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目的建立柱前衍生化LC-MS/MS法测定Beagle犬血浆中美沙拉嗪(5-ASA)及其主要代谢物N-乙酰-5-氨基水杨酸(N-Ac-5-ASA)的浓度,并将此方法用于美沙拉嗪肠溶片在Beagle犬体内的药物动力学研究。方法血浆样品经丙酸酐衍生后,以乙酸乙酯进行液液萃取。色谱分离采用Thermo ODS-2HYPERSIL(150mm×4.6mm,5μm)色谱柱,流动相为体积分数为0.2%的甲酸水溶液-甲醇(体积比为60∶40),流速为1.0mL·min-1(柱后分流比为5∶3)。质谱检测采用选择反应监测(SRM)模式,在电喷雾电离(ESI)源负离子化方式下,监测反应分别为m/z208→107(美沙拉嗪衍生物),m/z194→107(N-乙酰-5-氨基水杨酸),和m/z192→148(内标物对氨基苯甲酸衍生物)。测定6只Beagle犬分别单次给予美沙拉嗪肠溶片500mg后不同时刻血浆中美沙拉嗪及代谢物N-乙酰-5-氨基水杨酸的浓度,采用DAS 2.1.1软件以非房室模型计算药物动力学参数。结果血浆中美沙拉嗪和N-乙酰-5-氨基水杨酸的线性范围分别为0.05~50.00mg·L-1和0.01~1.00mg·L-1,定量下限分别为0.05mg·L-1和0.01mg·L-1;低、中、高3个质量浓度质控样品的日内,日间精密度均小于14.6%,准确度分别为95.8%~96.4%和93.3%~98.4%,提取回收率均在70.2%~80.8%之间。美沙拉嗪的主要药物动力学参数:t1/2为(3.9±1.3)h,tmax为(3.7±0.5)h,ρmax为(27.8±6.3)mg·L-1,AUC0t-为(144.4±39.4)mg·h·L-1,AUC0-∞为(150.8±33.3)mg·h·L-1。结论本法适用于血浆中美沙拉嗪及N-乙酰-5-氨基水杨酸的浓度测定。
OBJECTIVE To establish a pre-column derivatization LC-MS / MS method for the determination of mesalamine (5-ASA) and its major metabolite N-acetyl-5-aminosalicylic acid Concentration, and this method was used to study the pharmacokinetics of mesalazine enteric-coated tablets in Beagle dogs. Methods Plasma samples were derivatized with propionic anhydride and extracted with liquid ethyl acetate. The chromatographic separation was performed on a Thermo ODS-2HYPERSIL (150 mm × 4.6 mm, 5 μm) column with a mobile phase of 0.2% formic acid in methanol (60:40 by volume) at a flow rate of 1.0 mL · min -1 After the split ratio of 5: 3). The mass spectrometry detection was performed using the selective reaction monitoring (SRM) mode. In the negative ionization mode of electrospray ionization (ESI), the monitoring reactions were m / z208 → 107 (mesalazine derivatives), m / z194 → 107 -5-aminosalicylic acid), and m / z 192 → 148 (internal standard p-aminobenzoic acid derivative). The concentrations of mesalazine and its metabolite N-acetyl-5-aminosalicylic acid in plasma were determined at 6 different times after single administration of 500 mg of mesalazine enteric-coated tablets to 6 Beagle dogs respectively. Using DAS 2.1.1 software, The model calculates pharmacokinetic parameters. Results The linear ranges of mesalazine and N-acetyl-5-aminosalicylic acid in plasma were 0.05-50.00 mg · L -1 and 0.01-1.00 mg · L -1, respectively. The lower limits of quantitation were 0.05 mg · L -1 And 0.01 mg · L-1, respectively. The precision of the control samples with low, medium and high quality were both less than 14.6% and 95.8% -96.4% and 93.3% -98.4%, respectively. Rates are between 70.2% ~ 80.8%. The main pharmacokinetic parameters of mesalazine were as follows: t1 / 2 was (3.9 ± 1.3) h, tmax was (3.7 ± 0.5) h, ρmax was (27.8 ± 6.3) mg · L -1 and AUC0t- was (144.4 ± 39.4) ) mg · h · L-1 and AUC0-∞ (150.8 ± 33.3) mg · h · L-1. Conclusion This method is suitable for the determination of mesalazine and N-acetyl-5-aminosalicylic acid in plasma.