甜樱桃果实GA_3的测定技术研究A study of the measurement method of gibberellic acid in sweet cherry
张倩;贝峰;孙欣;王明林;辛力;
摘要(Abstract):
【目的】寻求建立甜樱桃果实中赤霉酸的定性定量检测方法,为研究赤霉素在大棚甜樱桃的使用安全性上提供方法参考。【方法】样品经乙腈提取,液-液分配法进行净化,采用高效液相色谱-串联质谱(HPLC-MS/MS)法在SRM模式下对目标物质进行定性和定量检测。【结果】方法的最低检出限(S/N=3)为1.21μg·kg~(-1),定量限(S/N=10)为4.03μg·kg~(-1),在5.0~100.0μg·L~(-1)内呈现良好的线性关系,相关系数r2为0.999 3,添加浓度为0.001、0.002、0.01 mg·kg~(-1)时赤霉素加标回收率分别为89.69%,91.84%和95.37%(n=6),相对标准偏差分别为7.46%,6.21%和4.85%,方法的不确定度为0.13μg·kg~(-1),5份大棚樱桃样品中GA_3含量在21.3~71.1μg·kg~(-1)。【结论】本方法简单快速,灵敏度高,定量准确,可用于樱桃果实中的赤霉素残留量测定。
关键词(KeyWords): 甜樱桃;赤霉酸;高效液相色谱-串联质谱;定性定量检测
基金项目(Foundation): 山东省现代农业产业技术体系水果创新团队建设(SDAIT-06-13);; 山东省农业重大应用技术创新项目——樱桃采后商品化处理设备研制与试验示范;; 泰安市科技计划重大专项(201340629)
作者(Author): 张倩;贝峰;孙欣;王明林;辛力;
Email:
DOI: 10.13925/j.cnki.gsxb.20160298
参考文献(References):
- [1]张晓娜,卢明华,徐林芳,校瑞,蔡宗苇.赤霉素类植物激素分析方法研究进展[J].色谱,2015,33(8):786-791.ZHANG Xiaona,LU Minghua,XU Linfang,XIAO Rui,CAI Zongwei.Recent advances in the analysis of gibberellins plant hormones[J].Chinese Journal of Chromatography,2015,33(8):786-791.
- [2]MEDEIROS M J,OLIVEIRA M T,WILLADINO L,SANTOS M G.Overcoming seed dormancy using gibberellic acid and the performance of young Syagrus coronata plants under severe drought stress and recovery[J].Plant Physiology and Biochemistry,2015,97(4):278-286.
- [3]BOSE S K,YADAV R K,MISHRA S,SANGWAN R S,SINGH A K,MISHRA B,SRIVASTAVA A K,SANGWAN N S.Effect of gibberellic acid and calliterpenone on plant growth attributes,trichomes,essential oil biosynthesis and pathway gene expression in differential manner in Mentha arvensis L.[J].Plant Physiology and Biochemistry,2013,66(5):150-158.
- [4]DOMINGOS S,NOBREGA H,RAPOSO A,CARDOSO V,SOARES I,RAMALHO J C,LEIT?O A E,OLIVEIRA C M,GOULAO L F.Light management and gibberellic acid spraying as thinning methods in seedless table grapes(Vitis vinifera L.):cultivar responses and effects on the fruit quality[J].Scientia Horticulturae,2016,201:68-77.
- [5]EROGUL D,SEN F.Effects of gibberellic acid treatments on fruit thinning and fruit quality in Japanese plum(Prunus salicina Lindl.)[J].Scientia Horticulturae,2015,186:137-142.
- [6]DAGAR A,WEKSLER A,FRIEDMAN H,LURIE S.Gibberellic acid(GA3)application at the end of pit ripening:effect on ripening and storage of two harvests of‘September Snow’peach[J].Scientia Horticulturae,2012,140:125-130.
- [7]ZANG Y X,CHUN I J,ZHANG L L,HONG S B,ZHENG W W,XU K.Effect of gibberellic acid application on plant growth attributes,return bloom,and fruit quality of rabbiteye blueberry[J].Scientia Horticulturae,2016,200:13-18.
- [8]刘芳,袁华招,沈欣杰,廖雄,李天红.外源GA3和PP333对甜樱桃新梢生长及赤霉素代谢关键基因表达的影响[J].核农学报,2013,27(3):272-278.LIU Fang,YUAN Huahao,SHEN Xinjie,LIAO Xiong,LI Tianhong.Effects of GA3and PP333on shoot growth and gene expression of gibberellins metabolism in Prunus avium[J].Journal of Nuclear Agricultural Sciences,2013,27(3):272-278.
- [9]丛庆,张琪,宋丽莉,陈月,蔡洪生,郭长虹.激素在植物冷胁迫应答中的角色[J].核农学报,2016,30(3):614-619.CONG Qing,ZHANG Qi,SONG Lili,CHEN Yue,CAI Hongsheng,GUO Changhong.Roles of phytohormones on cold response in plants[J].Journal of Nuclear Agricultural Sciences,2016,30(3):614-619.
