高效液相色谱-串联质谱法检测蜂花粉中 赭曲霉毒素A残留量

Determination of ochratoxin A residue in bee pollen by high performance liquid chromatography-tandem mass spectrometry

中文关键词:  赭曲霉毒素A  蜂花粉  固相萃取  高效液相色谱-串联质谱法

英文关键词:ochratoxin A  bee pollen  solid phase extraction  high performance liquid chromatography- tandem mass spectrometry

中文摘要:

      目的 建立蜂花粉中赭曲霉毒素A(ochratoxin A, OTA)的高效液相色谱-串联质谱(HPLC-MS/MS)检测方法。方法 样品经乙腈提取, 固相萃取柱富集净化后, 采用HPLC-MS/MS对目标物进行定性确证和定量分析。在Leapsil C18 (100 mm×3.0 mm, 2.7 μm)上以0.005 mol/L乙酸铵甲酸水溶液和乙腈为流动相进行梯度洗脱分离; 质谱采集模式为电喷雾负离子监测模式。结果 本方法的赭曲霉毒素A的定量限(以S/N=10计)为5 μg/kg; 在5～100 μg/L的质量浓度范围内呈现良好的线性关系, 相关系数(r)大于0.99。本底空白的玉米花粉、茶花粉、荷花粉、油菜花粉4种基质中5、10、20 μg/kg添加水平下的加标回收率范围为84.6%～103.6%, 精密度范围为5.1%～12.2％。结论 本方法准确可靠、灵敏度高、经济实用, 可替代较为昂贵的免疫亲和柱, 较大降低检测成本, 且适用于大部分蜂花粉基质的测定。

英文摘要:

      Objective To establish a method for the determination of ochratoxin A in pollen based on high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). Methods The sample was extracted by acetonitrile and cleaned up by solid phase extraction cartridge. The separation was carried out on a Leapsil C18 (100 mm×3.0 mm, 2.7 μm) with a gradient elution using 0.005 mol/L ammonium acetate formic acid solution and acetonitrile as mobile phases. The analysis of ochratoxin A was performed under electrospray negative ionization mode. Results The limits of quantification (LOQ, S/N=10) was 5 μg/kg. A good linearity (r >0.99) was achieved for the target compounds over the range of 5~100 μg/L. The recoveries at the three spiked levels (5, 10, 20 μg/kg) in the blank matrices such as corn pollen, camellia pollen, lotus pollen, and rape pollen, were varied from 84.6% to103.6% with the relative standard deviations varied from 5.1% to 12.2%. Conclusion The method is accurate, efficient, sensitive and practical. The cost of detection is obviously reduced by replacing immunoaffinity column with HLB column which has the same purification effect. It can be applied to most of pollen which may be contaminated by Ochratoxin A.