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Talanta,Vol. 29, pp. 535 to 537, 1982 0039-9140/82/060535-03$03.

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Printedin Great Britain.All rightsreserved Copyright 0 1982 Pergamon Press Ltd

SPECTROPHOTOMETRIC DETERMINATION OF
ATROPINE, PILOCARPINE AND STRYCHNINE
WITH CHLORANILIC ACID
M. ABDEL-HADY ELSAYED and SURAI P. AGARWAL*

Faculty of Pharmaceutical Sciences, University of Nigeria,


Nsukka, Nigeria

(Received 4 August 1980. Revised 3 August 1981. Accepted 22 October 1981)

Summary-In dioxan-chloroform medium the acceptor chloranilic acid forms 1: 1 molecular complexes
with the alkaloids atropine, pilocarpine and strychnine, with maximum absorption at 535, 527.5 and
535 nm respectively. Conformity with Beer’s law allows the use of the complexes for the assay of these
drugs.

Molecular complexes are formed by interaction Chloranilic acid solution. p-Chloranilic acid dissolved in
between electron donors and electron acceptors.’ 1,4_dioxan to give 0.005M and 0.2% solutions, which were
kept in the dark when not in use and under these con-
Donors fall into two major categories, lone-pair
ditions were stable for up to 6 weeks.
donors such as amines (including alkaloids), alcohols, Other reagents and solvents were analytical grade and
sulphoxides etc., and x-donors such as aromatics, par- used as such.
ticularly polycyclic systems. Some compounds such
as azoaromatics and aromatic amines may behave as Standard solutions
lone-pair donors towards some acceptors and A- Atropine and strychnine bases. An accurately weighed
amount of the base was dissolved in chloroform to give
donors towards others. Acceptors may be of Q- and of
0.2% and 5.0 x lo-“M solutions.
x-type. The former include iodine, recently employed Pilocarpine hydrochloride. An accurately weighed amount
in the assay of alkaloids’-’ and antihistamines.” K- (0.1173 g) was dissolved in about 15 ml of water in a
Acceptors are aromatic systems containing electron- 100-m] separatory funnel. The solution was alkalinized to
withdrawing substituents such as nitro, cyano or litmus with a few drops of dilute ammonia solution and
extracted with 15, 10, 10 and lo-ml portions of chloroform,
halogen groups, e.g., p-benzoquinone,g dichlorobenzo- each extract being washed with the same 15 ml of water in
quinone,” fluoranil,” chloranil,” tetracyano-pbenzo- another separatory funnel. The washed extracts were fil-
quinone13 and 7,7’,8,8’-tetracyanoquinodimethane. l4 tered through anhydrous sodium sulphate in a filter paper
p-Chloranilic acid has been largely used in the deter- in a small funnel, into a 50-m] standard flask. The funnel
and paper were washed with l-2 ml of chloroform and the
mination of metal ions,15,‘6 but not hitherto in the
volume was made up with the same solvent to 50 ml.
assay of alkaloids. A 5.0 x 10e3M solution of pilocarpine base in chloro-
The present work describes the spectrophotometric form was similarly prepared from 0.2447g of pilocarpine
determination of atropine, pilocarpine and strychnine hydrochloride, with final dilution to 200 ml.
by complefation with chloranilic acid. These alka- Preparation of assay solutions
loids are usually present in pharmaceutical dosage
Atropine sulphate, pilocarpine hydrochloride and strychnine
forms in very small amounts (1 mg or less per unit hydrochloride powders. About 0.12 g, accurately weighed,
dose). Such small amounts preclude the use of many was dissolved in about 15 ml of water in a lOO-ml separa-
methods for assay of these alkaloids. tory funnel and alkalinized. The bases were extracted with
chloroform and collected in a 50-ml standard flask as
described for pilocarpine hydrochloride.
EXPERIMENTAL Atropine sulphate eye-drops. A 5.0-m] portion was trans-
ferred into a lOO-ml separatory funnel containing 10 ml of
Reagents water and alkalinized. The base was extracted with chloro-
Alkaloids. Atropine base (BDH), atropine sulphate
form and collected in a 50-ml standard flask as above.
Atropine sulphate ampoules. The contents of 12 ampoules
(Merck), pilocarpine hydrochloride (Merck), strychnine
base (Merck), strychnine hydrochloride (Sigma). were pooled in a dry 20-ml conical flask, and mixed; 10.0
Alkaloid dosage forms. Atropine sulphate eye-drops, 1%
ml were measured into a 50-ml separatory funnel, alkalin-
solution containing 0.002’A phenylmercuric nitrate (Evans ized to litmus with ammonia and extracted with 4, 2, 2 and
Medical). Atropine sulphate injections B.P., 0.1% 2 ml of chloroform, the extracts being collected in a lo-ml
(E.G.Y.T.) and atropine sulphate injections B.P., 0.06% standard flask.
(Evans Medical). General procedure
An aliquot of assay solution was transferred into a 20-m]
* Author to whom reprint requests should be addressed. standard flask, 15 ml of 0.2% chloranilic acid solution were

