摘要
目的 建立测定狼疮性肾炎（LN）患儿血浆中环磷酰胺含量的高效液相色谱（HPLC）方法,为制订个体化治疗方案提供依据.方法 色谱柱为Hypersil ODS-2 C18柱,流动相为乙腈-水（体积比为30：70）,流速为1.0 ml/min,柱温为30℃,检测波长为195 nm,进样量为20μl.以异环磷酰胺为内标,血浆样品经甲醇沉淀蛋白、离心取上清液进样检测.结果 血浆中环磷酰胺的质量浓度在0.5～20.0 mg/L范围内,环磷酰胺与内标的峰面积比呈良好的线性（r=0.9994）,最低检出限为0.1 mg/L;日内、日间精密度和稳定性实验的相对标准偏差（RSD）均＜8％;加样回收率为100.60％～101.05％,结果满意.结论 该法简便、快速、准确、重现性好,可用于LN患儿血浆中环磷酰胺的含量测定及免疫抑制治疗的临床药学监护.
Objective To establish an high performance liquid chromatography（HPLC） method for the determination of cyclophosphamide in plasma of children with lupus nephritis（LN）,and to provide basis for the individualized drug therapy.Methods HPLC was performed on the column of Hypersil ODS-2 C18 with the mobile phase of acetonitrile-water （30：70,V/V） at a flow rate of 1.0 ml/min.The column temperature was 30 ℃,and the detection wavelength was 195 nm.The injection volume was 20 μl.The plasma samples were deproteinized by methol with ifosfamide as the internal standard,and the centrifugal supernatant was injected for analysis.Results The linear range of plasma cyclophosphamide of children with LN was 0.5-20.0 mg/L （r=0.999 4）.The detection limit of cyclophosphamide was 0.1 mg/L.Relative standard deviation （RSD） of stability,the intra-day and inter-day precisions were all less than 8%.The recovery rate was 100.60%-101.05%,and the result was satisfactory.Conclusions The method is simple,rapid,accurate,convenient and with good reproducibility.It can be used for the determination of plasma cyclophosphamide concentration of children with LN and clinical pharmaceutical care of immunosuppressive therapy.
出处
《国际生物医学工程杂志》
CAS
2016年第5期294-298,共5页
International Journal of Biomedical Engineering
基金
天津市卫生局科技基金（2014KZ032）
关键词
高效液相色谱法
血药浓度
环磷酰胺
狼疮性肾炎
High performance liquid chromatography
Plasma concentration
Cyclophosphamide
Lupus nephritis
