Journal ofEthnopharmacoZogy, 13 (1985) 289-300 289

Elsevier Scientific Publishers Ireland Ltd.

ANTI-INFLAMMATORY ACTIONS OF TANNINS ISOLATED FROM THE

BARK OF ANACARDIUM OCCIDENTALE L.

M.L.R. MOTA, G. THOMAS* and J.M. BARBOSA FILHO

Laborat6rio de Tecnologia FarmacButica, Universidade Federal de Paraiba, 58.000,

Joa’0 Pessoa, Paraiba (Brazil)

(Accepted March 20, 1985)

Summary

A mixture of tannins (hydrolysable and non-hydrolysable) obtained from

the bark of Anacardium occidentale L., on i.p. injection, demonstrated
apparent anti-inflammatory activity in carrageenan- and dextran-induced rat
paw oedemas, cotton pellet granuloma test and adjuvant-induced polyarthri-
tis in rats. At higher doses orally administered tannins also had activity in
carrageenan paw oedema and adjuvant arthritis experiments. The tannins i.p.
also inhibited acetic acid-induced “writhing responses” in mice and were
found to antagonise the permeability-increasing effects in rats of certain
mediators of inflammation and to inhibit the migration of leucocytes to an
inflammatory site. While not appearing to act by the release of adrenal
hormones, tannins may produce effects in a non-specific manner by their
astringent properties on cell membranes thus affecting cell functions. The
above results should be considered while studying the anti-inflammatory
actions of plant extracts which contain tannins.

Introduction

Anacardium occidentale Linn (Anacardiaceae) commonly known as the

cashew tree grows in many tropical countries including Brazil. In addition
to its well known industrial and nutritional applications, parts of the tree
are also used in Brazil for their diuretic (Braga, 1960), antidiabetic (Correa,
1969; Lewis and Elvin-Lewis, 1977) and anti-inflammatory properties
(Coimbra, 1958; Gomes, 1973). In laboratory animals, extracts of the bark
of the tree have been reported to reduce blood sugar levels (Aguiar et al.,

*To whom all correspondence should be addressed.

0378-8741/85/$04.55 o 1985 Elsevier Scientific Publishers Ireland Ltd.

Published and Printed in Ireland
290

1959) and anacardic acid obtained from the cashew nut shell has been re-
ported to have antibacterial and antipyretic activities (Eichbaum, 1949).
In earlier studies an aqueous fraction obtained from the eth~~olic extract
of the trunk bark of A. occidentde was found to have anti-infl~mato~~
action in dextran- and carrageenan-induced rat paw oedemas when admini-
stered in doses of 10-80 mg/kg i.p. 300-900 mg/kg p.o., respectively (Mota
and Thomas, 1981). As the phyto~hemic~ screening of this fraction demon-
strated the presence of large quantities of tannins, a group of polyphen~ls
extensively distributed in the plant kingdom, it was considered wo~hwhile
to investigate whether these agents were responsible for the observed effects.

Materials and methods

Plant material

The trunk bark was collected, after identification by the botany depart
ment, from mature trees growing around the University Campus during
January-June, 1982. Voucher specimens are preserved in the herbarium of
the biology department of the University (catalog no. JPB 4.177).

The method described by IIaslam (1966) was employed. Briefly, it in-

volved maceration of small pieees of freshly cut bark with a solution of 10%
acetone in water for 72 h. The resulting extract was subjected to liquid-
liquid extraction with ethyl acetate and the ethyl acetate fraction was evapo-
rated to dryness under reduced pressure using a rota-evaporator, The residue
obtained was pulverized and mixed with silica gel. The mixture was placed
on a ~hromato~aphic column (70 cm X 4 cm) packed with silica gel in
methanol/water (60 : 40) and eluted with the solvent mixture. The first 8
fractions (50 ml each) were found to be tannins when analysed using paper
chromato~aphy. The fractions were joined and evaporated to dryness as
before to obtain a reddish brown residue, the nature of which was confirmed
by generally used chemical (Rosenth~er, 1930; Salem et al., 1969; Costa,
1972) and physical methods (Arshad et al., 1969) for the identification of
tannins.

