LC-MS/MS法测定对氨基水杨酸浓度及在大鼠组织分布研究中的应用

中国临床药理学与治疗学 ›› 2017, Vol. 22 ›› Issue (3): 272-280.

LC-MS/MS法测定对氨基水杨酸浓度及在大鼠组织分布研究中的应用

袁雅文¹，李岩²，杨劲¹

¹ 中国药科大学药学院，药物代谢动力学研究中心，南京 210009，江苏； ² 新疆医科大学药学院，药剂物化教研室，乌鲁木齐 830000，新疆

收稿日期:2016-11-03 修回日期:2016-12-15 出版日期:2017-03-26 发布日期:2017-03-29
通讯作者: 杨劲，男，博士，教授，博士生导师，主要从事体内药物分析研究。 Tel: 025-83271386 E-mail: yjcpu@yahoo.com
作者简介:袁雅文，女，硕士研究生，主要从事体内药物分析研究。 E-mail: 942767671@qq.com
基金资助:
国家自然科学基金资助项目（81460570）

Development and validation of a sensitive LC-MS/MS method for the determination of para-aminosalicylic acid in different tissues in rats

YUAN Yawen¹, LI Yan ², YANG Jin ¹

¹ Center of Drug Metabolism and Pharmacokinetics, China Pharmaceutical University, Nanjing 210009, Jiangsu, China; ²Department of Pharmaceutical Sciences, School of Pharmacy,Xinjiang Medical University, Urumqi 830011, Xinjiang, China

Received:2016-11-03 Revised:2016-12-15 Online:2017-03-26 Published:2017-03-29

摘要/Abstract

摘要：

目的:开发HPLC-ESI-MS/MS方法测定大鼠组织对氨基水杨酸浓度以改善以往方法的复杂前处理过程及低灵敏度的问题，并应用于研究其在大鼠体内的分布。方法: 采用一步蛋白沉淀的前处理，选择菲诺门Synergi 4u Polar-RP 80A (150 mm×4.6 mm;4 μm)色谱柱以及梯度法进行色谱分离。内标为氯唑沙宗。采用API4000液相色谱串联质谱（LC-MS/MS）法测定。扫描方式采用多重反应监测（MRM），负离子模式。对氨基水杨酸及内标（氯唑沙宗）监测离子对为m/z 152.0→108.1，m/z 167.9→131.9。结果:总分析运行时间为6.5 min。对氨基水杨酸在0.05～50 μg/mL内线性关系良好，定量下限低至0.05 μg/mL。本实验方法学所有验证结果都符合FDA生物样品分析指导原则要求。结论:本分析方法准确可靠，并成功应用于临床前组织分布研究。

关键词: 结核病, 对氨基水杨酸, HPLC-ESI-MS/MS, 组织分布

Abstract:

AIM: To develop a novel HPLC-ESI-MS/MS method to determine P-aminosalicylic acid (PAS) in different rat tissues to overcome the weaknesses of the conventional complicated pretreatment process and its low sensitivity. METHODS: The assay was performed with a one-step pre-column process of protein precipitation for extraction and the use of a Phenomenex Synergi 4u Polar-RP 80A (150 mm×4.6 mm; 4 μm) column with gradient elution for separation. Chlorzoxazone was selected as an internal standard (IS). LC-MS/MS (API4000) was applied for determination. PAS and IS were detected by tandem mass spectrometer using multiple reaction monitoring mode (MRM) with negative electrospray ionization. The ion transitions recorded in multiple reaction monitoring mode were m/z 152.0→108.1 for PAS and m/z 167.9→131.9 for the internal standard. RESULTS: The total analytical run time was short of 6.5 min. Calibration curve was linear over the range of 0.05-50 μg/mL and the lower limit of quantification (LLOQ) was low up to 0.05μg/mL. All validation criteria were fulfilled in compliance with the guidelines of the United States Food and Drug Administration (FDA). CONCLUSION: This HPLC-MS/MS method is accurate and reliable. The described assay method has been successfully applied for the measurement of PAS in preclinical distribution study.

Key words: tuberculosis, PAS, HPLC-ESI-MS/MS, tissue distribution

中图分类号:

R965.2

袁雅文，李岩，杨劲. LC-MS/MS法测定对氨基水杨酸浓度及在大鼠组织分布研究中的应用[J]. 中国临床药理学与治疗学, 2017, 22(3): 272-280.

YUAN Yawen, LI Yan, YANG Jin. Development and validation of a sensitive LC-MS/MS method for the determination of para-aminosalicylic acid in different tissues in rats[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2017, 22(3): 272-280.

