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Likaroloana tse nang le porous silica li ne li lokiselitsoe ke mokhoa oa sol-gel o nang le liphetoho tse ling ho fumana likaroloana tsa macroporous. litšiea tsa tšepe tse sa hloekang (id ea 100 × 1.8 mm) li ne li tletse ka ho paka ka slurry. E ile ea hlahlojoa karohano ea kholomo ea PMP ea motsoako oa peptide e nang le li-peptide tse hlano (Gly-Tyr, Gly-Leu-Tyr, Gly-Gly-Tyr-Arg, Tyr-Ile-Gly-Gly-Ser-Argmatography) ts'ebetso ea ts'ebetso ea Tyr-Ile-Gly-Gly-Ser-Arglin tšilo ea li-albumin tsa serum ea batho (HAS) . Tlas'a maemo a holimo a ho hlakisa, palo ea poleiti ea theoretical ea motsoako oa peptide e phahame ho feta 280,000 plates/m².Ho bapisa ts'ebetso ea karohano ea kholomo e ntlafalitsoeng le kholomo ea khoebo ea Ascentis Express RP-Amide, ho ile ha hlokomeloa hore ts'ebetso ea karohano ea kholomo ea khoebo ea PMP e ne e le ea boleng bo holimo le ho feta.
Lilemong tsa morao tjena, indasteri ea biopharmaceutical e fetohile 'maraka o ntseng o hola oa lefats'e ka keketseho e kholo ea karolo ea mmaraka. ho khetholla liprotheine ka marong a 'mele, lisele le lisele ke mosebetsi o boima haholo ka lebaka la palo e kholo ea mefuta e ka bonoang ka sampuli e le 'ngoe.Le hoja boima ba spectrometry ke sesebelisoa se sebetsang sa peptide le protheine sequencing, haeba disampole tse joalo li kenngoa ka har'a spectrometer ea boima ka nako e le' ngoe, karohano e ke ke ea e-ba molemo. fokotsa palo ea bahlahlobisisi ba kenang spectrometer ea boima ka nako e fanoeng4,5,6. Ho phaella moo, nakong ea karohano ea mohato oa mokelikeli, li-analyte li ka tsepamisoa libakeng tse moqotetsane, ka tsela eo li tsepamisa maikutlo litlhahlobo tsena le ho ntlafatsa kutloisiso ea ho lemoha ha MS.Liquid chromatography (LC) e tsoetse pele haholo lilemong tse leshome tse fetileng 'me e fetohile mokhoa o tummeng oa ho hlahloba proteomic,9,8,17.
Reversed-phase liquid chromatography (RP-LC) e sebelisoa haholo bakeng sa tlhoekiso le karohano ea metsoako ea peptide e sebelisang octadecyl-modified silica (ODS) e le phase11,12,13. mekhahlelo e emeng e hlokeha ho hlahloba li-peptide le liprotheine tse nang le likarolo tsa polar le tse sa polar ho sebelisana le ho boloka li-analyte tsena16.Mixed-mode chromatography, e fanang ka litšebelisano tse ngata, e ka ba mokhoa o mong oa RP-LC bakeng sa karohano ea li-peptide, liprotheine le metsoako e meng e rarahaneng. karohano ea peptide le liprotheine17,18,19,20,21.Mekhahlelo e emeng e kopaneng (WAX/RPLC, HILIC/RPLC, polar intercalation/RPLC) e loketse karohano ea peptide le protheine ka lebaka la boteng ba lihlopha tsa polar le tse seng tsa polar22,23,24,25,26,26,26,26. lihlopha tsa polar tse kopanetsoeng li bonts'a matla a karohano e ntle le khetho e ikhethang bakeng sa li-analyte tsa polar le tse seng tsa polar, kaha karohano e itšetlehile ka tšebelisano pakeng tsa mohlahlobi le mohato o emeng. Litšebelisano tse ngata 29, 30, 31, 32. Haufinyane tjena, Zhang et al. 30 e lokiselitse karolo ea dodecyl e felisitsoeng ea polyamine 'me e atlehile ho arola li-hydrocarbon, li-antidepressants, flavonoids, nucleosides, estrogens, le li-analyte tse ling tse' maloa. The polar intercalator e na le lihlopha tsa polar le tse seng tsa polar, kahoo e ka sebelisoa ho arola li-peptide le liprotheine tse nang le hydrophobic le hydrophilic hydrophilic. Likholomo tsa C18 tse kentsoeng ka amide) li fumaneha khoebong tlas'a litšiea tsa Ascentis Express RP-Amide, empa litšiea tsena li sebelisetsoa tlhahlobo ea amine 33 feela.