Male albino mice and rats bred and maintained in the animal house
attached to the laboratory were used. Free access to food and water were
allowed prior to and during the experiments.

Acute toxicity studies

Mice (20-30 g) and rats (126-150 g) in groups of 10 were administered

 291

i.p. or p.o. with different doses of tannins. The animals were observed for
24 h and the number of deaths noted. LD5,, values and their 95% confidence
ranges were calculated from log-dose/mortality (probit) graphs using the
methods of Litchfield and Wilcoxon (1949).

Carrageenan-induced paw oedema

Rats weighing 120-150 g in groups of 6 were used. Doses of tannins were
administered i.p. or p.o. 30 or 60 min, respectively, before the S.C. injections
of 0.1 ml of a 1% suspension of carrageenan into the plantar region of one of
the hind paws of each animal. Percent inhibition of oedema relative to a
control group receiving 0.9% saline was calculated as described by Winter et
al. (1962).
In a separate experiment, the inhibitory activity of tannins in carrageenan
oedema was compared in sham-operated and adrenalectomized rats. The
animals were operated and maintained with the usual precautions for 7 days
before the test.

Dex tran paw oedema

The method described above for carrageenan was used except that a
0.1 ml solution of 1% clinical dextran was the oedema-inducing agent and
that the anti-inflammatory action was assessed 2 h after pedal injection.

Cotton pellet granuloma test in rats

The method of D’Arcy et al. (1960) with minor modifications was fol-
lowed. Sterilized cotton wool pellets of known weights (10 + 1.5 mg) were
implanted S.C. in groups of 7 rats (120-150 g) under ether anaesthesia. Two
pellets, one in each groin were placed in each animal. Tannins were admini-
stered daily i.p. or p.o. at various doses for 7 days with the first dose given
1 h before the implantation. On the eighth day the rats were killed and the
pellets dissected free of extraneous tissue and dried at 60°C to constant
weights. The weight of granulation tissue in each pellet was calculated and
the mean values for treated groups were compared with that of the saline-
treated group. For both the control and tannin-injected (50 mg/kg i.p.)
groups, the weights of adrenal and thymus glands were determined at the
end of the experiment.

Adjuuant-induced polyarthritis in rats

Polyarthritis was induced in rata (130-150 g) by i.d. injections of 0.05 ml
of a 5 mg/ml suspension of heat-killed Mycobacterium tuberculosis in liquid
paraffin into the plantar surface of the hind paws (Newbould, 1963). The
secondary lesions in the ears, tail and paws (more or less established by the
15th day) were assessed on an arbitrary scale (O-3) according to the method
of Rosenthale (1970) with slight changes. Animals with pronounced arthritis
on the last day were separated into groups of 7, so that each group had
approximately similar mean arthritic scores. Oral or i.p. administration of
tannins were made daily from this day for the next 7 days. Mean arthritic
292

scores obtained for each day for treated groups were compared with appro-
priate scores of the control group.

Writhing test in mice

Tannins (i.p. and p.o.) or acetylsalicylic acid (i.p.) were given 30 min
before i.p. injections of a 0.75% acetic acid solution (0.1 ml/10 g) to groups
of 5 mice (18-24s). The number of abdominal constrictions produced in
each group for the succeeding 15 min were counted and compared with the
response in the control group as described by Koster et al. (1959).

