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1、國際藥學(xué)研究雜志2013年4月第4O卷第2期JIntPharmRes,V01.40,No.2,April,2013·213·脫毒蓖麻籽粕中蓖麻毒素的提取及測定李倩,董娜。,武軍華,王玉霞,賈培媛,張任鵬,睢大貰[摘要]目的優(yōu)化雙夾心ELISA法檢測蓖麻籽粕中蓖麻毒素含量檢測方法,并對方法學(xué)進(jìn)行必要的驗(yàn)證。方法以蓖麻籽粕中蓖麻毒素的含量為指標(biāo),對提取溶劑、蓖麻籽粕與溶劑體積比、提取時間、提取過程是否勻漿進(jìn)行單因素實(shí)驗(yàn),選取最佳提取條件。在最佳提取條件下對方法的精密度和準(zhǔn)確度進(jìn)行驗(yàn)證。結(jié)果優(yōu)化出雙夾心ELISA法檢測蓖麻籽粕中蓖麻毒素
2、的最佳提取溶劑為含0.1%Tween20的磷酸鹽緩沖液(PBST),提取溶劑體積為1:lO(質(zhì)量/體積),勻漿后提取浸泡時間為1h。該方法特異性高,蓖麻籽粕中雜質(zhì)成分對測定無干擾。蓖麻毒素在3.91—250晷/L濃度范圍內(nèi)線性關(guān)系良好,13內(nèi)和日間精密度測定的相對標(biāo)準(zhǔn)偏差(RSD)值分別小于3.2%和11.8%。同一樣品重復(fù)測定日間RSD值小于5.5%。結(jié)論蓖麻籽粕中的殘余蓖麻毒素可采用優(yōu)化后的實(shí)驗(yàn)方法進(jìn)行有效提取,應(yīng)用雙夾心ELISA方法進(jìn)行毒素含量的定量分析,該方法為蓖麻籽粕脫毒工藝的確定及脫毒樣品的質(zhì)量控制提供了技術(shù)支持。[
3、關(guān)鍵詞]蓖麻籽粕;蓖麻毒素;酶聯(lián)免疫吸附分析[中圖分類號]R965.2[文獻(xiàn)標(biāo)識碼]A[文章編號]1674-0440(2013)02-0213-06ExtractionanddeterminationofricinindetoxifiedmealsofcastorbeansLIQian,DUNGNa,WUJun.hua,WANGYu—xia,JIAPei.yuan,ZHANGRen.pengr.SUIDa.yun(1.DepartmentofMilitaryToxicologyandBiochemicalPharmacology,
4、InstituteofPharmacologyandToxicology,AcademyofMilitaryMedicalSciences,Beng100850,China;2.SchoolofPharmaceuticalScience,JilinUniversity,Changchun130021,China,3.ChinesePEAGeneralHospital,Bering100853,China)[Abstract]ObjeetiveTooptimizeextractionconditionsofricinfromcast
5、orbeanmealsviaasandwichenzyme—linkedimmu—nosorbentassay(sandwichELISA).MethodsSandwichenzyme—linkedimmunosorbentassay(ELISA)wasusedtodetectricininthecastorbeanmeals.Thesignalfactortestwasdesignedtooptimizethemethodofricinextraction,includingtheextractionsolvents,thera
6、tiobetweenbeanmealandliquidvolume,treatmenttimeandwithorwithouthomogenization.ResultsItwasshownthatphos·phatebufercontaining0.1%Tween20(PBST)wasthebestbuferforricinextraction.TheoptimumextractionratioofbeanmealVSliquidwas1:10(weight/volume),andtheidealtimewas1hforrici
7、nreleasing.Theoptimizedmethodhadahighspecificityandgoodprecisionwithoutthedisturbancefromimpuritiesinthemeals.Goodlinearcurverelationshipwaspresentedbetweentheconcentrationofricinfrom3.91to250g/L.Therelativestandarddeviation(RSD)within—daywas3.2%.Whenthesamesamplewasd
8、etectedrepetitivelytheRSDinter—dayswas5.5%.ConclusionThericinincastorbeanmealcouldbewellextractedbytheoptimizedmethodestabli