535
536 SHORT COMMUNICATIONS

yellow H,A at very low pH, the dark violet HA-


0.8 which is most stable at pH 2, and the pale violet A2-,
stable at high pH. It gives a purple colour in water,
0.7
t
acetonitrile, dimethylformamide and ammonia. As the
OS-
reaction products in non-aqueous medium are purple,
we conclude that HA- is the form of chloranilic acid
involved in the complexes. Previous reports on anal-
ogous systems’4~‘7~19-2’ support this finding.
The colour of the complexes is stable for at least
24 hr if the solutions are kept in the dark. Doubts
have been voiced about the utility of chloroform as
the solvent’4.22,23 since it may give hydrogen-bond-
ing with some alkaloids. Chloroform did not interfere
in the present system and was preferred to dioxan
because it gave lower experimental error.

Properties of the complexes


Wavelength, nm The continuous-variation24 and molar-ratio2’
Fig. 1. Absorption spectra of chloranilic acid (H*A, 0.35 methods both showed that 1: 1 complexes are formed
mg/ml) and its complexes with atropine (0.15 mg/ml, At), as expected from the single donor centre in the
pilocarpine (0.05 mg/ml, broken line, Pi), and strychnine alkaloids.
(0.16 mg/ml, solid line, St) in 1:3 chloroform-dioxan The mechanism is similar to that” for amino-acid
mixture.
complexes with chloranilic acid.
However, both methods of investigation indicate
added, and the solution was mixed and diluted to 20 ml that a 2: 1 donor:acceptor complex is also formed
with dioxan. The absorbance was measured at the corre- under certain conditions.
sponding EL,,, against a blank solution prepared by dilut- Hence for quantitative formation of the 1: 1 com-
ing 15 ml of chloranilic acid solution with dioxan to
plexes an excess of chloranilic acid is needed. Ad-
20.0 ml.
dition of 15 ml of 0.27; chloranilic acid solution in
RESULTS AND DISCUSSION dioxan gives maximal complex formation with I in
the concentration range 0.025-0.200 mg/ml, II at
Molecular complex formation 0.01-0.08 mg/ml and III at 0.03-0.24 mg/ml (as free
In dioxan-chloroform medium, atropine (I), pilo- base in the final solution) and the solutions conform
carpine (II) and strychnine (III) react instanta- with Beer’s law. Plots of absorbance (A) IX. concen-
neously with chloranilic acid (CA) to give purple tration of free base (C) are linear but give small nega-
products having similar spectra (Fig. 1). Chloranilic tive intercepts on the absorbance axis.
acid in dioxan-chloroform mixture is golden yellow, The accuracy of the method was tested by recovery
with i.,,;,, at 428 nm and a minimum near i,,,;,, of the experiments on known amounts of the salts of the
complexes. The similarity of i.,,,;,, for all three com- alkaloids (Table 1). The error (957; confidence limits)
plexes is probably due to the reaction mechanisms was between 0.8 and 1.87: relative.
being the same with the alkaloid as donor and chlora- Various atropine dosage forms were also assayed.
nilic acid as acceptor. The phenylmercuric nitrate preservative in some of
Chloranilic acid exists in three forms,” the neutral them (0.002%) did not interfere. The results are pre-

Table 1. Recovery results of alkaloids from their salts

Atropine Pilocarpine Strychnine

Added. Recovery, Added, Recovery, Added, Recovery,


“/
m&l 0,
10 my/ml % m0il ,‘o

0.100 100.1 0.036 99 0.100 loo.9


0.120 102.2 0.057 100 0.120 loo.7
0.140 100.4 0.063 100 0.140 101.1
0.160 99.6 0.068 100 0.160 100.1
0.185 100.0 0.078 101 0.180 loo.7
0.200 100.6 0.078 101 0.200 101.4

Mean 100.5 100 100.8


Standard
+ 0.9 +0.8 +0.4
deviation
SHORT COMMUNlCATlONS 537

Table 2. Assay results for atropine dosage forms

Eye-drops (1%) Injections (0.1%) Injections (0.06%)

Taken,* Recovered, Taken,* Recovered, Taken,* Recovered,


me/ml % mglml % melml %

0.125 105.3 0.120 105.4 0.108 110.2


0.150 105.7 0.140 102.5 0.114 109.5
0.160 105.1 0.150 102.8 0.120 109.2
0.170 105.0 0.160 101.6 0.126 107.6
0.180 105.2 0.170 101.4 0.132 110.3
0.190 1043 0.180 100.8 0.138 109.1
0.200 104.2 0.190 102.0 0.144 108.1
0.210 106.5 0.200 104.0 0.150 111.4
0.220 105.5
0.230 104.3

