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Porous silica particles yakagadzirwa ne-sol-gel nzira ine mamwe magadzirirwo kuti awane macroporous particles.Zvikamu izvi zvakatorwa ne reversible addition fragmentation chain transfer (RAFT) polymerization neN-phenylmaleimide-methylvinylisocyanate (PMI) uye styrene kugadzirira N-phenylmaleidhi yesimbi yesimbi (MPI-station) ye-polymaleimide. s (100 × 1.8 mm id) dzaive dzakatakurwa ne slurry packing.Kuongorora PMP koramu kupatsanurwa kwepeptide musanganiswa unosanganisira mashanu peptides (Gly-Tyr, Gly-Leu-Tyr, Gly-Gly-Tyr-Arg, Tyr-Ile-Gly-Ser-Arnkeg, leucineps yemunhu album digesinch etryucineps) digestion yemunhu. .Pasi pemamiriro ekugadzirisa akajeka, iyo theoretical plate count yepeptide musanganiswa yakakwira kusvika ku280,000 plates/m².Kuenzanisa kupatsanurwa kwekoramu yakagadziridzwa neyekutengesa Ascentis Express RP-Amide column, yakaonekwa kuti kupatsanurwa kwekoramu ye PMP kwaive kwakakwirira kune mbiru yekutengeserana maererano nekugadzirisa kwekuparadzanisa.
Mumakore achangopfuura, indasitiri yebiopharmaceutical yave kuwedzera musika wepasirese nekuwedzera kukuru kwemugove wemusika.Nekuwedzera kuri kuita indasitiri yebiopharmaceutical indasitiri1,2,3, kuongororwa kwepeptides nemapuroteni kunodiwa zvakanyanya.Pamusoro pechinangwa chepeptide, tsvina yakati wandei inogadzirwa panguva yepeptide synthesis, nekudaro ichida kucheneswa kweprotein yehushu. mvura dzemuviri, matishu uye masero ibasa rakaoma zvikuru nekuda kwenhamba huru yezvipenyu zvinogona kuonekwa mune imwe sampuli.Kunyange mass spectrometry chinhu chinoshanda che peptide neprotein sequencing, kana masampuli akadaro akaiswa mu spectrometer yakawanda mune imwe pass, kupatsanurwa hakuzove kwakanaka.Dambudziko iri rinogona kudzikiswa nekushandisa (kudzikisa huwandu hwehuwandu hweLC), hunoita kuti hugadzirise huwandu hweLC, hunoita kuti hugadzirise hunyoro hweLC, hunoita kuti hugadzirise hutsananguro hwehuwandu hweLC. spectrometer panguva yakapiwa4,5,6.Kuwedzera, panguva yekuparadzaniswa kwechikamu chemvura, vaongorori vanogona kutarisa munzvimbo dzakamanikana, nokudaro vachiisa pfungwa idzi dzekuongorora nekuvandudza MS kuonekwa senitivity.Liquid chromatography (LC) yakafambira mberi zvakanyanya mumakore gumi apfuura uye yava nzira yakakurumbira muproteomic analysis7,8,9,10.