- [10]商桑,王健,丁晓雯.赤霉素GA3急性毒性和遗传毒性[J].食品科学,2015,36(17):236-239.SHANG Sang,WANG Jian,DING Xiaowen.Acute toxicity and genetic toxicity of GA3[J].Food Science,2015,36(17):236-239.
- [11]史玉琴.赤霉素GA3的高效液相色谱检测法及对小鼠机体的氧化损伤作用研究[D].保定:河北农业大学,2011.SHI Yuqin.Study on detection of GA3by HPLC method and on oxidation damage in mice[D].Baoding:Agricultural University of Hebei,2011.
- [12]BO?A A,BINOKAY S,SERTDEMIR Y.The toxicity and teratogenicity of gibberellic acid(GA3)based on the frog embryo teratogenesis assay-Xenopus(FETAX)[J].Turkish Journal of Biology,2009,33(3):181-188.
- [13]TROUDI A,SAMET M A,ZEGHAL N.Hepatotoxicity induced by gibberellic acid in adult rats and their progeny[J].Experimental and Toxicologic Pathology,2010,62(6):637-642.
- [14]YILMAZ Z,CELIK I.Neurotoxic and immunotoxic effects of Indole-3-butyric acid on rats at subacute and subchronic exposure[J].Neurotoxicology,2009,30(3):382-385.
- [15]ERIN N,AFACAN B,ERSOY Y,ERCAN F,BALCI M K.Gibberellic acid,a plant growth regulator,increases mast cell recrutment and alters substance plevels[J].Toxicology,2008,254:75-81.
- [16]TULUCE Y,CELIK I.Influence of subacute and subchronic treatment of abcisic acid and gibberellic acid on serum marker enzymes and erythrocyte and tissue antioxidant defense systems and lipid peroxidation in rats[J].Pestic Biochem Physiol,2006,86(2):85-92.
- [17]崔娟.影响南京地区甜樱桃坐果的关键时期及其机制[D].郑州:中国农业科学院,2015.CUI Juan.Critical period affecting fruit setting of sweet cherry in Nanjing and its mechanism[J].Zhengzhou:Chinese Academy of Agricultural Sciences,2015.
- [18]刘丙花.花期喷激素对红灯樱桃坐果率的影响[J].落叶果树,2007,39(2):10-11.LIU Binghua.Effect of GA3treatment on the fruit setting rate of‘Hongdeng’sweet cherry[J].Deciduous Fruits,2007,39(2):10-11.
- [19]袁琳,许大全.外源赤霉素GA3对大豆光合作用的促进和叶片内源赤霉素GA1+3水平[J].植物生理与分子生物学学报,2002,28(4):317-320.YUAN Lin,XU Daquan.Stimulatory effect of exogenous GA3on photosynthesis and the level of endogenous GA1+3in soybean leaf[J].Journal of Plant Physiology and Molecular Biology,2002,28(4):317-320.
- [20]付磊,陶燕飞.气相色谱法测定草莓中的赤霉素[J].武汉科技大学学报,2002,25(4):359-361.FU Lei,TAO Yanfei.Determination of gibberellin in strawberry by gas chromatography[J].Journal of Wuhan University of Science and Technology,2002,25(4):359-361.
- [21]GUPTA V,KUMAR M,BRAHMBHATT H,REDDY C,SETH A,JHA B.Simultaneous determination of different endogenetic plant growth regulators in common green seaweeds using dispersive liquid-liquid micro extraction method[J].Plant Physiology and Biochemistry,2011,49(11):1259-1263.
- [22]GE L Y,PEH C Y,YONG J W,ONG E S.Analyses of gibberellins by capillary electrophoresis-mass spectrometry combined with solid-phase extraction[J].Journal of Chromatogr A,2007,1159(1-2):242-249.
- [23]中华人民共和国国家卫生和计划生育委员会,农业部,国家食品药品监督管理总局.GB2763-2016.食品中农药最大残留限量[S].北京:中国标准出版社,2016:1-253.National Health and Family Planning Commission of the People's Republic of China,Ministry of Agriculture of the People's Republic of China,China Food and Drug Administration.GB2763-2016.National food safety standard-Maximum residue limits for pesticides in food[S].Beijing:China Standards Press,2016:1-253.
- [24]农业部农药检定所,山西省农药重点实验室.NY/T 788-2004.农药残留试验准则[S].北京:中国标准出版社,2004:5.Institute for the Control of Agrochemicals,Key Laboratory of Pesticides of Shanxi.NY/T 788-2004.Guideline on pesticide residue trials[S].Beijing:China Standards Press,2004:5.
- [25]吴凤琪,靳保辉,陈波,陈沛金,肖锋.水果中8种外源性植物生长调节剂的液相色谱—串联质谱测定[J].中国农学通报,2010,26(15):115-119.WU Fengqi,JIN Baohui,CHEN Bo,CHEN Peijin,XIAO Feng.Determination of eight exogenous plant growth regulator in fruit by liquid chromatography-tandem mass spectrometry[J].Chinese Agricultural Science Bulletin,2010,26(15):115-119.
