高效液相串联质谱法检定活性污泥中N高丝氨酸内酯罗梦

ENVIＲONMENTALCHEMISTＲY38220192Vol．38No．2February2019201837ＲeceivedMarch72018．*513784922017ZX07106003．SupportedbytheNationalNaturalScienceFoundationofChina51378492andtheMajorScienceandTechnologyProgramforWaterPollutionControlandTreatmentofChina2017ZX07106003．＊＊[e-mail][e-mail]10．7524/j．issn．0254-6108．2018030704．N-J．2019382281-286．LUOMengYIHaoCHENJingetal．DeterminationofN-acyl-homoserinelactonesinactivatedsludgebyhighperformanceliquidchromatography-tandemmassspectrometryJ．EnvironmentalChemistry2019382281-286．N-*1234121212＊＊1．1000852．1000493．5106554．710055N-AHLs．HPLC-MS/MS7AHLs．-10min．1—1000μg·L－11．0—2．0μg·L－12．0—5．0μg·L－180．0%—113．4%．SBＲAHLs．N-．DeterminationofN-acyl-homoserinelactonesinactivatedsludgebyhighperformanceliquidchromatography-tandemmassspectrometryLUOMeng12YIHao3CHENJing4ZHANGWenzhe12HANYunping12XIAOBenyi12＊＊1．ＲesearchCenterforEco-EnvironmentalSciencesChineseAcademyofSciencesBeijing100085China2．UniversityofChineseAcademyofSciencesBeijing100049China3．SouthChinaInstituteofEnvironmentalSciencesMinistryofEnvironmentalProtectionGuangzhou510655China4．SchoolofEnvironmentalandMunicipalEngineeringXi'anUniversityofArchitectureandTechnologyXi'an710055ChinaAbstractN-homoserinelactonesAHLsarevitalquorum-sensingsignalmoleculesofgram-negativebacteria．Ahighperformanceliquidchromatography-tandemmassspectrometryHPLC-MS/MSmethodwasestablishedtodeterminetheAHLsinactivatedsludge．Amixtureofacetonitrileandwaterservedasthemobilephaseandanelectrospraypositiveionmodewasset．Afterseperationbymobilephasewithgradientelutionthesamplewasmeasuredinmulti-reactionmonitoringmodewithadetectiontimeof10min．Theresultsshowedthatthemethodhadagoodlinearityintherangeof1—1000μg·L－1．Theinstrumentdetectionlimitinstrumentquantificationlimitandaveragerecoveryefficiencywere1．0—2．0μg·L－12．0—5．0μg·L－1and80．0%—113．4%respectively．Whenthismethodwasusedtoanalyzetheactivatedsludgefromwastewatertreatmentplantandlab-scalesequentialbatchactivatedsludgereactorSBＲsystemdifferenttypesofAHLsweredetectedwithagoodpeakshapesandseparationefficiency．ThereforethismethodcansatisfytheactualrequirementforthemeasurementofAHLsinactivatedsludge．28238KeywordsactivatedsludgeN-homoserinelactonesHPLC-MS/MSdetection．1-3．AHLs4-5．、AHLs6-8．．AHLs、9．10．AHLs．AHLsAHLs．AHLs．SBＲHPLC-MS/MS7AHLsAHLs．1Experimentalsection1．1AgilentHPLC-MS/MS1200Agilent1290-6460YDCY-24SL、、＞99%ThermoScientificN-C4-HSL、N-C6-HSL、N-C8-HSL、N-C10-HSL、N-C12-HSL、N-OXO-3-oxo-C8-HSLN-OXO-3-oxo-C10-HSL7AHLs≥98%Sigma500mg·L－1－20℃．