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目的:建立超高效液相色谱-串联质谱测定血浆中新型酪氨酸酶抑制剂UP302的方法,并用于研究UP302在大鼠、狗、猴和人血浆中的稳定性。方法:血浆中加入内标大豆苷元经2倍体积甲醇沉淀后进行分析。采用Hypersil Gold C18(50 mm×2.1 mm,1.9μm)色谱柱,流动相为甲醇(A)-5 mmol.L-1甲酸铵水溶液(B),梯度洗脱,梯度流速恒定为0.2 mL.min-1,柱温为30℃,整个分析时间为6 min。采用负离子电喷雾离子化电离源和选择反应监测模式进行检测。结果:在5～2000 ng.mL-1的浓度范围内,标准曲线线性关系良好(r=0.9998);血浆中UP302最低定量下限为5 ng.mL-1;本方法日内日间准确度在99.2%～107.3%,日内日间精密度均小于9.3%。结论:本方法灵敏度高,重现性好,操作简便,可用于血浆中UP302浓度的测定。
OBJECTIVE: To establish a method for the determination of UP302, a novel tyrosinase inhibitor in plasma, using ultra performance liquid chromatography-tandem mass spectrometry and to study the stability of UP302 in rat, dog, monkey and human plasma. Methods: The internal standard daidzein was added into the plasma and analyzed by 2-fold volume methanol precipitation. The mobile phase was methanol (A) -5 mmol.L-1 ammonium formate aqueous solution (B) with gradient elution using a Hypersil Gold C18 column (50 mm × 2.1 mm, 1.9 μm) with a constant flow rate of 0.2 mL · min -1, the column temperature was 30 ℃, the whole analysis time was 6 min. Negative ion electrospray ionization ionization source and selective reaction monitoring mode were used for detection. Results: The standard curve showed good linearity (r = 0.9998) within the range of 5-2000 ng · mL-1, and the lowest limit of quantification of UP302 in plasma was 5 ng.mL-1. The daytime and daytime accuracy of this method was 99.2 % ~ 107.3%, the intra-day and inter-day precision were less than 9.3%. Conclusion: The method has high sensitivity, good reproducibility, easy operation and can be used for the determination of UP302 in plasma.