- [26]XIE W,HAN C,ZHENG Z Q,CHEN X,QIAN Y,DING H Y,SHI L,LV C H.Determination of GA3residue in fruit samples by liquid chromatography–tandem mass spectrometry[J].Food Chemistry,2011,127(2):890-892.
- [27]ROGIS C,GIBSON L R,KNAPP A,HORTON R.Enhancing germination of eastern gamagrass seed with stratification and gibberellic acid[J].Crop Science,2004,44(44):549-552.
- [28]LEHOTAY S J,SON K A,KWON H,KOESUKWIWAT U,FU W S,MASTOVSKA K,HOH E,LEEPIPATPIBOON N.Comparison of Qu ECh ERS sample preparation methods for the analysis of pesticide residues in fruits and vegetables[J].Journal of Chromatography A,2010,1217(16):2548-2560.
- [29]黄红林,刘实,张桃芝.固相萃取—高效液相色谱测定番茄中的赤霉素GA3残留[J].分析科学学报,2005,21(1):75-77.HUANG Honglin,LIU Shi,ZHANG Taozhi.Determination of gibberellin residue in tomato by solid phase extraction-high performance liquid chromatography[J].Journal of Analytical Science,2005,21(1):75-77.
- [30]姜楠,刘思洁,崔勇,李青.2015年吉林省市售水果中植物生长调节剂残留量监测结果分析[J].食品安全质量检测学报,2016,7(1):33-38.JIANG Nan,LIU Sijie,CUI Yong,LI Qing.Monitoring results of plant growth regulators residues of fruits in Jilin in 2015[J].Journal of Food Safety and Quality,2016,7(1):33-38.
- [31]林海丹,谢守新,吴思彦,王岚,邹志飞.广东地区果蔬中植物生长调节剂应用现状调查[J].广东农业科学,2015(12):82-85.LIN Haidan,XIE Shouxin,WU Siyan,WANG Lan,ZOU Zhifei.Investigation application situation of plant growth regulators in fruits and vegetables in Guangdong[J].Guangdong Agricultural Sciences,2015(12):82-85.
- [32]蔡铮,汪雯,陆剑飞,张志恒,杨桂玲.浙江特色水果中植物生长调节剂使用现状与对策[J].浙江农业科学,2015,56(9):1427-1430.CAI Zheng,WANG Wen,LU Jianfei,ZHANG Zhiheng,YANG Guiling.The using situation and countermeasures of plant growth regulators on the characteristic fruit in Zhejiang provence[J].Journal of Zhejiang Agricultural Sciences,2015,56(9):1427-1430.
- [33]戴莹,王纪华,韩平,马帅,冯晓元.农产品中6种植物生长调节剂残留限量标准及检测方法研究进展[J].食品安全质量检测学报,2016,7(2):425-432.DAI Ying,WANG Ji Hua,HAN Ping,MA Shuai,FENG Xiao Yuan.Research progress of 6 kinds of plant growth regulator residue limits standards and detection methods in agricultural products[J].Journal of Food Safety and Quality,2016,7(2):425-432.
- [34]吴凤琪,靳保辉,陈波,陈沛金,肖锋.植物源性食品中赤霉素(GA3)分析方法的研究进展[J].食品工业科技,2011,32(3):435-437.WU Fengqi,JIN Baohui,CHEN Bo,CHEN Peijin,XIAO Feng.Research progress in Gibberellins(GA3)in analysis methods of plant-derived foods[J].Science and Technology of Food Industry,2011,32(3):435-437.
- [35]中华人民共和国浙江出入境检验检疫局.SN/T 0350-2012出口水果中赤霉素残留量的测定液相色谱-质谱/质谱法[S].北京:中国标准出版社,2012:1-14.Zhejiang Entry-Exit Inspection and Quarantine of the People’s Republic of China.SN/T 0350-2012 Determination of gibbrellic acid residues in fruit for export—LC-MS/MS[S].Beijing:China Standards Press,2012:1-14.
- [36]吉林出入境检验检疫局检验检疫技术中心,珲春出入境检验检疫局,中国医科大学临床医药学院.DB22/T 1677-2012豆芽中赤霉素残留量的测定液相色谱-质谱/质谱法[S].北京:中国标准出版社,2012:1-6.Inspection and Quarantine Technology Center of Jilin Entry-Exit Inspection and Quarantine Bureau,Huichun Entry-Exit Inspection and Quarantine Bureau,Clinical Medicine College of China Medical University.DB22/T 1677-2012 Determination of gibbrellic acid residues in soybean sprout and merbean sprout LC-MS/MS[S].Beijing:China Standards Press,2012:1-6.
- [37]北京市海淀区产品质量监督检验所.DB11/T 379-2006豆芽中4-氯苯氧乙酸钠、6-苄基腺嘌呤、2,4-D、赤霉素、福美双的测定[S].北京:中国标准出版社,2006.Beijing Haidian District Product Quality Supervision and Testing Institute.DB11/T 379-2006 Determination of 4-chlorphenoxyacetic sodium,6-benzylam inopurine,2,4-D,gibberellic acid and thiram residues in soybean sprout and merbean sprout[S].Beijing:China Standards Press,2006.