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Ukusetshenziswa Kwe-SepaFlash Strong Anion Exchange Chromatography Columns ekuhlanzweni kwama-Acidic Compounds

Ukusetshenziswa kwe-SepaFlash Strong

Rui Huang, Bo Xu
Isicelo se-R&D Center

Isingeniso
I-Ion exchange chromatography (IEC) iyindlela yechromatographic evame ukusetshenziswa ukuhlukanisa nokuhlanza izinhlanganisela ezethulwa ngendlela ye-ionic njengesixazululo.Ngokwezimo zokushaja ezihlukene zama-ion ashintshanayo, i-IEC ingahlukaniswa ngezinhlobo ezimbili, i-cation exchange chromatography kanye ne-anion exchange chromatography.Ku-chromatography yokushintshisana nge-cation, amaqembu ane-asidi ahlanganiswe phezulu kwemidiya yokuhlukanisa.Isibonelo, i-sulfonic acid (-SO3H) iyiqembu elivame ukusetshenziswa ekuhwebeni okuqinile kwe-cation (SCX), ehlukanisa i-H + kanye neqembu elikhokhiswa kabi -SO3- ngaleyo ndlela lingakwazi ukukhangisa amanye ama-cations kusixazululo.Ku-anion exchange chromatography, amaqembu ane-alkaline ahlanganiswe phezulu kwemidiya yokuhlukanisa.Isibonelo, i-quaternary amine (-NR3OH, lapho i-R iyiqembu le-hydrocarbon) ivame ukusetshenziswa ekushintsheni okuqinile kwe-anion (SAX), ehlukanisa i-OH- kanye neqembu elikhokhiswa kahle -N+R3 lingakwazi ukukhangisa amanye ama-anion esixazululweni, okuholela ku-anion. ukushintshana umphumela.

Phakathi kwemikhiqizo yemvelo, ama-flavonoid adonse ukunaka kwabacwaningi ngenxa yendima yabo ekuvimbeleni nasekwelapheni izifo zenhliziyo.Njengoba ama-molecule e-flavonoid ene-acidic ngenxa yokuba khona kwamaqembu e-phenolic hydroxyl, i-ion exchange chromatography ingenye inketho ngaphezu kwesigaba esivamile esivamile noma i-chromatography yesigaba ehlehlisiwe yokuhlukaniswa nokuhlanzwa kwalezi zinhlanganisela ezine-asidi.Ku-flash chromatography, imidiya ehlukanisayo evame ukusetshenziswa ekushintshisaneni kwe-ion i-silica gel matrix lapho amaqembu okushintshanisa ama-ion ehlanganiswa khona phezulu.Izindlela zokushintshaniswa kwe-ion ezisetshenziswa kakhulu ku-flash chromatography yi-SCX (imvamisa iqembu le-sulfonic acid) kanye ne-SAX (imvamisa iqembu le-quaternary amine).Encwadini yesicelo eshicilelwe ngaphambilini enesihloko esithi "Ukusetshenziswa Kwe-SepaFlash Strong Cation Exchange Chromatography Columns in the Purification of Alkaline Compounds" yi-Santai Technologies, amakholomu e-SCX asetshenziselwa ukuhlanzwa kwezinhlanganisela ze-alkaline.Kulokhu okuthunyelwe, inhlanganisela yamazinga angathathi hlangothi kanye ne-acidic yasetshenziswa njengesampula ukuhlola ukusetshenziswa kwamakholomu e-SAX ekuhlanzweni kwama-acidic compounds.

Isigaba Sokuhlola

Umfanekiso 1. Umdwebo wohlelo lwesigaba esimile esiboshelwe ebusweni bemidiya yokuhlukanisa i-SAX.

Kulokhu okuthunyelwe, ikholomu ye-SAX efakwe ngaphambili nge-quaternary amine bonded silica isetshenzisiwe (njengoba kuboniswe kuMfanekiso 1).Ingxube ye-Chromone ne-2,4-dihydroxybenzoic acid isetshenziswe njengesampula okufanele ihlanzwe (njengoba kuboniswe kumfanekiso 2).Ingxube yachithwa ku-methanol futhi yalayishwa ku-flash cartridge ngomjovo.Ukusethwa kokuhlolwa kokuhlanzwa kwe-flash kufakwe kuhlu kuThebula 1.

