Kwilabhoratri yohlalutyo yanamhlanje, ichromatography ingaphezulu lee kuneprotocol yesiqhelo; ngumdaniso ontsonkothileyo wonxibelelwano lweemolekyuli. Nokuba ulinganisa ubumdaka bamayeza okanye ufumanisa ungcoliseko lokusingqongileyo kumanqanaba omkhondo, ukuqonda isiseko sefiziksi yokwahlula kwakho kuko okwahlula igcisa kwichromatographer eyinkosi.
Impumelelo kwilebhu ayiveli nje kwisethingi yesixhobo esiphezulu. Ihlala kwintsebenziswano phakathi kwendlela yakho yekhemistri kunye nezinto ezisetyenziswayo ezikumgangatho ophezulu ezikhusela ingqibelelo yakho yesampulu. Kwabo bajonga ubunzima be
I-HPLC vs LC-MS: Yeyiphi omawuyikhethe
, okanye ukulwa neziseko ezingazinzanga, esi sikhokelo sabelana ngeemfihlo zebhentshi-side ukuze ufezekise idatha yeklasi yehlabathi kunye nokwandisa izixhobo zokusebenza.
Ukuqonda i-Handshake ye-Hydrophobic: Yintoni i-Reverse Phase Chromatography?
Ukuphendula umbuzo osisiseko-yintoni i-reverse phase chromatography-umntu kufuneka ajonge ngaphaya kwenkcazo yencwadi. Kwisiseko sayo, kukwahlula okusekelwe "kuloyiko lwamanzi" (i-hydrophobicity). Kwinqanaba eliguqukayo loxinzelelo oluphezulu lwekhromatography yolwelo (RP-HPLC), sisebenzisa isigaba sokumisa esingesopolar kunye nesigaba esiphathwayo sepolar ukuhlela iimolekyuli ngophawu lwazo lwe-hydrophobic.
IKhemistri yeC18 (Octadecyl) Chain
Inqanaba eliqhelekileyo lokumisa libandakanya iinqununu ezisekelwe kwi-silica eziboshwe kunye ne-C18 (octadecyl) amatyathanga. Ngokusuka kumava am, ukuxinana kwala matyathanga e-C18 kunye nomgangatho wenkxaso ye-silica umisela indlela yakho enzima. Abahlalutyi abaninzi badibana nencopho yomsila xa behlalutya iikhompawundi ezisisiseko. Oku kuhlala kubangelwa ngamaqela e-silanol ashiyekileyo kumphezulu we-silica asebenza njengeziza ezingafunekiyo zotshintshiselwano nge-ion.
Ukuyicombulula le nto, sisebenzisa ikholamu ene-end-capped, apho ii-silanes ezincinci "zifihla" ezi ndawo zisebenzayo. Nangona kunjalo, neyona kholamu igqwesileyo ayikwazi ukulungisa inaliti engalunganga. Ukuba incopho yakho yechromatographic ibonakala igqwethiwe ngaphambi kokuba ibethe ikholamu, jonga ivial yakho. Ukusebenzisa ubunyulu obuphezulu
9mm Iingqayi zoMsonto oMfutshane ezineFit yeHlabathi
iqinisekisa ukuba umhlalutyi wakho akabhengezi kwiindonga zevial phambi kokuba inaliti iyichole.
UbuGcisa beModyuli ye-Organic kuPhuhliso lweNdlela ye-HPLC
Ixesha lokugcinwa kwabahlalutyi bakho lilawulwa yi-organic modifier HPLC yoxinaniso. Ngokulungelelanisa umlinganiselo we-Acetonitrile okanye iMethanol kwisithinteli sakho samanzi, ulungisa "amandla elution" wesigaba esiphathwayo.
