液相色谱-质谱联用方法测定大鼠血浆、尿液及结肠组织中美沙拉嗪及其代谢产物N-乙酰-5-ASA的浓度

Development of LC-MS /MS Method for Determination of Mesalazine and N-Ac-5-ASA in Rat Plasma,Urine and Colon

目的建立应用LC-MS/MS进行大鼠血浆、尿液、结肠中美沙拉嗪及其体内代谢物N-乙酰-5-ASA浓度的测定方法,并将其应用于美沙拉嗪及N-乙酰-5-ASA在大鼠体内过程的研究。方法血浆、尿液及结肠样品均用甲醇沉淀蛋白进行提取,以ESI电离源在负离子模式下检测,用Agilent HC-C18色谱柱,甲醇-醋酸水溶液为流动相梯度洗脱,流速为1.0 mL·min-1。结果美沙拉嗪及N-乙酰-5-ASA在大鼠血浆、尿液及结肠中的线性范围及定量限均符合测定要求,日内、日间精密度及基质效应均<15%。结论本方法快速、灵敏、专属性强且重现性好,可用于大鼠血浆、尿液、结肠中美沙拉嗪及N-乙酰-5-ASA测定。

OBJECTIVE To develop an LC-MS/MS method for determination of mesalazine and its metabolite, N-Ac-5-ASA, in rat plasma, urine and colon for the pharmaeokinetic study of mesalazine and N-Ac-5-ASA in rats. METHODS The plasma, urine and colon samples were treated with methanol to precipitate protein. An Agilent HC-CI8 column was used as the analytical column, with methanol and acetic acid aqueous solution as mobile phase eluted by a gradient program at a flow rate of 1.0 mL · min-1. The detection was accomplished with ESI source operated in negative ionization mode. RESULTS The linearity and LLOQs of mesalazine and N-Ac-5-ASA in rat plasma, urine and colon both met the requirements, and the inter- and intra-day RSDs and matrix effect were less than 15%. CONCLUSION This assay is a rapid, sensitive, selective and reliable method for determination of mesalazine and N-Ac- 5-ASA in rat plasma, urine and colon.