IHongcheng Wang, Bo xu
Isicelo se-R & D
Intshayelelo
Ngokwemvelo yesigaba senqanaba lesikhululo kunye nesigaba seselula, i-chromatography inokwahlulwa ibe yi-chromatography eqhelekileyo (NPC) kunye neSigaba seSigaba se-chromatography se-chromatography (RPC). Kwi-RPC, i-polared yenqanaba leselula inamandla kunelo lesigaba sokuma. I-RPC iye yeyona isetyenziswa kakhulu kwi-chromatography ye-chromatography ye-chromatography ngenxa yokusebenza okuphezulu, isisombululo esifanelekileyo kunye nendlela yokugcina yokugcina. Ke ngoko i-RPC ilungele ukwahlukana nokuhlanjululwa kwemida yepolar okanye engafakwanga, kubandakanya i-Alkalods, i-carbohydrate, i-Acids, i-Acino Acids, i-STETYEAC, iProtet, C8, Phenyl, I-Syano, i-Amino, njl njl. Kuqikelelwa ukuba ngaphezulu kwe-80% ye-RPC ngoku isebenzisa i-C18 iSigaba esiHloniphekileyo. Ke ngoko c18 Ikholamu yeCromatography iye yeyokuba yikholamu yendalo iphela kwilebhu nganye.
Nangona ikholamu ye-C18 inokusetyenziswa kuluhlu olubanzi kakhulu lwezicelo, nangona kunjalo, kwezinye iisampulu ezikwi-polar okanye i-hydrophilic kakhulu, i-chddrophilic, i-C18 ye-chddrophilic, i-C18 inokuba neengxaki xa kusetyenziswa iisampulu ezinjalo. Kwi-RPC, i-SORTE isetyenziswe ngendlela esetyenziswayo i-owel inokuyalelwa ngokwe-polady yayo: amanzi <mecethanol <icethanol <e-ecelanol <tirahydrofuran <isopropanol. Ukuqinisekisa ukugcinwa okuhle kwikholamu kwezi zisampulu (polar ezomeleleyo okanye i-hydrophilic), umda ophezulu wenkqubo eninzi kufuneka isetyenziswe njengesigaba sefowuni. Nangona kunjalo, xa usebenzisa inkqubo yamanzi amsulwa (kubandakanya amanzi amsulwa okanye isisombululo esisulungekileyo) njengesigaba sekhavane, umxokelelwane wekhabhoni we-Carbon kwiSigaba seC18 siye sancipha kwikholamu yekholamu okanye ukungasebenzi. Le phenomenon ibizwa ngokuba yi "Hydrophobic Spanipse" (njengoko kubonisiwe kwinxalenye yomzobo 1). Nangona le meko iguqukile xa ikholamu ihlanjululwe ngezinto eziphilayo ezifana ne-methanol okanye i-acetonitrile, isenokubangela ukonakala kwikholamu. Ke ngoko, kuyimfuneko ukuthintela le meko ukuba yenzeke.
Umzobo 1. Umzobo we-Schematic wezigaba ezinamafutha kumphezulu we-lelica gel kwikholamu ye-C18 (ngasekhohlo) kunye nekholamu ye-C18aq
Ukulungisa ezi ngxaki zikhankanywe apha ngasentla, abavelisi bezinto zokupakisha kwe-chromatographic yokupakisha benze uphuculo lobugcisa. Olunye lwezi kuphuculwa zenza uhlengahlengiso kumphezulu we-matrix ye-silica matrix, njengokuqalisa kwe-hydrophilic cydrophilic amaqela (njengoko kubonisiwe kwinxalenye ye-1), ukwenza umphezulu we-slica gel gerophilic. Yiyo loo nto ke imixokelelwane ye-C18 kumphezulu we-SILICA inokwandiswa ngokupheleleyo phantsi kweemeko ezininzi kakhulu kwaye i-hydrophobic syowip inokuthintelwa. Ezi khola eziguqulweyo ze-C18 zibizwa ngokuba ziikholamu ze-C18, ezizezi-C18aq Iikholamu ze-C18aq I-C18aq Iikholamu zisetyenziswe ngokubanzi ngokwahlukeneyo kunye nokucocwa kwamanzi awomeleleyo, kubandakanywa i-acids ye-organ, i-peptiossides, i-nuckiossides kunye neevithamini ze-nucitisi.
