高效液相色谱法同时测定人血浆三种视黄酸及临床应用

Simultaneous Determination of 3 1somers of Retinoic Acid in Human Plasma by HPLC and Its Clinical Application in the investigation of chronic bronchitic

目的建立同时测定血浆中全反式(all-trans)、13-顺式(13-cis)、9-顺式(9-cis)-视黄酸(RA)的方法。方法采用反相高效液相色谱(HPLC)法同时测定3种视黄酸同分异构体。用该法测定29名慢性支气管炎患者和30名健康人群血浆中3种视黄酸同分异构体的含量。结果3种视黄酸同分异构体的线性范围均为3.9～500ng/ml;最低检测限均为2ng/ml;批内和批间变异系数均小于13.9%。临床应用研究发现:经近似正态秩和检验(T1=608.5,T2=1163.5,ν=57),慢性支气管炎患者与正常对照人群血浆中all-trans-RA的含量差异有统计学意义(P<0.01)。结论所建方法可用于3种视黄酸同分异构体的血浆浓度监测和研究;临床研究结果也显示,慢性支气管炎患者支气管上皮细胞的炎症反应可能与血浆视黄酸同分异构体中all-trans-RA的含量减少有着密切的关系。

Objective To establish a sensitive and specific method for simultaneous determination of 13-cis-,all-trans-,9-cis-retinoic acid(RA)in human plasma.Method The three RA were simultaneously determined by reversed phase high-performance liquid chromatography.The method was applied to plasma specimens collected from two groups:chronic bronchitis group(n=29);normal control group(n=30).Result The linearity of the method was in the range of 3.9 ～500 ng/ml.The limit of quantification was 2 ng/ml for each of the compounds.The within-day and between-day CV were less than 13.9%.Statistical analysis revealed that there was significant difference as compared to the all-trans-RA of two groups(P< 0.01).Conclusion The method can be used to monitor the level of drug in clinical and research.The clinical study revealed:All-trans-RA might play an important role in the airway inflammation of chronic bronchitis and providing experimental evidence for clinical therapy.