Umfanekiso 2. Isakhiwo samakhemikhali sezingxenye ezimbili zengxube yesampula.

Insimbi

SepaBean™ umshini T

Amakhatriji

4 g SepaFlash Standard Series flash cartridge (silica engajwayelekile, 40 - 63 μm, 60 Å, Inombolo ye-oda: S-5101-0004)

4 g I-SepaFlash Bonded Series SAX flash cartridge (i-silica engajwayelekile, 40 - 63 μm, 60 Å, inombolo ye-oda:SW-5001-004-IR)

Ubude begagasi

254 nm (ukutholwa), 280 nm (ukuqapha)

Isigaba seselula

I-solvent A: I-N-hexane

I-solvent B: I-Ethyl acetate

Izinga lokugeleza

30 mL/min

20 mL/min

Isampula iyalayisha

20 mg (inhlanganisela Yesakhi A kanye Nesakhi B)

I-Gradient

Isikhathi (CV)

I-solvent B (%)

Isikhathi (CV)

I-solvent B (%)

0

0

0

0

1.7

12

14

100

3.7

12

/

/

16

100

/

/

18

100

/

/

Imiphumela nengxoxo

Okokuqala, ingxube yesampula yahlukaniswa yi-cartridge yesigaba esivamile efakwe ngaphambili nge-silica evamile.Njengoba kubonisiwe kuMfanekiso 3, izingxenye ezimbili zesampula zakhishwa ku-cartridge ngokulandelana.Okulandelayo, kwasetshenziswa i-flash cartridge ye-SAX ukuze kuhlanzwe isampula.Njengombukiso kuMfanekiso 4, i-acidic Component B igcinwe ngokuphelele kukhatriji ye-SAX.Ingxenye A engathathi hlangothi yakhishwa kancane kancane kukhathriji ngokuchazwa kwesigaba seselula.

Umfanekiso 3. I-flash chromatogram yesampula kukhatriji yesigaba esivamile.

Umfanekiso 4. I-flash chromatogram yesampula kukhatriji ye-SAX.
Uma kuqhathaniswa Umfanekiso 3 kanye noMfanekiso 4, Ingxenye A inomumo ophakeme ongaguquki kuma-flash cartridge amabili ahlukene.Ukuqinisekisa ukuthi ukuphakama kwe-elution kuyahambisana yini nengxenye, singasebenzisa isici esigcwele sokuskena se-wavelength esakhelwe kusofthiwe yokulawula yomshini we-SepaBean™.Vula idatha yokuhlola yokuhlukaniswa okubili, hudula uye kumugqa wenkomba ku-eksisi yesikhathi (CV) ku-chromatogram uye endaweni ephakeme kakhulu kanye nephoyinti lesibili eliphakeme kakhulu le-elution peak elihambisana ne-Component A, kanye nobubanzi obugcwele be-wavelength balezi zinto ezimbili. amaphuzu azovezwa ngokuzenzakalelayo ngaphansi kwechromatogram (njengoba kukhonjisiwe kuMfanekiso 5 kanye noMfanekiso 6).Uma kuqhathaniswa idatha ye-spectrum ye-wavelength egcwele yalokhu kuhlukaniswa okubili, Ingxenye A ine-spectrum yokumunca engashintshi ekuhlolweni okubili.Ngenxa yesizathu seNgxenye A inokwakheka okuphezulu okungahambisani kuma-flash cartridges amabili ahlukene, kucatshangwa ukuthi kukhona ukungcola okuthile Engxenyeni A enokugcinwa okuhlukile kukhatriji yesigaba esivamile kanye nekhatriji ye-SAX.Ngakho-ke, ukulandelana kwe-luting kuhlukile Kungxenye A kanye nokungcola kulawa ma-flash cartridges amabili, okuholela ekumeni kwesiqongo okungahambisani kuma-chromatogram.

