近红外荧光传感法测定中药材中赭曲霉毒素A
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摘要
以近红外荧光碳量子点为探针,以赭曲霉毒素A(OTA)适配体为特异性结合体,建立了近红外荧光核酸适体OTA传感检测平台。研究显示,所建立的近红外荧光核酸适体OTA传感对OTA具有高特异性,其他真菌毒素无明显干扰。研究了传感体系组成、p H值、孵化时间等因素对近红外荧光传感性能的影响,确定了最优检测条件。在最优检测条件下,OTA浓度在0.015～1.5 ng·m L~(-1)范围与体系荧光强度恢复呈良好线性关系,检测限为8 pg·m L~(-1)。该方法用于连翘、葛根、莲子、麦芽、陈皮、甘草等中草药中痕量OTA的测定,发现这些中药材都存在赭曲霉毒素A的污染。
A novel near-infrared fluorescence aptasensor was developed using near-infrared fluorescence carbon quantum dots as probes,ochratoxin A(OTA)aptamers as specific binding OTA.The other mycotoxins were no interferences in the determination of OTA.The influence factors on the performance of the aptasensors were investigated including components of aptasensors,pH of the solution,incubation period.The optimum conditions were determined.Under the optimum conditions,a linear fluorescence signal response to OTA concentration was obtained over a wide OTA concentration range of 0.015-1.50 ng·m L~(-1)with a detection limit of 8 pg·m L~(-1).The proposed method was employed in the determination of trace OTA in forsythia suspense,radix puerariae,semen nelumbinis,malt,pericarpium citri reticulatae and radix liquiritiae successfully.
A novel near-infrared fluorescence aptasensor was developed using near-infrared fluorescence carbon quantum dots as probes,ochratoxin A(OTA)aptamers as specific binding OTA.The other mycotoxins were no interferences in the determination of OTA.The influence factors on the performance of the aptasensors were investigated including components of aptasensors,pH of the solution,incubation period.The optimum conditions were determined.Under the optimum conditions,a linear fluorescence signal response to OTA concentration was obtained over a wide OTA concentration range of 0.015-1.50 ng·m L~(-1)with a detection limit of 8 pg·m L~(-1).The proposed method was employed in the determination of trace OTA in forsythia suspense,radix puerariae,semen nelumbinis,malt,pericarpium citri reticulatae and radix liquiritiae successfully.
引文
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[13]李满秀,李永霞,张媛,等.氨基化碳量子点荧光猝灭法测定焦性没食子酸的研究[J].发光学报,2017,38(1):117-123.LI M X,LI Y X,ZHANG Y,et al.. Determination of pyrogallic acid in amino-functional carbon quantum dots fluorescence quenching[J]. Chin. J. Lumin.,2017,38(1):117-123.(in Chinese)
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[20]曾云龙,王天伦,元晓云,等.以考马斯亮蓝为碳源制备近红外碳量子点的方法:中国,CN104479674A[P]. 2015-04-01.ZENG Y L,WANG T L,YUAN X Y,et al.. Method of preparing near-infrared carbon quantum dots by taking Coomassie brilliant blue as carbon source:China,CN104479674A[P]. 2015-04-01.(in Chinese)
[21]WANG B,ZHU Q K,LIAO D L,et al.. Perylene probe induced gold nanoparticle aggregation[J]. J. Mater. Chem.,2011,21(13):4821-4826.
[22]JIN R C,WU G S,LI Z,et al.. What controls the melting properties of DNA-linked gold nanoparticle assemblies[J]. J.Am. Chem. Soc.,2003,125(6):1643-1654.
[23]ZHANG X,SERVOS M R,LIU J W. Instantaneous and quantitative functionalization of gold nanoparticles with thiolated DNA using a p H-assisted and surfactant-free route[J]. J. Am. Chem. Soc.,2012,134(17):7266-7269.
[24]SAHA K,AGASTI S S,KIM C,et al.. Gold nanoparticles in chemical and biological sensing[J]. Chem. Rev.,2012,112(5):2739-2779.
[25]GAO N,YANG W,NIE H L,et al.. Turn-on theranostic fluorescent nanoprobe by electrostatic self-assembly of carbon dots with doxorubicin for targeted cancer cell imaging,in vivo hyaluronidase analysis,and targeted drug delivery[J]. Biosens.Bioelectron.,2017,96:300-307.
[2]KOSALEC I,CVEK J,TOMIC'S. Contaminants of medicinal herbs and herbal products[J]. Arh. Hig. Rada. Toksikol.,2010,60(4):485-501.
[3]张鑫,王福,陈鸿平,等.中药材真菌及真菌毒素污染研究现状[J].世界科学技术—中医药现代化,2015,17(11):2381-2388.ZHANG X,WANG F,CHEN H P,et al.. Research progress on fungi and mycotoxin infection of Chinese medicinal materials[J]. World Sci. Technol. Modern. Trad. Chin. Med. Mater. Med.,2015,17(11):2381-2388.(in Chinese)
[4]BUGNO A,ALMODOVAR A B,PEREIRA T C,et al.. Occurrence of toxigenic fungi in herbal drugs[J]. Braz. J. Microbiol.,2006,37(1):47-51.
[5]HITOKOTO H,MOROZUMI S,WAUKE T,et al.. Fungal contamination and mycotoxin detection of powdered herbal drugs[J]. Appl. Environ. Microbiol.,1978,36(2):252-256.
