Hongcheng Wang, Bo Xu
Kopo ea R & D Center
Kenyelletso
Ho ea ka seratsoana se lekanang sa karolo ea boemo bo phahameng le karolo ea mehala e nang le metsi, metsi chromatography (NPC) le ho fetolela li-cordgraphy tsa li-chromatography tse tloaelehileng tsa mekhahlelo. Bakeng sa RPC, Polaring ea Mohlophatsi oa Mobile e bohloko ho feta karolo ea terate. RPC e se e le ntho e sebelisoang ka ho fetisisa ea li-chromatography tse nang le metsi a marang-rang ka lebaka la katleho ea eona e phahameng, qeto e ntle le ts'ebetso e hlakileng ea ho thibela. Ka hona rpc e loketse karohano le tlhoekiso ea metsoako e fapaneng ea polar kapa e se nang mafura, liprotheine tse tloaelehileng tse sebelisoang RPC, liprotheine tse sebelisoang haholo CHANO, Amino, joalo-joalo har'a lihlopha tsena tse sebetsang, e sebelisoang haholo ke C18. Ho hakanngoa hore e feta 80% ea RPC e ntse e sebelisa karolo ea C18 e tlotsitsoeng. Ka hona, kholima ea c18 chromagraphy e se e le - e be khokahanyo ea bokahohleng ka leng.
Le ha C18 kholomo e ka sebelisoa likopanong tse fapaneng haholo tsa lits'ebetsong tse ling tsa li-sarpole tse ling tse nang le hydrophic, li ka ba le mathata ha li sebelisoa ho hloekisa sampole tse joalo. RPC, hangata li-solvel tse sebelisitsoeng ho latela ho latela Polarity ea tsona: metsi <methanol <ethanol <ethanol <tetrahydrofufufuran <isopropanol. Ho netefatsa ho boloka hantle khorong bakeng sa sampole tsena (polasetiki e matla kapa haholo. Leha ho le joalo, ha u sebelisa sistimi e hloekileng ea metsi (ho kenyelletsa le metsi a hloekileng kapa ka lebaka la khang ea letsoai Tsela ena e bitsoa "Hydrophobic Mokhasi oa" Hydrophobic "(joalo ka ha ho bontšitsoe ka lehlakoreng le letšehali la setšoantšo sa 1). Le ha boemo bona bo fetoha ha kholomo e hlatsuoa ka li-solvent tse kang methanol kapa aceonitrole, e ntse e ka baka tšenyo ea kholomo. Ka hona, hoa hlokahala ho thibela maemo ana ho etsahala.
Setšoantšo sa 1. Setšoantšo se thellang se nang le metheo ea metheo ea silica sereleng sa silica se tloaelehileng ho C18 (ka ho le letšehali) le C18AQ kholomo (ka ho le letona).
Ho sebetsana le mathata a boletsoeng ka holimo, baetsi ba li-chromatograph ba ile ba etsa liphetoho tsa tekheniki. E 'ngoe ea lintlafatso tsena e etsa liphetoho tse holimo holim'a semolao sa silile Matrix ea Hydrophilic, joalo ka ha ho bontšitsoe karolo e nepahetseng ea setšoantšo sa Siisa Kahoo liketane tsa C18 tse fumanehang ka har'a bokaholimo li ne li atolosoa ka botlalo tlasa maemo a thata haholo le moqha oa hydrophobic o ka qojoa. Likholomo tsena tse fetotsoeng C18 li bitsoa likholomo tsa C18, e leng Clipns tsa C18, tse etselitsoeng tsamaiso ea lintho tse mashome a mararo ebile li ka mamella 100% e ka mamellang 100% e ka mamellang 100%. C18Aq likholomo li sebelisitsoe hohle karolong le tlhoekiso ea metsoako e matla, ho kopanyelletsa le li-acid tse matla, li-a li-li-vicleos tse qhalaneng.
Ho nyahama ke e 'ngoe ea likopo tse tloaelehileng tsa C18AQ li-coloumber tsa Flash bakeng sa sampole bakeng sa li-sampole kapa lipelaelo lithutong tse latelang. Ka poso ena, FCF e bohlale e boputsoa e nang le polasetiki e matla e ile ea sebelisoa e le sampole ebile e hloekisitsoe kholomong ea C18Aq. Mohlala oa samputer o ile oa nkeloa sebaka ke solvent e entsoeng ka tharollo ea buffer, ka hona e tsamaisa mouoane e latelang ea robong le nako ea ho boloka solt. Ntle le moo, bohloeki ba sampole bo ile ba ntlafala ka ho tlosa litšila ka sampole.
Karolo ea liteko
Setšoantšo sa 2. Sebopeho sa lik'hemik'hale sa mohlala.