Vascular permeability studies

Groups of 7 rats (150-200 g) had their backs depilitated 48 h before the
experiments. Various doses of tannins were administered i.p. 30 min before
the animals were anaesthetized with ether followed by i.v. injections of
Evan’s blue solution (20 mg/kg). Intradermal injections of histamine (25 pg),
5-hydroxytryptamine (100 ng), bradykinin (5 pg) and prostaglandin E,
(1 pg) in dose volumes of 0.1 ml in Tyrode’s solution were then made into
the shaved areas. Forty five minutes later the rats were killed and the
amount of plasma protein-bound dye leaked/site was measured spectro-
photometrically (Harada et al., 1971). In each rat the quantity of dye exuded
after i.d. injection of 0.1 ml Tyrode’s solution was subtracted from all the
other values before calculations were made. The effect of tannins on agonist-
induced dye leakage was assessed by comparing the responses in control and
treated groups. The doses of agonists were such that these caused approxi-
mately similar (15-20 pg/site dye exudation) submaximal responses in pre-
liminary studies. However, prostaglandin El was used at 1 pg/site, the dose
which produced the maximum response of 8-12 pg/site of dye leakage
obtained by this agent.

Leucocy te migration studies

The method described by Ford-Hutchinson et al. (1975) was followed.
Sterilized-preweighed polyvinyl sponges (1.0 X 1.0 X 0.3 cm, 10 f 1 mg)
were implanted S.C. into the dorsal region of Wistar rats (6/group) under
ether anaesthesia. The animals received i.p. injections of various doses of
tannins or saline 30 min before the implantation. Five hours later, the rats
were killed and the sponge from each rat was placed in a 5-ml plastic
syringe, the fluid squeezed gently into a volumetric flask and the process
repeated after successive additions of 0.5-l .O ml Turk’s solution into the
syringe to make the final volume of 5 ml. Total and differential leucocyte
counts of the exudate were made using standard histological techniques.
Inhibitory activities of tannins were assessed by comparing the mean number
of cells present in the exudates of control and treated groups of rats.

Prostaglandin synthesis inhibitory activity

Rat fundic strip preparation was set up according to the method of Vane
 293

(1957). The bathing fluid (Kreb’s) contained antagonists such as atropine,

mepyramine and methylsergide, all at a concentration of 10 ng/ml. Various
concentrations of arachidonic acid were added into the bath to obtain a
concentration which would produce a submaximal contraction (60--80%
of the maximal response) of the tissue. The selected concentration of
arachidonic acid was repeated in the absence and presence of various con-
centrations of tannins to observe whether these would inhibit the responses
to arachidonic acid. Indomethacin was used as the standard drug for com-
parison. When tested, both tannins and indomethacin were allowed to act
for 5 min before the addition of arachidonic acid.

Results

Isolation and identification of tannins

From the freshly cut trunk bark, the extraction procedure gave a yield of
1.4% tannins. It was a mixture of hydrolysable (14%) and non-hydrolysable
(86%) types.

Acute toxicity tests

The LD,,, values in mice for tannins were 118.8 (95% confidence range
73.5-192.9) and 944.1 (95% confidence range 810.9-1099.5) mg/kg for
i.p. and p.o. administrations, respectively. The value in rats was 245.0 (95%
confidence range 187.5-320.3) mg/kg after i.p. dosing. As only 10% deaths
occurred with the highest attempted dose of 4.0 g/kg p.o. the LDSOvalue
could not be determined for this route.

Carrageenan and dex tran paw oedemas

A summary of the results obtained with tannins in these tests is presented

in Table 1. When given i.p. the tannins were highly active in inhibiting the
carrageenan oedema, the ED,,, value calculated from dose-response curve
(method of least squares analysis) being 11.2 mg/kg (95% confidence range
6.2-20.2). A much greater dose of 1.0 g/kg had to be given orally to pro-
duce significant inhibition. Tannins injected i.p. also caused significant dose-
dependent suppression of dextran oedema, the inhibitions were, however,
less than that for carrageenan. Oral dosing of tannins up to 1.0 g/kg was
inactive in this test. Adrenalectomy did not appear to reduce the tannin
effect.

