反相高效液相色谱法定量检测基因重组胸腺素α_1

RP-HPLC Determination of Recombination Thymosin Alpha 1 (Tα_1)

目的:采用反相高效液相色谱法(RP-HPLC)测定胸腺素α_1含量。方法:在 Waters Alliance 2690高效液相色谱仪上,以 Waters Xterra C_(18)柱(150mm×3.9mm,5μm)为分离柱,996检测器,紫外检测波长215 nm,选择0.05%三氟乙酸-水溶液作流动相 A、0.05%三氟乙酸-乙腈作流动相 B 进行线性梯度洗脱,B 相0,30,35 min 比例依次为0%,60%,100%。结果:胸腺素α_1(Tα_1)在5.76～13.44μg·mL^(-1)浓度范围内呈现良好的线性关系(r=0.9997,n=5),平均回收率大于95%。结论:RP-HPLC 法测定胸腺素α_1的方法准确、快速、可行。

Objective ：To develop a method to determine the contents of Recombination Thymosin alpha 1 （Tα1） by RP- HPLC. Methods： RP- HPLC method, Waters Xterra C18 （150 mm × 3.9 mm,5 μm） ;mobile phase A was 0. 05 % trifluoracetic - water, mobile phase B was 0. 05 % trifluoracetic - acetonitrile, the ratio of phase B was 0%, 60%, 100% in 0,30,35 minutes respectively;detecting wavelength was 215 nm;the flow rate was 0. 5 mL/min. Resuits ：The calibration curve was linear in the concentrations Conclusion ：This method is simple and reliable, which can ranged from 5.76 to 13.44 μg · mL^- 1 （ r = 0. 9997 ）. be applied to the determination of Tα1.