基質(zhì)固相分散_高效液相色譜法測定水稻中痕量異丙威

1、 基金項(xiàng)目河北省教育廳資助項(xiàng)目作者簡介楊容(1962-,女,教授,從事農(nóng)藥殘留分析。【研究報(bào)告】基質(zhì)固相分散-高效液相色譜法測定水稻中痕量異丙威楊容,馬晶軍,宋雙居(河北農(nóng)業(yè)大學(xué)理學(xué)院化學(xué)系,河北保定071001摘要研究出一種應(yīng)用基質(zhì)固相分散(MSPD -高效液相色譜法測定水稻中痕量異丙威殘留物的新方法。并討論了應(yīng)用MSPD 技術(shù)進(jìn)行前處理與傳統(tǒng)殘留分析的區(qū)別。本實(shí)驗(yàn)將MSPD 技術(shù)用于水稻中異丙威殘留物的高效液色譜分析,并對定量檢測條件作了詳細(xì)的研究。關(guān)鍵詞高效液相色譜;基質(zhì)固相分散;異丙威;水稻中圖分類號(hào)O65717+2文獻(xiàn)標(biāo)識(shí)碼A 文章編號(hào)1004-8685(200501-0048-02

2、Matrix solid -phase dispersion (MSPDand high performance liquid chromatographic de 2termination of trace isoprocarb in riceYang Rong ,M a Jing 2jun ,Song S huang 2ju(Department of Basic Courses ,Agricultrure University Hebei ,Baoding 071001,China AbstractA simple and reliable method for the determin

3、ation of trace isoprocarb in rice by matrix solid -phase dispersion (MSPD and liquid chromatography was presented 1The MSPD -HPLC technique used for assay of trace isoprocarb in rice was discussed and the conditions of quantitative analysis investigated in detail 1K ey w ordsHPLC ;MSPD ;Isoprocarb ;

4、Residue ;Rice異丙威(isoprocarb 是氨基甲酸酯類殺蟲劑,具有較強(qiáng)的觸殺作用,常用于水稻害蟲的防治1。近十幾年來,食品中農(nóng)藥殘留物在世界范圍內(nèi)引起了人們的關(guān)注,但是對異丙威殘留物的研究較少,尤其是利用高效液相色譜法(HPLC 23。一般應(yīng)用HPLC 對食物中某種痕量物質(zhì)進(jìn)行測定時(shí),樣品的前處理都比較復(fù)雜。美國Austin RL 等4報(bào)道的基質(zhì)固相分散(MSPD 前處理提取分離技術(shù),樣品吸附到很大面積上(500m 2/g C 18,然后依靠所選定的溶劑依次洗脫,可大大簡化許多殘留物前處理方式,最大限度地避免被測殘留物的流失。本試驗(yàn)將MSPD 技術(shù)用于水稻中異丙威殘留物的HPL

5、C 分析,并對定量檢測條件作了詳細(xì)的研究。1試驗(yàn)材料與方法111儀器及試劑高效液相色譜儀由島津(Shimadzu LC -6A 高壓輸液泵,島津SPD -6AV 紫外可見分光檢測器,7125注射進(jìn)樣閥,PEHS -5C 18(5m ,416mm ×125mm 色譜柱,臺(tái)式自動(dòng)平衡記錄儀組成。100140目ODS 按文獻(xiàn)5,6方法合成,產(chǎn)品經(jīng)碳元素分析測定符合要求。環(huán)己烷、丙酮、甲苯、甲醇均為分析純試劑,使用前經(jīng)過蒸餾提純。甲醇經(jīng)0145m 濾膜過濾。水經(jīng)3次蒸餾。異丙威標(biāo)樣由農(nóng)業(yè)部農(nóng)藥檢測所提供。112色譜條件流動(dòng)相:甲醇/水(v/v ,65/35;流量:110ml/min ;記錄紙

6、速:4mm/min ;進(jìn)樣體積:10l ;紫外檢測波長:254nm 。113基質(zhì)柱的制備取1g C 18鍵合相與0125g 樣品及異丙威標(biāo)準(zhǔn)溶液(25l 質(zhì)量濃度為300mg/L 異丙威混合均勻,得到ODS/大米粉混合物,稱其為基質(zhì)。取1支2ml 注射器中,于注射器底部墊一圓濾紙片,將基質(zhì)填入后在注射器頂部再加一圓濾紙片以防止基質(zhì)流出。然后用注射器塞加壓至最后體積為115ml ,制得基質(zhì)柱。2結(jié)果與討論211洗脫劑用量的選擇首先用810ml 分析純環(huán)已烷淋洗按前述方法制備的基質(zhì)柱。在所有情況下洗脫液的流動(dòng)是重力控制。如果洗脫液流動(dòng)不暢,則在柱頂用吸液球加壓,以推動(dòng)重力流動(dòng)。當(dāng)洗脫液流出停止后,