Cotton pellet granuloma test

Tannins administerd daily for 7 days in doses of 6.25-50 mg/kg i.p. pro-
duced dose-dependent reduction of the granuloma tissue formation in rats.
294

TABLE 1
INHIBITORY ACTIONS OF TANNINS ON CARRAGEENAN- AND DEXTRAN-
INDUCED PAW OEDEMAS
-_________.._
Rats Dose Percent inhibition of oedema * S.E.M.
l_l- _..... ____- -
;z:y,
Carrageenan Dextran
_- ___. ___~_ _--~_-. _._._ _
Normals 3.12 i.p. 35.7 t 5.8* 7.4 k 1.8
Normals 6.25 i.p. 51.8 f 4.9** 12.6 * 1.9
Normals 12.50 i.p. 65.4 f 4.6** 29.7 f 2.8*
Normals 25.00 i.p. 68.4 i 5.1** 31.0 i 3.8*
Normals 50.00 i.p. 80.4 f 5.7*** 40.4 f 3.6*
Normals 200.00 p.0. 7.3 * 1.6 NDa
Normals 500.00 p.0. 23.9 * 2.8 ND
Normals 1000.00 p.0. 38.1 i 3.6* 0.0

Sham-operated 6.25 i.p. 47.5 k 5.0* ND

control 25.00 i.p. 70.0 1: 6.1** ND
Adre~ectomized 6.25 i.p. 42.0 * 5.0* ND
control 25.00 i.p. 65.5 i 6.51 ND
-___ -. _--. - --_II_._--- .-
aND, Note done.
Significance relative to untreated animals with pedal edemas *P C; 0.05; **f < 0.01;
***p < 0.001.

However, only the highest inhibition of 43.3% produced by the 50 mg/kg

dose was statistically significant (P< 0.05). While the mean (+S.E.M).
adrenal and thymus weights of control rats were 23.0 + 1.6 mg and 441.0 f
23.6 mg, respectively, the corresponding values in the tannin-treated group
(50 mg/kg i.p.) were 26.0 f 2.1 and 420 f 30.0 mg, respectively. The results
of the test group were not statistically different from those of the control
group. No inhibitory activity was observed when tannins were given orally
in daily doses of 500 mg/kg for 7 days.

Studies on polyarthritic rats

The results presented in Table 2 show that tannins injected i.p. or p.o. at
a higher dose, reduced the severity of secondary lesions in rats with estab-
lished arthritis. Statistically significant (P < 0.05) inhibitions of the severity
of the lesions in test groups when compared with appropriate scores in the
control group were obtained after a few day’s treatment with tannins. While
control animals on average lost 2 g/rat during the experimental period,
tannin-treated groups gamed between 4-8 g/rat, the gain depending on the
route and doses administered.

~~r~thing test in mice

Tannins given i.p. in graded doses of 6.25,12.5 and 25.0 mg/kg inhibited
the writhing response by 34.2%, 42.6% and 59.2% respectively. The EDs0
TABLE 2
EFFECT OF TANN~S ON ~OL~~T~R~~C RATS
Tannins were administered daily from days 15-21.
-. -I_
Agent DOSt! Mean arthritic score f S.E.M. for days
(mg/kgI, I___- --
Route 15 16 17 18 IQ 20 21 22
-._~
Saline lOml/kg, i.p. 13.2 * 1.3 13.7 +_1.3 14.0 * 1.5 14.6 * 1.3 14.8 + 1.4 15.0 t 1.4 15.0 t 1.4 15.0 f 1.4
Tannins 12.5 i.p. 13.5 f 1.6 13.5 f 1.7 13.8 s 1.7 13.5 r 1.0 13.0 f 0.8 11.6 f 0.7* X1.6 * 0.6* 11.0 f ox*
Tannins 25.0 i.p. 13.0 f 1.4 X2,5 f 1.4 12.0 f 1.1 11.0 * 1.0” 11.0 f. l.O* 10.0 I O.Q** 9.6 ). 0x$*** 9.2 i 0.8***
Tannins 500 p-o. 12.8 1: 1.6 12.9 f 1.6 11.9 i 1.0 11.5 * 1.0 11.2 * l.O* 10.7 +-0.8** 10.0 * 0.8*” 9.9 2 0.8***

Significance *P < 0.05,

**P< 0.025, ***P < 0.01 when compared with the mean arthritic score of corresponding days in control rats.
value was calculated (method of least squares analysis) as 15.8 mg (95% con-
fidence range 7.4-33.7) while in the same series of experiments, the i.p.
ED,, for aspirin was found to be 177.8 mg/kg (95% confidence range 92.3-
342.7). Tannins were without effect up to 300 mg/kg p.o.

