基于微透析技术及HPLC-ESI-MS测定大鼠关节腔透析液中双氯芬酸钠浓度及其应用

Determination of diclofenac sodium in dialysate of joint cavity in rats based on microdialysis and HPLC-ESI-MS and its application

目的:利用微透析技术为采样平台,以双氯芬酸钠为工具药,建立大鼠膝关节腔透析液中双氯芬酸钠的HPLC-ESI-MS测定方法,满足非甾体类抗炎药物靶部位药物采集及测定的需求。方法:采用HPLC-MS单极四极杆联用技术,通过优化色谱、质谱条件,建立快速灵敏的双氯芬酸钠的液-质测定方法,并将该法应用于大鼠关节腔透析液中双氯芬酸钠浓度的测定。本法所用色谱柱为Waters Symmetry-C18(dp 5μm,150mm×2.1mm ID),流动相为甲醇-0.1%甲酸(80∶20,V/V)。结果:本实验建立的大鼠膝关节腔透析液中双氯芬酸钠的HPLC-ESI-MS测定方法的最低定量限为1.95ng/mL,在1.95～125ng/mL范围内线性关系良好,批内批间精密度均小于10%。介质效应在99.9%～113.4%之间。结论:本实验成功地将微透析技术引入到大鼠关节腔部位的取样过程中,建立的大鼠关节腔透析液中双氯芬酸钠的HPLC-ESI-MS测定方法快速,灵敏。两者的结合有助于实现对以双氯芬酸钠为代表的非甾体类抗炎药物作用靶部位的取样及其药动学研究。

AIM： To develop and validate a high liquid chromatography-electrospray ioniza- tion-mass spectrometry （HPLC-ESI-MS） meth- od for the quantification of diclofenac sodium in dialysate of joint cavity in rats in order to meet the requirement of collection and determination of non-steroidal anti-inflammatory drugs （NSAIDs） in target site based on microdialysis and use diclofenac sodium as paradigm drug. METHODS： A sensitive and specific liquid chro matography-electrospray ionization-mass spec- trometry （LC-ESI-MS） method using a unipolar quadrupole technology by optimizing chromatog raphy and mass spectrometry conditions has been applied to determine the concentration of diclofenac sodium in dialysate of joint cavity in rats. Dialysate samples were separated by HPLC on a reversed phase C18 column [Waters Symme try-C18（dp 5 /μm, 150 mm×2.1 mm ID）]with a mobile phase of methanol- 0.1% formic acid （80 ： 20, V/V）. RESULTS：The method was val-idated over the concentration range of 1.95-125 ng/mL with the low limit of quantitation of di clofenac sodium in dialysate was 1.95 ng/mL. Within-batch and between-batch precision （RSD. %） were all within 10% while matrix effect were between 99.9% - 113.4%. CON CLUSION. The microdialysis technology was successfully applied in the collection of dialysate samples in joint cavity of rats, the determination method described in this paper is rapid also has a high sensitivity. The combination of two technol ogies were suitable for the collection of synovial samples, also will be helpful to study the phar macokinetic profile of non-steroidal anti-inflam- matory drugs （NSAIDs） in target site.