高效液相色谱法检测肾病综合征患者泼尼松血药浓度及其临床意义

摘要

目的:建立一种高效液相色谱(HPLC)法测定泼尼松(PS)血药浓度的方法,并探讨肾病综合征PS血药浓度与蛋白尿消减关系.方法:以乙酰苯胺为内标,采用HP1100系列高效液相色谱仪,分析柱HPLichrosphere Ca柱(250 mm×4 mm,5 μm);流动相为:甲醇:四氢呋喃:水=25:25:50(V/V),流速为1.0 ml/min;检测波长为240 nm;室温25 ℃.血清标本以甲醇沉淀蛋白.结果:泼尼松浓度与峰面积比值之间的线性关系良好,回归方程为Y=6.84×10-4×-39.4×10-4,相关系数为0.999 93,回收率为98.70%,批间CV3.76%,批内CV2.25%.线性范围:10～500 μg/ml;检测限:5 μg/ml.NS组与对照组比较血液平均SP浓度,差异有统计学意义(P<0.05);男女组别比较,差异无统计学意义(P>0.05);激素抵抗组与激素敏感组血药浓度比较,抵抗组患者SP血药低于敏感组血药浓度,浓度差异有统计学意义(P<0.05).结论:高效液相色谱法检测SP血药浓度,色谱峰分离良好,无干扰,是一种可靠、便捷的检测血清PS浓度的方法,对监测长期应用泼尼松治疗类疾病和肾病综合征的血药浓度监测(TDM),了解用药个体化差异,规范用药方法,为临床治疗提供科学、合理用药理论依据和重要参考,对药效学研究具有重要临床意义.%Objective: To develop a high - performance liquid chromatography method to determine the concentration of prednisone( PS )in human serum,and to assess the relationship hetween serum concentration of PS and changes in urinary protein.Methods : The HP1100 high performance liquid chromato - graphic( HPLC )system was used.The analytic column was a HP Lichrosphere C8 column( 250 mm ×4 mm,5 μm ).The mobile phase was methanol,tetrahydrofuran and water with the ratio 25: 25: 50 The flow rate was 1.0 ml/min The detective UV wavement was 240 nm The serum samples, added acetanilide as internal standard, deposited down the protein with methanol.Results : The chromatography was good and not interfered by the components of the serum.Good Linearity was observed in 10 ～ 500 μg/L range for prednisone, the detection limit was 5 mg/ml.The serum concentration of NS group was different from that in the control group( P ＜ 0.05 ), there was no significant difference between the females and males groups( P ＞0.05 ) , the serum concentration of hormone resistant group was lower significantly compared with hormone sensitive group( P ＜ 0.05 ).Conclusion: This is an accurate and convenient method to determine the concentration of PS in serum , it can monitor the plasma concentrations of PS in patients with long - term PS treatment and nephrotic syndrome, and help to know Individualized differences, normalize the preparation method,lt holds significant implications in the Pharmacodynamic Study.