty-four to forty-eight hours by the local application of penicillin. The results for other infections so far have been inconclusive, although preliminary reports (2-5) indicate that Group A strepto- cocci may be reduced in number and even eliminated by local applications of peni- cillin. However, more satisfactory results may be obtained by parenteral administra- tion of the drug. Parenteral administration of penicillin has also proved to be of value in Ludwig’s angina (6, 7).
The treatment of acute ulcerative stoma-