UPLC-MS法测定胸腺肽注射剂中胸腺肽α₁的含量

目的:建立UPLC-MS法测定胸腺肽注射剂中胸腺肽α₁(Tα₁)的含量。方法:选用ACQUITY UPLC BEH Shield RP18色谱柱(150 mm×2.1 mm,1.7 μm),以0.1%甲酸-水(v/v)为流动相A,0.1%甲酸-25%乙腈(v/v)为流动相B,梯度洗脱,流速0.2 mL·min^-1,选用Waters Acquity UPLC-三级串联四极杆色谱质谱联用仪的选择离子监测(SIM)扫描方式进行监测,电喷雾离子源,正离子方式,选择监测离子为m/z 778.5。结果:胸腺肽α₁的线性范围为0.05~0.5 μg·mL^-1(r>0.99),检出限为0.02 μg·mL^-1,定量限为0.05 μg·mL^-1;平均加样回收率为102.0%,RSD小于5%;各种贮存条件下注射剂溶液中胸腺肽α₁均较稳定。结论:该方法适用于胸腺肽注射剂中胸腺肽α₁的测定。

Objective:To establish a UPLC-MS method for thymosin α₁(Tα₁) determination in thymopolypeptide injections.Methods:An ACQUITY UPLC BEH Shield RP18 column(150 mm×2.1 mm,1.7 μm) was adopted;The mobile phase A:0.1% (v/v) formic acid solution,B:0.1% (v/v) formic acid in 25% (v/v) acetonitrile solution was applied with gradient elution at flow rate of 0.2 mL·min^-1.Determination was carried out by electrospray positive ionization mass spectrometry in the selective ion monitoring(SIM) mode.The ion m/z 778.5 was used to quantify thymosin α₁.Results:The linearity was ranged from 0.05-0.5 μg·mL^-1(r>0.99) with the LOD 0.02 μg·mL^-1 and LOQ 0.05 μg·mL^-1.The average recovery was 102.0% in six samples with intra and inter day RSD both below 5%.Thymosin α₁ in injection solutions was found to be stable under various storage conditions.Conclusion:The established method can be used for determination of thymosin α₁ in thymopolypeptide injections.