Mean 105.1 102.6 109.4


Standard +1.2
kO.7 f1.5
deviation

* According to label claim.

sented in Table 2. The recovery values are based on 6. H. Abdine, M. A. Elsayed and Y. M. Elsayed, Analyst,
the amounts found and those calculated to be present 1978.103.354.
7. M. A. Elsayed, Pharmazie, 1979, 34, 115.
from the nominal concentration of the preparations.
8. M. A. Elsayed and M. A. Abdel-Salam, Acta Pharm.
The reproducibility was rather poorer than that for Jugosl., 1978, UI, 75.
the pure salts. 9. T. Okan and K. Uekoma, Yakugaku Zasshi, 1967, 87,
1231; Chem. Abstr., 1968,68,591826.
Interferences 10. H. Abdine, M. A. Elsayed, I. Chaaban and M. E.
Abdel-Hamid, Analyst, 1978, 103, 1227.
Substances having no basic centre are not expected 11. N. M. D. Brown, R. Foster and C. A. Fyfe, J. Chem.
to interfere, since extraction of the alkaloid base pre- Sot. (B), 1967,406.
cedes the colour reaction. However, the method will 12. S. F. Belal, M. A. Elsayed, M. E. Abdel-Hamid and H.
not differentiate between the alkaloids investigated, Abdine, Analyst, 1980,~105,774.
13. K. Wallenfels. G. Bachman. D. Hofmann and R. Kern.
and amines will interfere. Nevertheless, the method is Tetrahedron, 1965, 21, 2239.
useful for routine analysis and quality control, and 14. A. Taha and J. Rticker, Arch Pharm. Weinheim, 1977,
the non-specificity could be overcome by using a 310, 485.
preliminary chromatographic separation. 15. D. D. Perrin, Organic Complexing Reagents, p. 189.
As the acceptor, chloranilic acid is superior to Interscience, New York, 1964.
16. J. F. Verchere, J. Chem. Research (S), 1978, 178.
chloranil, reaction with which is s~ow*~-~~ and gives 17. M. A. Slitkin, B. M. Smith and R. H. Walmsley,
complexes which are stable only for about 2 hr. Spectrochim. Acta, 1969,25A, 1479.
whereas chloranilic acid gives immediate formation of 18. G. Schwarzenbach and H. Suter, He/u. Chem. Acta,
a colour which is stable for at least 24 hr. 1941.24.617.
19. J. Bolard, J. Chim. Phys., 1969. 66, 221.
Acknowledgemenrs-The authors are grateful to Professor 20. H. J. Shine and R. D. Goodwin, J. Org. Chem., 1970,
P. Iwe Akubue, Dean of the Faculty, for providing facili- 35, 949.
ties, to Mr. C. U. Ogbuehi, Chief Pharmacist, University 21. P. G. Farrell and R. K. Wojtowski, J. Chem. Sot. C.,
Medical Centre, for providing dosage forms used in the 1970, 1394.
analysis, and to Senate Research Grants Committee, Uni- 22. J. Lascombe, J. Devanre and M. L. Josien, J. Chim.
versity of Nigeria, Nsukka, for funds (Grant No. 00354/79). Phys., 1964, 61, 1271.
23. M. D. Johnston, F. P. Gasparro and T. D. Kuntz, J.
Am. Chem. Sot., 1969, 91, 5715.
REFERENCES 24. P. Job, Ann. Chim., Paris, 1928, 9, 113; 1936, 16, 97.
25. J. H. Yoe and A. L. Jones, Ind. Eng. Chem., Anal. Ed.,
1. R. S. Mulliken, J. Chim. Phys., 1964, 61, 20. 1944, 16, 111.
2. A. M. Taha, A. K. S. Ahmad, C. S. Gomaa and H. 26. T. S. Al-Gabsha, S. A. Rahim and A. Townshend, Anal.
El-Fatatry, J. Pharm. Sci., 1974, 63, 1853. Chim. Acta, 1976,85, 189.
3. C. Gomaa and A. Taha, ibid., 1975,64, 1398. 27. Y. Tashima, H. Hasegawa H. Yaki and K. Takuira,
4. A. M. Taha and C. S. Gomaa, ibid., 1976, 65, 986. Bunseki Kagaku, 1970, 19, 43.
5. M. A. Elsayed, M. A. Abdel-Salam, N. A. Abdel-Salam 28. F. Alsulimany and A. Townshend, Anal. Chim. Acta,
and Y. A. Mohamed, Planta Medica, 1978, 34,430. 1973,66, 195.

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