腌腊肉制品中13-HODE、9,10-DHODE、9,10-EPODE、9,10,13-THODE的HPLC-MS/MS检测

采用高效液相色谱-串联质谱法对腌腊肉制品中13-羟基-9,11-十八碳二烯酸(13-hydroxy-9,11-octadecadienoic acid,13-HODE)、9,10-二羟基-12-十八碳烯酸(9,10-dihydroxy-12-octadecenoic acid,9,10-DHODE)、9,10-环氧-12-十八碳烯酸(9,10-epoxyoctadec-12-enoic acid,9,10-EPODE)及9,10,13-三羟基-11-十八碳烯酸(9,10,13-trihydroxy-11-octadecenoic acid,9,10,13-THODE)进行检测。腌腊肉制品中13-HODE、9,10-DHODE、9,10-EPODE、9,10,13-THODE用甲醇提取,经固相萃取柱去净化浓缩,然后以0.1%甲酸与乙腈为流动相,在XBridge色谱柱上进行梯度分离,采用电喷雾源负离子多反应监测模式进行分析。结果表明,13-HODE、9,10-DHODE、9,10-EPODE、9,10,13-THODE分别在质量浓度为0.05~2.0、0.01~0.5、0.05~1.0、0.01~0.5μg/mL的范围内具有较好的线性关系(R20.999);检出限分别为0.120、0.008、0.200、0.016μg/g;不同添加水平的平均回收率分别为95.1%、85.2%、86.8%、86.2%。对26种腌腊肉制品含量分析表明,所有样本都含有13-HODE、9,10-DHODE、9,10,13-THODE,含量分别在1.4~100.7、0.1~3.9、0.4~10.2μg/g范围内,21种样本中含有9,10-EPODE,含量范围为0.8~6.9μg/g。结果表明,腌腊肉制品中存在目标分析物的异构体,羟基脂肪酸实际含量可能大大高于检测的量。

Simultaneous Determination of 13-HODE, 9,10-DHODE, 9,10-EPODE and 9,10,13-THOD in Cured Meat Products by HPLC-MS/MS

13-Hydroxy-9,11-octadecadienoic acid (13-HODE), 9,10-dihydroxy-12-octadecenoic acid (9,10-DHODE),
9,10-epoxyoctadec-12-enoic acid (9,10-EPODE) and 9,10,13-trihydroxy-11-octadecenoic acid (9,10,13-THODE) in cured
meat products were detected with high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS).
The analytes, extracted with methanol and cleaned by solid phase extraction, were separated on an XBridge C18 column with
a mobile phase consisting of 0.1% formic acid in water and acetonitrile, followed by ionization and quantification with an
electrospray ionization source in negative ion mode and multiple reaction monitoring mode (MRM). The results indicated
that the calibration curves for 13-HODE, 9,10-DHODE, 9,10-EPODE and 9,10,13-THODE exhibited good linearity
(R2 > 0.999) in the ranges of 0.05–2.0, 0.01–0.5, 0.05–1.0 and 0.01–0.5 μg/mL, respectively. The limits of detection (LODs)
for the four analytes were 0.120, 0.008, 0.200, and 0.016 μg/g, and the recoveries were 95.1%, 85.2%, 86.8% and 86.2%,
respectively. The method was successfully employed to detect the analytes in 26 cured meat products and the results
showed that all the samples contained 13-HODE, 9,10-DHODE and 9,10,13-THODE ranging from 1.4–100.7, 0.1–3.9 and
0.4–10.2 μg/g, respectively, and out of these samples, 21 contained 9,10-EPODE ranging from 0.8–6.9 μg/g. Since some
isomers of the analytes were detected in samples as well, the real content of fatty acids with hydroxyl group (s) from
oxidation of linoleic acid (LA) might be much higher.