CARBAMAZEPINE AND THIOTRIAZOLINE SIMULTANEOUS DEFINITION IN MODEL MIXTURE BY HPLC Message 1: phase selection for the simultaneous determination of carbamazepine and thiotriazolin in model mixture by high performance liquid chromatography

Abstract

CARBAMAZEPINE AND THIOTRIAZOLINE SIMULTANEOUS DEFINITION IN MODEL MIXTURE BY HPLC

Message 1: phase selection for the simultaneous determination of carbamazepine and thiotriazolin in model mixture by high performance liquid chromatography

L. I. Kucherenko¹², Н. R. Nimenko^1,2, O. V. Vashchenko¹, V. V. Vashchenko¹

Scientific-and-Production Corporation «Pharmatron»¹

Zaporizhian State Medical University²

Epilepsy is a chronic disease of the brain, the only or dominant feature of which is recurring seizures. Number of patients with epilepsy in the world is more than 40-50 million people inUkraine- about 500 thousand. People on the basis of antiepileptic elaborated by experts of the International League (MPEL) standards and recommendations of the treatment main forms of focal epilepsy, the drug of first choice is carbamazepine (chemical name 5 -karbonoil-5H-dibenzo (b, f) azepin).

However, the proven efficacy of carbamazepine is only for large seizures and apparent side effects that limit its use in the clinic such as hepatotoxicity, nephrotoxicity, hastrotoksychnist, hematotoksychnist, cardiotoxicity. Solution to that problem was the creation of new, more effective antiepileptic drug that shows pronounced antidepressant, nootropic, neuroprotective and antioxidant properties based on fixed combination with carbamazepine Thiotriazoline, allowing also significantly reduce the side effects. And this drug is karbotril. It is important that it is planned to expand the scope of assignments of karbotril by small forms of epilepsy, benign partial epilepsy with Center-temporal spikes, as well as other disorders such as alcoholism, drug addiction, gambling, pain syndromes, to prevent seizures in traumatic brain injury , multiple sclerosis.

The aim of our study is the selection of items for the joint determination of carbamazepine and Thiotriazoline in a modular mixture by high performance liquid chromatography (HPLC).

Materials and methods: For new combined drug containing carbamazepine and thiotriasolin best methods of standardization have to be developed. For this model, we prepared a mixture consisting of carbamazepine and thiotriasolin in a ratio of 1.5: 1. Based on the fact that standardization of most dosage forms as tablets is made by physicochemical methods, and one of the most modern techniques are HPLC, so the task was to conduct determination of active substances by this method. To perform this task, it was necessary to make an attempt of eluent selection in the first case it was methanol and phosphate buffer, and in the second methanol and0.01 M district tetrabutylammoniy hydrosulphates (TBAHS).

Results and discussion.

In connection with joining ofUkraineto the EU and the introduction of the State Pharmacopoeia of Ukraine and its four amendments modern technologies of standardization go to the forefront. Increasingly, to determine organic active ingredients like mono or in combination dosage forms HPLC method is used. This method in appropriate conditions can simultaneously identify and quantify the active ingredients in medicines, so for the definition of active ingredients in the new fixed combination medical product "Karbotryl" we proposed this method. Firstly we drew their attention to the existing method of Thiotriazoline standardization by HPLC. Studies have been conducted on inverse phase (C18) under isocratic elution using eluent, which is water-methanol mixture. The study found that the simultaneous determination of active ingredients in a model mix of Thiotriazoline and carbamazepine complicated by:

1. The difference in the solubility of these substances, carbamazepine, unlike Thiotriazoline, is insoluble in water but is readily soluble in alcohol;

2. The big difference in chromatographic mobility analytes - very polar Thiotriazoline and little polar carbamazepine.

Based on the above, it was assumed that eluent for a common definition of Thiotriazoline and carbamazepine should contain more methanol (50%) compared with eluent for determining Thiotriazoline (phosphate buffer and methanol in a ratio of 90:10), but we know that such eluent greatly reduce retention time of Thiotriazoline, which also complicates the selection conditions determining the active substances in the modular mix.

Conclusion: during the study selection phase for the joint determination of carbamazepine and Thiotriazoline by HPLC were conducted. This was used by various inverse phase and eluent, the possibility of chromatographic determination of active substances in the model mix. Found that Thiotriazoline’s volume retention in terms of isocratic elution is between 2.6 - 2.9 ml dead volume near the column, making it difficult, and in some cases makes it impossible to conduct pharmaceutical analysis.

References

1. Badalyan O.L. Nerobkova L.N., Voronyna T.A., Avakyan H.N., Markyna N.V., Burd S.H., Stoyko M.Y., Oleynykova O.M., Mytrofanov A.A. Vlyyanye karbamazepyna na strukturno- funktsyonalʹnye svyazy v razvytyy épyleptycheskoy systemy // Éksperym. y klyn. farmakol. – 2002. – 65, №2. – S. 6-8.

2. Badalyan O.L. Vlyyanye kombynyrovannoho prymenenyya karbamazepyna y antyoksydantov na épyleptycheskuyu systemu: Avtoref.dys. kand.med.nauk. M. 1998.
3. Derzhavna Farmakopeya Ukrayiny / Derzhavne pidpryyemstvo «Naukovo-ekspertnyy farmakopeynyy tsentr». – 1-e vyd. – Kharkiv: RIREH, 2001. – Dopovnennya 1. – 2004. – 520 s.

4. Mazur Y.A., Voloshyn N.A., Chekman Y.S. Tyotryazolyn. - Zaporozhʹe, Lʹvov : Nautylus, 2005.- 156 s.

5. Pakhomov V. P. Khromatohrafiya u khimiko-farmatsevtychnykh doslidzhennyakh / Pakhomov V. P. // Khym. Far mats. zhurn. – 2003 – T. 37 , № 8. – S. 55-56.

6. EL, Johansen U, Christophersen AS. Drug screening of preserved oral fluid by liquid chromatography-tandem mass spectrometry. Clin Chem. 2007;53:300–9.

7. Heorhyevskyy H.V. Razrabotka kompleksa fyzyko-khymycheskykh metodyk, obespechyvayushchykh sozdanye y kontrolʹ kachestva oryhynalʹnykh otechestvennykh preparatov, proyzvodnykh 1,2,4 – tryazola// Zaporozhskyy medytsynskyy zhurnal. – 2011, tom 13. - №1. – s.58-69.

8. Pidbir optymalʹnykh umov analizu shtuchnoyi sumishi izoniazydu ta tiotryazolinu metodom vysokoefektyvnoyi ridynnoyi khromatohrafiyi / L. I. Kucherenko, O. V. Khromylʹova, Z. B. Moryak, H. I. Tkachenko // Zaporozh. med. zhurn. – 2014. – № 2. – S. 118–120.

9. Pidvyshchennya efektyvnosti likuvannya epilepsiyi / Kucherenko L. I., Byelenichev I. F., Mamchur V.Y., Opryshko V.I., Portna O.O., Nimenko H.R.– K., 2015. – 4 s. – (Inform. lyst pro novovvedennya v systemi okhorony zdorov'ya / Ukrmedpatentinform; № 225-2015, Vyp. 19 z problemy «Farmatsiya»).