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Porous silica particles yakagadzirwa ne-sol-gel nzira ine mamwe magadzirirwo kuti awane macroporous particles.Izvi zvikamu zvakatorwa ne reversible addition fragmentation chain transfer (RAFT) polymerization neN-phenylmaleimide-methylvinylisocyanate (PMI) uye styrene kugadzirira N-phenylmaleidhi yepolyboocyanate (MPI). mbiru dzesimbi dzisina tsvina (100 × 1.8 mm id) dzaive dzakaputirwa ne slurry packing.Ikaongororwa PMP column kupatsanurwa kwepeptide musanganiswa unosanganisira peptide shanu (Gly-Tyr, Gly-Leu-Tyr, Gly-Gly-Tyr-Arg, Tyr-Ile-Gly-Gly-Ser-Ser-Argrographic) mashandiro digestion of human serum albumin (HAS) .Pasi pemamiriro ekugadzirisa ekugadzirisa, iyo theoretical plate count yepeptide musanganiswa yakakwira kusvika ku280,000 plates/m².Kuenzanisa kupatsanurwa kwekoramu yakagadziridzwa neyekutengeserana Ascentis Express RP-Amide column, yakaonekwa kuti kupatsanurwa kwekushanda kwePMP yekoramu yekutengeserana kwepamusoro-soro uye kugadzirisa kwekoramu kwepamusoro.
Mumakore achangopfuura, indasitiri yebiopharmaceutical yave kuwedzera musika wepasirese nekuwedzera kukuru kwemusika.Nekukura kuri kuwedzera kweiyo biopharmaceutical indasitiri1,2,3, kuongororwa kwepeptides nemaprotein kunodiwa zvakanyanya.Kuwedzera kune inotarirwa peptide, tsvina yakati wandei inogadzirwa panguva yepeptide synthesis, nekudaro ichida kutsvagisa chishuwo chekutsvagisa. hunhu hwemapuroteni mumvura dzemuviri, matishu uye masero ibasa rakaoma zvakanyanya nekuda kwenhamba huru yezvipenyu zvinogona kuonekwa mune imwe sampuli.Kunyange mass spectrometry chiri chishandiso chinoshanda che peptide neprotein sequencing, kana sampuli dzakadaro dzakabaiwa mu mass spectrometer mune imwe pass, kupatsanurwa hakuzove kwakanakira. kuderedza nhamba yeanalytes inopinda mu spectrometer yakawanda panguva yakapiwa4,5,6.Mukuwedzera, panguva yekuparadzaniswa kwechikamu chemvura, vaongorori vanogona kutarisa munzvimbo dzakamanikana, nokudaro vachiisa pfungwa idzi dzekuongorora nekuvandudza kunzwisisika kweMS.
Reversed-phase liquid chromatography (RP-LC) inoshandiswa zvakanyanya kucheneswa uye kupatsanurwa kwepeptide misanganiswa uchishandisa octadecyl-modified silica (ODS) seyakamira phase11,12,13.Zvisinei, RP stationary phases haipei kunogutsa kupatsanurwa kwepeptides nemaproteins nekuda kweamphiphi5 yakaomesesa chimiro uye mapuroteni ayo akaomesesa. zvimiro zvakamira zvinodiwa kuti uongorore peptides uye mapuroteni ane polar uye asiri-polar moieties kuti adyidzane nekuchengetedza aya analytes16.Mixed-mode chromatography, iyo inopa multimodal interactions, inogona kuva imwe nzira yeRP-LC yekuparadzaniswa kwepeptides, mapuroteni, uye mamwe masanganiswa akaoma. peptide neprotein separations17,18,19,20,21.Mixed-mode stationary phases (WAX/RPLC, HILIC/RPLC, polar intercalation/RPLC) akakodzera peptide nemaprotein separations nekuda kwekuvapo kwepolar and non-polar groups22,23,24,25,25,26,26,26,26,26,24,25,25,26. Covalently bonded polar mapoka anoratidza simba rekuparadzanisa rakanaka uye yakasarudzika sarudzo yepolar 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 kuparadzanisa peptides nemapuroteni ane zvose hydrophobic uye hydrophilic molar. amide-embedded C18 columns) inotengeswa pasi pezita rekutengeserana Ascentis Express RP-Amide makoramu, asi aya makoramu anoshandiswa pakuongorora amine 33 chete.
