黄鳝肌肉中的20种性激素同时测定的气质联用检测方法

Stimultaneous Determination of Residue of Sex Hormones in Eel Muscle by Gas Chromatography-Mass Spectrometry

建立了用安捷论GC-MS5975对黄鳝肌肉中20种性激素能同时进行筛选、定量和确证的灵敏、特异可靠的检测方法。对5.0g肌肉样品,采用β-葡糖苷酸酶(来自Helix pomatia)对提取物水解后,经叔丁基甲醚提取,异辛烷脱脂,经脱脂棉过滤后进入C18小柱,再经NH2小柱净化后七氟丁酸酐对其进行衍生,最后利用GC-MS进行检测。采用外标法定量,通过对空白样品添加标准品,使之理论含量为1、2.5和5μg/kg,追宝龙按相应水平的5倍添加,以S/N≥3,甲睾酮、去氢睾酮和甲孕酮,检出限为0.25μg/kg,除了追宝龙的检测限为5μg/kg外,其他的检出限均为0.5～1μg/kg。对待测物三个水平的添加回收实验表明,平均回收率范围为79.8%～106.3%,平均变异系数的范围为11.6%～17.9%。当添加水平为1μg/kg(追宝龙为结果5μg/kg),本方法不同批次回收率的相对偏差范围为9.3%～22.1%,重复性好。因此,本方法对目前动物源食品中的性激素监督检验有参考价值。

A highly sensitive, specific and credible method for the simultaneous determination of 20 sex hormones in muscle of swamp eel performed by GC-MS（Agilent 5975） was studied. Muscle tissues of swamp eel （5.0 g） were hydrolyzed by glucuronidase from Helixpomatia juice, extracted by tert-butyl methyl ether, defatted by iso-octane, and cleaned-up through dual columns of C18 and NH2. The extracts obtained were derivatized with heptafluorobutyric anhydride, following formation of the di-heptafluorobutyryl （di-HFB） derivative, and then identification and quantification were carded out by means of capillary gas chromatography-mass spectrometry. For quantitation, the external standards were used and the method was validated with blank samples fortified at 1, 2.5 and 5μ g/kg with 5.0 g sample（trenbolone spiked 5 times as others）. Results showed that the detection limits are 0.25 -5μg/kg for all analytes. Average recovery （%） and coefficient of variance range from79.8% to 106.3%, and from 11.6% to 17.9%, respectively. The relative standard deviation （RSD） ranges from 9.3% to 22.1% at 1 mg/kg spiked level. The study results showed that the recovery and repeatability are good. Therefore, the method is very useful for the animal food quality surveillance at present.