摘要
通过以细菌纤维素(Bacterial cellulose,BC)为流动相添加剂,提高毛细管电泳的分离能力,构建基于蜂蜜中氨基酸的指纹图谱。系统考察了电泳缓冲液种类、离子强度、pH值、分离电压及BC添加量对氨基酸分离效果的影响。结果表明:以30 mmol/L,p H 9.8硼砂-磷酸盐缓冲液为电泳缓冲液,BC添加量为0.5%,25 kV作为分离电压时,17种常见氨基酸的异硫氰酸荧光素的衍生物能在25 min内实现基线分离。最优电泳条件下,17种氨基酸的异硫氰酸荧光素的衍生物在相应浓度范围内线性关系良好,相关系数大于0.993,检测限为2.5×10~(-4)μmol/L~30.0×10~(-4)μmol/L。精密度试验表明:17种氨基酸衍生物峰面积相对标准偏差(Relative standard deviations,RSDs)(n=11)分别为1.84%~5.50%(日间,n=5)和0.94%~3.10%(日内,n=11)。洋槐蜜中17种氨基酸添加回收率在81.10%~96.15%,RSDs(n=3)小于8.22%。
Fingerprint of amino acids in honey was constructed by capillary electrophoresis. Herein,bacterial cellulose(BC)was used as additive in running buffer to improve the separation efficiency. Experimental parameters such as the p H and concentration of the running buffer,separation voltage as well as content of BC in running buffer were systematically investigated. The results showed that 17 amino acids derivatized with fluorescein isothiocyanate(FITC)could be baseline separated within 25 min using Na_2B_4O_7-NaH_2PO_4 buffer(30 mmol/L,pH 9.8)containing 0.5 % BC for background electrolyte,25 kV for the separation voltage. Under the optimized conditions,17 amino acid derivatives showed good linearities with correlation coefficients greater than 0.993.The limit detections of this developed method for 17 amino acid derivatives were ranging from 2.5×10~(-4) to 30.0×10~(-4)μmol/L. The relative standard deviations(RSDs)for peak areas were 1.84 %-5.50 %(inter-day)and 0.94 %-3.10 %(intra-day),respectively. The developed method was further applied to determine the amino acids in Acacia honey with the satisfactory recovery in the range of 81.10 %-96.15 % with RSDs(n=3)below 8.22 %.
引文
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