Muchidzidzo chemazuva ano, polar-embedded stationary phase (N-phenylmaleimide-embedded polystyrene) yakagadzirirwa uye yakaongororwa kuti iparadzanise peptides uye trypsin digests yeHSA.Chikamu chekumira chakagadzirirwa uchishandisa nzira inotevera.Porous silica particles yakagadzirirwa maererano nemaitiro akapiwa mukudhindwa kwedu kwekare nemamwe magadzirirwo. TMOS, mvura acetic acid yakagadziridzwa kugadzirira silica particles nehombe pore size.Chechipiri, new ligand, phenylmaleimide-methyl vinyl isocyanate, yakagadzirwa uye yakashandiswa kubvisa silica particles kugadzirira polar embedded stationary phase.Iyo yakaguma yakamira chikamu chakanga chakazara mu optimized simbi 100 mm × 100 simbi yesimbi. scheme.The column packing inobatsirwa ne mechanical vibration kuti mubhedha une homogeneous waumbwa mukati me column.Ongorora packed column separation ye peptide misanganiswa inosanganisira peptides shanu; (Gly-Tyr, Gly-Leu-Tyr, Gly-Gly-Tyr-Arg, Tyr-Ile-Gly-Ser-Arg, Leucine Enkephalin) uye Trypsin digest ye human serum albumin (HAS) .Musanganiswa wepeptide uye trypsin digest yeHSA yakaonekwa kuti iparadzanise nekugadzirisa kwakanaka uye kushanda zvakanaka kweiyo yeP. RP-Amide column.Zvose peptides uye mapuroteni akaonekwa kuti akanyatsogadziriswa uye anobudirira pane PMP column, iyo yakanyatsoshanda 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, xy-POH, MCTEM, Perfecta, Benzol-Chloride, MCTEM, Benzol-Chloride (BPO), HPLC Grade Acetonitrile (ACN), Methanol, 2-Propanol, uye Acetone Yakatengwa kubva kuSigma-Aldrich (St. Louis, MO, USA).
Musanganiswa weurea (8 g), polyethylene glycol (8 g), uye 8 mL ye0.01 N acetic acid yakasimudzwa kwemaminitsi gumi, uye ipapo 24 mL yeTMOS yakawedzerwa ipapo pasi pemamiriro echando-inotonhora. Musanganiswa wekuita wakapisa pa40 ° C kwemaawa matanhatu uye ipapo pa120 ° C yakadururwa mumvura isina mvura kwemaawa masere. zvinhu zvakasara zvakaomeswa pa 70 ° C kwemaawa 12. Iyo yakaomeswa yakapfava yakapfava yakanyorovesa pasi muchoto uye calcined pa 550 ° C kwemaawa 12. Zvikwata 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 mu toluene yakaoma, uye 0.1 mL ye 2,2'-azoisobutyronitrile (AIBN) yakawedzerwa kune reaction flask kugadzirira phenylmaleimide-methyl vinyl isocyanate copolymer pa 3 C ° musanganiswa pa 3 C ° musanganiswa pa 60 PM ° musanganiswa. yakasvibiswa uye yakaomeswa muchoto pa40 ° C kwemaawa matatu.
Dried silica particles (2 g) yakaparadzirwa mu toluene yakaoma (100 mL), yakakonzerwa uye yakasvibiswa 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 wakadzoserwa pa 100 ° C uye wakaomeswa pa 100 ° C uye wakaomeswa sefa ye100 ° C. 60 ° C kwemaawa 3. Zvadaro, PMCP-bonded silica particles (100 g) yakanyungudutswa mu toluene (200 ml) uye 4-hydroxy-TEMPO (2 mL) yakawedzerwa pamberi pe 100 µL ye dibutyltin dilaurate sechinhu chinokonzera. Musanganiswa wakakonzerwa ne0 ° C uye wakaomeswa pa50 ° C kwemaawa 50, 3 ° C yakaomeswa pa50 ° C. maawa.
Styrene (1 mL), benzoyl peroxide BPO (0.5 mL), uye TEMPO-PMCP-yakabatanidzwa silica particles (1.5 g) yakaparadzirwa mu toluene uye yakacheneswa nenitrogen.Ipolymerization ye styrene yakaitwa pa 100 ° C kwemaawa 12. Chigadzirwa chacho chakagezwa ne methanol muusiku hwe Firiji uye Firiji yakaomeswa pahusiku. 1.
