反相HPLC定量检测人体泪液溶菌酶水平的方法研究  

Quantitative determination of lysozyme levels in human tears by reversed phase HPLC

作　　者：史瑾 杨小琳 赵金礼 SHI Jin;YANG Xiaolin;ZHAO Jinli(ShaanXi HuiKang Bio-tech Co.,Ltd.Xi′an,Shaanxi 710054,China)

机构地区：[1]陕西慧康生物科技有限责任公司,陕西西安710054

出　　处：《检验医学与临床》2018年第A01期85-87,共3页Laboratory Medicine and Clinic

摘　　要：目的 建立一种反相高效液相色谱法(RP-HPLC)检测人体泪液中溶菌酶水平的方法。方法 采用WeLch Ultimate XB-C8色谱柱(4.6 mm×250.0 mm,填料孔径300,粒径5μm),以0.9%氯化钠溶液,0.1%三氟乙酸为流动相A,0.9%氯化钠溶液:乙腈(40∶60),0.1%三氟乙酸为流动相B,检测波长215 nm,流速1.0 mL/min,线性梯度洗脱。结果 本方法专属性良好;重复性RSD为0.65%,定量限0.50 ng、检测限0.05 ng;在进样量10~50μg范围内,线性关系良好(r=0.9996);准确度验证平均回收率(n=9)98.91%,RSD为0.76%。分析16例人体天然泪液的样本,其中溶菌酶的水平在1.80~3.12 mg/mL。结论 本法简单快速,具有进样量少,准确度高,重复性好等特点,可用于临床及科研上人体泪液中溶菌酶水平的检测。Objective To establish a reversed phase high performance liquid chromatography(RP-HPLC)method for the determination of lysozyme in human tears.Methods Using WeLch Ultimate XB-C8 column(4.6 mm×250.0 mm,packing aperture size 300,particle size 5μm).Preparing 0.9%sodium chloride solution and 0.1%trifluoroacetic acid as mobile phaseA,0.9%sodium chloride solution-acetonitrile(40∶60)and 0.1%trifluoroacetic acid as mobile phaseB.The detection wavelength was 215 nm,with linear gradient elution at a flow rate of 1.0 mL/min.Results The method is good in specificity.Repetitive RSD was 0.65%,quantitative limit was 0.50 ng and detection limit was 0.05 ng.In a sample range of 10-50μg,linearity is good(r=0.999 6).The average recovery rate with accuracy verification was 98.91%(n=9),RSD was 0.76%.The samples of 16 human natural tears were analyzed,and the lysozyme content was 1.80-3.12 mg/mL.Conclusion This method is simple and rapid,which also has advantages such as less sample introduction,high accuracy and good repeatability.It can be applied to clinical and scientific research on the detection of lysozyme content in human tears.

关 键 词：人体泪液 溶菌酶 反相高效液相色谱 定量检测 

分 类 号：R781.4[医药卫生—口腔医学]