FCF ea boputsoa bo botle e sebelisitsoe e le sampole ka poso ena. Bohloeki ba sampole e tala e ne e le 86% le sebopeho sa lik'hemik'hale sa sampole se ile sa bontšoa ho Setšoantšo sa 2 m. Tharollo ea sampole e ne e kenngoe kholomong ea flash ke enjene. Sebopeho sa Tlhahlobo ea Flash ea Flash e thathamisitsoe tafoleng ea 1.
| Sesebelisoa | Mochini oa karohano ea karohano2 | |||
| Mabokose | 12 g e belaetsangFlash c18 RP Flash cartridge (Spilecal silica, 20 - 45 μm, 100 \ | 12 g e belaetsangFlash c18Aq rp flash cartridge (Spirical silica, 20 - 45 μm, 100 \)) | ||
| Wavelength | 254 nm | |||
| Karolo ea mobile | Solvent a: metsi Solvent B: Methanol | |||
| Sekhahla sa ho phalla | 30 ml / min | |||
| Sampole Lodding | 300 mg (bo-'mōtoana bo boputsoa bo boputsoa bo hloekileng bo hloekile ka boea ba 86%) | |||
| Gradient | Nako (CV) | Solvent b (%) | Nako (CV) | Solvent b (%) |
| 0 | 10 | 0 | 0 | |
| 10 | 10 | 10 | 0 | |
| 10.1 | 100 | 10.1 | 100 | |
| 17.5 | 100 | 17.5 | 100 | |
| 17.6 | 10 | 17.6 | 0 | |
| 22.6 | 10 | 22.6 | 0 | |
Liphetho le puisano
Koloi ea Uniflash C18Aq Rp Flash cartridge e sebelisetsoa mohlala bakeng sa ho nyalisoa le ho hloekisoa. Gratient gratient e sebelisitsoe ka nose e hloekileng e ne e sebelisoa e le karolo ea mobile ho qala ka ho hlahella le ho balehela malinyane a 10 Colum. Joalokaha ho bontšitsoe ho Setšoantšo sa 3, ha u sebelisa metsi a hloekileng joaloka mohato oa mobile, sampole e ile ea bolokoa ka botlalo katseteng ea flash. Ka mor'a moo, methanol ka karolo ea mobile e ile ea eketseha ka kotloloho ho 100% le Grandient e bolokiloe ka 7.5 CV CV. Mohlala o ile oa phahamisoa ho tloha ho 11.5 ho isa ho 13.5 CV. Likarolong tse bokelitsoeng, tharollo ea sampole e ile ea nkeloa sebaka ho tsoa ho tharollo ea Nah2Po4 Buffer tharollo ea methanol. Ho bapisa le tharollo e telele haholo, methanol e ne e le bonolo haholo ho tlosoa ka lebaka la ho pota-potiloe ke ho palama bohatong bo latelang, bo lekang lipatlisisong tse latelang.
Setšoantšo sa 3. The Samromatogram ea sampole ho lebokose la C18AQ.
Ho bapisa boitšoaro ba ho ts'oaroa ha cardidge ea C18AQ le cartridge ea kamehla ea C18 bakeng sa mehlala e tiileng, tlhahlobo e matla ea papiso e neng e bapisoa. Setsebi sa Koloi ea Khetla Ka hona, gratient ea ho qala e ne e behiloe ho 10% methanol ka metsi a 90%. Joalokaha ho bontšitsoe ho Setšoantšo sa 4, ka lebaka la karolo ea litulo tsa hydrophobic ea litulo tsa C18 tse bakiloeng ke katse e phahameng ea C18 ebe e felisitsoe ka kotloloho ke karolo ea modimo. Ka lebaka leo, ts'ebetso ea ho nyahlatsa mohlala kapa tlhoekiso e ke ke ea phethoa.
Setšoantšo sa 4. The Flash of the sampole ho lebokose le tloaelehileng la C18.
Ha e bapisoa le gratient gratient, tšebeliso ea gradient gradient e na le melemo e latelang:
1. QETELLO SEBAKA LE MOKHATLO OA HO FIHLA MAHALA.
2. Sehlahisoa se lebisitsoeng se fetolela tlhoro e kholo e ommeng e bokellang likaroloana tse bokelletseng mme ka hona e fa seva ea ho boloka bokaholimo.
3. Sehlahisoa se bokelletsoeng se ka methanool tse bonolo ho se mo cheseha, kahoo ho omisa nako se fokotsehile.
Qetellong, hore tlhoekiso ea mohlala e nang le hydrophilic, e arola ea hydrophilic, e arohana haholo c18Aq.
About Series Series C18 RP Flash cartridges
Ho na le letoto la li-Aroflash C18Aq rp flash ka litlhaloso tse fapaneng le theknoloji ea Santai (joalo ka ha ho bontšitsoe tafoleng ea 2).
| Nomoro ea ntho | Sekhahla sa Column | Sekhahla sa ho phalla (ml / min) | Max.Pressure (PSI / BAR) |
| SW-5222-004-SP (AQ) | 5.4 g | 5-15 | 400 / 27,5 |
| SW-5222-012-SP (AQ) | 20 g | 10-25 | 400 / 27,5 |
| SW-5222-025-SP (AQ) | 33 g | 10-25 | 400 / 27,5 |
| SW-5222-040-SP (AQ) | 48 g | 15-30 | 400 / 27,5 |
| SW-5222-080-SP (AQ) | 105 g | 25-50 | 350 / 24.0 |
| SW-5222-120-SP (AQ) | 155 g | 30-60 | 300 / 20.7 |
| SW-5222-220-SP (AQ) | 300 g | 40-80 | 300 / 20.7 |
| SW-5222-330-Sp (AQ) | 420 g | 40-80 | 250 / 17.2 |
Tafole ea 2. Searaflash c18alash cardidges.
Lisebelisoa tsa Packing: Lisebelisoa tse phahameng tsa sepakapaka
logy (joalo ka ha ho bonts'itsoe tafoleng ea 2).
Nako ea poso: Aug-27-2018