Permeability studies

Previous i.p. administration of tannins in rats caused dose-related inhibi-

tion of the increased dye leakage induced by several mediators of inflamma-
tion. The antagonism was found to be non-specific as the effects of the
agonists were reduced approximately to a similar degree even though the
action of bradykinin appeared to be somewhat less affected (Table 3).

Effect on leucocyte migration

Approximately 90% of the cells present in the 5 h sponge exudates were

polymorphonuclear leucocytes (PMNL), and further, the number of mono-
nuclear cells present varied widely among different samples. Therefore, only
the effects of tannins on total leucocyte counts were determined. While the
mean (*S.E.M.) total cell count for the control group was 8.1 t 1.2 millions,
tannins injected i.p. in doses of 6.25, 12.5, 25.0 or 50 mg/kg significantly
(P < 0.05 or less) reduced the number of cells by 30.0%, 49.5%, 77.0% and
81.2%, respectively, the ED5,, being 12.6 mg/kg (95% confidence range 8.7-
18.3).

Prostaglandin synthetase inhibitory activity

Depending on the preparations, additions of l-8 @g/ml of arachidonic

acid in the bath produced submaximal tissue contractions of the required
magnitude. Presence of tannins in concentrations of up to 100 pg/ml did not
affect the response to arachidonic acid. However, indomethacin (l-10 ng/
ml) produced concentrationdependent inhibitions or arachidonic acid-

TABLE 3

EFFECTS OF 30 min PREMEDICATION OF TANNINS i.p. ON DYE LEAKAGE

INDUCED BY i.d. INJECTIONS OF HISTAMINE (H), 5-HYDROXYTRYPTAMINE
(5-HT), BRADYKININ (BK) AND PROSTAGLANDIN E,, IN RATS

Dose of Mean percent inhibition r S.E.M. of dye leakage induced by

tannin -
(mgkg) H 5-HT BK PGE,
-- -___
6.25 18.2 * 2.3 30.2 * 3.8* 14.3 +z2.6 35.1 f 4.0*
12.5 35.7 f 3.0* 45.4 t 3.9* 33.3 f 2.7* 50.9 i 6.1**
25.0 66.2 i 5.0** 58.0 f 4.0** 42.8 r 3.5* 66.7 * 8.5**

Significance relative to untreated animals: *P < 0.05; **P < 0.01.

 297

induced stimulation of the fundic strips. These concentrations of indometha-

tin failed to modify prostaglandin E2 caused contractions of the prepara-
tions.

