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UPLCMS/MS !"#$%&'()*+,-./012 !"# $%& '() *+& ,- ./0 !12 345 678 1. !"#$%&'()*+,-./0 100081 2. #$1()234567!8/0 100081 34 9:;<=>?@ABCDEFGCDHIGJDKDLMNOPQRST:'U。[ 'UVWXYZ[\]^ _`a^ UPLCMS/MS ?@ABCDEFGLMNOPbc。[ deVWfg hihjklUmlMNOPWatersBEHn\ 18 2.1mm×50.0mm 1.7 μ Toa^VWpqrsot ESI pounvwAxyz{MRM |A} ~CDEFG RL|AMNOPST} 0.1~20.0 μ g/L R\; 0.9994 \RSD =6 ; 2.4%|LOD S/N=3 ; 0.06~0.15 μ g/L BcLOQ S/N=10 ; 0.3~0.4 μ g/L 。[ d'U?@|ACDE FGLMNOPQRST567 XYZ[\]^ _`a^UCDEFGSTMNOP *8.'9 S635  :;<=> A  :?@9 05176611 2015 36-01-03 DeterminationofBioactiveCompoundSulforaphaneinBrassica VegetablesbyUPLCMS/MS LIZhansheng LIUYumei FANGZhiyuanetal  InstituteofVegetablesandFlowers ChineseAcademyofAgriculturalSciences Beijing100081 Abstract Objective Theaimwastoestablisharapidmethodforthedeterminationofsulforaphanein Brassica vegetables cabbage brocco li Chinesekaleandkohlrabi Method Ultrahighperformanceliquidchromatographycoupledwithtandemmassspectrometry UPLCMS/ MS wasdevelopedtodeterminethesulforaphanecontentin Brassica vegetables. Result Sulforaphanewasextractedbythesolventofethylac etate.ThecompoundwasseparatedonWatersBEHC 18 2.1mm×50.0mm 1.7μ column andmassspectrum wasanalyzedbyelectro sprayionizationinpositivemodeESI undermultiplereactionmonitoring MRM mode. Brassica vegetablescontainedsulforaphane and thecalibrationcurveshowedagoodlinearintherangeof0.1-20.0 μ g/Lwiththecorrelationcoefficientof0.9994.Therelativestandarddevi ation RSD was2.4% =6 .Thedetectionlimit LOD S/N=3 wasinrangeof0.060.15 μ g/L andthequantitationlimit LOQ S/N= 10 was0.30.4 μ g/L. Conclusion Themethodwasaccurateandreliable soitcouldquicklydetecttheanticancercompoundofsulfora phanein Brassica vegetables. Keywords UPLCMS/MS Brassica vegetables Anticancercompound Sulforaphane ABCD  ¡¢£¤¥¦931372067 ); §¨©ª«ª¬-® =¯°¦±¥¦9CARS-25-A ); ¢¬²³´µ ¶· 2013BAD01B04 );Y¸¬-¹º´µ 2012AA100105 ); ©ª»¼½¾¿À¿£Á~aÂÃÄÅ ÆÇÈL©ª¢£É¢¬Â¸ÊËCAASASTIPIVF CAAS )。 EFGH !"#1982-), ÌÍÎÏÐÑÒÓÔÕÖ×ØÙÚ FGÛÜÝ~ÁÀ¿¬-'Þ:ÔÕßà¾áÔÕ Ö×ØÙÚFGÛÜÝ~ÁÀ¿¬-'Þ:ÔÕIJKL 20151113 9:*%;<=()Brassicaoleracea >?@A)B*)*C)D@E@FGHIJKLMNOA3N PQ RSTUV3>WXYZF[!;<=?@ \()>B*)?@]^?@F_GHS`abcde f———ghijSulforaphane SF )( k )。ghijflm n 177.29 oS`piqrs\V3tu()!v wx yz{|{i}Glucoraphanin GRA~ RAA Ku V!GRAPVcdH {3!DlPPeghi j 1-2 ,-ghij ¡¢£¤`b¥K¦ §>¨b©bª«b¬-bR®bF¯°tu±² ³´µ¶·¸R8¹º»¼¦½¾¿HSÀKÁ 3-10 ghijK65½ÃHÄÅÆÇGC )、 ÄÅÆÇ 3 ÇGCMS RÈÂÅÆÇHPLC 11-12 ÉÊËÌ¤Í Î HPLC Ã5ÀghijGmÏghijuÈÐÑÒÓÀ ÔÕ¢.v GCR GCMS ÃÖ×Ø HPLCÙÚÛ 8 1 /012MNO HPLC ÃÜÜÝÞßàKáâã~äáã65åßæ 13 ç¯èéS`êëìí65()!ghijK½ ÃîïÈðñ§Ròóôõöæ÷HøÞùúûü ÍÎ ýþÃïþghijvwÿRn!"Åè éS`êë65ghijK#ÈÂÅÆÇ $%3Çà UPLCMS/MS )。 1 PQRST 1.1 UVPQ ghij&ì'2()" LKT TM *+ , 98%- 10mg &ì2ÎwÿHPLC À. 10ml / Æ.m0!12e 1000mg/L K3u -20℃£Ð4 5678ÝÞ9þSÀmK3-[f:;<e 0.1 0.5 1.0 5.0 10.0 R 20.0 μ g/L =>&ì1.2 WXYZ ?@\()Ø?@!"#$%&'() *+,-.ô7{Af:î?@\()BCd1fD2 EFGHþ'I?@JKLB*)*KLD @*<LRE@KMLþ'N-[OPQRSN78ÝÞ-SÀm'2GHTUVWXYZR[1.3 WX[\] -WXYZR[\K'2]S^_` ae`bcídþR['2 0.5g efgån 1∶30 ^_`abcJournalofAnhuiAgri.Sci.2015 43 36 ): 1-3 de@f $âã degh äåå