参考文献

［1］Lehmann J. Twenty years afterward historical notes on the discovery of the antituberculosis effect of paraaminosalicylic acid (PAS) and the first clinical trials［J］. Am Rev Respir Dis, 1964, 90:953-956.
［2］Gandhi NR,Nunn P,Dheda K,et al.J. Multidrug-resistant and extensively drug-resistant tuberculosis:a threat to slobal control fo tuberculosis［J］.Lancet(London,England),2013,375:1830-1843.
［3］Dye C, Williams BG, Espinal MA, et al. Erasing the world's slow stain: strategies to beat multidrug-resistant tuberculosis ［J］. Science (New York, N.Y.), 2002, 295(5562):2042-2046.
［4］Recommends EMA. Two new tuberculosis treatments［J］. Drug Discov Devel, 22, 2013.
［5］Falzon D, Jaramillo E, Schunemann HJ, et al. WHO guidelines for the programmatic management of drug-resistant tuberculosis: 2011 update ［J］. Eur Respir J, 2011, 38(3): 516-528.
［6］Para-aminosalicylic acid［J］. Tuberculosis, 2008, 88(2): 137-138.
［7］Liu P, Derendorf H. Antimicrobial tissue concentrations［J］. Infect Dis Clin North Am, 2003, 17(3): 599-613.
［8］Pentikainen PJ, Wan SH, Azarnoff DL. Bioavailability of aminosalicylic acid and its various salts in humans IV: comparison of four brands of the sodium salt［J］. J Pharm Sci, 1974, 63(9): 1431-1434.
［9］Peloquin CA, Zhu M, Adam RD, et al. Nix, pharmacokinetics of para-aminosalicylic acid granules under four dosing conditions［J］. Ann Pharmacother, 2001, 35(11): 1332-1338.
［10］Hong L, Jiang W, Zheng W, et al. HPLC analysis of para-aminosalicylic acid and its metabolite in plasma, cerebrospinal fluid and brain tissues［J］. J Pharm Biomed Anal, 2011, 54(5): 1101-1109.
［11］Cummins CL, O'Neil WM, Soo EC, et al. Determination of p-aminosalicylic acid and its N-acetylated metabolite in human urine by capillary zone electrophoresis as a measure of in vivo N-acetyltransferase 1 activity［J］. J Chromatogr B Biomed Sci Appl, 1997, 697(1/2): 283-288.
［12］Lianidou ES, Ioannou PC. Simple spectrofluorometric determination of p-aminobenzoic and p-aminosalicylic acids in biological fluids by use of terbium-sensitized luminescence［J］. Clin Chem, 1996, 42(10): 1659-1665.
［13］Gennaro MC, Calvino R, Abrigo C. Ion interaction reagent reversed-phase high-performance liquid chromatography determination of anti-tuberculosis drugs and metabolites in biological fluids［J］. J Chromatogr B Biomed Sci Appl, 2001, 754(2): 477-486.
［14］关欣，徐雅娟，解生旭，等. HPLC 法测定芪丹颗粒剂中黄芪甲苷的含量［J］. 长春中医药大学学报, 2015,31(2): 245-247.
［15］解朝霞, 张孟. 高效液相色谱法测定糖尿康胶囊中葛根素的含量［J］. 长春中医药大学学报, 2013, 29(5): 932-933.

[1]	郭璐, 钟振东, 韩奕岑, 石毅, 杨勇, 边原, 刘晓姝, 张静, 郎琴, 钟甜. 不同给药方式在大鼠内对利巴韦林的药代动力学和组织分布的影响[J]. 中国临床药理学与治疗学, 2022, 27(1): 25-32.
[2]	陈爱瑛, 程敏, 繆云萍, 叶小弟, 田雪君, 郑高利. 舒尼替尼药代动力学和脑肾组织分布特征[J]. 中国临床药理学与治疗学, 2020, 25(10): 1105-1110.
[3]	邢晗，宁晨，孔德璇，蔡卉，周时雨，任畅，孔颖，程钰洁，陈西敬. 高抗肿瘤活性汉黄芩素衍生物GL-V9在大鼠体内的组织分布和排泄研究[J]. 中国临床药理学与治疗学, 2019, 24(8): 889-895.
[4]	姚天怡, 尹少平, 刘雯, 赵炎磊, 王广基, 李娟. 紫杉醇聚合物胶束的组织分布和生物学评价[J]. 中国临床药理学与治疗学, 2017, 22(11): 1207-1214.
[5]	唐志华, 王玮, 丁洁卫. 雾化吸入氨茶碱在犬体内的组织分布[J]. 中国临床药理学与治疗学, 2014, 19(2): 150-155.
[6]	许秋霞, 王寅, 马丽焱. Na⁺偶联单羧酸转运蛋白及其在药物转运中的作用[J]. 中国临床药理学与治疗学, 2009, 14(1): 103-109.
[7]	黄秀旺, 许建华, 陈元仲. 雷公藤内酯醇在大鼠体内的组织分布[J]. 中国临床药理学与治疗学, 2008, 13(7): 764-767.
[8]	李晓天, 张丽容, 王天奎, 王红娟, 贾欣. 马钱子碱在小鼠体内的组织分布[J]. 中国临床药理学与治疗学, 2006, 11(3): 342-344.
[9]	师少军, 李忠芳, 顾世芬, 陈汇, 曾繁典. 蝙蝠葛苏林碱在兔体内的药代动力学及组织分布[J]. 中国临床药理学与治疗学, 2005, 10(6): 622-626.
[10]	范益, 丁建花, 刘苏怡, 张徐宁, 刘强, 章静, 胡刚. α-与β-甘草酸在小鼠体内分布的研究[J]. 中国临床药理学与治疗学, 2004, 9(6): 619-622.