Discussion

The anti-inflammatory activities of plant constituents such as flavonoids,

triterpenoids, coumarins, have been recently reviewed (Gabor, 1979). In
addition, colchicine, cryogenine, tomatine and other alkaloids have also been
reported to have anti-inflammatory or antirheumatic properties (Shen,
1974). Tannins are extensively distributed in the plant kingdom and tea
tannin has been shown to inhibit rat paw oedemas induced by formalin or
hyaluronidase, when administered along with vitamin C, but having no effect
on its own (see Gabor, 1979).
The present results suggest that the tannins from Anacardium occidentale
apparently have some anti-inflammatory activity as these agents given i.p.
inhibited the acute inflammatory responses of carrageenan and dextran paw
oedemas and demonstrated activity in chronic inflammatory models such
as cotton pellet-granuloma test and adjuvant-induced arthritis in rats. In
higher oral doses, they also were effective against carrageenan-induced pedal
edema and adjuvant-induced polyarthritis. In the carrageenan test tannins
were approximately equiactive in normal and adrenalectomized rats and in
the cotton pellet test the adrenal and thymus weights of animals were not
different in control and treated groups. These results indicate that the
action of the tannins is not mediated through the release of adrenal hor-
mones. Preliminary studies with the rat fundic strip suggest that tannins
may have no prostaglandin synthesis inhibitory activity.
It is probable that the anti-inflammatory actions of these agents are due
to an effect on leucocyte migration and antagonism of the phlogistic actions
of mediators of inflammation as observed in these studies. Tannins may do
so by their well known astringent properties causing precipitation of cell
membrane proteins, thus affecting cell functions, for example, cell move-
ments and membrane permeability changes. The analgesic action in the
acetic acid-induced writhing test may also be explained in a similar manner
as a non-specific effect as tannins in preliminary studies were inactive when
heat was used to induce pain (unpublished results); further, throughout the
pharmacological studies, dosing with tannins never demonstrated the
presence of analgesia in the animals.
There are many reports that irritant substances inhibit experimental in-
flammation by a counter-irritant mechanism (Laden et al., 1958; Benitz
and Hall, 1963; Robinson and Robson, 1964; Jori and Bernardi, 1966) the
nature of which is still unresolved (Atkinson and Hicks, 1971; Ford-
Hutchinson et al., 1973; Doherty and Robinson, 1976; Maguire and Wallis,
1977). To minimize the possibility of counter-irritation from parenteral
injection Benitz and Hall (1963), originally recommended oral dosing for
anti-inflammatory testing as the gastrointestinal tract was found to be less
298

sensitive to irritation than the peritoneal cavity. Since then others have
continued to emphasize oral administration for anti-inflammatory screening
of agents (Kaplan et al., 1967; Malone and Trottier, 1973). The present
results with tannins demonstrating considerably higher activity by the i.p.
than by the oral route also indicate that the irritant properties of these
substances might have contributed to their anti-inflammatory actions.
It would appear from these studies that the popular use of the decoction
of the bark of Anacardium occidentale for rheumatic diseases may be due
to its tannin content, which use must be discouraged as the toxic effects of
tannin in animals (Glick and Joslyn, 1969; Peaslee and Einhelling, 1973;
Pradhan et al., 1974; Kapadia et al., 1976) and humans (Krezanoski, 1966;
Eschar and Friedman, 1974) are well documented. As water-soluble sub-
stances, tannins are likely to be present in aqueous or aqueous-ethanol
extracts of plants and produce non-specific effects including anti-inflamma-
tory and analgesic actions. This may present further difficulties in addition
to the general problems already discussed by others (Farnsworth and Bingel,
1976; Malone, 1980) in evaluating the biological actions of plant extracts.

Acknowledgement

We thank Prof. Delby F. Medeiros for his encouragement, Prof. F. Agra

for identifying and supplying the plant materials, Mr. J.C. Duarte, Mrs. C.G.
de Oliveira and Mr. Gilmario Moreira Lima for technical help. Financial
assistance was provided by CEME and CNPq.

References

Aguiar, F.J.C., Cardoso, J.V. and Azoubel, R. (1959) Novas considera@es sobre o efeito
hipoglicemiante do Anacardium occidentale L. Anais da Faculdade de Medicina
da Universidade do Recife 19, 353-367.
Arshad, M., Beg, A. and Siddiqui, Z.A. (1969) Infrared spectroscopic investigation of
tannins. Angewandte Makromolekulare Chemie 7, 67-78.
Atkinson, D.C. and Hicks, R. (1971) The possible occurrence of endogenous antiinflam-
matory substances in the blood of injured rats. British Journal of Pharmacology 53,
85-91.
Benitz, K.F. and Hall, L.M. (1963) The carrageenin-induced abscess as a new test for
antiinflammatory activity of steroids and nonsteroids. Archives Internationales de
Pharmacodynamie 144,185-195.
Braga, R. (1960) Plantas do Nordeste, Especialmente do Ceara, 2nd edn., Imprensa
Official, Fortaleza, Brazil, pp. 104-110.
Coimbra, R. (1958) Notas de Fitoterapia, Laboratorio Clfnico Carlos da Silva Araujo
S.A., Rio de Janeiro, Brazil, p. 85.
Correa, P.M. (1969) Dictionario das Plantas cteis do Brazil e Exbticas Cultivadas, Vol. 1,
Ministerio da Agricultura, IBDF, pp. 400-401.
Costa, A.F. (1972) Farmacognosia, 2nd edn., FundacXo Calouste Gulbenkian. Lisboa,
Portugal, pp. 638-641.
D’Arcy, P.F., Howard, E.M., Muggleton, P.W. and Townsend, S.B. (1960) The antiin-
flammatory actions of griseofulvin in experimental animals. Journal of Pharmacy and
Pharmacology 12, 659-665.
 299

