Hongcheng Wang, Bo Xu
Aikace-aikacen R & D Cirini
Shigowa da
Dangane da dangin polarid na tsattsauran lokaci da kuma lokaci-lokaci na zamani, ruwa cromatography na yau da kullun na al'ada (NPC) da kuma juyawa Chromatography Recompography (RPC). Don RPC, polarity Pay Mobile ya fi ta ƙarfi fiye da na matakin farko. RPC ta zama ruwan da aka fi amfani da shi a cikin ruwa Chramoography rabon-rabuwa saboda ingancinsa, ƙuduri mai kyau da bayyananniyar ƙuduri. Saboda haka rpc ya dace da rabuwa da tsarkakewa daban-daban ko polar, ciki da siloids, carbohyd, siloids, pendes, serioids, phenyl, Cyano, Amino, da sauransu a cikin waɗannan rukunin ayyukan da aka yi, wanda aka fi amfani da shi shine C18. An kiyasta cewa sama da kashi 80% na RPC yanzu amfani da tsarin C18. Saboda haka C18 Chromatography shafi ya zama dole ne ya zama mai-da-din-kowa na duniya ga kowane dakin gwaje gwaje.
Kodayake ana iya amfani da shafi na C18 a cikin kewayon aikace-aikace sosai, duk da haka, ga wasu samfurori waɗanda ke da kyau sosai ko hydrophilic sosai, alamomin C18 na yau da kullun suna da matsaloli yayin da ake amfani da su don tsarkake irin waɗannan samfuran. A RPC, ana iya ba da umarnin da ake amfani da su na ƙirƙira gwargwadon ƙarfinsu: ruwa <methanol <acetonitrile <ethhydrile <isopropancanol. Don tabbatar da riƙe mai kyau a cikin shafi na waɗannan samfurori (polar polar ko kuma hydrophilic sosai, babban rabo na tsarin ruwa wajibi ne don amfani dashi azaman Mobile Pas. Koyaya, lokacin amfani da tsarkakakken tsarin ruwa (gami da tsarkakakken ruwa ko tsarkakakken sarkar a cikin tsarin riƙewar ko da aka yi shi a cikin tsarin riƙewar ko kuma ga riƙewa da kai tsaye. Wannan sabon abu ana kiransa "rushewar ruwa mai ruwa mai ruwa" (kamar yadda aka nuna a bangaren hagu na Hoto 1). Kodayake wannan yanayin yana juyawa lokacin da aka wanke shafi tare da abubuwan da ke cikin kwayoyin cuta methanol ko acetonitrile, har yanzu na iya haifar da lalacewar shafi. Saboda haka, ya zama dole don hana wannan yanayin daga faruwa.
Hoto 1
Don magance matsalolin da aka ambata a sama, masana'antun chromatographic masana'antu sun yi haɓaka fasaha. Ofayan ɗayan waɗannan haɓakar shine samar da wasu gyare-gyare a saman rukunin Silica, kamar gabatarwar ƙungiyoyin Hydrophilic (kamar yadda aka nuna a cikin ɓangaren silica. Don haka ne C18 Chain a kan silica saman za a iya cikakken fa'ida a karkashin yanayin ruwa mai ruwa mai ruwa da rushewar ruwa na hydrophobic za a iya. Wadannan kayan haɗin C18 ana kiransu da ginshiƙan C18 na ruwa-ruwa, wato C18aq ginshiƙai, waɗanda aka tsara don yanayin ƙa'idar ƙa'idodi kuma suna iya jure tsarin 100%. Anyi amfani da ginshiƙan C18AQ da yawa a cikin rabuwa da tsarkakewa na karfi polar, ciki har da Opt acid acidic, peptides da bitamin mai narkewa.
Walwing shine ɗayan aikace-aikacen aikace-aikacen C18AQ a cikin tsarkakawar filasha don samfurori ko kuma abubuwan haɗin da ke cikin samfin ɗin a cikin binciken mai zuwa. A cikin wannan post, blue blf fcf tare da mai karfi polarity an yi amfani da shi azaman samfurin da tsarkake a shafi na C18aQ. An maye gurbin samfurin samfurin ta hanyar kwayar halitta daga maganin buffer, saboda haka yana gyara wadataccen fitsari mai zuwa da kuma adana abubuwan sha da lokacin aiki. Bugu da ƙari, tsarkin samfurin an inganta ta hanyar cire wasu ƙazanta a cikin samfurin.
Sashi na gwaji
Hoto na 2. Tsarin sunadarai na samfurin.
