The HPLC analysis of urinary aminolevulinic acid after chemical derivatization and purification derivatives on hypercross-linked polystyrene (Purosep-200)
- 1 Dr.Sc. (Med.) Senior Research, Laboratory of Experimental and Clinical Biochemistry and Immunology, Research Institute of Molecular Medicine, Chita State Medical Academy
- 2 Head of the Laboratory of Chemical Analysis, Zabaikalsky State University, Chita
- 3,4,5 Post-graduate Student, Department of Chemistry, Zabaikalsky State University, Chita
- 6 Physician Ophthalmologist, Regional Hospital № 2, Chita
- 7 Physician Ordinator, Department of Neurology, State Medical Academy, Chita
Abstract
We proposed HPLC method of analysis urinary aminolevulinic acids on the basis of updating method [1]. To aliquot daily urine, which the stabilized acetic acid, added acetylacetone and formaldehyde. Heated up during 10 min at 100ºС and after cooling to a room temperature, cleared on a cartridge packed 30 mg hypercross-linked polystyrene (Purosep-200). Derivatives separation on a monolithic column Chromolith Performance RP-18e 100×4.6 mm and fluorimetric detection at ex370-em460 nm. Mobile phase consisted of 30 % acetonitrile and 0.025 % trifluoroacetic acid. Flow rate was 1.4 ml/min. Fast clearing of derivatives on cartridges without evaporation, has allowed to receive pure extracts. Full separation was reached less than for 2 minutes. Simplicity and sufficient sensitivity of a method, allow to use it in a clinical practice for diagnostics porphyries. Tomokuni et all. [1].
How to cite
Дутов А.А., Никитин Д.А., Сверкунова А.В., Мартынова А.В., Коновалова О.Н., Ермолина А.В., Лукьянова Ю.Л. ВЭЖХ-анализ аминолевулиновой кислоты в моче после химической дериватизации и очистки дериватов на сверхсшитом полистироле (Purosep-200) // Вопросы биологической, медицинской и фармацевтической химии. 2014. № 4. С. 25–29.
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