超高效合相色谱法分离和测定氟苯尼考对映体  

Separation and Determination of Florfenicol Enantiomers Using Ultra-performance Convergence Chromatography

作　　者：张文华 王英 侯建波 陶红霞 胡起立 王良莉 谢文 徐敦明 ZHANG Wenhua;WANG Ying;HOU Jianbo;TAO Hongxia;HU Qili;WANG Liangli;XIE Wen;XU Dunming(The Technical Center of Hangzhou Customs,Hangzhou 310016,China;Zhejiang Academy of Science and Technology for Inspection and Quarantine,Hangzhou 310016,China;The Technical Center of Xiamen Customs,Xiamen 361026,China)

机构地区：[1]杭州海关技术中心,浙江杭州310016 [2]浙江省检验检疫科学技术研究院,浙江杭州310016 [3]厦门海关技术中心,福建厦门361026

出　　处：《现代食品科技》2022年第8期280-287,137,共9页Modern Food Science and Technology

基　　金：国家重点研发计划(2019YFC1605400);海关总署科研项目(2020HK196);浙江省科技计划项目(2018C37014)。

摘　　要：建立一种超高效合相色谱法(UPC^(2))快速分离和测定氟苯尼考对映体的方法。采用Acquity Trefoil CEL2(150 mm×3.0 mm,2.5μm)手性色谱柱,CO_(2)和甲醇为流动相,梯度洗脱,流速1.0 mL/min,检测波长为224 nm,进样体积5μL,系统背压17.2 MPa,柱温40℃。实验结果表明:两种氟苯尼考对映体在4.00-400.00 mg/L线性范围内与峰面积呈良好线性关系,相关系数均(r^(2))在0.9993以上,仪器检出限(LOD,S/N=3)为2.0 mg/L。连续重复进样10.0 mg/L混合标准工作溶液6次,(-)、(+)-氟苯尼考的峰面积相对标准偏差值(RSD,n=6)分别为0.65%、0.81%。应用该方法对市售氟苯尼考标准品进行了分离及测定,结果显示,3份标准品均未检出(+)-氟苯尼考,均检出(-)-氟苯尼考,含量为9.20-10.00 mg/L,在6.0 min内完成了两个氟苯尼考对映体的分离及含量测定,该方法检测速度快、分离度高、绿色环保,可以满足氟苯尼考对映体的分离检测要求。A new method was established for the separation and determination of two florfenicol enantiomers using ultra-performance convergence chromatography(UPC^(2)). The chromatographic separation conditions were optimized, and the stability of standard solutions of(-)-florfenicol and(+)-florfenicol enantiomers were observed. The separation process was carried out in Waters Acquity Trefoil CEL2(150×3.0mm, 2.5 μm) chiral columns. The mobile phase used carbon dioxide and methanol at a flow rate of 1.0 mL/min with gradient elution. The UV detection wavelength was set to 224 nm, the injection volume was 5 μL, the system backpressure was 17.2 MPa, and the column temperature was maintained at 40 ℃. The method validation results indicate that the limit of detection(LOD, S/N=3) for both enantiomers was 2.00 mg/L,and the calibration linear range was between 4.00 and 400.00 mg/L, with correlation coefficients(r^(2)) > 0.9993. Repeated measurements of the peak area using 10 mg/L standard solutions of each florfenicol enantiomer showed relative standard deviations(RSD, n=6) of 0.65 and 0.81%,respectively. This method was then used to separate and measure commercially available standard florfenicol racemates. The results indicate that(+)-florfenicol was not detected in the three standard florfenicol racemates, but(-)-florfenicol(9.20-10.00 mg/L) was detected in all of them.The separation and content determination of the two florfenicol enantiomers were completed within 6.0 min. Therefore, this is a rapid,high-resolution, eco-friendly method that can be used for the separation and determination of florfenicol enantiomers.

关 键 词：超高效合相色谱 氟苯尼考 对映体 手性分离 

分 类 号：S859.84[农业科学—临床兽医学] O657.72[农业科学—兽医学]