Milli-Q．1．2A2OSBＲ．10-1335mL4℃9000r·min－130min30mL0．45μm0．5%3100μL．1．3C1850mm×2．1mm3．5μmWatersSunfire0．3mL·min－110μL10min．ESI+MＲM3．0kV30V99．999%180L·h－1300℃99．999%．2Ｒesultsanddiscussion2．112-14--1．1-7AHLs2N-283．--．10．1Fig．1ChormatogramsofN-acyl-homoserinelactonesAHLsstandardswithtwotypesofmobilephases71．7AHLs2710-11．AHLsAHLsAHLsAHLsAHLs13-14．1Table1ParametersforgradientelutionTime/minAMobilephaseA/%BMobilephaseB/%FlowＲate/mL·min－1Maximumcolumnpressure/bar09550．3600250500．3600440600．3600540600．36005．15950．360075950．36007．19550．360099550．360027N-Table2Ｒetentiontimeof7N-acyl-homoserinelactonesAHLsCompoundMolecularformulaＲetentiontime/sC4-HSLC8H13NO31．480C6-HSLC10H17NO32．356C8-HSLC12H21NO33．095C10-HSLC14H25NO34．143C12-HSLC16H29NO35．7573-oxo-C8-HSLC12H21NO42．5843-oxo-C10-HSLC14H25NO43．322．2ESI+．SCANSIMProductionscan．2．2．3．284382Fig．2MS/MSfragmentsof3AHLs3MＲM7AHLsTable3MultiplereactionmonitoringMＲMconditionsof11AHLsCompoundPrecursorionm/zProductionm/zFragmentor/VCollisionenergy/eVC4-HSL172．1102*/71．1757C6-HSL200．2102*/99955C8-HSL228．1102．1*/127．2854C10-HSL256．1102．1*/155．1954C12-HSL284．2102*/183．17563-oxo-C8-HSL242．1102*/183．18073-oxo-C10-HSL270．2102*/169．2907*Qualitativeion．AHLsM+H+m/z102m/zM+H－101+AHLsAHLs．MＲM．．2．3AHLsAHLs1、10、20、50、100、250、500、1000μg·L－1m/z102．S/N310AHLs4．41—1000μg·L－17AHLs0．991．0—2．0μg·L－12．0—5．0μg·L－1．2．4710μg·L－1100μg·L－165．10μg·L－17AHL2N-28585．2%—113．4%ＲSD4．8%—10．3%．100μg·L－180．0%—105．4%4．2%—7．3%．ＲSD80%—120%ＲSD10%15．．47、Table4LinearequationdetectionlimitLODandquantitationlimitLOQof7AHLscompoundsCompoundLinearequationCorrelationcoefficientＲ2LOD/μg·L－1LOQ/μg·L－1C4-HSLy=132．74x+2462．450．9941．03．0C6-HSLy=92．92x+1677．600．9951．03．0C8-HSLy=60．96x+1139．010．9962．05．0C10-HSLy=54．84x+360．740．9991．02．0C12-HSLy=49．19x+870．170．9971．03．03-oxo-C8-HSLy=123．85x+1260．160．9981．03．03-oxo-C10-HSLy=104．26x+886．270．9991．02．05AHLsn=6Table5Averagerecoveriesandrelativestandarddeviationsof7AHLsn=6Compound10μg·L－1100μg·L－1Ｒecovery/%ＲSD/%Ｒecovery/%ＲSD/%C4-HSL85．210．380．05．2C6-HSL110．27．898．67．1C8-HSL92．38．1102．34．8C10-HSL105．44．8105．27．3C12-HSL93．27．692．35．23-oxo-C8-HSL107．110．392．35．73-oxo-C10-HSL113．47．2105．44．22．5SBＲAHL．1．23．3Fig．3ChromatogramsofAHLsextractedfromsludgesamples286384Fig．4ConcentrationofAHLsintwotypesofactivatedsludge32AHLs．4．4SBＲC4-HSLSBＲAHLsC10-HSLAHLs．3ConclusionAHLs--AHLs1．0—2．0μg·L－12．0—5．0μg·L－17AHLs．Ｒeferences1ＲEVGMAALIUWetal．BacterialsignalsN-acylhomoserinelactonesinducethechangesofmorphologyandetha