高效液相色谱法在贻贝黏蛋白检测中的应用  

The Determination of the Mussel Adhesive Protein by HPLC

作　　者：柯林楠[1] 宋茂谦 高敏 陆云龙 母瑞红[1] KE Lin-nan;SONG Mao-qian;GAO Min;LU Yun-long;MU Rui-hong(National Institutes for Food and Drug Control,Beijing 102629;Jiangyin USUN Biochemical Technology Co.Ltd.,Jiangsu Wuxi 214437)

机构地区：[1]中国食品药品检定研究院,北京100050 [2]江阴贝瑞森生化技术有限公司,江苏无锡214437

出　　处：《中国医疗器械信息》2022年第11期21-23,33,共4页China Medical Device Information

基　　金：国家重点研发计划(项目编号:2016YFC1103200)。

摘　　要：目的:建立高效液相色谱法(High Performance Liquid Chromatography,HPLC)鉴别检测贻贝黏蛋白(Mussel Adhesive Protein,MAP)的方法。方法:采用Agilent Zobax SB 300A C8(4.6×250mm,5μm)色谱柱,流动相为0.1%三氟乙酸水溶液(A)和含0.1%三氟乙酸乙腈溶液(B)梯度洗脱,检测波长280nm,洗脱温度25℃,流量为0.9mL/min。进样量20μL。考察方法的系统适用性、线性、精密度、检测限及定量限。结果:贻贝黏蛋白供试样品与对照品的HPLC图谱一致。贻贝黏蛋白在0.01~9.79mg/mL范围内峰面积与其浓度呈良好的线性关系(r=1),峰面积及保留时间RSD均小于5%,最低定量限(LOQ)为0.03mg/mL,最低检出限(LOD)为0.01mg/mL。结论:该方法专属性强,稳定性好,简单易用,适用贻贝黏蛋白的检测。Objective:A High Performance Liquid Chromatography(HPLC)method was established for the determination of the Mussel Adhesive Protein(MAP).Methods:Agilent Zorbax SB 300A C8 column(4.6×250mm,5μm)was used.The mobile phase was gradient elution with 0.1%trifluoroacetic acid aqueous solution(A)and acetonitrile solution containing 0.1%trifluoroacetic acid(B).The column temperature was 25℃ and the detection wavelength was performed at 280 nm.Its flow rate was 0.9 mL/min.The injection volume was 20μL.The system applicability,linearity,precision,detection limit and quantitation limit were investigated.Results:There was no difference in the HPLC spectrum of MAP between the sample and the control.The peak area of MAP in the range of 0.01mg/mL to 9.79mg/mL showed a good linear relationship with its concentration(r=1).The RSD of peak area and retention time were all less than 5%.The limits of quantification(LOQ)were 0.03mg/mL and the limits of detection(LOD)were 0.01mg/mL.Conclusion:This HPLC method is simple,specific and stable.It can be used for the quality control of MAP.

关 键 词：高效液相色谱法(HPLC) 贻贝黏蛋白(MAP) 检测 

分 类 号：TH776[机械工程—仪器科学与技术]