Umfanekiso 5. Ububanzi obugcwele be-wavelength beSakhiwo A kanye nokungcola okuhlukaniswa yikhatriji yesigaba esivamile.

Umfanekiso 6. Ububanzi obugcwele be-wavelength beSakhiwo A kanye nokungcola okuhlukaniswa yikhatriji ye-SAX.

Uma umkhiqizo ohlosiwe ozoqoqwa uyiNgxenye A engathathi hlangothi, umsebenzi wokuhlanza ungaqedwa kalula ngokusebenzisa i-cartridge ye-SAX ngokuqondile ngemva kokulayisha isampula.Ngakolunye uhlangothi, uma umkhiqizo oqondiwe ozoqoqwa Uyingxenye B ene-acidic, indlela yokukhishwa kokuthwebula ingase yamukelwe ngokulungiswa okuncane kuphela ezinyathelweni zokuhlola: lapho isampuli ilayishwa kukhatriji ye-SAX kanye Nengxenye Ephakathi nendawo A. yakhishwa ngokuphelele ngezincibilikisi ze-organic zesigaba esivamile, shintsha isigaba seselula sibe isixazululo se-methanol esiqukethe i-acetic acid engu-5%.Ama-acetate esigabeni esihambayo azoqhudelana neNgxenye B ngokubophezela kumaqembu e-amine e-quaternary esigabeni esimile se-cartridge ye-SAX, ngaleyo ndlela akhiphe Ingxenye B kukhathriji ukuze kutholwe umkhiqizo oqondiwe.I-chromatogram yesampula ehlukaniswe ngemodi yokushintshanisa i-ion iboniswe kuMfanekiso 7.

Umfanekiso 7. I-flash chromatogram Yengxenye B ikhishwe kumodi yokushintshanisa i-ion kukhatriji ye-SAX.

Sengiphetha, isampula eline-asidi noma elingathathi hlangothi lingahlanzwa ngokushesha ngekhatriji ye-SAX ehlanganiswe nekhatriji yesigaba esivamile kusetshenziswa amasu ahlukene okuhlanza.Ngaphezu kwalokho, ngosizo lwesici esigcwele sokuskena se-wavelength esakhelwe kusofthiwe yokulawula yomshini we-SepaBean™, i-spectrum yokumuncwa kwesici sezingxenyana ezikhishiwe ingase iqhathaniswe kalula futhi iqinisekiswe, kusize abacwaningi basheshe banqume ukwakheka nobumsulwa bezingxenyana ezihlutshiwe futhi ngaleyo ndlela kuthuthukiswe. ukusebenza kahle.

Inombolo Yento

Usayizi Wekholomu

Isilinganiso sokugeleza

(mL/min)

Ubukhulu.Ingcindezi

(i-psi/ibha)

I-SW-5001-004-IR

5.9g

10-20

400/27.5

I-SW-5001-012-IR

23 g

15-30

400/27.5

I-SW-5001-025-IR

awu 38g

15-30

400/27.5

I-SW-5001-040-IR

awu 55g

20-40

400/27.5

I-SW-5001-080-IR

awu 122g

30-60

350/24.0

I-SW-5001-120-IR

180 g

40-80

300/20.7

I-SW-5001-220-IR

awu 340g

50-100

300/20.7

I-SW-5001-330-IR

awu 475g

50-100

250/17.2

 

Ithebula 2. I-SepaFlash Bonded Series SAX cartridges flash.Izinto zokupakisha: I-silica eboshwe nge-SAX ehlanzekile kakhulu engajwayelekile, 40 - 63 μm, 60 Å.

Ukuze uthole ukwaziswa okwengeziwe mayelana nokucaciswa okuningiliziwe kwe-SepaBean™umshini, noma imininingwane yoku-oda ku-SepaFlash series flash cartridges, sicela uvakashele iwebhusayithi yethu.


Isikhathi sokuthumela: Nov-09-2018