[6]EFUNTOYE M O. Mycotoxins of fungal strains from stored herbal plants and mycotoxin contents of Nigerian crude herbal drugs[J]. Mycopathologia,1999,147(1):43-48.
[7]HALT M. Moulds and mycotoxins in herb tea and medicinal plants[J]. Eur. J. Epidemiol.,1998,14(3):269-274.
[8]MALLY A,HARD G C,DEKANT W. Ochratoxin a as a potential etiologic factor in endemic nephropathy:lessons from toxicity studies in rats[J]. Food Chem. Toxicol.,2007,45(11):2254-2260.
[9]BAYMAN P,BAKER J L. Ochratoxins:a global perspective[J]. Mycopathologia,2006,162(3):215-223.
[10]TRUCKSESS M,WEAVER C,OLES C,et al.. Determination of aflatoxins and ochratoxin a in ginseng and other botanical roots by immunoaffinity column cleanup and liquid chromatography with fluorescence detection[J]. J. AOAC Int.,2006,89(3):624-630.
[11]THIRUMALA-DEVI K,MAYO M A,REDDY G,et al.. Occurrence of ochratoxin A in black pepper,coriander,ginger and turmeric in India[J]. Food Addit. Contam.,2001,18(9):830-835.
[12]HAN Z,ZHENG Y L,LUAN L J,et al.. Analysis of ochratoxin A and ochratoxin B in traditional Chinese medicines by ultra-high-performance liquid chromatography-tandem mass spectrometry using[13C20]-ochratoxin A as an internal standard[J]. J. Chromatogr. A,2010,1217(26):4365-4374.
[13]李满秀,李永霞,张媛,等.氨基化碳量子点荧光猝灭法测定焦性没食子酸的研究[J].发光学报,2017,38(1):117-123.LI M X,LI Y X,ZHANG Y,et al.. Determination of pyrogallic acid in amino-functional carbon quantum dots fluorescence quenching[J]. Chin. J. Lumin.,2017,38(1):117-123.(in Chinese)
[14]苏安梅,钟青梅,余姝轶,等.碳量子点荧光猝灭法测定饮料中日落黄[J].发光学报,2017,38(4):530-534.SU A M,ZHONG Q M,YU S Y,et al.. Fluorescence quenching method for Determination of sunset yellow in drinks with carbon quantum dots[J]. Chin. J. Lumin.,2017,38(4):530-534.(in Chinese)
[15]苏安梅,余姝轶,覃秀,等.基于碳量子点荧光恢复的三聚氰胺测定方法[J].发光学报,2017,38(7):967-972.SU A M,YU S Y,QIU X,et al.. Determination of melamine based on fluorescence recovery of carbon quantum dots[J].Chin. J. Lumin.,2017,38(7):967-972.(in Chinese)
[16]WANG B,CHEN Y F,WU Y Y,et al.. Aptamer induced assembly of fluorescent nitrogen-doped carbon dots on gold nanoparticles for sensitive detection of AFB1[J]. Biosens. Bioelectron.,2016,78:23-30.
[17]GUO X D,WEN F,ZHENG N,et al.. Development of an ultrasensitive aptasensor for the detection of aflatoxin B1[J]. Biosens. Bioelectron.,2014,56:340-344.
[18]CHROUDA A,SBARTAI A,BARAKET A,et al.. An aptasensor for ochratoxin A based on grafting of polyethylene glycol on a boron-doped diamond microcell[J]. Anal. Biochem.,2015,488:36-44.
[19]MEJRI-OMRANIN,MIODEK A,ZRIBI B,et al.. Direct detection of OTA by impedimetric aptasensor based on modified polypyrrole-dendrimers[J]. Anal. Chim. Acta,2016,920:37-46.
[20]曾云龙,王天伦,元晓云,等.以考马斯亮蓝为碳源制备近红外碳量子点的方法:中国,CN104479674A[P]. 2015-04-01.ZENG Y L,WANG T L,YUAN X Y,et al.. Method of preparing near-infrared carbon quantum dots by taking Coomassie brilliant blue as carbon source:China,CN104479674A[P]. 2015-04-01.(in Chinese)
[21]WANG B,ZHU Q K,LIAO D L,et al.. Perylene probe induced gold nanoparticle aggregation[J]. J. Mater. Chem.,2011,21(13):4821-4826.
[22]JIN R C,WU G S,LI Z,et al.. What controls the melting properties of DNA-linked gold nanoparticle assemblies[J]. J.Am. Chem. Soc.,2003,125(6):1643-1654.
[23]ZHANG X,SERVOS M R,LIU J W. Instantaneous and quantitative functionalization of gold nanoparticles with thiolated DNA using a p H-assisted and surfactant-free route[J]. J. Am. Chem. Soc.,2012,134(17):7266-7269.
[24]SAHA K,AGASTI S S,KIM C,et al.. Gold nanoparticles in chemical and biological sensing[J]. Chem. Rev.,2012,112(5):2739-2779.
[25]GAO N,YANG W,NIE H L,et al.. Turn-on theranostic fluorescent nanoprobe by electrostatic self-assembly of carbon dots with doxorubicin for targeted cancer cell imaging,in vivo hyaluronidase analysis,and targeted drug delivery[J]. Biosens.Bioelectron.,2017,96:300-307.