Doherty, N.S. and Robinson, B.V. (1976) Some biological and pharmacological proper-
ties of inflammatory exudates. Journal of Pharmacy and Pharmacology 28,859.-864.
Eichbaum, F.W. (1949) Biological properties of anacardic acid (O-penta-decadienyl-
salicylic acid and related compounds. Memorias do Znstituto Butanti 19, 119--138.
Eshchar, J. and Friedman, G. (1974) Acute hepatotoxicity of tannic acid added to
barium enemas. American Journal of Digestive Diseases 19,825-829.
Farnsworth, N.R. and Bingel, A.S. (1977) Problems and prospects of discovering new
drugs from higher plants by pharmacological screening. In: H. Wagner and P. Wolff
(Eds.), New Natural Products and Plant Drugs with Pharmacological, Biological or
Therapeutical Activity, Springer-Verlag, New York, pp. l-22.
Ford-Hutchinson, A.W., Insley, M.Y., Elliot, P.N.C., Sturgess, E.A. and Smith, M.J.H.
(1973) Antiinflammatory activity in human plasma. Journal of Pharmacy and Phar-
macology 25, 881-886.
Ford-Hutchinson, A.W., Smith, M.J.H., Elliot, P.N.C., Bolam, J.P., Walker, J.R., Lobo,
A.A., Badcock, J.K., Colledge, A.J. and Billimoria, F.J. (1975) Effects of a human
plasma fraction on leucocyte migration into inflammatory exudates. Journal of
Pharmacy and Pharmacology 27,106-112.
Gabor, M. (1979) Antiinflammatory substances of plant origin. In: J.R. Vane and S.H.
Ferriera (Eds.), Handbook of Experimental Pharmacology, Vol. 50, II, Springer-
Verlag, Berlin, Heidelberg. pp. 698-739.
Glick, Z. and Joslyn, M.A. (1969) Food intake depression and other metabolic effects
of tannic acid in the rat. Journal of Nutrition, 100, 509-515.
Gomes, P.O. (1973) Cajueiro (Anacardium occidentale L.). Fruticultura Brasileira.
Livraria Noel, S?Io Paulo, Brazil, pp. 136-148.
Harada, M., Takaeuchi, M., Fukao, T. and Katagiri, K. (1971) A simple method for the
quantitative extraction of dye extravasated into skin. Journal of Pharmacy and Phar-
macology 23, 218-219.
Jori, A. and Bernardi, D. (1966) Presence of a general irritation and inhibition of a local
inflammation. Medicina et Pharmacologia Experimentalis 14, 500-506.
Kapadia, G.J., Paul, B.D., Chung, E.B., Ghosh, B. and Pradhan, S.N. (1976) Carcinogeni-
city of Camellia sinesis (Tea) and some tannin-containing folk medicinal herbs adminis-
tered subcutaneously in rats. Journal of the National Cancer Institute 57, 207-209.
Kaplan, H.R., Wolke, R.E. and Malone, M.H. (1967) Antiinflammatory evaluation of
Cryogenine. Journal of Pharmaecutical Sciences 56, 1385-1392.
Koster, R., Anderson, M. and de Beer, E.J. (1959) Acetic acid for analgesic screening.
Federation Proceedings 18,412.
Krezanoski, J.Z. (1966) Tannic acid: chemistry, analysis and toxicology. Radiology 87,
655-657.
Laden, C., Quentin Blackwell, R. and Fosdick, L.S. (1958) Antiinflammatory effects of
counter-irritants. American Journal of Physiology 195, 712-7 18.
Lewis, W.H. and Elvin-Lewis, M.P.F. (1977) Medical Botany: Plants Affecting Man’s
Health. John Wiley & Sons, New York, p. 218.
Litchfield, J.T. and Wilcoxon, F. (1949) A simplified method of evaluating dose-effect
experiments. Journal of Pharmacology and Experimental Therapeutics 96, 99-118.
Maguire, E.D. and Wallis, R.B. (1977) The role of bacterial contamination in the isolation
of apparent antiinflammatory factors from rabbit antilymphocytic serum. British
Journal of Pharmacology 59,261-268.
Malone, M.H. (1980) Common problems encountered in ethnopharmacological investi-
gations and how to solve them. Ciencia e Cultura (Sio Paulo, Brazil), 33 Supplement,
19-31.
Malone, M.H. and Trottier, Jr., R.W. (1973) Evaluation of cryogenine on rat paw thermal
oedema and rat isolated uterus. British Journal of Pharmacology 48, 255-262.
Mota, M.L.R. and Thomas, G. (1981) AFoes antiinflamatorias do extrato aquoso da
casda do Anacardium occidentale L. Z’roc. 33rd Reuniio da Sociedade Brasileira para
o Progress0 da Ciencia (Salvador, Brazil), p. 730.
300

