中空纤维离心超滤-高效凝胶排阻色谱法测定青霉素V钾中蛋白杂质  

Hollow Fiber Centrifugal Ultrafiltration-High Performance Gel Exclusion Chromatography for the Determination of Protein Impurities in Phenoxymethylpenicillin Potassium

作　　者：杨中华[1] 张立娜[1] 张锦[1] 高敬林 庞海英 王薇 王聪聪 蒋晔[1] YANG Zhong-hua;ZHANG Li-na;ZHANG Jin;GAO Jing-lin;PANG Hai-ying;WANG Wei;WANG Cong-cong;JIANG Ye(Department of Pharmacy, Hebei Medical University, Shijiazhuang 050017)

机构地区：[1]河北医科大学药学院,河北石家庄050017

出　　处：《分析科学学报》2018年第2期209-213,共5页Journal of Analytical Science

摘　　要：建立了一种采用中空纤维离心超滤(HFCF-UF)-高效凝胶排阻色谱法(HPSEC)对青霉素V钾中的残留蛋白进行检测的方法。样品用HFCF-UF进行富集,采用HPSEC进行分析,色谱柱为TSK gel Super SW2000凝胶色谱柱(300×4.6mm,4μm),流动相为0.06mol/L NaCl-0.02mol/L NaH_2PO_4(pH=7.0),流速为0.3mL/min,紫外检测波长为220nm,柱温为室温。对照品牛血清白蛋白(BSA)在1.0~100.0μg/mL内线性关系良好(R2=0.999),回收率均大于87%,检测限为3.03ng/mL,经HFCF-UF富集处理后,富集倍数(EF)高达99倍。结果表明,该方法操作简便、结果准确、灵敏度高,为同类药物中的大分子蛋白检测提供了有效手段。A hollow fiber centrifugal ultrafiltration（HFCF-UF）-high performance gel exclusion chromatography （HPSEC） method was established for the determination of the residual protein in phenoxymethylpenicillin potassium. A TSK gel Super SW2000 column（300 X 4. 6 mm, 4 μm） was used. The mobile phase consisted of phosphate buffer solution（0.06 mol/L NaC1-0.02 mol/L NaH2PO4 ,pH=7.0） at a flow rate of 0.3 mL/min,and the detection wavelength was at 220 nm. The column temperature was at ambient temperature. The linear range of bovine serum albumin（BSA） was 1.0-100.0 μg/mL（R2 =0. 999）,with the average recovery more than 87%. The limit of detection was 3. 03 ng/mL, the enrichment factor （EF） could reach 99 after concentration by HFCF-UF. The proposed method was proven to be simple, accurate and sensitive, providing a new approach for the analysis of residual protein in other similar drugs.

关 键 词：青霉素V钾 蛋白杂质 高效凝胶排阻色谱法 中空纤维离心超滤法 

分 类 号：O657.7[理学—分析化学]