Qian Guang-sheng, Han Hai, Li Zhang-wan, Weng Ling-ling, Zheng Hu
West China School of Pharmacy, Sichuan University, Chengdu 610041, China.
Sichuan Da Xue Xue Bao Yi Xue Ban. 2004 Sep;35(5):711-4.
To establish an HPLC method and a non-aqueous titration for the determination of piperazinylethylestrone drug substance, and an HPLC method for the determination of piperazinylethylestrone in dog plasma.
Anhydrous acetic acid as solvent, 0.1 mol/L perchloric acid as titrant, crystal violet solution as indicator to establish non-aqueous titrations and ODS column as stationary phase, methanol and a mixture of 0. 025 mol/L sodium phosphate monobasic and 0.02 mol/L sodium dedecyl sulfate (80:20) [adjusted with phosphoric acid to a pH (4.8 +/- 0.1)] as mobile phase, 220 nm as detective wavelength to establish HPLC-UV method for determination of piperazinylethylestrone drug substance; FMOC-CL as a derivatization reagent, FD as a detector to establish an HPLC method with derivatization and column switching for determination of piperazinylethylestrone in dog plasma.
The RSD of non-aqueous titrations within-day and between-day were 0.28% and 0.21%, respectively; the established HPLC-UV method had good linearity and precision within the range of 1.001-5.005 microg of piperazinylethylestrone, the detection limit was 4 ng (S/N=3); the linearity of HPLC method with derivatization and column switching was within the range of 8.4-420 ng/ml, the detection limit was 1 ng (S/N=3). The clean-up recoveries were from 77.24% to 83.10%, and the method recoveries were 98.33%-103.3%. The RSD of within-day and between-day were less than 7.7% and 7.3%, respectively.
The above three methods are simple, accurate and precise for the determination of piperazinylethylestrone drug substance and for the determination of piperazinylethylestrone in dog plasma.
建立高效液相色谱法（HPLC）和非水滴定法测定哌嗪基乙基雌酮原料药，以及建立HPLC法测定犬血浆中哌嗪基乙基雌酮。
以无水乙酸为溶剂，0.1 mol/L高氯酸为滴定剂，结晶紫溶液为指示剂建立非水滴定法；以ODS柱为固定相，甲醇与0.025 mol/L磷酸二氢钠和0.02 mol/L十二烷基硫酸钠混合溶液（80∶20）[用磷酸调至pH（4.8±0.1）]为流动相，220 nm为检测波长建立HPLC-UV法测定哌嗪基乙基雌酮原料药；以9-芴甲氧羰基氯（FMOC-CL）为衍生化试剂，荧光检测器（FD）为检测器，建立衍生化-柱切换HPLC法测定犬血浆中哌嗪基乙基雌酮。
非水滴定法日内、日间RSD分别为0.28%和0.21%；所建立的HPLC-UV法在1.0015.005 μg哌嗪基乙基雌酮范围内线性关系良好，精密度高，检测限为4 ng（S/N=3）；衍生化-柱切换HPLC法线性范围为8.4420 ng/ml，检测限为1 ng（S/N=3）。净化回收率为77.24%83.10%，方法回收率为98.33%103.3%。日内、日间RSD分别小于7.7%和7.3%。
上述三种方法用于测定哌嗪基乙基雌酮原料药及犬血浆中哌嗪基乙基雌酮，方法简便、准确、精密。
