固相萃取-高效液相色谱法测定蜂蜜中的有机酸

建立了固相萃取-高效液相色谱(SPE-HPLC)同时测定蜂蜜中5种有机酸(L-苹果酸、马来酸、琥珀酸、柠檬酸、D-苹果酸)含量的方法。蜂蜜经制样后过Bond Elutes SAX固相萃取(SPE)小柱净化,用C18-MS-II反相色谱柱(250 mm×4.6 mm, 5 μm)进行分离,流动相为2%偏磷酸溶液,流速为0.7 mL/min,检测波长为210 nm。在此条件下5种有机酸在相应的线性范围内其线性相关系数均大于0.9967;方法的回收率为86.0%～103.9%,相对标准偏差为5.7%～9.8%(n=6),检出限为0.06～9.4 mg/kg。所建立的方法可用于蜂蜜样品中有机酸的测定。

Determination of organic acids in honey by solid phase extraction-high performance liquid chromatography

A method for the simultaneous determination of 5 organic acids (L-malic acid, maleic acid, succinic acid, citric acid, D-malic acid) in honey was developed by solid-phase extraction and high performance liquid chromatography (SPE-HPLC). The sample was purified by a Bond Elutes SAX SPE cartridge after pretreatment, then separated using a reversed-phase C18-MS-IIcolumn. A 2% metaphosphoric acid solution was used as the mobile phase at a flow rate of 0.7 mL/min. The detection was performed by a diode array detector at 210 nm. Under these conditions, the linear ranges of the method for L-malic acid was 5.0~500 mg/L, for maleic acid was 0.1~500 mg/L, and for succinic acid, citric acid, D-malic acid were 18.0~500 mg/L. The correlation coefficients of 5 organic acids were more than 0.9967. The average recoveries of 5 organic acids were 86.0%~103.9% with the relative standard deviations of 5.7%~9.8% (n=6). The detection limits ranged from 0.06 to 9.4 mg/kg. The method can be used for determining organic acids in honey samples.