【摘要】：苯巴比妥鈉(Phenobarbital Sodium)是農(nóng)業(yè)部176號(hào)公告規(guī)定的禁止在飼料和動(dòng)物飲用水中使用的藥物品種。目前,尚沒(méi)有針對(duì)飼料中苯巴比妥鈉的檢測(cè)方法和標(biāo)準(zhǔn),這就為飼料質(zhì)量監(jiān)管和人民飲食健康埋下了隱患,制定飼料中苯巴比妥鈉的檢測(cè)方法刻不容緩。苯巴比妥鈉是弱酸強(qiáng)堿鹽,在酸性環(huán)境中會(huì)發(fā)生水解以苯巴比妥的形式存在。飼料樣品經(jīng)過(guò)正己烷預(yù)處理后,加鹽酸甲醇溶液提取,其中的苯巴比妥鈉被以苯巴比妥(Phenobarbital)的形式提取出來(lái)。提取液經(jīng)Waters Oasis HLB固相萃取小柱(SPE)凈化,經(jīng)甲醇+水(2+98)溶液淋洗后,用5%氨化甲醇洗脫。洗脫液經(jīng)氮吹濃縮后,用甲醇+水(2+8)復(fù)溶,上高效液相色譜儀(HPLC)檢測(cè)。所用色譜柱是粒徑5μm,250 mm×4.6 mm的C8柱,柱溫箱控制溫度為30℃,流動(dòng)相流速為1 mL/mi,梯度洗脫,紫外檢測(cè)器于215 nm波長(zhǎng)檢測(cè)。經(jīng)過(guò)公式計(jì)算和轉(zhuǎn)化,得到苯巴比妥鈉的含量。分析表明,該液相方法的檢測(cè)限(LOD)為0.5mg/kg,定量限(LOQ)為1mg/kg,該方法具有良好的準(zhǔn)確度和精密度,且對(duì)配合飼料、預(yù)混合飼料、濃縮飼料以及精料補(bǔ)充料均有較好的適用性,回收率均在在80%-120%之間,相對(duì)偏差(RSD)均在10%以?xún)?nèi)。標(biāo)準(zhǔn)曲線取點(diǎn)0.5μg/mL、1.0μg/mL、2.0μg/mL、5.0μg/mL、10.0μg/mL、20.0μg/mL、50.0μg/mL,曲線相關(guān)系數(shù)達(dá)到0.999,具有良好的線性回歸。在確定的檢測(cè)條件下,地西泮、司可巴比妥等鎮(zhèn)定類(lèi)藥物無(wú)干擾。液相方法取得了良好的實(shí)驗(yàn)效果。苯巴比妥鈉的液相色譜-串聯(lián)質(zhì)譜法(LC-MS-MS)是在液相方法的基礎(chǔ)上深入研究的。飼料樣品中苯巴比妥鈉的提取、凈化、濃縮在已有液相方法的基礎(chǔ)上進(jìn)一步調(diào)整和優(yōu)化。液相色譜分離柱選擇100mm×2.1 mm的C18柱,在流速為0.2 mL/min,柱溫為40℃時(shí)苯巴比妥被分離出來(lái),然后經(jīng)串聯(lián)質(zhì)譜儀檢測(cè)信號(hào)。在質(zhì)譜儀上,苯巴比妥的定性定量監(jiān)測(cè)離子對(duì)為231.084.9,231.0188.1,毛細(xì)管出口端電壓為90V,碰撞能分別為5eV和3eV。苯巴比妥標(biāo)準(zhǔn)系列為0.05μg/mL,0.1μg/mL,0.2μg/mL,0.5μg/mL,1.0μg/mL,2.0μg/mL,相關(guān)系數(shù)達(dá)到0.997,具有良好的線性相關(guān)。經(jīng)過(guò)公式計(jì)算和轉(zhuǎn)化,得到苯巴比妥鈉的含量。分析表明,方法的檢測(cè)限為0.1mg/kg,定量限為0.2mg/kg,測(cè)試方法具有良好的準(zhǔn)確度和精密度,且對(duì)配合飼料、預(yù)混合飼料、濃縮飼料以及精料補(bǔ)充料均有較好的適用性,回收率均在60%-120%之間,相對(duì)偏差均在20%以?xún)?nèi)。本方法建立的飼料中苯巴比妥鈉液相色譜-串聯(lián)質(zhì)譜測(cè)定方法,為飼料中苯巴比妥鈉的定性定量檢測(cè)提供了方法依據(jù)。
[Abstract]:Phenobarbital sodium (Phenobarbital Sodium) is a drug banned from feed and animal drinking water as stipulated by the Ministry of Agriculture's Proclamation No. 176. At present, there are no methods and standards for the determination of phenobarbital sodium in feed, which is a hidden danger for the quality supervision of feed and the healthy diet of people. It is urgent to establish a method for the determination of phenobarbital sodium in feed. Phenobarbital sodium is a weak acid strong base salt, and hydrolysis occurs in acid environment in the form of phenobarbital. The feed sample was pretreated with n-hexane and extracted with methanol hydrochloride solution. The sodium phenobarbital was extracted in the form of phenobarbital (Phenobarbital). The extract was purified by Waters Oasis HLB solid-phase extraction column (SPE) and eluted with 5% ammoniated methanol after elution with methanol water (298) solution. The eluent was condensed by nitrogen blowing, then redissolved with methanol and water (28), and detected by (HPLC) with high performance liquid chromatography (HPLC). The chromatographic column was C8 column with a diameter of 5 渭 m of 250 mm 脳 4.6 mm. The column temperature was controlled at 30 鈩，

本文編號(hào)：2253892