HPLC-MS/MS法测定胸腺肽肠溶片中胸腺素α_1的含量

Content Determination of Thymosin α_1 in Thymopolypeptides Enteric-coated Tablets by HPLC-MS/MS

目的:建立测定胸腺肽肠溶片中胸腺素α_1含量的方法。方法:采用高效液相色谱-串联质谱法。色谱柱为Luna C_(18)(2),流动相为0.1%甲酸-乙腈(梯度洗脱),流速为0.7 mL/min,柱温为30℃,进样量为20μL;离子化模式为电喷雾电离,喷雾电压为4.5kV,鞘气流速为60 arb,辅助气流速为30 arb,吹扫气流速为10 arb,毛细管温度为320℃,工作模式为正离子监测模式。结果:胸腺素α_1检测质量浓度线性范围为1~1 000 ng/mL(r=0.999 9);定量限为1 ng/mL,检测限为0.1 ng/mL;精密度、稳定性、重复性试验的RSD<2.0%;加样回收率为95.0%~98.0%(RSD=1.2%,n=6)。结论:该方法简单快速、灵敏度高、结果准确,适用于胸腺肽肠溶片中胸腺素α_1含量的测定。

OBJECTIVE：To establish a method for content determination of thymosin α_1 in Thymopolypeptides enteric-coated tablets. METHODS：HPLC-MS/MS method was adopted. The determination was performed on Luna C_（18）（2）with mobile phase consisted of 0.1% formic acid-acetonitrile（gradient elution）at the flow rate of 0.7 mL/min. The column temperature was set at 30 ℃,and sample size was 20 μL. ESI was used with ion spray voltage of 4.5 kV,sheath gas flow rate of 60 arb,aux gas flow rate of30 arb,sweep gas flow rate of 10 arb and capillary temperature at 320 ℃.The working mode was positive ion monitoring mode.RESULTS：The linear range for thymosin α_1 was 1-1 000 ng/mL（r=0.999 9）. The limit of quantitation was 1 ng/mL. The limit of detection was 0.1 ng/mL. RSDs of precision,stability and reproducibility tests were all lower than 2.0%. The recoveries were95.0%-98.0%（RSD=1.2%,n=6）,respectively. CONCLUSIONS：The method is simple,rapid,sensitive and accurate,and can be suitable for simultaneous determination of thymosin α_1 in Thymopolypeptides enteric-coated tablets.