Mienzaniso yakasvibiswa pa 393 K kweawa ye1 kuti iwane kumanikidzwa kwakasara kwepasi pe10-3 Torr. Nhamba yeN2 yakashambadzirwa pane imwe dzvinyiriro yeP / P0 = 0.99 yakashandiswa kugadzirisa huwandu hwepore.Iyo morphology ye bare uye ligand-bonded silica particles yakaongororwa neTechnologies electron, Tokyo scanning ne Tokyo scanning. Japan) .Dried samples (isina silica uye ligand-bonded silica particles) yakaiswa paaluminium column uchishandisa adhesive carbon tape.Goridhe yakaputirwa pamasampuli uchishandisa Q150T sputter coater, uye 5 nm Au layer yakaiswa pane samples.Izvi zvinovandudza kushanda kwemaitiro uchishandisa low voltages uye inopa zviyo zvakanaka, MAWaspult, USA, inotonhora Eleham. Flash EA1112 elemental analyzer yakashandiswa pakuongorora kwekutanga.A Malvern (Worcestershire, UK) Mastersizer 2000 particle size analyzer yakashandiswa kuwana particle size distribution.Naked silica particles uye ligand-bonded silica particles (5 mg imwe neimwe) yakaparadzirwa mu 5 mL ye isopropanol, yakaiswa pa 5, sonicated pa min5, sonicated for the min. bhenji reMastersizer.Kuongorora kweThermogravimetric kwakaitwa 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. . yakashandiswa se slurry solvent pamwe chete nepropelling solvent.Zadza mbiru zvakatevedzana nekushandisa zvinomanikidza zve 100 MP kwemaminitsi gumi, 80 MP kwemaminitsi gumi nemashanu, uye 60 MP kwemaminitsi 30. Panguva yekurongedza, mechanical vibration yakashandiswa neGC column shakers mbiri (Alltech, Deerfield) kuti uone yunifomu yeClurry , USA packing IL, USA. uye usunungure kudzvinyirira zvishoma nezvishoma kudzivirira chero kukanganisa mukati membiru.Bvisa mbiru kubva kune slurry packing unit uye ubatanidze imwe yakakodzera kune inlet 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 mudziyo detector (UV-2075) uye girazi-lined yekubatanidza madiki tulum uye mapfupi madiki madiki madiki madiki. extra column band wideening yakaongororwa neOriginPro8 (Northampton, MA).
Albumin kubva muserum yevanhu, lyophilized powder, ≥ 96% (agarose gel electrophoresis) 3 mg yakasanganiswa 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 67, ° C mushure memaawa 67 0.1% TFA.Sefa mhinduro uye chengeta pasi 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 .SEM mifananidzo yevasina silica particles (A, B) inoratidza kuti, kusiyana nezvidzidzo zvedu zvekare, zvimedu izvi zvine spherical umo zvidimbu zvakarebeswa kana kuti zvine symmetry isina kukwana.Pamusoro peiyo ligand-bonded silica particles (C, D) inotsvedzerera kupfuura iyo ye-silica isina chinhu inogona kuva ye-poly particles pamusoro pe-coly particles. silica particles.
Kutarisa maerekitironi maikorosikopu mifananidzo yeisina silica particles (A, B) uye ligand-bonded silica particles (C, D).
