Lebedin A.M., Mamina О.О., Kovalenko Z.I.

Isolation of chlorohyramine from liver tissue of corpses

Ukrainian National University of Pharmacy,t.Kharkov

The list of drugs of domestic and foreign production offered on the pharmaceutical market of Ukraine is constantly spreading, so the development of methods of chemical-toxicological and forensic analysis potent drugs is an actual problem. In recent years, in medical practice is observed a significant increase in acute poisoning agents antihistamine action, which includes chloropyramine hydrochloride.

According to the literary sources of data on the systematic study chloropyramine hydrochloride in biological objects are not available, not studied methods of isolation of chloropyramine hydrochloride from biological objects, removal of impurities, identification and quantitative determination in the extracts, the distribution in the organs of animals, storage in biological material and metabolism.

The aim of this work is development of chemical and toxicological analysis of chloropyramine hydrochloride in liver tissue of corpses, based on effective and express technique its water isolation from biological material.

Materials and method. Extraction of chloropyramine hydrochloride from model mixture of 10,0 g of minced liver tissue of corpses of animals and 1,0 mg substance, and control samples were performed using water acidified with 10% oxalic acid solution (pH 2-2,5) [1,2].

After alkalinization aqueous phase of 25% ammonium hydroxide solution to pH 9-10 chloropyramine-base was extracted with chloroform.

Purification of the extracts was performed using centrifuge 6000 r / min for 10 min. After drying the organic extract at room temperature and dissolving the dry residue in 20 ml of 0,1 M solution of hydrochloric acid extraction was performed three times with hexane impurities 5 ml.

For quantification of chloropyramine in extracts used the UV- spectrophotometric method: spectrophotometer SF-46, the cell thickness of 10 mm; chloropyramine hydrochloride solution in 0,1 M hydrochloric acid solution, λ max = 312 ± 2 nm; reference solution obtained from the control sample [3].

For quantification of chloropyramine using calibration curve constructed in coordinates: A ( optical density solutions) - С, µg / ml (concentration of solutions of the substance in 0,1 M solution of hydrochloric acid ): C = -0,018 + 0,024 A.

Interval linear calibration curve was in the range of concentrations of 5,0 – 35,0 µg / ml; lower limit of detection – 5,0 µg / ml [3].

Results. These data in Table are shown reproducibility of determination of the substance by the developed pattern analysis, as evidenced by metrological characteristics.

Table

Results chloropyramine extraction with water, acidified with oxalic acid liver tissue corpses of animals (n = 5, Р = 95%)

Contributed chloro-pyramine, mg	Allocated chloro-pyramine, %	Metrological characteristics
			S^2	S	S
1,0	69,6 – 65,2	67,4	3,03	1,74	0,78	2,16	3,2

The study found that the use of water to isolate chloropyramine is efficient, economical and expressive, due to lower losses substances when cleaning extracts from impurities.

The technique can be proposed for implementation in practice of forensics, toxicology centers, clinical laboratories for the study of drugs in biological objects.

Conclusions

1. As a result of researches chloropyramine analysis of biological material, which consists of stages: extraction of chloropyramine with water acidification oxalic acid from the liver corpse, cleaning hexane the extracts and assay chloropyramine by UV- spectrophotometry.

2. Established that chloropyramine extraction from biological material with water, acidified with oxalic acid to determine – 67,4 ± 3,2% substance.

References

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

2. Burchynsky V.G. General methods for the isolation of toxic and potent substances from biological material: method. recommendations /V. G. Burchynsky, F.M. Kahanovsky, K. I. Kahanovska and others. - Odessa: Astroprint, 2010. - 44 p.

3. Lebedin A.M. Selection of optimal conditions chloropyramine analysis by UV-spectrophotometry, suitable for chemical and toxicological studies / A. М. Lebedin, О.О. Mamina, L.I. Boryak // Proceedings of employees NMAPE named P.L.Shupyk - 2012. - Vol. 21.- Kn. 4. - P. 313-318.