A METHOD FOR DETERMINATION OF UKRAIN IN BLOOD PLASMA FOR MONITORING AND PHARMACOKINETIC STUDY

DOROSHENKO Y.M.,¹ HODYSH Y.Y.,² UGLYANITSA K.N.,³ NEFYODOV L.I.¹

1) Institute of Biochemistry, National Academy of Sciences, Grodno, Belarus.
2) Department of Inorganic Chemistry, Technical University, Vienna, Austria.
3) Grodno Medical Institute, Grodno, Belarus.

Address for correspondence: Y.M. Doroshenko, Institute of Biochemistry, Belorussian National Academy of Science, 50 BLK, Grodno 230017, Belarus.
Tel: (0152) 33-62-71 Fax: (0152) 33-41-21
E-mail: nef@biochem.belpak.grodno.by

Summary: We developed a method using high-performance liquid chromatography for the determination of the main fluorescent component of Ukrain, a novel antitumor and immune-stimulating drug. Our method was based on ion-pair separation of Ukrain from perchloric acid extracts using reversed-phase column, buffer with high molarity (0.5 M potassium phosphate, pH 2.65), high concentration of ion-pair reagent in the mobile phase (10 mM octylsulfonic acid), controlled temperature of the separation (45°C) and detection by fluorescence (360/455 nm). Under the above conditions a peak of the main Ukrain compound was resolved from fluorescent peaks of the sum of alkaloids of Chelidonium majus L. although several peaks of alkaloids were retained in Ukrain as traces. The height of this main peak was nearly constant, while the alkaloid peaks varied depending on the series of the preparation; chelidonine and thio-triethylenephosphoramide gave no peaks. Analytical recovery for Ukrain from human plasma was 98.0 ± 4.5%. Therefore, Ukrain possesses neither significant stable binding to plasma proteins nor adsorption in blood cells.