高效液相法同时测定血清中卡马西平、苯妥英钠、苯巴比妥的实验方法学研究

【摘要】目的建立高效液相色谱／二极管阵列同时测定血清中卡马西平、苯妥英钠、苯巴比妥含量的检测法。方法以600μl乙酸乙酯为萃取溶剂；样品用量0．2ml血清，萃取时间3min，以3500r／min速度离心4min，取400txl萃取剂于75℃水浴中挥发至干；用1．0ml流动相溶解残留物，10000r／min离心20min后进样分析。分析条件：柱温30℃，流动相（甲醇：水=60：40），波长254nm，实现了3种药物的有效分离。结果在优化条件下，3种待测组分在1．52～120mg／L范围内呈线性，相关系数（r）≥0．999，检出限（S／N=3）为0．4—1．5mg／L，样品的平均加标回收率为91．3％～111％，相对标准偏差（RSD）〈5％。结论该方法确定了最佳样品预处理条件，优化了色谱分离及检测条件，灵敏、准确，能够满足血清中抗癫痫药物浓度的监测要求。

Experimental methodology of simultaneous determination of carbamazepine,phenytoin, and pheno- barbital in serum by high-performance liquid chromatography

Abstract] Objective To establish a high-performance liquid chromatography （HPLC） method with diode array detection to simultaneously determine carbamazepine, phenytoin, and phenobarbital in ser- um. Methods Extraction solvent （8001xl ethylene acetate） and sample （0. 2 ml） was mixed, extracted for 2 min, and centrifuged （3500 r/min, 4 minutes）. A volume （600 μl） of extract liquor was volatilized to dryness in water bath with the volatilization temperature 75 ℃, then was redissolved with 1.0 ml mobile phase. Analysis conditions was column temperature 30, mobile phase （methanol： water =40：60 ）, and detection wavelength of 254 nm. Three metabolites were effectively separated. Results Under the opti- mized condition, calibration curves of three metabolites were linear in the ranges of （ 1.52 - 120 mg/L） and the correlation coefficients were not less than 0. 999. The detection limits （ S/N = 3 ） were in the range of O. 4 - 1.5 mg/L. The spiked recoveries were in the range of 91.3% - 111% with relative standard devia- tions （RSD） less than 5%. Conclusions The optimal pretreatment condition for the sample was estab- lished. The chromatographic separation and the detection condition were optimized. The method was sensi- tive and accurate, and could meet the need of monitoring serum drug concentration.