Reversed-phase liquid chromatography (RP-LC) inoshandiswa zvakanyanya kucheneswa uye kupatsanurwa kwepeptide musanganiswa uchishandisa octadecyl-modified silica (ODS) seyakamira phase11,12,13.Zvisinei, RP stationary phases haipe kupatsanurwa kunogutsa kwepeptides nemaproteins nekuda kweamphiphi1 dhizaini yakasarudzika uye mapuroteni akasarudzika. inodiwa kuongorora mapeptide nemapuroteni ane polar uye asiri-polar moieties kuti awirirane uye kuchengetedza aya analytes16.Mixed-mode chromatography, iyo inopa kupindirana kwema multimodal, inogona kuva imwe nzira kune RP-LC yekuparadzaniswa kwepeptides, mapuroteni, uye mamwe masanganiswa akaoma.Several mixed-mode-mode chromatography yakashandiswa, mapuroteni aya akamira uye zvikamu7 zvakagadzirirwa, mapuroteni aya akashandiswa uye zvikamu zve1 zvakagadzirirwa, mapuroteni aya akaiswa uye zvikamu zve1 zvakagadzirirwa, mapuroteni aya akaiswa uye zvikamu zve1. 8. selectivity ye polar uye isiri-polar analytes, sezvo kupatsanurwa kunoenderana nekudyidzana pakati peanalyte uye yakamira chikamu.Multimodal kupindirana 29, 30, 31, 32.Recently, Zhang et al.30 yakagadzira dodecyl-terminated polyamine stationary phase uye yakabudirira kupatsanura hydrocarbons, antidepressants, flavonoids, nucleosides, estrogens, uye mamwe akawanda analytes.Iyo polar intercalator ine mapoka maviri epolar uye asiri-polar, saka inogona kushandiswa kupatsanura peptides nemapuroteni ane zvose hydrophobic uye hydrophilicembed hydrophilicembed, hydrophilicembed, hydrophilicembed, hydrophilicembed. columns) inotengeswa pasi pezita rekutengeserana Ascentis Express RP-Amide makoramu, asi makoramu aya anoshandiswa kuongororwa kweamine 33 chete.
Muchidzidzo chemazuva ano, polar-embedded stationary phase (N-phenylmaleimide-embedded polystyrene) yakagadzirwa uye yakaongororwa kuti iparadzaniswe peptides uye trypsin digests yeHSA.Chikamu chekumira chakagadzirirwa uchishandisa nzira inotevera.Porous silica particles yakagadzirirwa maererano nemaitiro akapiwa mukudhinda kwedu kwekare nemamwe magadzirirwo. cetic acid yakagadziridzwa kugadzirira silica particles ine hombe pore size.Chechipiri, new ligand, phenylmaleimide-methyl vinyl isocyanate, yakagadzirwa uye yakashandiswa kuburitsa silica particles kugadzirira polar embedded stationary phase.Iyo yakakonzera stationary phase yakarongedzerwa mustainless steel packing ne optimized column packing icolumn i 100 mm inotsigira i. mechanical vibration kuti uone kuti mubhedha une homogeneous waumbwa mukati me column.Ongorora packed column separation ye peptide misanganiswa inoumbwa neshanu peptides;(Gly-Tyr, Gly-Leu-Tyr, Gly-Gly-Tyr-Arg, Tyr-Ile-Gly-Ser-Arg, Leucine Enkephalin) uye Trypsin digest yehuman serum albumin (HAS) .Musanganiswa wepeptide uye trypsin digest yeHSA yakaonekwa kuti iparadzanise nekugadzirisa kwakanaka uye kubudirira.Iyo mutsara weAMP-AMP yakaenzaniswa neiyo Pth-AMP-column ye-separation. peptides uye mapuroteni akaonekwa kuti akanyatsogadziriswa uye anobudirira pane PMP column, iyo yainyanya kushanda kupfuura Ascentis Express RP-Amide column.
PEG (Polyethylene Glycol), Urea, Acetic Acid, Trimethoxy Orthosilicate (TMOS), Trimethyl Chlorosilane (TMCS), Trypsin, Human Serum Albumin (HSA), Ammonium Chloride, Urea, Hexane Methyldisilazane (HMDS), Methacryloyl Chloride, PO-YL oxide, PO-TEMH oksidi (MC) HPLC Giredhi Acetonitrile (ACN), Methanol, 2-Propanol, uye Acetone Yakatengwa kubva kuSigma-Aldrich (St. Louis, MO, USA).
Musanganiswa we urea (8 g), polyethylene glycol (8 g), uye 8 mL ye 0.01 N acetic acid yakamutswa kwemaminitsi gumi, uye ipapo 24 mL yeTMOS yakawedzerwa ipapo pasi pemamiriro echando-inotonhora. Musanganiswa wekuita wakapisa pa40 ° C kwemaawa matanhatu uye zvino pa 120 ° C yemvura yakaomeswa uye yakaomeswa nesimbi isina mvura kwemaawa masere. 70 ° C kwemaawa 12. Iyo yakaomeswa yakanyorova yakanyorovesa pasi muchoto uye yakaverengerwa pa550 ° C kwemaawa 12. Zvitatu zvitatu zvakagadzirirwa uye zvakaratidzwa kuti zviongorore kuberekana muhukuru hwechidimbu, hukuru hwepore uye nzvimbo yepamusoro.
