Mamina О.О.

The extraction of Suprastin from biological material by amphiphilic solvent

Ukrainian National University of Pharmacy,t.Kharkov

For the isolation of drugs of the basic nature from biological objects recommended methods of extraction by organic solvents, including quite often used amphiphilic solvents. When performing chemical-toxicological studies most commonly used amphiphilic solvent – acetonitrile [1-3].

Suprastin and some first-generation antihistamines can cause a substance abuse, heavy intoxication, affect the central nervous system, respiratory system and disrupt the function of the liver and kidneys. In severe cases of intoxication antihistamine drugs perhaps offensive coma and paralysis of respiratory center [2-4]. Due to the wide application in medical practice Suprastin, its toxicity and the absence of systematic research in the biological objects the development of methods of chemical-toxicological analysis Suprastin is an actual task.

The aim of this work is to develop an algorithm of Suprastin analysis in biological material consisting of effective methods of extraction by acetonitrile from liver tissue of dead animals, purification of extracts from impurities and application of highly sensitive methods for identification and quantification of Suprastin.

Materials and method. The prepared model mixture 10,0 g of liver tissue with 1000,0 μg Suprastin and a control sample was left for 24 h at room temperature. After a day conducted investigation according to developed technique acetonitrile extraction: to the model mixture added 10% solution hydrochloric acid for destroying bonds "protein-substance" to pH 2,0-2,5 and extracted twice Suprastin-base by acetonitrile solvent volumes – 25,0 and 10,0 ml for 10 minutes each extraction.

For cleaning aqueous-acetonitrile extract was filtered through a paper filter in the separating funnel with 70,0 ml of 2,5% solution sodium sulfate. The extraction of Suprastin-base was conducted twice using 10,0 ml chloroform at pH 2,0-2,5. The aqueous-acetonitrile extract pH 2,0-2,5 basified with 25% solution ammonia to pH 9,0-10,0 by universal indicator paper and extracted twice Suprastin-base by chloroform (portions -10,0 ml).

Chloroform extracts were purified by extraction of impurities with hexane and TLC-method: chromatographic plates Sorbfіl PSTH-AF-A, the system of organіc solvents - ethylacetate-methanol-25% solution ammonia hydroxide (85:10:5), location reagent - Dragendorff's reagent for Mounier (sensitivity of location reagent -1,0-3,0 μg substance); R_{f Suprastin} = 0,60-0,63[5].

Quantitative determination of Suprastin carried out by UV spectrophotometric method after extraction and TLC-purification. Optical density values were measured on an SF-46, cell thickness 10 mm; λ_max 312 ± 2 nm, reference solution - 0,1 M solution of hydrochloric acid. Submission law Bouguer-Lambert-Beer is observed within concentrations (5,0 – 35,0 μg / ml); lower limit of detection (5,0 μg / ml). Suprastin concentration in solution was calculated from the equation of the linear dependence of concentration (C, µg / ml) and absorbance (А) (С = -0,018 + 0,024 А) [6]. Results of Suprastin extraction by acetonitrile from liver tissue presented in the Tab.

Table

Results of Suprastin extraction by acetonitrile from liver tissue (n = 5, Р = 95%)

The method of quantitative determination	Allocated Suprastin, %	Metrological characteristics, %
			S^2	S	S
UV spectro-photometric method	56,6 – 64,8	60,7	3,96	1,99	0,89	2,47	4,07

Results: the extraction of Suprastin by acetonitrile from biological material allows to determine – 60,7+ 4,1% of the substance. Methods can be proposed for implementation in practice for forensic medical examination and toxicology centers.

Conclusions:

1. The extraction of Suprastin by acetonitrile from the liver of a corpse, purification of extracts with hexane and by thin-layer chromatography, quantitative determination of Suprastin by UV spectrophotometry were carried out.

2. The extraction of Suprastin by acetonitrile from biological material allows to determine – 60,7+ 4,1% of the substance.

References:

1. General methods for isolating toxic and potent substances from biological material / B. G. Burchynsky, F.M. Kahanovsky, K.I. Kahanovskaya, T.V. Hoholeva. - Odessa: Astroprint, 2010. - 44 p.

2. Mashkovsky, M.D. Medicines – Moscow: LLC "Publishing House "New Wave", 2010.- 1216 p.

3. Clarke, E.J.C. Isolation and Identification of Drugs in Pharmaceuticals, Body Fluids and Postmortem Material.- London: The Pharm.Press, electronic version, 2005.

4. Secrets of Toxicology / Dzh. Ling L., R. F. Clark, T.B. Erickson, D.H. Trestreyl. - Moscow: LLC "Publishing house" Binom", 2006. – 376 p.

5. The investigation of antihistamines by thin layer chromatography / O.O. Mamina, A.M. Lebedin, E.L. Bondarenko, N.O. Shum // XI Abstracts оf Scientific papers "The issue of forensic medicine and expert practice" - Donetsk, 2013.- P.101-102.

6. Lebedin, A.M. The choice of optimal conditions of chloropyramine analysis by UV spectrophotometry suitable for chemical-toxicological studies /A.M. Lebedin, O.O. Mamina, L.I. Boryak // Collection of Scientific Papers of the employees NMAPE named P.L. Shupik - 2012.- Vol. 21, №.4. - P. 313-318.