Umngxunya oqhelekileyo endiwubonayo kugqithiselo lwendlela kukungaqondwa kakuhle kwegophe elution. Ukuba unqakraza isilungisi sakho sendalo ngokukhawuleza, uphulukana nesisombululo; ngokucothayo kakhulu, kwaye unengxaki yokwandiswa kwencopho kunye nokumoshakala kwenyibiliko. Xa utshintsha phakathi
Uhlalutyo vs Preparative HPLC
, ukulawula ukuthambeka kwethambeka kuba yeyona nto iphambili ekunyuseni umgangatho wempumelelo. Khumbula, i-Acetonitrile inikezela nge-viscosity ephantsi kunye namandla aphezulu e-lution, ngelixa i-Methanol inokubonelela ngokukhetha okuhlukeneyo kwiikhompawundi ze-polar ezinokuphoswa yi-ACN.
Ukucacisa uMyalelo we-GC we-Elution: Ukuqikelela uKwahlulwa kweCompound
Ukuqikelela i-gc elution order ngomnye weyona mingeni inomvuzo kwi-chromatographer yerhasi. Ngokungafaniyo ne-HPLC, apho i-solvent chemistry iyi-lever ephambili, ukuhlukana kwe-GC kuchazwe "nguBathathu Emnye wokwahlula": indawo yokubilisa, i-molecular polarity, kunye neprogram yokushisa ikholomu.
Thermodynamics vs. Dipole Interactions
Kwinqanaba lokumiswa kwe-non-polar, i-gc elution order ilandela ngokungqongqo iqondo lokubilisa. Iikhompawundi eziguquguqukayo ngakumbi ziphuma kuqala. Nangona kunjalo, umdlalo uyatshintsha xa usebenzisa ikholamu yepolar njengePEG\/Wax. Kutshanje ndibambe iprojekthi ebandakanya i-isomers apho iindawo zokubilisa ziphantse zafana. Ngokutshintshela kwisigaba se-polar, ukusebenzisana kwe-dipole-dipole kwasivumela ukuba sihlukanise ezi zixhobo ngokusekelwe kwisakhiwo sabo sombane kunobukhulu bawo.
Kumsebenzi onobuzaza obuphezulu njengohlalutyo lokulandela umkhondo wokusingqongileyo, ukuthembeka kwesampulu yakho yesikhongozeli yeyona nto ibalulekileyo. Ukusebenzisa
I-20mm ye-Crimp Top Headspace Vials
inqanda ilahleko yabahlalutyi abaguquguqukayo, iqinisekisa ukuba i-massspectral deconvolution yakho isekelwe kumelo lokwenyani lwesampulu yakho, hayi iqhezu elivuzayo.
I-HPLC vs LC-MS: Ukukhangela i-Sensitivity Shift
Umbuzo oqhelekileyo kwilebhu yethu ngulo: "Simele sihambe nini ukusuka kwi-HPLC vs LC-MS?" Impendulo ilele kubuntununtunu obufunekayo. Ngelixa i-HPLC enokubonwa kwe-UV igqwesile kwi-QC yesiqhelo kumanqanaba e-microgram, i-LC-MS iyafuneka xa ufuna ukufumanisa amanqanaba epicogram okanye ukuchonga okungaziwayo kwiimatriki ezinzima.
Ukuba ufudukela kwi-LC-MS, ukhetho lwakho lweengqayi luba lubaluleke ngakumbi. Iicaps eziqhelekileyo zinokungenisa iiplasticizers kwisigaba sakho seselfowuni, zenza "iiNcopho zeGhost" ezikhathaza ichromatogram yakho. Kungenxa yoko le nto uAijiren's
Bonded Screw Caps
ziyi-staple kwiilebhu eziqinisekisiweyo ze-MS-ziphelisa umngcipheko we-septa ewela kwi-vial okanye i-leaching contaminants kwi-detector.
IAnatomy yeChromatographic Peak egqibeleleyo
Yonke incopho yechromatographic sisixhobo sokuxilonga. Incopho ebukhali, elinganayo ye-Gaussian ikuxelela ukuba inkqubo yakho ilungiselelwe. "Ihlombe" okanye "umsila" ukuxelela ukuba kukho into engalunganga, ehlala ihambelana ne-hardware okanye ivolumu efile.