Nekugadziriswa kwepamusoro kwe silica particles ne pre-synthesized ligand phenylmaleimide-methylvinylisocyanate (PCMP) inoteverwa neradial polymerization ne styrene, polar-containing compound yakagadzirwa.Kumira chikamu cheaggregates uye polystyrene cheni.Maitirwo ekugadzirira anotsanangurwa pazasi.
N-phenylmaleimide (200 mg) uye methyl vinyl isocyanate (100 mg) yakanyungudutswa mutoluene yakaoma, uye 0.1 mL ye2,2'-azoisobutyronitrile (AIBN) yakawedzerwa kune reaction flask kuti igadzirire phenylmaleimide-methyl vinyl isocyanate copolymer yakaomeswa pamaawa 60, C ° musanganiswa wakaomeswa muC 3, C ° musanganiswa wakaomeswa mumaawa 60. ovheni pa40°C kwemaawa matatu.
Yakaomeswa silica particles (2 g) yakaparadzirwa mu toluene yakaoma (100 mL), yakavhenganiswa uye sonicated mu 500 mL round bottom flask for 10 min.PMCP (10 mg) yakanyungudutswa mu toluene uye yakawedzerwa dropwise kune reaction flask kuburikidza nefunnel inodonha. Musanganiswa wacho wakagadziriswa pa 100 ° C uye wakaomeswa pa 100 ° C uye 100 ° C yakaomeswa sefa ye 100 ° C uye 100 ° C yakaomeswa. maawa.Zvino, PMCP-bonded silica particles (100 g) yakanyungudutswa mu toluene (200 ml) uye 4-hydroxy-TEMPO (2 mL) yakawedzerwa muhupo hwe100 µL ye dibutyltin dilaurate sechinhu chinokonzera.Musanganiswa wakakonzerwa pa50 ° C uye wakaomeswa kwemaawa 8, 0 ° .
Styrene (1 mL), benzoyl peroxide BPO (0.5 mL), uye TEMPO-PMCP-yakabatanidzwa silica particles (1.5 g) yakaparadzirwa mu toluene uye yakacheneswa ne nitrogen.Iyo polymerization ye styrene yakaitwa pa 100 ° C kwemaawa 12. Chigadzirwa chinoguma chakashambidzwa ne methanol mu 60 ° C yose yakaratidzwa usiku.
Mienzaniso yakasvibiswa pa 393 K kweawa ye1 kuti iwane kumanikidzwa kwakasara kwepasi pe 10-3 Torr. Nhamba yeN2 yakashambadzirwa pane imwe dzvinyiriro yeP / P0 = 0.99 yakashandiswa kuona iyo yakazara pore volume.The morphology ye bare and ligand-bonded silica particles, High tachies electron scanning, Tokyo electron scanning Japan scanning, Tokyo Tokyo electron scanning. (isina silica uye ligand-bonded silica particles) yakaiswa pamusoro pealuminium column uchishandisa adhesive carbon tape.Gold yakaputirwa pamienzaniso uchishandisa Q150T sputter coater, uye 5 nm Au layer yakaiswa pane samples.Izvi zvinovandudza kushanda kwemaitiro uchishandisa low voltages uye inopa zviyo zvakanaka, inotonhora sputtering, EA2 Elementary sputtering, USA1 The USA Eleham1, USA Eleham1 Eleron. yakashandiswa pakuongorora kwekutanga.A Malvern (Worcestershire, UK) Mastersizer 2000 particle size analyzer yakashandiswa kuwana chidimbu chekukura kwekugovera.Naked silica particles uye ligand-bonded silica particles (5 mg imwe neimwe) yakaparadzirwa mu 5 mL ye isopropanol, sonicated for 10 min, vortexed, vortexed and the 5 min ormotric analysis for the Master's or gratis. inoitwa pamwero we5 °C paminiti pane tembiricha ye30 kusvika 800 °C.
