Cai Qinren, Feng Jiawang, Zhang Yi, Peng Yufen, Xue Liangchen, Du Zhifeng
Technical Center of Zhuhai Entry-Exit Inspection and Quarantine Bureau, Zhuhai 519015, China.
Se Pu. 2011 Aug;29(8):712-7.
A method was developed for the simultaneous determination and identification of 12 steroid hormone residues in pig tissues, including stanolone, aldosterone, boldenone, danazol, metandienone, methyltestosterone, nadrolone, norethindrone, progesterone, stanozolol, testosterone and testosterone propionate, using liquid chromatography-tandem triple-quadrupole linear ion trap mass spectrometry(LC-MS/MS). Homogenized pig tissue samples were purified with a Waters MCX solid phase extraction column after enzymatic hydrolysis by beta-glucuronidase, then separated on a Venusil MP C18 column (100 mm x 2, 1 mm, 3 microm) using gradient elution with the mobile phases of acetonitrile and water with 0.1% (v/v) formic acid. A multiple reaction monitoring (MRM) as survey scan and an enhanced product ion (EPI) scan as dependent scan were performed in an information dependent acquisition (IDA) experiment. The compound identification was carried out by library search with a newly developed MS/MS library based on EPI spectra at three different collision energies in positive mode. The results showed that the limits of detection (LODs, S/N = 3) were in the range of 0.2 - 0.5 microg/kg for the steroid hormones, and with a good linearity (r > 0.99) ranged from 0.5 to 100.0 microg/L. The average recoveries (n = 6) of the 12 steroid hormones spiked in pig tissue samples at 5.0 microg/kg ranged from 72.0% to 98.1% with the relative standard deviations (RSDs) between 3.1% and 12.5%. The method was applied for the qualitative and quantitative determination of steroid hormone residues in pig tissues with sensitive and accurate characteristics.
建立了一种使用液相色谱 - 串联三重四极杆线性离子阱质谱法（LC-MS/MS）同时测定和鉴定猪组织中12种甾体激素残留的方法，这些激素包括司坦唑醇、醛固酮、苯丙酸诺龙、达那唑、美雄酮、甲基睾酮、诺乙雄龙、炔诺酮、孕酮、司坦唑醇、睾酮和丙酸睾酮。猪组织匀浆样品经β-葡萄糖醛酸酶酶解后，用Waters MCX固相萃取柱进行净化，然后在Venusil MP C18柱（100 mm×2.1 mm，3μm）上分离，流动相为乙腈和含0.1%（v/v）甲酸的水，采用梯度洗脱。在信息依赖采集（IDA）实验中，进行多反应监测（MRM）作为全扫描，增强产物离子（EPI）扫描作为依赖扫描。通过在正模式下基于三种不同碰撞能量的EPI光谱的新开发的MS/MS库进行库检索来进行化合物鉴定。结果表明，甾体激素的检测限（LOD，S/N = 3）在0.2 - 0.5μg/kg范围内，线性良好（r > 0.99），范围为0.5至100.0μg/L。在猪组织样品中添加5.0μg/kg的12种甾体激素的平均回收率（n = 6）在72.0%至98.1%之间，相对标准偏差（RSD）在3.1%至12.5%之间。该方法用于猪组织中甾体激素残留的定性和定量测定，具有灵敏、准确的特点。
