LC-MS/MS快速测定大鼠血浆中HHY-002浓度

目的: 建立用于测定治疗慢性心律失常药物HHY-002的大鼠血浆测定法。方法: 流动相为乙腈-0.1%甲酸(65∶35),流速 0.200 mL/min,色谱柱Phenomenex Luna C₁₈柱(150 mm×2.00 mm,5 μm,5 micron),进样量为 10 μL,柱温 35 ℃,以地西泮为内标。结果: HHY-002在 1.98~1013.50 ng/mL 的范围内呈良好的线性关系(r=0.998),最低定量限达 1.98 ng/mL,绝对回收率高于80%,日内和日间误差小于10%,方法回收率大于(81.4±4.5)%,符合生物样品分析要求。结论: 建立的LC-MS/MS方法专属性强,灵敏度高,可用于HHY-002的体内定量分析。

LC-MS/MS determination of HHY-002 in rat plasma

AIM: A highly sensitive and reliable LC-MS/MS method was validated for the determination of HHY-002 in rat plasma, a novel antiarrhythmic. METHODS: After addition of diazepam as internal standard(IS); LC separation with a Phenomenex Luna C₁₈ analytical column (150 mm×2.00 mm, 5 μm, 5 micron) and quantification by MS/MS after positive ESI. The mobile phase consisted of acetonitrile∶water∶formic acid (65∶35∶0.1, V/V/V) in an isocratic elution mode pumped at 0.200 mL/min, and with the injection volume of 10 μL and column oven temperature of 35 ℃. RESULTS: The calibration curve ranging from 1.98 to 1013.50 ng/mL. Lower limit of quantification (LLOQ) was 1.98 ng/mL in 100 μL of plasma. Extraction recovery was above 80%, inter- and intra- day accuracy and precise was below 10%.The method recovery was over(81.4±4.5)%.This method was shown to meet all requirements in bioanalytical determination.CONCLUSION: The method was successfully applied to the determination of HHY-002 in rat plasma.