Girazi-lined Stainless simbi yakatetepa-yakaborewa makoramu ezviyero (100 × 1.8 mm id) yaive yakarongedzwa vachishandisa slurry kurongedza nzira, vachishandisa nzira imwechete inoshandiswa muRef.. slurry solvent pamwe chete nepropelling solvent.Zadzai mbiru sequentially nekushandisa zvinodzvinyirira zve 100 MP kwemaminitsi gumi, 80 MP kwemaminitsi gumi nemashanu, uye 60 MP kwemaminitsi 30. Panguva yekutakura, mechanical vibration yakashandiswa neGC column shakers mbiri (Alltech, Deerfield, IL kutakura yunifomu yekudzivirira mukati mekuvhara yunifomu inodzivirira zvishoma nezvishoma .Clurry kutakura mukati mekuvhara yunifomu. the column.Bvisa mbiru kubva kune slurry packing unit uye ubatanidze imwe yakakodzera kune inopinza uye kune LC system kuti uone kushanda kwayo.
Pombi yeLC (10AD Shimadzu, Japan), jekiseni (Valco (USA) C14 W.05) ine 50nL jekiseni loop, membrane degasser (Shimadzu DGU-14A), UV-VIS capillary hwindo rakagadzirwa Special µLC device detector (UV-2075) uye girazi-lined inosungirira madiki madiki madiki madiki madiki madiki madiki madiki simbi. Ening.Mushure mekurongedza, macapillaries (50 μm id 365 uye kuderedza mubatanidzwa capillaries (50 μm) akaiswa pa1/16 ″ kubuda kwemubatanidzwa unoderedza. Northampton, MA).
Albumin kubva muserum yevanhu, lyophilized powder, ≥ 96% (agarose gel electrophoresis) 3 mg yakavhenganiswa netrypsin (1.5 mg), 4.0 M urea (1 mL), uye 0.2 M ammonium bicarbonate (1 mL). Mushonga wacho wakamutswa kwemaminetsi gumi uye wakachengetwa mumvura yekugeza kwemaawa 60% ne 60 %. Sefa mushonga wochengeta pazasi pe4 °C.
Kupatsanurwa kwepeptide musanganiswa uye HSA trypsin digests zvakaongororwa zvakasiyana pamakoramu ePMP.Tarisa kupatsanurwa kwepeptide musanganiswa uye trypsin digest yeHSA nePMP column uye enzanisa mibairo kune Ascentis Express RP-Amide column.The theoretical plate number inoverengwa sezvinotevera:
SEM mifananidzo yevasina silica particles uye ligand-bonded silica particles inoratidzwa muFIG.2 .Sse mifananidzo yechiSilica maseru (a, b) ratidza kuti zvidzidzo zvepamberi.
Kutarisa maerekitironi maikorosikopu mifananidzo yeisina silica particles (A, B) uye ligand-bonded silica particles (C, D).
Iyo particle size distributions of bare silica particles uye ligand-bonded silica particles inoratidzwa muMufananidzo 3 (A) .Volume-based particle size distribution distribution curves yakaratidza kuti ukuru hwemasikirwo emasikirwo akawedzera mushure mekugadzirisa kemikari (Fig. 3A) .The particle size distribution distribution data of the silica particles kubva pachidzidzo chemazuva ano uye chidzidzo chekare cheP particle , chidimbu cheP chidimbu , chiyero cheP . 3.36 μm, zvichienzaniswa nekudzidza kwedu kwekare ne ad (0.5) kukosha kwe 3.05 μm (polystyrene-bound silica particles) 34.Iri batch raive nediki diki rekugovera saizi yekugovera kana ichienzaniswa nekudzidza kwedu kwekare nekuda kwekusiyana kwakasiyana kwePEG, urea, TMOS, uye acetic acid mukupindura kwechikamu chikuru chePlycarene yakadzidzwa chikamu chikuru chesilili. .Izvi zvinoreva kuti kushanda kwepamusoro kwe silica particles ne styrene kunongoisa polystyrene layer (0.97 µm) panzvimbo ye silica, nepo muchikamu chePMP ukobvu hwemasanji hwakanga huri 1.38 µm.
