高效液相色谱中流动相pH值对氢化可的松琥珀酸钠保留行为的影响

目的:考察高效液相色谱中流动相pH值的变化对氢化可的松琥珀酸钠保留行为的影响。方法:色谱柱为VarianC18,流速为1.0mL.min-1,柱温为40℃,检测波长为242nm,进样量为20μL,流动相为甲醇-磷酸盐缓冲液(8mmol.L-1磷酸二氢钾液,43∶57,临用前用8mmol.L-1磷酸氢二钾溶液调节pH值分别为5.0、5.2、5.6、6.0)。结果:流动相pH值的变化仅改变氢化可的松琥珀酸钠和游离氢化可的松的保留时间,不改变其出峰顺序,当pH值为5.2时氢化可的松琥珀酸钠峰形及与游离氢化可的松分离较好,对称度在0.95~1.05之间。结论:本方法可为常用药物色谱行为规律的基础性系统研究提供依据。

Effect of pH Value of Mobile Phase in HPLC on the Retention Behavior of Hydrocortisone Sodium Succinate

OBJECTIVE:To study the effect of pH value of mobile phase in HPLC on the retention behavior of hedrocortisone succinate sodium.METHODS:The separation was performed on Varian C18 with the column temperature at 40℃. The mobile phase consisted of methanol-phosphate buffered solution(43∶57) with(8mmol·L-1 KH2PO4 solution,the pH value of the mobile phase was adjusted to 5.0, 5.2, 5.6 and 6.0,respectively with 8mmol·L-1 K2HPO4 solution) flow rate at 1.0mL·min-1,detection wavelength at 242nm and sample size at 20μL. RESULTS: The pH value of mobile phase could only change the retention time of hydrocortisone sodium succinate and hydrocortisone, but not their peak occurring order.At a pH value of 5.2, both hydrocortisone sodium succinate and free hydrocortisone separated well and their peaks were in good shape,with symmetry value at 0.95～1.05.CONCLUSION:This method can provide bases for the basic systematic study of chromatographic behaviors of commonly used drugs.