超高效液相色谱-三重四极杆质谱法同时测定饲料中8种类固醇激素

建立了同时测定饲料中勃地酮、甲睾酮、诺龙、群勃龙、甲羟孕酮乙酸酯、美仑孕酮、乙酸甲地孕酮和17α-羟基孕酮8种类固醇激素的超高效液相色谱-三重四极杆质谱(UPLC-MS/MS)检测方法。考察了提取溶剂,除蛋白和除脂条件对8种类固醇激素回收率的影响,最终选择乙腈为提取溶剂,用三氯乙酸和氢氧化钠除蛋白,用氯化镁和正己烷除脂,样品经Athena C18-WP(2.1×150 mm,5 μm)色谱柱分离,以5 mmol/L乙酸铵(1‰甲酸)和乙腈(1‰甲酸)进行梯度洗脱,正离子电喷雾电离多反应监测模式测定,内标法进行定量分析。结果显示,8种化合物在0~10 μg/L范围内线性相关系数(r)≥0.9995;检出限为0.10~0.34 μg/kg(S/N≥3),定量限为0.35~0.98 μg/kg(S/N≥10)。方法的平均回收率范围为70.4%~109%,相对标准偏差为0.38%~10.3%(n=6)。该方法操作简单,灵敏度高,重复性好,适用于饲料中8种类固醇激素的快速检测。

Simultaneous Determination of 8 Steroid Hormones in Feed by Ultra Performance Liquid Chromatography-tandem Mass Spectrometry

A method was developed for the screening and quantification of 8 steroid hormones including( boldenone,17-alpha- Methyltestosterone,19 - Nortestosterone,trenbolone,medroxyprogesterone,melengestrol,megestrol - 17 - acetate,17 - alpha - Hydroxyprogesterone) in feed by ultra performance liquid chromatography- tandem mass spectrometry( UPLC-MS /MS).The effects of extraction solvent,protein removal and degreasing conditions on the recovery of 8 steroid hormones were investigated. Finally,acetonitrile was selected as the extraction solvent,protein was removed with trichloroacetic acid and sodium hydroxide, and fat was removed with magnesium chloride and n- hexane.In the chromatographic analysis,the 8 target compounds were separated by Athena C18-WP( 2.1 × 150 mm,5 μm) column and gradient elution with 5 mmol /L ammonium acetate( 1‰ formic acid) and acetonitrile( 1‰ formic acid),detected in positive multiple reaction monitoring( MRM) and quantified with internal standards.The results showed that the 8 steroid hormones exhibited good linear relationships in the range of 0~ 10 μg /L with correlation coefficients( r) of more than 0.9995;the limits of detection( LODs,S /N≥3) were 0.10~0.34 μg /kg and the limits of quantification( LOQs,S /N≥10) were 0.35~0.98 μg /kg.Average recoveries were 70.4%~109% with relative standard deviation of 0.38%~ 10.3%( n = 6).The method was simple,rapid,sensitive and reproducible,and was suitable for rapid screening and detection of 8 steroid hormones in feed.