Kamińska Adrianna, Głowacka Iwona E, Pasternak Beata, Głowacki Rafał, Chwatko Grażyna
University of Lodz, Faculty of Chemistry, Department of Environmental Chemistry, 163 Pomorska Str., 90-236 Łódź, Poland.
Medical University of Lodz, Faculty of Pharmacy, Laboratory of Bioorganic Chemistry, 1 Muszyńskiego Str., 90-151 Łódź, Poland.
Bioanalysis. 2019 Jul;11(14):1359-1373. doi: 10.4155/bio-2019-0011. Epub 2019 Aug 1.
The first method on urinary excreted amounts of lipoyllysine (LLys) after lipoic acid (LA) supplementation was developed and validated. The suggested procedure allowed simultaneous determination of LLys and LA. After the conversion of analytes into their reduced forms with tris(2-carboxyethyl)phosphine and derivatization via thiol group with 1-benzyl-2-chloropyridinium bromide, separation of analytes derivatives was performed on C18 column using a gradient mobile phase consisting of acetic acid and acetonitrile. The calibration curves for LA and LLys were linear (R > 0.999) in the range of 0.4-12 μM concentration and all validation results were acceptable, according to the US FDA bioanalytical method guidelines. This method was effectively applied for LA and LLys quantification in human urine after oral LA supplementation.
开发并验证了第一种测定补充硫辛酸（LA）后尿中硫辛酰赖氨酸（LLys）排泄量的方法。所建议的程序能够同时测定LLys和LA。在用三（2-羧乙基）膦将分析物转化为还原形式并通过1-苄基-2-氯吡啶溴化物对硫醇基团进行衍生化后，使用由乙酸和乙腈组成的梯度流动相在C18柱上对分析物衍生物进行分离。根据美国食品药品监督管理局（US FDA）生物分析方法指南，LA和LLys的校准曲线在0.4 - 12 μM浓度范围内呈线性（R > 0.999），所有验证结果均可接受。该方法有效地应用于口服LA补充后人尿中LA和LLys的定量分析。
