HPLC-ELSD测定硫酸卷曲霉素原料及其制剂中硫酸根含量

Determination of sulfate in capreomycin sulfate raw materials and its preparation by HPLC-ELSD

目的:建立高效液相色谱-蒸发光散射检测法(HPLC-ELSD法)测定硫酸卷曲霉素及其制剂中硫酸根含量。方法:釆用GRACE Apollo C_(18)色谱柱(250 mm×4.6 mm,5μm),柱温35℃,流动相为0.2 mol·L^(-1)三氟乙酸溶液-甲醇(94:6),流速0.6 mL·min^(-1),进样量20μL;漂移管温度为110℃,载气流量为2.8 L·min^(-1),增益为1。结果:硫酸线性范围为0.015~0.300 mg·mL^(-1)(r=0.997 8),日内RSD为0.3%,精密度为0.4%~1.0%,平均回收率为101.6%(RSD=1.3%,n=9)。原料及其制剂的硫酸根含量为19.3%~23.3%,结合硫酸分子数为1.7~2.0。结论:经方法学验证本法能测定硫酸卷曲霉素原料及其制剂的硫酸盐成分含量,可为其质量标准提高与质量控制提供参考。

Objective： To develop a rapid and simple method for the separation and quantitation of sulfate in eapreomycin sulfate and its preparation using HPLC coupled with evaporative light scattering detection （ HPLC- EKSD ）. Methods： A GRACE Apollo Cis （ 250 mm× 4.6 mm, 5 μm ）column was adopted. The column temperature was set at 35 ℃. The mobile phase consisted of 0.2 mol·L-1 trifluoroacetic acid-methanol （ 94 ： 6 ） at a flow rate of 0.6 mL·min-1, and the injection volume was 20 μL; The drift tube temperature of Alltech 2000ES was 110 ℃, the flow rate of cartier gas was 2.8 Lmg·min 1, and the gain was 1. Results： The content determination of sulfate incapreomycin sulfate and its preparation by HPLC-EI,SD was achieved. The linear range of sulfate was 0.015-0.300 mg· mL-1 （ r=0.997 8 ）with average recovery of 101.6% （ RSD=1.3%, n=9 ）, the inter-day RSD was 0.3%, and the precision was 0.4%-1.0% ）. The content of sulfate in capreomycin sulfate and its preparations was 19.3%-23.3%. The molecular number of sulfuric acid was 1.7-2.0. Conclusion： The new established HPLC-ELSD method shows good repeatability, linearity and robustness, which can be used as a rerence of the improvement of quality control of sulfate in capreomycin sulfate and its preparations.