Ukupheliswa ngolunye losetyenziso oluqhelekileyo lwe-C18aq Iikholamu zokuhlanjululwa kweesampulu, ezisusa izinto zetyuwa okanye ze-buffer kwisampulu yesampulu yokuqhuba isampulu kwizifundo ezilandelayo. Kule posi, i-FCF eblowu eblibo eblowu ebomvu enomhlaba oqinileyo isetyenziswa njengesampulu kwaye ihlanjululwe kwikholam ye-C18AQ. Imodeli yesandla ithathelwe indawo yiSolgan kwiSisombululo se-Buffer, ngaloo ndlela siququzelela oku kulandelayo kulandelayo kunye nokugcina izinyithisi kunye nexesha lokusebenza. Ngapha koko, ukucoceka kwesampulu kwaphuculwa ngokususa ukungcola kwisampulu.
Icandelo lovavanyo
Umzobo 2. Isakhiwo sekhemikhali sesampulu.
I-FCF ebukhali i-fcf isetyenziswa njengesampulu kwesi sithuba. Ukucoceka kwesampulu ye-RAW yayingama-86% kwaye isakhiwo sekhemikhali sesampulu yaboniswa kumzobo 2. Isisombululo sesampulu emva koko satyalwa kwikholamu yokuKhanya yintsholongwane. Ukuseta uvavanyo lovavanyo lokuhlanjwa kweFlash kudweliswe kwiTheyibhile 1.
| Isixhobo | Umatshini we-wandbean ™2 | |||
| Iikhatriji | I-12 g eyahlula i-clash cartridge (20 - 45 μm, 100 μ, inombolo ye-oda: sw-5222-012-SP) | I-12 g eyahlula i-closh ye-caltridge (20 - 45 μm, 100 μ, iNombolo: Inombolo ye-WO-5222-012-SP)) | ||
| Wavelength | I-254 NM | |||
| Isigaba sefowuni | I-Solvent A: Amanzi I-Solvent B: Metanol | |||
| Umgangatho wohambo | I-30 ml / min | |||
| Isampulu yokulayisha | I-300 mg (i-risling fcf ngococeko lwe-86%) | |||
| Igradient | Ixesha (cv) | I-Solvent B (%) | Ixesha (cv) | I-Solvent B (%) |
| 0 | 10 | 0 | 0 | |
| 10 | 10 | 10 | 0 | |
| 10.1 | I-100 | 10.1 | I-100 | |
| 17.5 | I-100 | 17.5 | I-100 | |
| 17.6 | 10 | 17.6 | 0 | |
| 22.6 | 10 | 22.6 | 0 | |
Iziphumo neengxoxo
I-PopffLash C18aq RP ye-Cartridge isetyenziselwe isampulu yesampulu kunye nokucocwa. Inyathelo leGradient lalisetyenziswa apho amanzi acocekileyo asetyenziswa njengenqanaba leselula ekuqaleni kwe-LELTER kwaye iqhutywa i-10 yekholamu (CV). Njengoko kubonisiwe kumzobo 3, xa usebenzisa amanzi acocekileyo njengesigaba esiselula, isampulu yagcinwa ngokupheleleyo kwi-carridge ye-flash. Emva koko, i-methanol kwisigaba seefowuni yanda ngokuthe ngqo ukuya kwi-100% kwaye igrafient yagcinwa i-7.5 cv. Isampulu yayiphuma ku-11.5 ukuya kwi-13.5 cv. Kumaqhezu aqokelelweyo, isisombululo sesampulu sathatyathelwa indawo ukusuka kwi-Nah2Po4 isisombululo se-buffer kwi-methanol. Ukuthelekisa isisombululo esinamandla kakhulu, i-methanol yayikukulula kakhulu ukuba isuswe kukujikeleza kwereyitha kwinyathelo elilandelayo, eliququzelela kolu phando lulandelayo.