Particle size distribution (A) uye pore size distribution (B) yevare silica particles uye ligand-bound silica particles.
Kukura kwepore, pore volume uye nzvimbo yepamusoro ye silica particles yezvidzidzo zvemazuva ano zvinopiwa muTebhu 1 (B) PSD profiles ye bare silica particles uye ligand-bonded silica particles inoratidzwa mumufananidzo 3 (B) . ukuru hunoderera ne69 mushure mekugadziriswa kwemakemikari, sezvinoratidzwa muTebhu 1 (B), uye kuchinja kwevheji kunoratidzwa muFig. 3 (B) .Saizvozvowo, pore volume ye silica particles yakaderera kubva ku0.67 kusvika ku0.58 cm3 / g mushure mekugadzirisa kemikari.Iyo chaiyo nzvimbo yepamusoro yeiyo ikozvino yakadzidzwa silica 11 m / 2 m / 2g yekudzidza yakapfuura 11 m / 2 m / 2g yekudzidza yakapfuura .Sezvinoratidzwa muTebhenekeri 1 (B), nzvimbo yepamusoro (m2 / g) ye silica particles yakadererawo kubva 116 m2 / g kusvika 105 m2 / g mushure mekugadzirisa kemikari.
Mhedzisiro yekuongorora kwekutanga kwechikamu chekumira inoratidzwa muTable 2. Iyo carbon loading yezvino yakamira yechikamu ndeye 6.35%, iyo yakaderera pane iyo carbon loading yekudzidza kwedu kwekare (polystyrene bonded silica particles, 7.93% 35 uye 10.21%, maererano) 42. The carbon loading of the current station in the polar styling, SP , SP, 35% 35 uye 10.21%. nds yakadai se phenylmaleimide-methylvinylisocyanate (PCMP) uye 4-hydroxy-TEMPO yakashandiswa.Iyotrojeni inorema muzana yezvino inomira yechikamu ndeye 2.21%, ichienzaniswa ne0.1735 uye 0.85% nekurema kwe nitrogen mune zvidzidzo zvekare, maererano. 4) uye (5) vaiva 2.7% uye 2.9%, maererano, nepo carbon loading yechigadzirwa chekupedzisira (6) chaiva 6.35%, sezvakaratidzwa muTebhurari 2. Kurasikirwa kwehuremu kwakaongororwa nePMP stationary phase, uye TGA curve inoratidzwa muMufananidzo 4.TGA curve inoratidza kurasikirwa kwehuremu hwe3 iyo inotakura 5% muchibvumirano che 8.6 chete, iyo carbon 6 ine chibvumirano che 8.6 chete. uyewo N, O, uye H.
The phenylmaleimide-methylvinylisocyanate ligand yakasarudzwa kuti igadziriswe pamusoro pekugadzirisa kwesilica particles nokuti ine polar phenylmaleimide mapoka uye vinylisocyanate mapoka.Vinyl isocyanate mapoka anogona kuwedzera kuita ne styrene nekurarama radical polymerization.Chikonzero chechipiri ndechokuisa boka rine mwero wepakati nepakati neanalyte yemagetsi emagetsi uye pasina analyte yemagetsi emagetsi, uye pasina analyte emagetsi e-analyte. mide moiety haina virtual charge at normal pH.The polarity of the stationary phase inogona kudzorwa nehuwandu hwakakwana hwe styrene uye nguva yekuita yemahara radical polymerization.Nhanho yekupedzisira yemhinduro (free-radical polymerization) inokosha uye inogona kuchinja polarity yechikamu chakamira.Elemental analysis yakaitwa kuti itarise carbon loading yeiyi yakawedzera nguva yekabhoni iyo yakawedzera dhizha rekuita uye kuwedzera kwekabhoni kunowedzera nguva yekabhoni inowedzera. vice versa.SPs dzakagadzirirwa nemhando dzakasiyana-siyana dze styrene dzine carbon loadings dzakasiyana.Zvakare, takura zvikamu izvi zvakamira mumitsipa yesimbi isina tsvina uye tarisa maitiro avo echromatographic (selectivity, resolution, N value, etc.) .Kubva pane izvi zvidzidzo, kuumbwa kwakagadziridzwa kwakasarudzwa kugadzirira PMP stationary phase kuti ive nechokwadi chekudzora polarity uye yakanaka anate anate.
