HPLC-MS/MS法测定人血浆中烟酸及其代谢物

Determination of Nicacid and Its Metabolites in Human Plasma by HPLC-MS/MS

目的建立人血浆中烟酸及其代谢物的高效液相色谱-质谱联用（HPLC-MS／MS）测定法。方法血浆样品加乙腈沉淀蛋白，取上清液挥干，用流动相溶解，二氯甲烷反洗后进样分析。使用Genimi C18柱（50×3．00mm，3μm），柱温35℃，流动相为0．1％乙酸水溶液；甲醇：异丙醇（98：1：1），流速为0．2mL／min，以正离子MRM模式检测，烟酸、烟酰胺、烟脲酸和内标6-甲基烟酸的检测离子对质荷比（m/z）分别为124．1→80．0、123．1→80．0、181．1→135．0和138．1→92．0。结果烟酸线性范围为1．25→320μg／L，烟酰胺和烟脲酸线性范围均为1．25→1280μg／L，最低检测限均为1．25μg／L，日内和日间RSD均小于9％，方法回收率为89％～105％。结论该法具有快速、灵敏、准确、简便等特点，适用于人血浆烟酸浓度测定及药代动力学和生物利用度研究。

Objective To develop a sensitive HPLC-MS/MS method for the determination of nicacid and its metabolites in human plasma. Methods The assay was conducted with an API 3000 LC-MS/MS system comprising of a Genimi C18 column （50 × 3.00 mm, 3 μm）, and an eluate of 0. 10% acetic acid-methanol-isopropyl alcohol （98 1 ： 1）, and flow rate was 0.2 mL/min. Acetonitrile was used to precipitate protein from the plasma samples. The loading samples contained the residue from the supernatant that were dissolved ix/the eluate solution and rinsed by diehloromethane was used as the loading samples. The ion pairs of m/z 124. 1→80. 0, m/z 123. →80. 0, m/z 181.1→135.0 and m/z 138. 1→92.0 were used to quantify nicaeid, niacinamide, nicotinuric acid and 6-methyl nicotinic acid （IS）, respectively. Results The standard curves of nicacid, niacinamide and nieotinuric were linear in the range of 1.25-320 μg/L,1.25-1280 μg/L and 1.25-1280μg/L, respectivly. All with a low determination limits of 1.25 μg/L and a less than 9% within-day and inter-day RSD. The recovery rates reached 89% to 105%. Conclsuion The method is simple, rapid, sensitive, and suitable for the determination of nicaeid and its metabolites in human plasma.