CAJ | 학술논문

采用超高效液相色谱系统,并配有大体积流通池的荧光检测器快速检测奶及奶制品中的黄曲霉毒素M1.在样品中按 m(牛奶)∶V (乙腈)=1∶2.5的比例加入乙腈,采用涡旋及超声辅助液液萃取法对样品中的黄曲霉毒素M 1进行提取,经离心后,取上清液用磷酸盐缓冲液稀释,过黄曲霉毒素M1免疫亲和柱进行净化及浓缩;采用V (甲醇)∶V (乙腈)=50∶50及纯水作为流动相,经UPLC BEH C18柱(100 mm ×2.1 mm ,1.7μm)分离.结果表明:该方法的定量限为0.03μg/kg ,低于现行食品安全国家标准对食品中黄曲霉毒素M1的最低限量标准,符合检测要求;同时,在0.06~1.2μg/kg范围内具有良好的线性,其线性相关系数大于0.999;3个加标浓度的回收率在81.95%~94.20%之间,效果较好;采用大体积流通池检测的灵敏度较普通流通池提高了近3倍;经用于奶及其制品中黄曲霉毒素M1的检测结果与现行标准方法一致;另外,在前处理中,用乙腈对样品中的黄曲霉毒素M 1进行提取,可以有效地沉淀样品中的蛋白质,从而得到澄清的提取液,在过免疫亲和柱净化时只需利用溶液自身重力作用即能过柱.综上,此定量方法具有样品前处理简单、检测速度快和灵敏度高等优点,适用于对牛奶中黄曲霉毒素M1的检测.
채용초고효액상색보계통,병배유대체적류통지적형광검측기쾌속검측내급내제품중적황곡매독소M1.재양품중안 m(우내)∶V (을정)=1∶2.5적비례가입을정,채용와선급초성보조액액췌취법대양품중적황곡매독소M 1진행제취,경리심후,취상청액용린산염완충액희석,과황곡매독소M1면역친화주진행정화급농축;채용V (갑순)∶V (을정)=50∶50급순수작위류동상,경UPLC BEH C18주(100 mm ×2.1 mm ,1.7μm)분리.결과표명:해방법적정량한위0.03μg/kg ,저우현행식품안전국가표준대식품중황곡매독소M1적최저한량표준,부합검측요구;동시,재0.06~1.2μg/kg범위내구유량호적선성,기선성상관계수대우0.999;3개가표농도적회수솔재81.95%~94.20%지간,효과교호;채용대체적류통지검측적령민도교보통류통지제고료근3배;경용우내급기제품중황곡매독소M1적검측결과여현행표준방법일치;령외,재전처리중,용을정대양품중적황곡매독소M 1진행제취,가이유효지침정양품중적단백질,종이득도징청적제취액,재과면역친화주정화시지수이용용액자신중력작용즉능과주.종상,차정량방법구유양품전처리간단、검측속도쾌화령민도고등우점,괄용우대우내중황곡매독소M1적검측.
@@@@Summary Aflatoxins ( AFT ) , whose basic structure is composed of difuran and coumarin , have 17 kinds of derivatives including B1 , B2 , G1 , G2 , M1 , M2 , etc . AFT M 1 was firstly separated from milk . AFT M 1 and AFT M2 were the derivatives of AFT B1 and AFT B2 through the animal metabolism . Especially , AFT B1 and AFT M 1 have been defined as a category A and 2B carcinogen by the International Agency for Research on Cancer ( IARC) from World Health Organization ( WHO) in 1993 , respectively . Moreover , AFT B 1 and AFT M 1 are regarded as strong carcinogens , the carcinogenic mechanism of which is achieved via affecting the pericellular membrane ,inhibiting the synthesis of RNA and interfering the inductive style of specific enzymes . In December 2011 , the General Administration of Quality Supervision , Inspection and Quarantine of the People s Republic of China ( AQSIQ) announced the selective examination results of 200 kinds of liquid milk products . The aflatoxin M 1 in parts of milk products were over ranging the maximum residue limits ( MRLs) of M 1 in milk and milk products , of which the maximum superscalar were 140% exceeded . Moreover , the contents of aflatoxin M 1 were found exceeded in milk powder again in July 2012 . The method used now was to first heat the milk and milk products in water bath , then samples were clean‐up and concentrated by immunoaffinity column after filtered or centrifuged . In this method , no clear sample solutions were obtained , when passing through the immunoaffinity column , sometimes the immunoaffinity column would be blocked , and the recovery would not in expectation . So a better method was needed for determinating the aflatoxin M 1 in milk and milk products . The present study developed an improved analytical method for the fast determination of aflatoxin M 1 in milk and milk products by ultra‐performance liquid chromatography ( UPLC ) combined with large volume flow cell fluorescence detection ( FLD) . The milk sample was extracted by acetonitrile , and the ratio of the milk sample to acetonitrile was 1 to 2 .5 in mass to volume . Then , the sample was extracted by vortex and ultrasound assisted liquid‐liquid extraction ( ULLE) , and was cleaned‐up and concentrated by aflatoxin M 1 immunoaffinity column . The analyte was separated by UPLC BEH C18 column (100 mm × 2 .1 mm , 1 .7μm) , and was eluted with acetonitrile‐methanol ( 50∶50) and pure water . The results showed that the limit of quantitation ( LOQ) of aflatoxin M 1 was 0 .03 μg/kg , which was lower than the national criteria on determination of the minimum level of aflatoxin M 1 in milk and milk products . Meanwhile , high correlation coefficient ( R2 >0 .999) was obtained within linear range from 0 .06 to 1 .2 μg/kg , and reasonable recoveries (81 .95%‐94 .20% ) were in different spike level . In addition , acetonitrile could effectively precipitate protein in milk during the pretreatment to obtain clear extraction which could rapidly pass through immunoaffinity column only by gravity . When using the large volume flow cell , the sensitivity was increased , which was three times than the standard flow cell . The results obtained from this method were similar to the classical method . In conclusion , this quantitative method has many advantages including simple pretreatment , rapid determination and high sensitivity , which can be applied to the determination and quantification of aflatoxin M 1 in milk sample .