Mishonga mishanu yepeptide (Gly-Tyr, Gly-Leu-Tyr, Gly-Gly-Tyr-Arg, Tyr-Ile-Gly-Ser-Arg, leucine enkephalin) yakaongororwawo uchishandisa PMP column uchishandisa chikamu chefoni;60/40 (v/v) acetonitrile/mvura (0.1% TFA) pamwero wekuyerera we80 μL/min. Pasi pemamiriro akakwana eelution, nhamba yetioretical plate number (N) pakoramu (100 × 1.8 mm id) i20,000 ± 100 (mapuleti anobatika 3/m² eP 200 uye 0MP yeP 000). ma chromatogram anoratidzwa muMufananidzo 5A.Kukurumidza kuongorora paPMP pambiru pakuyerera kwepamusoro (700 μL/min), mapeptide mashanu akaburitswa mukati meminiti imwe, N tsika dzaive dzakanaka kwazvo, 13,500 ± 330 pakoramu (100 × 1.8 mm id), inoenderana ne135 maplateredhi / 0,500 maplateredhim0. (100 × 1.8 mm id) yakanga yakazara nemijenya mitatu yakasiyana yePMP yakamira yechikamu chekutarisa kuberekana.The analyte concentration yekoramu yega yega yakanyorwa uchishandisa yakakwana elution mamiriro uye nhamba yezvinyorwa zvidhinha N uye nguva yekuchengetedza kuti iparadzanise musanganiswa wekuedzwa pakoramu yega yega.Iyo yekuberekazve kweiyo PMPble mutsara wakaderera wePMPble mutsara wakaderera% mucolumn column 4 inoratidzwa muPMPble mutsara wakaderera% mucolumn column 4 inoratidzwa. RSD kukosha, sezvakaratidzwa muTable 3.
Kuparadzaniswa kwepeptide musanganiswa paPMP column (B) uye Ascentis Express RP-Amide column (A);mobile phase 60/40 ACN/H2O (TFA 0.1%), PMP column dimensions (100 × 1.8 mm id);analytical The elution order of the compounds: 1 (Gly-Tyr), 2 (Gly-Leu-Tyr), 3 (Gly-Gly-Tyr-Arg), 4 (Tyr-Ile-Gly-Ser-Arg) uye 5 (leucine) acid enkephalin)).
A PMP column (100 × 1.8 mm id) yakaongororwa kupatsanurwa kwetryptic digests of human serum albumin in high performance liquid chromatography.Chromatogram iri mumufananidzo 6 inoratidza kuti sampuli yakanyatsopatsanurwa uye kugadziriswa kwakanaka kwazvo.HSA digests yakaongororwa pachishandiswa kuyerera kwe10min/totriri yemvura ye100/letri3µ 1% TFA.Sezvinoratidzwa muchromatogram (Mufananidzo 6), iyo HSA digestion yakakamurwa kuita 17 peaks inoenderana ne17 peptides.Kugona kwekuparadzanisa kwepamusoro pese muHSA digest kwakaverengerwa uye kukosha kwakapihwa muTable 5.