7、柱內(nèi)剩余的環(huán)已烷用上述加壓法從柱上除去。然后用8ml 甲苯洗脫異丙威,收集洗脫液,每1ml 1份,共收集8份,分別測定各份中異丙威的含量(以峰高表示。實(shí)驗(yàn)結(jié)果表明,6ml 甲苯就可將異丙威提取完全。用8ml 環(huán)已烷清洗基質(zhì)柱,可除去大米中的油質(zhì)、蠟質(zhì)及色素。對清洗液作高效液相色譜分析,結(jié)果表明環(huán)已烷中無異丙威存在。然后用6ml 甲苯洗脫,洗脫液用無油壓縮空氣流緩緩吹干。這一步驟很重要,否則在進(jìn)行測定時(shí)會(huì)出現(xiàn)雜峰。吹干后的農(nóng)藥殘留物用015ml 甲醇溶解,1700r/min 離心10min ,取上清液(10l 進(jìn)行液相色譜分析。212流動(dòng)相的確定實(shí)驗(yàn)表明,以甲醇-水為流動(dòng)相洗脫效果好。進(jìn)一步檢驗(yàn)

8、了不同配比的甲醇/水二元流動(dòng)相的洗脫效果,結(jié)果以體積比為65/35(v/v 的甲醇/水作流動(dòng)相為宜。213檢測波長的選擇取適當(dāng)濃度的異丙威的標(biāo)準(zhǔn)溶液在200350nm 范圍內(nèi)(下轉(zhuǎn)第123頁的效率。蛋白質(zhì)組學(xué)的出現(xiàn)雖然只有幾年時(shí)間,但已逐步形成產(chǎn)業(yè)和市場。2003年生命科學(xué)產(chǎn)業(yè)著名市場研究公司美國加州前線戰(zhàn)略管理咨詢公司預(yù)測2008年全球蛋白質(zhì)組市場將達(dá)到2618億美元。未來幾年,蛋白質(zhì)組市場綜合年增長率將達(dá)到12%。總之,蛋白質(zhì)組分析技術(shù)將越來越多地應(yīng)用于醫(yī)學(xué)研究中,為探索生命的奧秘和疾病的發(fā)生機(jī)理做出貢獻(xiàn)。參考文獻(xiàn)1OFarrell PH1High resolution two-dimen

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17、d proteinsin human nasal lavage fluid from non-smokers and smokers using two-dimensional gel electrophoresis and peptide mass flngerprinting J.Proteomics,2002,2:112-1201(收稿日期:2004-06-02(上接第48頁進(jìn)行紫外吸光度掃描,觀察到異丙威的最大吸收峰分別在208nm和254nm處,由于208nm接近甲醇的紫外光截止波長,故選擇254nm作為檢測波長。214工作曲線、檢出限和回收率用甲醇作溶劑,準(zhǔn)確配制一系列異丙威的標(biāo)準(zhǔn)

18、溶液,按選定的條件實(shí)驗(yàn)。當(dāng)異丙威的進(jìn)樣量為30900ng時(shí),以異丙威峰高(cm對其進(jìn)樣量X(ng進(jìn)行線性回歸,求得線性回歸方程為Y=01315+01950X,相關(guān)系數(shù)為01995;檢出限(以2倍信噪比計(jì)算為0186ng。于一定體積的提取液中添加適量的標(biāo)準(zhǔn)品后進(jìn)行回收率試驗(yàn),結(jié)果見表1。表1異丙威的回收率農(nóng)藥添加濃度(ng/g回收率(%(x±s,n=5平均回收率(%異丙威15030045060075090088107±413089100±510088125±412087109±511488150±417588135±411088121±4158215樣品測定對不同地區(qū)的大米樣品進(jìn)行測定,每個(gè)樣品測定6次,樣品中異丙威的殘留量分別為:天津小站米1190ng/g,浙江億禾大米2180ng/g,安徽梗稻米0180ng/g,黑龍江七河源大米未檢出。3討論上述試驗(yàn)表明,應(yīng)用