Newbould, B.B. (1963) Chemotheraphy of arthritis induced in rats by mycobacterial

adjuvant. British Journal of Pharmacology and Chemotherapy 21, 127-136.
Peaslee, M.H. and Einhelling, F.A. (1973) Tannic acid-induced alterations in mouse
growth and pituitary melanocyte-stimulating hormone activity. Toxicology and
Applied Pharmacology 25, 507-514.
Pradhan, S.N., Chung, E.B., Ghosh, B., Paul, B.D. and Kapadia, G.J. (1974) Potential
carcinogens. 1. Carcinogenicity of some plant extracts and their tannin containing
fractions in rats. Journal of the National Cancer Institute 52, 1579-1582.
Robinson, B.B. and Robson, J.M. (1964) Production of an antiinflammatory substance at
a site of inflammation. British Journal of Pharmacology and Chemotherapy 23,
420-432.
Rosenthale, M.E. (1970) A comparative study of the Lewis and Sprague Dawley rat in
adjuvant arthritis. Archives Internationales de Pharmacodynamie 188, 14-22.
Rosenthaler, L. (1930) The Chemical Investigation of Plants. G. Bell and Sons, Ltd,
London, U.K., p. 117.
Salem, N.A.M., El-Sherbeiny, A.E.A. and El-S&i, H.I. (1969) Local plants as potential
sources of tannins in Egypt, Part IV (Aceraceae to Flacourtiaceae). Qualitas Plantarum
et Materiae Vegetabiles 17(4), 384-394.
Shen, T.V. (1974) Nonacidic antirathritic agents and the search for new classes of agents.
In: R.A. Scherrer and M.W. Whitehouse (Eds.), Antiinflammatory Agents, Chemistry
and Pharmacology, Academic Press, New York, pp. 179-207.
Vane, J.R. (1957) A sensitive method for assay of 5-hydroxytryptamine. British Journal
of Pharmacology and Chemotherapy 12, 344-345.
Winter, C.A., Risely, E.A. and Nuss, G.W. (1962) Carrageenin induced oedema in hind
paw of the rats as an assay for antiinflammatory drugs. Proceedings of the Scoeity for
Experimen to1 Biology and Medicine 111, 544-547.