Ukuphelisa uMqulu oFakeleyo kunye noMqulu weKholamu eyoNgezelelweyo
Omnye wababulali abathuleyo besisombululo ngumthamo ofileyo. Ukuba kukho nawuphi na umsantsa phakathi kwenaliti yakho kunye nomzantsi we-vial, isampuli iyahlakaza. Kwisampulu encinci, ndihlala ndincoma ukusebenzisa iConical Insert. Ezi zifakelo zinyanzelisa isampuli kwindlela emxinwa, ethe nkqo, iqinisekisa inaliti "yeplagi ebukhali". Oku kukhokelela ngokuthe ngqo kwincopho ebukhali kunye nomlinganiselo ongcono wesignali ukuya kwingxolo, ngakumbi kwiinkqubo zoxinzelelo oluphezulu apho ukusasazwa lutshaba.
Ukucombulula iingxaki njengePro: Ukusuka kwiNgxolo esisiseko ukuya kwiSepta Coring
Kwiminyaka yam yokujongana neengxaki, ndifumene ukuba i-70% yemicimbi yezixhobo ngokwenene yimiba enokusebenziseka.
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Isiseko ingxolo kunye neeNcopho zoMoya: Ukuba ubona iincopho apho kungafanele kubekho, izihluzi zakho zinokuba ngunobangela. Ngalo lonke ixesha bhekisa ku
IsiKhokelo esiGqibeleleyo kwiiFilters ze-Micron ze-0.22
phambi kokukhetha izihluzo zakho zesirinji.
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I-Septa Coring: Ukuba ubona amasuntswana e-silicone kwi-vial yakho, inaliti yakho "i-coring" i-septa. Oku kwenzeka ngeekepusi ezikumgangatho ophantsi. I-ND11 yethu ye-Crimp Caps kunye ne-10-425 Screw Caps zichaneke-zobunjineli ukuze zimelane neepunctures ezininzi ngaphandle kokuqhekeka.
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Uzinzo kuGcino: Kwiindlela ze-EPA okanye ukugcinwa kwexesha elide, usebenzisa
24-400 EPA Vials
eyenziwe ngeglasi ye-borosilicate iqinisekisa ukulahleka kwe-solvent ye-zero kunye nokungcoliswa kwe-zero.
Kutheni Ukunyaniseka kweDatha kuQala nge-Consumable
Sihlala sichitha i-50,000 USD kwi-high-end system, emva koko sizame ukugcina iipennies kwi-vial. Kumava am, i-sub-par vial yeyona nto ibiza kakhulu kwilebhu yakho kuba ikhokelela "ekuqhutyweni kwakhona" kunye "nokungaphumeleli ukuqinisekiswa."
Nokuba ufuna i-1ml ye-Shell Vials yechromatography elula okanye i-18mm Screw Thread Headspace Vials ye-GC ezenzekelayo, i-Aijiren ibonelela ngokungaguquguqukiyo okufunwa ziingcali. Siqinisekisa ukuba iodolo yakho ye-gc elution iyaphinda iphindeke imihla ngemihla, kwaye incopho yakho yechromatographic ihlala ibukhali njengosuku owaphuhlisa ngayo indlela.
INkxaso yeNgcali kunye nemibuzo: Ngaba ujongene neziphumo ezingahambelaniyo okanye usokola nophuhliso lwendlela? Masiphucule ukuhamba kwakho kunye. Qhagamshelana nam xa ufuna uthethwano lobugcisa:
Isiphelo Ukwazi ukuba yintoni inqanaba elibuyela umva kunye nokuqonda iinuances zesixhobo sakho luhambo lokufunda oluqhubekayo. Ngokukhetha i-HPLC efanelekileyo ye-organic modifier kunye nezinto ezisetyenziswayo ezikumgangatho ophezulu, utyala imali kwinyani yedatha yakho. Ungavumeli i-sub-par vial yonakalise indlela eqaqambileyo ochithe iiveki ukhula.
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