An yi amfani da FCF mai launin shuɗi mai launin shuɗi azaman samfurin a cikin wannan post. Tsarkin albarkatun ƙasa shine kashi 86% kuma tsarin sunadarai na samfurin an narkar da shi a cikin 1 m Nah2po4 mafita don zama mafi kyawun bayani. Daga nan sai a allurar samfurin samfurin a cikin allon filasha ta hanyar bushewa. Saitin gwaji na Flash tsarkakakken an jera shi a cikin tebur 1.
| Kayan aiki | Inji na Satal2 | |||
| Kera | 12 G | 12 G)) | ||
| Igiyar ruwa | 254 NM | |||
| Aikin hannu | Don haka sauran: Ruwa Subvent b: methanol | |||
| Rate | 30 ml / min | |||
| Samfuran Loading | 300 mg (shuɗi mai launin shuɗi na fanko tare da tsarkakakken kashi 86%) | |||
| M | Lokaci (CV) | Da yawa b (%) | Lokaci (CV) | Da yawa 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 | |
Sakamakon da tattaunawa
An yi amfani da wata filayen Flash Carridge don samfurin abubuwan ban sha'awa da tsarkakewa. An yi amfani da gradient mataki a cikin abin da aka tsarkakakkiyar ruwa a matsayin wayar hannu a farkon ƙa'idodi kuma gudu don kundin shafi 10 (CV). Kamar yadda aka nuna a cikin Hoto na 3, lokacin amfani da tsarkakakken ruwa a matsayin lokacin hannu, samfurin an riƙe samfurin gaba ɗaya akan Flash Carridge. Next, da methanol a cikin wayar hannu ya karu kai tsaye zuwa 100% kuma an kiyaye gradient don 7.5 CV. An sanya samfurin daga 11.5 zuwa 13.5 CV. A cikin sassan da aka tattara, an maye gurbin samfurin samfurin daga Nah2po4 mai buffer mafita a Methanol. Idan aka kwatanta tare da ingantaccen bayani, methanol ya kasance mafi sauƙin cire ta hanyar lalacewa mai lalacewa a cikin mataki na baya, wanda ya sauƙaƙe binciken masu zuwa.
Hoto na 3. Flash Chromatogram na samfurin a kan C18aq Cartridge.
Don kwatanta halayen riƙewa na C18AQ na Clockge da na yau da kullun C18 na yau da kullun don samfuran ƙarfafa pelari, an yi gwajin a layi daya. Anyi amfani da C18 RP Flash Caringogram na samfurin an nuna a cikin Hoto na 4. Don katako na C18 na yau da kullun, mafi girman abin da ke cikin ruwa na yau da kullun shine kusan 90%. Saboda haka an saita farawa a kashi 10% methanol a cikin ruwa 90%. Kamar yadda aka nuna a cikin Hoto na 4, saboda rushewar ruwa na C18 wanda ke haifar da babban rabo a cikin C18 na yau da kullun. A sakamakon haka, ba za a iya kammala aikin whatsing ko tsarkakewa ba.
Hoto 4. Flash Chromatogram na samfurin a kan cartridge na yau da kullun.
Kwayar da gradient mai layi, amfani da matakai gradient yana da wadannan fa'idodi:
1. Amfanin amfani da lokacin da aka gudanar don tsarkakewar samfurin ya ragu.
2. Ainihin samfurin samfurin da aka yi amfani da shi a cikin babban ko ganiya mai ƙarfi, wanda ke rage yawan guntun lardi kuma saboda haka yana gyara ɓoyayyen lokaci mai zuwa.
3. Samfurin da aka tattara yana cikin methanol wanda yake da sauƙin shayarwa, don haka lokacin bushewa an rage.
A ƙarshe, don tsarkakewar samfurin wanda yake da ƙarfi polar ko hydrophils C18AQ RP Flash Carridgeroan ™ yana iya ba da mafi sauki da ingantaccen bayani.
Game da Series Subs C18 RP Flash Carrridges
Akwai jerin jeri na C18aQ RP filashin katako tare da bayanai daban-daban daga fasahar Santai (kamar yadda aka nuna a Tebur na 2).
| Lambar abu | Girman shafi | Rate (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-S (AQ) | 33 g | 10-25 | 400 / 27.5 |
| Sw-522-040-sp (AQ) | 48 g | 15-30 | 400 / 27.5 |
| Sw-522-080-sp (AQ) | 105 g | 25-50 | 350 / 24.0 |
| SW-5222-120-SP (AQ) | 155 g | 30-60 | 300 / 20.7 |
| SW-5222-20-SP (AQ) | 300 g | 40-80 | 300 / 20.7 |
| SW-522230-SP (AQ) | 420 g | 40-80 | 250 / 17.2 |
Tebur 2. Komawa C18AQ RP Flash Carridges.
Packing kayayyaki: ingantaccen feshin C18 (AQ) -Bonded silica, 20 - 45 μm, 100 å.
logy (kamar yadda aka nuna a Tebur na 2).
Lokaci: Aug-27-2018