9-芴甲氧羰酰氯柱前衍生化RP-HPLC法测定蒜氨酸含量及有关物质  

RP-HPLC determination of alliin and its related substances with pre-column derivatization of 9-fluorenylmethyl chloroformate

作　　者：徐晓燕[1] 宋敏[1] 杭太俊[1] 张正行[1] 陈坚[2] 

机构地区：[1]中国药科大学药物分析教研室,南京210009 [2]新疆医科大学药学院,乌鲁木齐830054

出　　处：《药物分析杂志》2010年第9期1604-1609,共6页Chinese Journal of Pharmaceutical Analysis

摘　　要：目的:采用9-芴甲氧羰酰氯(FMOC)为柱前衍生化试剂,建立了FMOC柱前衍生化反相高效液相色谱法用于测定蒜氨酸及其有关物质的含量。方法:采用KromasilC18柱(250mm×4.6mm,5μm),流动相为乙腈-0.1mol·L-1的醋酸铵缓冲溶液(冰醋酸调pH为4.2),线性梯度洗脱,流速1.0mL.min-1,紫外波长265nm检测,并采用LC-MS/MS法鉴定主要有关物质。结果:蒜氨酸及其有关物质衍生物色谱分离度良好,保留适宜;蒜氨酸浓度在1.0～200μg·mL-1范围内,线性关系良好(r=0.9995),检测限0.1μg·mL-1,进样精密度良好(RSD=0.28%,n=6);根据LC-MS/MS初步推断了蒜氨酸中杂质的结构。结论:方法专属性高、准确度和耐用性好,适用于蒜氨酸的含量测定及有关物质检测。Objective：To establish a reversed-phase high performance liquid chromatographic method with 9-fluorenylmethyl chloroformate pre-column derivatization for the determination of alliin and its related substances. Methods：The analysis was carried out on a Kromasil C18 column （250 mm×4.6 mm,5 μm） by using acetonitrile-0.1 mol·L-1 ammonium acetate （adjust to pH 4.2 with glacial acetic acid） as the mobile phase with linear gradient elution mode at a flow rate of 1.0 mL·min-1,and detection at 265 nm. LC-MS/MS method was employed for the identification of the major related substances. Results：Good retention and separations were achieved for derivatives of alliin and its related substances. The linear calibration was found for alliin within the range of 1.0-200 μg·mL-1（r=0.9995）.The limit of detection was 0.1 μg·mL-1. The precision was good with RSD of 0.28%（n=6）. The structures of major impurities in alliin were identified by LC-MS/MS. Conclusion：The established method is specific and accurate with very good ruggedness and suitable for the determination of alliin and its related substances.

关 键 词：蒜氨酸 含量测定 有关物质 9-芴甲氧羰酰氯(FMOC) 柱前衍生化 高效液相色谱法 

分 类 号：R917[医药卫生—药物分析学]