HPLC同时测定血液与尿液中的泼尼松和泼尼松龙

Simultaneous determination of Prednisone and Prednisolone in the blood and urine by HPLC

目的建立同时检测血、尿中泼尼松与泼尼松龙的方法。方法采用HPLC法，乙酸乙酯为样品萃取剂，以倍他米松作为内标，色谱柱为Zorbax SB—Aq柱，流动相为甲醇-0．01mol·L^-1KH2PO4（57：43）。结果泼尼松、泼尼松龙在血和尿中的线性范围分别为0．05～40．00ug·ml^-1（r≥0．9997）和0．04～50．00ug·ml^-1（r≥0．9991）；两者在血和尿中的检测限分别为0．012、0．008ug·ml^-1，平均方法回收率为92．9％～106．3％，日内、日间精密度≤7．8％，萃取回收率≥85．4％。结论所建方法准确、灵敏、重复性好，可同时测定血、尿中的泼尼松与泼尼松龙。

OBJECTIVE To establish a method for simultaneous determination of Prednisone and Prednisolone in the blood and urine. METHODS The compounds were extracted from blood and urine using ethyl acetate as the solvent for liquid - liquid extraction and detected by HPLC with Betamethasone as the internal standard. Separation was achieved on Zorbax SB - Aq column using methanol -0. 01 mol. L^-1 potassium dihydrogen phosphate （57 ： 43 ） as mobile phase. RESULTS The linear limits of Prednisone and Prednisolone ranged from 0.05 ug. ml^-1 to 40.00 ug. ml^-1 in the blood （ r ≥ 0. 9997 ） and 0.04ug. ml^-1 to 50.00ug. ml^-1 in urine （ r ≥ 0.9991 ） , respectively. The limits of detection for these two analytes were 0.012ug. ml^-1 and 0. 008ug. ml^-1 in the blood and urine, respectively. The intra and inter day assay precision of this method were no more than 7.8%. The mean recovery of Prednisone and Prednisolone in the blood and urine ranged from 92.9% to 106.3% , and the mean extraction recovery of these two analytes was no less than 85.4%. CONCLUSION The method is accurate, sensitive, well reproducible and suitable for simultaneous determination of Prednisone and Prednisolone.