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DeterminationofBioactiveCompoundSulforaphaneinBrassicaVegetablesbyUPLCMS/MSLIZhansheng,LIUYumei,FANGZhiyuanetal  (InstituteofVegetablesandFlowers,ChineseAcademyofAgriculturalSciences,Beijing100081)Abstract  [Objective]TheaimwastoestablisharapidmethodforthedeterminationofsulforaphaneinBrassicavegetables(cabbage,broccoli,Chinesekaleandkohlrabi).[Method]Ultrahighperformanceliquidchromatographycoupledwithtandemmassspectrometry(UPLCMS/MS)wasdevelopedtodeterminethesulforaphanecontentinBrassicavegetables.[Result]Sulforaphanewasextractedbythesolventofethylacetate.ThecompoundwasseparatedonWatersBEHC18(2.1mm ×50.0mm,1.7μm)column,andmassspectrumwasanalyzedbyelectrosprayionizationinpositivemodeESI(+)undermultiplereactionmonitoring(MRM)mode.Brassicavegetablescontainedsulforaphane,andthecalibrationcurveshowedagoodlinearintherangeof0.1-20.0μg/Lwiththecorrelationcoefficientof0.9994.Therelativestandarddeviation(RSD)was2.4%(n=6).Thedetectionlimit(LOD,S/N=3)wasinrangeof0.060.15μg/L,andthequantitationlimit(LOQ,S/N=10)was0.30.4μg/L.[Conclusion]Themethodwasaccurateandreliable,soitcouldquicklydetecttheanticancercompoundofsulforaphaneinBrassicavegetables.Keywords  UPLCMS/MS;Brassicavegetables;Anticancercompound;Sulforaphane

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[4]ABBAOUIB,RIEDLKM,RALSTONRA,etal.Inhibitionofbladdercancerbybroccoliisothiocyanatessulforaphaneanderucin:Characterization,metabolism,andinterconversion[J].Molecularnutrition&foodre

search,2012,56(11):1675-1687.[5]ABDULLRAZISAF,NOORNM.SulforaphaneissuperiortoglucoraphanininmodulatingcarcinogenmetabolisingenzymesinHepG2cells[J].AsianPacificjournalofcancerprevention,2013,14(7):4235-4238.

[6]AGYEMANAS,CHAERKADYR,SHAWPG,etal.TranscriptomicandproteomicprofilingofKEAP1disruptedandsulforaphanetreatedhumanbreastepithelialcellsrevealscommonexpressionprofiles[J].Breastcancerresearchandtreatment,2012,132(1):175-187.

[7]ANDELOVAH,RUDOLFE,CERVINKAM.InvitroantiproliferativeeffectsofsulforaphaneonhumancoloncancercelllineSW620[J].Actamedica,2007,50(3):171-176.

[8]ANNABIB,ROJASSUTTERLINS,LAROCHEM,etal.Thedietderivedsulforaphaneinhibitsmatrixmetalloproteinase9activatedhumanbrainmicrovascularendothelialcellmigrationandtubulogenesis[J].Molecularnutrition&foodresearch,2008,52(6):692-700.

[9]LIZ,GALLIU,BECKERLE,etal.Sulforaphaneprotectsheartsfromearlyinjuryafterexperimentaltransplantation[J].Annalsoftransplantation:Quarterlyofthepolishtransplantationsociety,2013,18:558-566.

[10]MYZAKMC,DASHWOODRH.Chemoprotectionbysulforaphane:KeeponeeyebeyondKeap1[J].Cancerletters,2006,233(2):208-218.

[11]LIANGH,YUANQP,DONGHR,etal.Determinationofsulforaphaneinbroccoliandcabbagebyhighperformanceliquidchromatography[J].JFoodComposAnal2006,19(5):473-476.

[12]CAMPASBAYPOLION,SANCHEZMACHADODI,BUENOSOLANOC,etal.HPLCmethodvalidationformeasurementofsulforaphanelevelinbroccolibyproducts[J].Biomedicalchromatography:BMC,2010,24(4):387-392.

[13]CELIKH,ARIBURNUE,BAYMAKMS,etal.ArapidvalidatedHPLCmethodfordeterminationofsulforaphaneandglucoraphanininbroccoliandredcabbagepreparedbyvariouscookingtechniques[J].AnalMethodsUk,2014,6(13):4559-4566.

[14]LIANGH,YUANQP,XIAOQ.Effectsofmetalionsonmyrosinaseactivityandtheformationofsulforaphaneinbroccoliseed[J].JMolCatalBEnzym,2006,43(1/2/3/4):19-22.

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