Iyo particle size distributions yevare 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 chinofananidzwa (A particle-based based). d (0.5), yePMP ndeye 3.36 μm, zvichienzaniswa nekudzidza kwedu kwekare ne ad (0.5) kukosha kwe 3.05 μm (polystyrene-bound silica particles) 34.Iri batch raive nekadiki diki kugovera saizi kana ichienzaniswa nekudzidza kwedu kwekare nekuda kwekusiyana kwereshiyo yePEG, urea, TMOS muchikamu chemusanganiswa musanganiswa mukuru wechikamu cheiyo acid. iyo ye polystyrene-bound silica particle phase yatakambodzidza.Izvi zvinoreva kuti pamusoro pekushanda kwe silica particles ne styrene inongoisa polystyrene layer (0.97 µm) pamusoro pe silica surface, asi muchikamu chePMP hupamhi hwehuputi hwaive 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 yekudzidza ikozvino inopiwa muTebhu 1 (B) PSD profiles ye bare silica particles uye ligand-bonded silica particles inoratidzwa mumufananidzo 3 (B) . inoratidza kuti ukuru hwepore hunoderera ne69 mushure mekugadziriswa kwemakemikari, sezvinoratidzwa muTebhu 1 (B), uye kuchinja kwevheji kunoratidzwa muFig. 3 (B) .Saizvozvo, pore volume ye silica particles yakaderera kubva ku0.67 kusvika 0.58 cm3 / g mushure mekugadzirisa kemikari. chidzidzo chedu chekare (124 m2 / g) .Sezvinoratidzwa muTebhu 1 (B), nzvimbo yepamusoro (m2 / g) ye silica particles yakadererawo kubva ku116 m2 / g kusvika 105 m2 / g mushure mekugadzirisa kemikari.
Mhedzisiro yekuongorora kwekutanga kwechikamu chekumira inoratidzwa muTable 2. Iyo carbon loading yezvino stationary phase ndeye 6.35%, iyo yakaderera pane carbon loading yekudzidza kwedu kwekare (polystyrene bonded silica particles, 7.93% 35 uye 10.21%, zvichienderana) 42. The carbon loading yeizvino, kugadzirwa kweSP, kugadzirwa kwekabhoni kune ikozvino, kugadzirwa kweSP, muchikamu chekare, SP. mamwe mapolar ligands akadai se phenylmaleimide-methylvinylisocyanate (PCMP) uye 4-hydroxy-TEMPO akashandiswa.Iyotrojeni inorema muzana yezvino yakamira yechikamu ndeye 2.21%, ichienzaniswa ne0.1735 uye 0.85% nekurema kwe nitrogen mune zvidzidzo zvekare, maererano.Izvi zvinoreva kuti iyo nitrogen wt yakakwirira muchikamu chezvino che nitrogen phenylmaleimide.Saizvozvo, iyo carbon loadings yezvigadzirwa (4) uye (5) yaiva 2.7% uye 2.9%, maererano, nepo carbon loading yechigadzirwa chekupedzisira (6) chaiva 6.35%, sezvakaratidzwa muTebhu 2.Kurasikirwa kwehuremu kwakatariswa nePMP stationary phase, uye TGA curve inoratidzwa mu 6. iri kubvumirana zvakanaka nekabhoni kurodha (6.35%) nekuti ligand haina C chete asiwo N, O, uye H.
Iyo phenylmaleimide-methylvinylisocyanate ligand yakasarudzwa kuti igadziriswe pamusoro pekugadzirisa silica particles nokuti ine polar phenylmaleimide mapoka uye mapoka evinylisocyanate.Mapoka eVinyl isocyanate anogona kuwedzera kuita ne styrene nekurarama radical polymerization.Chikonzero chechipiri ndechokuisa boka rine kuenzanisa kwepakati nepakati neanalyte uye pasina kupindirana kwemagetsi kubva kune analyte yemagetsi uye pasina kupindirana kwakasimba kweanalyte. phenylmaleimide moiety haina mutero chaiwo pane yakajairika pH.Iyo polarity yenzvimbo yakamira inogona kudzorwa nehuwandu hwakakwana hwe styrene uye nguva yekupindura yemahara radical polymerization.Nhanho yekupedzisira yekuita (free-radical polymerization) yakakosha uye inogona kushandura polarity yenzvimbo yakamira.Ongororo yeElemental yakaitwa kuti itarise iyo stationary loading yenguva yekuwedzera kwekabhoni kukwira kweiyo nguva yekukwira kwekabhoni. iyo carbon loading yenzvimbo yakamira uye vice versa.SPs dzakagadzirirwa nemhando dzakasiyana-siyana dze styrene dzine carbon loadings dzakasiyana. Zvakare, takura zvikamu izvi zvakamira mumakoramu esimbi isina tsvina uye tarisa maitiro avo echromatographic (selectivity, resolution, N value, etc.) .Kubva pane izvi zvidzidzo, kuumbwa kwakagadziridzwa kwakasarudzwa kugadzirira PMP inotungamirirwa nekugadzirisa danho rekugadzirisa zvakanaka.
