HPLC-ESI-MS联用法测定大鼠血浆中KJY-01浓度及其药动学研究

HPLC-ESI-MS determination of KJY-01 in rat plasma and pharmacokinetics research

目的建立测定KJY-01大鼠血浆中药物浓度的高效液相色谱。电喷雾离子化。质谱（HPLC—ESI-MS）联用的分析方法，对其进行大鼠体内的药动学研究。方法取大鼠血浆50μL，加入内标酮康唑，用甲醇提取后取上清液吹干，用80％甲醇100μL溶解，取2μL进行HPLC-MS测定。色谱柱为Agilent ZORBAX Eclipse plus C18（150mm×2．1mm，5μm），流动相为20mmol醋酸铵（甲酸调节PH为3．75）：甲醇（含1‰的甲酸）=22：78，流速为0．25mL／min，采用选择离子检测（SIM）法检测SQ109（m／z=331.3），KJY-01（m／z=475．4），酮康唑（内标，m／z=531．2）。结果SQ109的血药浓度在10～5000ng／mL范围内线性关系良好，最低检测限为10ng／mL，以质控样品计算，在各浓度水平下，此法的回收率均大于80％，日间和日内精密度小于20％，符合生物样品分析要求。结论该方法操作简便、快速、灵敏、专属性强，可用于KJY-01的大鼠体内大批量样品定量分析及早期药动学研究。

Objective To develop an HPLC-ESI-MS method for the determination of KJY-01 in rat plasma and the pharmacokinetic study of KJY-01. Methods The internal standard （Ketoconazole） was added to 50μL rat plasma,then extracted with methanol and evaporated,the residue was dissolved with 80% methanol and determined by injecting 2μLto HPLC-MS system.The Agilent ZORBAX Eclipse plus C18（150mm×2. 1mm, 5μm）column was adopted. The mobile phase was consisted of 20mmol ammonium acetate（pH 3.75 adjusted by formic acid）-methanol including 1‰ formic acid（22： 78）. Selected Ion Mass（SIM） was used to determine SQ 109（m/z=331.3）, KJY-01 （m/z=475.4）, Ketoconazole（IS, m/z=531.2）. Results The assay had a calibration range from 10 to 5000 ng/mLand the lowest limit of quantification （LOQ） of 10 ng/mL for SQ109. The recovery was above 80%. The intra-and inter-day RSDs were less than 20%. Conclusion This method is proved to be convenient, rapid, sensitive and suitable for pharmacokinetic study of KJY-01.