Umzobo 3. I-Clash Chromatogy yesampulu kwikhatriji ye-C18AQ.
Ukuthelekisa indlela yokuziphatha ye-C18AQ yeKartridge kunye ne-CARDRG ye-C18 ye-C18 ye-C18 yeesampulu zovavanyo oluqinileyo, lwenziwa uvavanyo oluthe tyaba. I-RP ye-RP18 RP ye-Cartridge isetyenzisiwe kwaye i-flash chromatom yesampulu yaboniswa kuMfanekiso we-4. Ke ngoko i-gradient ye-gradient icwangcisiwe kwi-10% methanol kuma-90% amanzi. Njengoko kubonisiwe kumzobo 4, ngenxa yokuwa kwinqanaba lesigaba se-h18 ebangelwa ngumlinganiso omkhulu we-C88, isampulu yayingayigcinile i-CARRIDRE ye-C18 eqhelekileyo kwaye yakhutshwa ngokuthe ngqo kwinqanaba lesibini. Ngenxa yoko, ukusebenza kwesampula yokuthathwa kwesampula okanye ukucocwa akunakugqitywa.
Umzobo 4. I-chromatogygram yesampulu kwi-cartridge ye-C18 eqhelekileyo.
Ukuthelekisa i-glodient ye-mgca, ukusetyenziswa kwenqanaba le-GAIGING inezi zinto zilandelayo:
I-1. Ukusetyenziswa kwesinyibilikisi kunye nokuqhuba ixesha lokuphocofa kwesampulu kuncitshiswa.
I-2. Imveliso ekujoliswe kuyo kwizibongozo ezibukhali, ezinciphisa ivolumu yamaqhezu aqokelelweyo kwaye ke ziququzelele ezi zinto zilandelayo zokujikeleza kunye nokugcina ixesha.
I-3. Imveliso eqokelelweyo eMetanol ekulula ukuba ikhutshiwe, ngaloo ndlela yomisa ixesha incitshisiwe.
Ukuqukumbela, ngenxa yokuhlanjululwa kwesampulu yepolar okanye i-hydrophilic, ye-hydflash c18aq rp ye-carridges ye-clash ye-Clash ye-Clash Brash Simahla
Malunga neNdawo ye-Dopfffflash Bonhring CART
Kukho uthotho lwezahlulo ze-P18aq rp carridges eneenkcukacha ezahlukeneyo ezivela kwiTekhnoloji ye-SAntai (njengoko kubonisiwe kwiTheyibhile 2).
| Inombolo yento | Ubungakanani bekholamu | Umgangatho wohambo (ml / min) | Max.pressored (i-psi / bar) |
| SW-5222-004-SP (AQ) | 5.4 g | 5-15 | I-400 / 27.5 |
| SW-5222-012-SP (AQ) | I-20 g | I-10-25 | I-400 / 27.5 |
| SW-5222-025-SP (AQ) | 33 g | I-10-25 | I-400 / 27.5 |
| SW-5222-040-SP (AQ) | 48 g | I-15-30 | I-400 / 27.5 |
| SW-5222-080-SP (AQ) | I-105 g | 25-50 | I-350 / 24.0 |
| SW-5222-120-SP (AQ) | I-155 g | I-30-60 | I-300 / 20.7 |
| SW-5222220-SP (AQ) | I-300 g | 40-80 | I-300 / 20.7 |
| SW-5222-330-SP (AQ) | I-420 g | 40-80 | I-250 / 17.2 |
Itheyibhile 2. Yahlula i-c18aq rp carridges.
Izinto zokupakisha: Ukusebenza kakuhle kwe-SP18 (AQ) ye-AICACA, 45 μm, 100 å.
i-logy (njengoko kubonisiwe kwiTheyibhile 2).
I-Post Time: Aug-27-2018