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, theoretical plate number (N) per column (100 × 1.8 mm id) i20,000 ± 100 (mapuleti matatu anokwana 3m² anopa iyo N²). PMP makoramu uye ma chromatograms anoratidzwa muMufananidzo 5A.Kukurumidza kuongororwa paPMP pachikamu chepamusoro chekuyerera (700 μL/min), mapeptide mashanu akaburitswa mukati meminiti imwe chete, N hunhu hwaive hwakanaka kwazvo, 13,500 ± 330 pakoramu (100 × 1.8 mm id), 0 inoenderana/Fire 0, 0 inopindirana/Fire 5B) Makoramu matatu akaenzana akakura (100 × 1.8 mm id) akanga akazara nemijenya mitatu yakasiyana yePMP yakamira kuti ione kuberekana.The analyte concentration yekoramu imwe neimwe yakanyorwa uchishandisa mamiriro ekugadzirisa ekugadzirisa uye nhamba yezvinyorwa zvinyorwa N uye nguva yekuchengetedza kuti iparadzanise musanganiswa wakafanana wekuedzwa mumutsara wePMP inoratidzwa pane imwe neimwe yePMP data. kuberekana kweiyo PMP column inoenderana zvakanaka neyakaderera% RSD kukosha, sezvakaratidzwa muTafura 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 kugadzirisa kwakanaka kwazvo.HSA digests yakaongororwa pachishandiswa kuyerera kwe100min/0 µ mobile phase L/7 acetonitrile/mvura uye 0.1% TFA.Sezvinoratidzwa muchromatogram (Mufananidzo 6), HSA digestion yakakamurwa kuita 17 peaks inoenderana ne17 peptides.Kugona kwekuparadzanisa kwepamusoro pepamusoro muHSA digest kwakaverengwa uye maitiro anopiwa 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 0. 1.8 mm id) yakanga yakafanana neyekudzidza kwedu kwekare Ref.34.Kupinda kwembiru yakarongedzwa nepamusoro-soro porous particles ndeiyi: 1.7 × 10-15 ye 1.3 μm particles, 3.1 × 10-15 ye 1.7 μm zvidimbu, 5.0-1 × 2 × 2 × 2 × 2 5.5 × 1. 2.6 μm zvidimbu Kune 5 μm zvikamu 43. Nokudaro, kupera kwechikamu chePMP kwakafanana ne5 μm core-shell particles.
apo 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 huremu hweSILICA PARTICLES 108 × 4 × 10 × 1.8 mm uye C18-Urea columns 31 dzatakambodzidza dzaive 0.63 uye 0.55, maererano.Izvi zvinoreva kuti kuvapo kweurea ligands kunoderedza kupera kwechikamu chakamira.Kune rumwe rutivi, iyo yose porosity yePMP id (100 × 1.8 mm id) ndeye 0.60 yakaderera P. C18-bonded silica particles nekuti muC18-type stationary phases maC18 ligands anonamirwa kune silica particles se linear cheni, uku ari mu polystyrene-type stationary phases, iyo A ikati gobvu polymer layer inoumbwa yakaitenderedza.
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 nhanho kuyerera kweiyo MP 80min inoyerera pachiyero cheHETP µ column uye Ascentis Express RP-Amide column yaive 2.6 µm uye 3.9 µm, zvichiteerana.The HETP values ​​​​inoratidza kuti kupatsanurwa kwekoramu yePMP (100 × 1.8 mm id) kuri nani pane inotengeswa Ascentis Express RP-Amide column (100 × id. kuti kuderera kweN kukosha nekuwedzera kuyerera hakusi kukosha kana kuenzaniswa nekudzidza kwedu kwekare.Kunyanya kuparadzaniswa kwepamusoro kwePMP column (100 × 1.8 mm id) kana ichienzaniswa neAscentis Express RP-Amide column inobva pakuvandudzika kwechikamu chechimiro, ukuru, uye nzira dzakaoma dzekutakura dzinoshandiswa mubasa remazuva ano34.
(A) van Deemter plot (HETP versus mobile phase linear velocity) yakawanikwa ichishandisa PMP column (100 × 1.8 mm id) mu 60/40 ACN/H2O ine 0.1% TFA.(B) van Deemter plot (HETP versus mobile phase linear velocity) inowanikwa uchishandisa Ascentis × 1. 60/40 ACN/H2O ine 0.1% TFA.
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