中央研究院-基因體研究中心-超高速藥物篩選系統

Agelient 1100 HPLC system with reverse‐phase C18 column Agilent Zorbanx 3.5um. XDB‐C18 (2.1x50nm) is used to determine purity based on UV absorbance at ... NRPB GRC SA 中文版 | English     藥物搜集與管理 實驗自動化 機械手臂系統 超高速讀盤儀器 資料儲存與數據分析 出版品 儀器設備 液體分注操控儀器       ●秘書   Ms.陳凌華   email:[email protected]   Phone:(02)2787-1285 ●計畫主持人   Dr.馬徹   email:[email protected]   Phone:(02)2787-1233 ●化合物收集與藥物庫開發   Dr.毛溪山   email:[email protected]   Phone:(02)2787-1303 ●篩選方法開發   Dr.鄭婷仁   email:[email protected]   Phone:(02)2787-1267 ●篩選系統操作與化學資訊學   Dr.吳盈達   email:[email protected]   Phone:(02)2787-1237 ●藥物化學合成   Dr.方俊民   email:[email protected]   Phnoe:(02)2787-1266              (02)3366-1663       Q1:WhocanhaveaccesstouHTSservices? A1:AnyonehavingaprojectapprovedandsupportedbyNSCorOtheragenciescanapplyforuHTS.ThisprogramissupportedbyNRPB,sowegiveserviceprioritytotheresearchprojectsofNRPB. Q2:HowtoarrangeforauHTSassay? A2:Asanewuser,youneedtofillinanaccountapplicationfirst.PleasehaveyourprojectinformationreadywhenworkingonboththeAccountandProjectapplicationforms.Onceapproved,youwillbegrantedaccesstoProjectandAssyManagements,whereyoucanaddnewprojectandnewassayrequests.Youmighthaveseveralassaysinoneproject. Note:Aprojectisatitledresearchsubject;anassayisoneofthejobsexecutedintheproject. Q3:Whatisthestatusofyourcompound?whatisthefinalDMSOconcentration? A3:Ourcompoundsaresolubilizedin100%DMSOandreformatedtobe1mMstockinonline1536-wellplate.       藥物搜集與管理   LoadingofCompoundPlatestoHTSFacilities A.Removeallrequiredplatesfromfreezers B.Transferwithaheavy‐dutycartandavoidtremblingonmoving. C.Warm‐uponbench‐topovernight. D.Centrifugeat1000rpmfor1min. E.Removesealingmembraneverycarefully. F.Coverwithsterilizedstainlessplatelids(GNFspeciallydesigned).Makesurefitnessofcoverandplate. G.Transferwithaheavy‐dutycarttoHTSsystemandavoidshakingonmoving CompoundAnalysesbyLC‐MS LC‐MSisgeneralacceptedasthestandardforcompoundanalysis.AlthoughcommercialcompoundssometimesprovidetheirLC‐MSinformation,wehaveperformedourownqualitycontrol(QC)onroughly2%ofthesamples.Generallyspeaking,twosamplesfromeach96‐wellparentsolutionsetsdescribedabovearerandomlypickedandanalyzed.Forcompoundscollectedfromcontributors,allnewsamplesincludingmixtureswillbetestedforourownrecord.LC‐MSprofileshavebeenusedforthe“finger‐print”ofbotanicaldrugsespeciallyfortheirmedicalapplicationapprovalinUSandEurope.Ourrecordshouldbeveryusefulforcomparinglaterbatcheswhenthe“hit”isidentified. MaterialsandInstrumentused: BrukermicroTOFisusedforMASSanalysisonpositiveions.Ifnoinformationon positiveionsappears,negativeionsdeterminationwillbeswitcheduponformass determinations. Agelient1100HPLCsystemwithreverse‐phaseC18columnAgilentZorbanx3.5um XDB‐C18(2.1x50nm)isusedtodeterminepuritybasedonUVabsorbanceat214nm. Fomicacid(reagentgrade,JTBaker) *replacestrifluoroaceticacidinrunningsolventstoavoiddamageoftheMSsystem Acetonitrile(HPLCgrade,JTBaker) LCauto‐samplerplasticvials(100ul,Agilent) PreparationofLC‐MSsamplesandLC‐MSprocess: A.Tenulofparentliquidsareaddedwith90ulDMSO. B.FortyulofsolutionsaretransferredtoLCsamplevial. C.LoadvialstoAgelientsystemusingauto‐sampler. D.Inject35ulintothesystemforanalysis E.Runwithtwobuffers(BufferA:waterwitho.1%formicacid(w/v),BufferB:90% acetonitrile/10%waterwith0.1%formicacid). F.HPLCrunningtimeissetat15minwithFlow‐rate:1ml/min. G.Gradientsusedtorun: Time(min) A(%) B(%) 0 90 10 2 90 10 9 0 100 11 0 100 12 90 10 15 90 10 Analysis: Singlecompounds: IntegrateareaunderthecurveofindividualpeakswithMASSinformation. Mixturesofnaturalproductextracts: HPLCprofileswithallMASSdatastored. Purityofsamplesisdeterminedbyintegrationofareaundercurve(at214nm)ofrightpeakwithcorrespondingMASSprofiles.Ingeneralthemajorityofsampleshavepurity>90%.Lowerpurityofcompoundswillbemarkedinthedatabaseforattention. Previous　1&nbsp&nbsp2&nbsp&nbsp3&nbsp&nbsp4&nbsp&nbsp　Next         關於我們 服務項目 科學技術 工作團隊 我們的任務 基因體研究中心超高速篩選系統 基因體中心化學藥物庫 使用者委員會 服務簡介 服務項目內容 計畫架構與執行流程 操作訓練與預約設備使用 實驗方法開發與驗證 藥物搜集與管理 實驗自動化 機械手臂系統 超高速讀盤儀器 資料儲存與數據分析 ..more　 ..more　 ..more　 Copyright©2013中央研究院基因體研究中心.AllRightReserved 台北市南港區115研究院路二段128號|電話:+886-2-2787-1306|傳真:+886-2-2789-9931 服務時間：週一至週五09:00-18:00|基因體研究中心網頁依循「中研院資訊安全規章」制訂與執行