固相萃取/液相色谱-串联质谱法测定麦卢卡蜂蜜特征化合物

Determination of Characteristic Compounds in Manuka Honey by Liquid Chromatography-Tandem Mass Spectrometry with Solid Phase Extraction

建立了固相萃取/液相色谱-串联质谱(SPE/LC-MS/MS)同时测定麦卢卡蜂蜜中4种特征化合物2'-甲氧基苯乙酮(2'-MAP)、2-甲氧基苯甲酸(2-MBA)、3-苯基乳酸(3-PA)和4-羟基苯基乳酸(4-HPA)的方法。蜂蜜样品用含1%甲酸水溶液溶解,经HLB固相萃取柱净化,Thermo Hypersil Gold C18(50 mm×2.1 mm,1.9μm)色谱柱分离,0.1%甲酸水溶液和0.1%甲酸乙腈为流动相进行梯度洗脱。4种化合物在电喷雾正/负离子多重反应监测模式(MRM)下同时进行检测,外标法定量。结果表明,2'-MAP、2-MBA和4-HPA在0.01~1.0 mg/L、3-PA在0.1~10.0 mg/L范围内线性关系良好,相关系数(r2)均大于0.99。方法检出限(LODs)和定量下限(LOQs)分别为0.1~1.0 mg/kg和0.5~5.0 mg/kg。蜂蜜样品的平均加标回收率为79.4%~98.3%,相对标准偏差(n=6)为3.2%~8.7%。利用该方法对采集于新西兰不同产地的12个品种的114个巢蜜样本和4个国家的50个商品化蜂蜜进行了验证。该法灵敏度和准确度高,分析速度快,可为判别单花种、多花种及非麦卢卡蜂蜜提供参考。

A solid phase extraction/liquid chromatography-tandem mass spectrometric （SPE/LC-MS/MS ） method was established for the simultaneous determination of 2′-methoxyacetophenone （2′-MAP ）,2-methoxybenzoic acid （2-MBA ）,3-phenyllactic acid （3-PA ） and 4-hydroxyphenyllactate （4-HPA ）.The honey sample was dissolved in 1 % formic acid solution,then purified with HLB SPE.The separation of the analytes was performed on a Thermo Hypersil Gold C 18 （50 mm×2.1 mm,1.9 μm ） column with 0.1 % formic acid solution and acetonitrile containing 0.1 % formic acid as mobile phases by gradient elution.The four compounds were detected simultaneously in electrospray negative and positive ionization modes under multiple reaction monitoring （MRM ） mode.The results showed that the calibration curves were linear in the range of 0.01-1.0 mg /L for 2′-MAP,2-MBA and 4-HPA,and 0.1-10.0 mg/L for 3-PA with correlation coefficients （ r 2 ） higher than 0.99.The limits of detection （LODs ） and quantitation （LOQs ） were in the ranges of 0.1-1.0 mg/kg and 0.5-5.0 mg/kg,respectively.The average spiked recoveries for honey samples ranged from 79.4 % to 98.3 % with relative standard deviations （ n =6 ） of 3.2 % -8.7 % .This method was used to validate 114 hive honey samples from 12 varieties collected in different locations in New Zealand and 50 commercial honey samples from four different countries.With the charateristics of sensitivity,accuracy and rapidness,the method could provid a reference for the identification of monoflower,multiflower and non-manuka honey.