A tryptic digest yeHSA (100 × 1.8 mm id) yakapatsanurwa pane PMP column;mwero wekuyerera (100 µL/min), mobile phase 60/40 acetonitrile/mvura ine 0.1% TFA.
apo L ndiyo kureba kwekoramu, η ndiyo viscosity yechikamu chekufambisa, ΔP ndiyo mutsara wekudzokera shure, uye u ndiyo mutsara wekufamba kwechikamu chekufambisa.Kupinda kwechikamu chePMP kwaiva 2.5 × 10-14 m2, kuyerera kwaiva 25 μL / min, uye 60 / 40 v / v / v ACN 0 ACN / mvura 1 × ACN 1 × mvura yakashandiswa PMP yemvura. yakanga yakafanana neyekudzidza kwedu kwekare Ref.34.The permeability yekoramu yakazara nepamusoro-soro porous particles ndeiyi: 1.7 × 10-15 ye 1.3 μm particles, 3.1 × 10-15 ye 1.7 μm particles, 5.2 × 10-5 × 2 μm 2 × 2 × 2 m. μm zvimedu 43. Nokudaro, kupera kwechikamu chePMP kwakafanana ne5 μm core-shell particles.
uko Wx huremu hwekoramu yakarongedzwa nechloroform, Wy huremu hwekoramu yakazara methanol, uye ρ ndiko kuwanda kwezvinonyungudutsa.Densities of methanol (ρ = 0.7866) uye chloroform (ρ = 1.484).Hwose porosity yeSILICA PARTICLES 108 × 18 × 18 × 18 × 8 × 8 × 8 × 1. Urea columns 31 dzatakambodzidza dzaive 0.63 uye 0.55, zvichiteerana.Izvi zvinoreva kuti kuvapo kweurea ligands kunoderedza kupera kwechikamu chakamira.Kune rumwe rutivi, porosity yese yePMP column (100 × 1.8 mm id) ndeye 0.60.The permeability yakaderera neC1 columns yeC1 bhondi yeCMP yeC1. 8-mhando yakamira nhanho iyo C18 ligands yakasungirirwa kune silica particles semutsara cheni, nepo mu polystyrene-mhando yakamira nhanho, iyo A yakati wandei polymer layer inoumbwa yakaitenderedza.Muchiyedzo chakajairika, iyo column porosity inoverengerwa se:
Mufananidzo 7A,B inoratidza PMP mbiru (100 × 1.8 mm id) uye Ascentis Express RP-Amide mbiru (100 × 1.8 mm id) vachishandisa elution mamiriro akafanana (kureva, 60/40 ACN/H2O uye 0.1% TFA).) yevan Deemter plot.Misanganiswa yepeptide yakasarudzwa (Gly-Tyr, Gly-Leu-Tyr, Gly-Gly-Tyr-Arg, Tyr-Ile-Gly-Ser-Arg, Leucine Enkephalin) yakagadzirwa mu20 µL/ Iyo yediki yekuyerera kwemakoramu ese ari 800 µL/min. Iyo shoma HETP mbiru inoyerera pachiyero cheMP / pfumo yeMP µminumu inoyerera pa800 yeHETP makoramu kuyerera kweiyo 80m yeHETP makoramu kuyerera kweiyo MP ye80m is Express RP-Amide column yaiva 2.6 µm uye 3.9 µm, zvichiteerana.The HETP values ​​​​inoratidza kuti kupatsanurwa kwePMP column (100 × 1.8 mm id) iri nani kudarika inotengeswa Ascentis Express RP-Amide column (100 × 1.8 mm Deemter id id) iyo inoyerera muchikamu cheN 7. kwete zvakakosha kana zvichienzaniswa nekudzidza kwedu kwekare.Kunyanya kuparadzaniswa kwepamusoro kwekona yePMP (100 × 1.8 mm id) kana ichienzaniswa neAscentis Express RP-Amide column inobva pakuvandudzwa kwechikamu chechimiro, ukuru, uye maitiro akaoma ekurongedza anoshandiswa mubasa razvino34.
(A) van Deemter plot (HETP versus mobile phase linear velocity) yakawanikwa uchishandisa PMP column (100 × 1.8 mm id) mu 60/40 ACN/H2O ne 0.1% TFA.(B) van Deemter plot (HETP versus mobile phase linear velocity) yakawanikwa uchishandisa Ascentis 6 × 1 × 1 . 0 ACN/H2O ine 0.1% TFA.
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