向志雄刘文丽（上海医药集团股份有限公司中央研究院 上海 201203）

UPLC-MS/MS 法测定比格犬血浆中双氯芬酸钠的浓度

向志雄*刘文丽
（上海医药集团股份有限公司中央研究院 上海 201203）

摘 要目的：建立灵敏的超高效液相色谱-质谱联用法测定比格犬血浆中的双氯芬酸钠的浓度。方法：选用BEH C18（2.1 mm×100 mm, 1.7 mm）色谱柱，以乙腈-醋酸铵（2 mmol·L-1）-甲酸（70∶30∶0.1，v/v/v）为流动相，采用等度洗脱进行分离，样品采用含0.1% 甲酸的乙腈进行蛋白沉淀后进样，通过电喷雾电离源, 以多重反应监测（MRM）方式进行负离子检测, 用于定量分析的离子对为m/z （293.9+293.9+296.1）→（249.9+213.7+251.8） (双氯芬酸)和m/z 283.0→239.0（内标, 大黄酸）测定双氯芬酸钠。结果：双氯芬酸钠的线性范围为：1.0 ~ 3 000.0 ng·ml-1；定量下限可达1.0 ng·ml-1；日内、日间精密度（RSD）均小于15 %。结论: 本方法同时选择3个碎片离子作为双氯芬酸钠定量产物离子，明显的改进了分析方法的灵敏度, 血浆用量少,适用于比格犬血浆样品中双氯芬酸钠的测定和药物动力学研究。

关键词超高效液相色谱-质谱联用法 双氯芬酸钠 比格犬血浆样品检测 血药浓度

文献标示码：A

UPLC-MS/MS determination of the diclofenac sodium (DS) in dog plasma

XIANG Zhixiong*, LIU Wenli
(Central Research Institute, Shanghai Pharmaceuticals Holding Co. Ltd., Shanghai 201203, China)

ABSTRACTObjective: To develop a UPLC-MS/MS method for determination of diclofenac sodium (DS) in dog plasma. Methods: DS and internal standard were extracted from plasma by acetonitrile containing 0.1% formic acid and separated on a column BEH C18（2.1 mm×100 mm, 1.7 mm）using acetonitrile-ammonium acetate (2 mmol·L-1)-formic acid (0.1%) (70∶30∶0.1, v/v/v) as a mobile phase. A tandem mass spectrometer equipped with electrospray ionization source was adopted as a detector. Multiple reaction monitoring (MRM) transitions of m/z (293.9+293.9+296.1) →(249.9+213.7+251.8), m/z 283.0→239.0 were measured in negative mode for DS and internal standard rhein, respectively. Results: The calibration curve for DS was linear over the range of 1.0~3 000.0 ng·ml-1and the lower limit of quantification was 1.0 ng·ml-1with the relative standard deviation of intra-day and inter-day over the entire concentration range less than 15%. Conclusion: This method is sensitive and suitable for pharmacokinetic study of DS in dog subjects.

KEY WORDSUPLC-MS/MS; diclofenac sodium; Beagle dog plasma sample testing; plasma concentration

双氯芬酸钠双释放肠溶胶囊（diclofenac sodium，DS）是一种非甾体抗炎药，它具有独特的双释放系统，主要通过抑制前列腺素合成而发挥抗风湿、消炎、镇痛及解热作用。临床上常用于类风湿性关节炎、肌腱炎、骨关节炎、滑囊炎及其他关节或关节周围疾患的消炎及镇痛[1]。有关双氯芬酸钠血药浓度的测定方法，国内外报道主要有 HPLC-UV、GC、LC-MS、LC-MS/MS 和UPLC-MS/MS法[2-8]，分析时间较长，前处理方法有蛋白沉淀、液-液萃取、固相萃取，血浆需求量一般比较大（0.1~2.0 ml），灵敏度较低（2.5～80 ng·ml-1）。……