柱前衍生高效液相色谱法测定复方氨基酸注射液中盐酸半胱氨酸含量  

Content Determination of the Cysteine Hydrochloride in Compound Amino Acid Injection by Pre-Column Derivatization HPLC

作　　者：陈娟 陈志永 孟雪[1] 崔小敏[1] 任慧[1] 乔园 陈瑞明[1] CHEN Juan;CHEN Zhiyong;MENG Xue;CUI Xiaomin;REN Hui;QIAO Yuan;CHEN Ruiming(Shaanxi Academy of Traditional Chinese Medicine,Xi′an,Shaanxi,China 710061)

机构地区：[1]陕西省中医药研究院,陕西西安710061

出　　处：《中国药业》2020年第17期74-76,共3页China Pharmaceuticals

基　　金：陕西省重点研发计划项目[2019SF-298];陕西省中医药研究院项目[2018-03]。

摘　　要：目的建立测定复方氨基酸注射液中盐酸半胱氨酸含量的柱前衍生高效液相色谱(HPLC)法。方法衍生化试剂选取2,4-二硝基氟苯(DNFB),色谱柱采用Kromasil C18柱(250 mm×4.6 mm,5μm),流动相为乙腈-0.014 mol/L磷酸氢二钾溶液(p H=8.2,35∶65,V/V),检测波长为360 nm,流速为1.0 m L/min,柱温为30℃。结果盐酸半胱氨酸的衍生化反应条件为衍生化试剂-盐酸半胱氨酸(80∶1,V/V),60℃水浴暗处反应5 min;盐酸半胱氨酸进样量在0.1320~0.6600μg(r=0.9997)范围内与峰面积线性关系良好;平均回收率为98.85%,RSD为0.50%(n=6)。结论该方法反应温度低,反应时间短,且简便易操作,结果准确,可用于复方氨基酸注射液中盐酸半胱氨酸含量的测定。Objective To establish a high performance liquid chromatography(HPLC)method with pre-column derivation for the determination of cysteine hydrochloride in compound amino acid injection.Methods 2.4-Dinitrochlorobenzene was used for the pre-column derivatization agent.The analysis was performed on a Kromasil C18 column(250 mm×4.6 mm,5μm),with acetonitrile:0.014 mol/L dipotassium hydrogen phosphate solution(p H=8.2,35∶65,V/V)as the mobile phase.The detection wavelength was set at 360 nm,the flow rate was 1.0 m L/min,and the column temperature was kept at 30℃.Results The derivatization reaction conditions of cysteine hydrochloride was that the ratio between the derivatization dosage and cysteine content was 80∶1,V/V,and the reaction in a dark bath at 60℃for 5 min.Cysteine hydrochloride showed good linearity in the range of 0.1320-0.6600μg(r=0.9997).The average recovery was 98.85%with RSD of 0.50%(n=6).Conclusion The method has low reaction temperature,short reaction time,simple operation and accurate results,which can be used for the content determination of cysteine hydrochloride in Compound Amino Acid Injection.

关 键 词：2 4-二硝基氟苯 柱前衍生 高效液相色谱法 盐酸半胱氨酸 复方氨基酸注射液 

分 类 号：R927.2[医药卫生—药学] R977.4