Phuputsong ea morao-rao, karolo ea polar-embedded stationary phase (N-phenylmaleimide-embedded polystyrene) e ile ea lokisoa le ho hlahlojoa bakeng sa ho arohana ha li-peptide le li-trypsin digests tsa HSA.Mokhahlelo oa ho emisa o ne o lokiselitsoe ho sebelisa leano le latelang.Likaroloana tsa silika tsa porous li ne li lokiselitsoe ho latela mokhoa o fanoeng khatisong ea rona e fetileng le tse ling tsa liphetoho, polyolthy ratio ea protocol ea u. TMOS, metsi a acetic acid e ile ea fetoloa ho lokisa likaroloana tsa silica tse nang le pore e kholo ea boholo. Ea bobeli, ligand e ncha, phenylmaleimide-methyl vinyl isocyanate, e ile ea etsoa 'me ea sebelisoa ho tlosa likaroloana tsa silica ho lokisetsa polar embedded stationary phase.The sephetho sa stationary phase was packed into a optimized the column 100 mm × . sekeme.The column packing is help with mechanical vibrations ho etsa bonnete ba hore bethe e tšoanang e thehoa ka har'a kholomo.Hlahloba karohano ea kholomo e pakiloeng ea metsoako ea peptide e nang le li-peptide tse hlano; (Gly-Tyr, Gly-Leu-Tyr, Gly-Gly-Tyr-Arg, Tyr-Ile-Gly-Ser-Arg, Leucine Enkephalin) and Trypsin digest of human serum albumin (HAS) .Motsoako oa peptide le trypsin digest tsa HSA li ile tsa hlokomeloa hore li arohane ka qeto e ntle le katleho ea ts'ebetso ea AMP ea karohano ea AMP e ne e bapisoa. Kholomo ea RP-Amide. Ka bobeli li-peptide le liprotheine li ile tsa hlokomeloa hore li rarollotsoe hantle ebile li sebetsa hantle holim'a kholomo ea PMP, e neng e sebetsa hantle ho feta kholomo ea Ascentis Express RP-Amide.
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, xyl-Chloride, Benzoyldroksidi, MC-TEMH (BPO), HPLC Grade Acetonitrile (ACN), Methanol, 2-Propanol, le Acetone Reka ho tloha Sigma-Aldrich (St. Louis, MO, USA).
Motsoako oa urea (8 g), polyethylene glycol (8 g), le 8 ml ea 0.01 N acetic acid e ile ea tsosoa ka metsotso e 10, 'me 24 mL ea TMOS e ile ea eketsoa moo tlas'a maemo a batang a leqhoa. lisebelisoa tse setseng li ne li omisitsoe ka 70 ° C bakeng sa lihora tsa 12. Boima bo bonolo bo omisitsoeng bo ne bo le fatše bo boreleli ka ontong 'me bo calcined ka 550 ° C bakeng sa lihora tse 12. Lihlopha tse tharo li ne li lokiselitsoe 'me li khetholloa ho hlahloba ho ikatisa ka boholo ba likaroloana, boholo ba pore le sebaka se ka holim'a metsi.
Ka ho fetola holim'a likaroloana tsa silica tse nang le ligand tse entsoeng pele ho li-phenylmaleimide-methylvinylisocyanate (PCMP) tse lateloang ke radial polymerization le styrene, motsoako o nang le sehlopha sa polar o ile oa lokisoa. Mokhahlelo oa ho emisa bakeng sa li-aggregates le liketane tsa polystyrene.Ts'ebetso ea ho lokisetsa e hlalositsoe ka tlase.
N-phenylmaleimide (200 mg) le methyl vinyl isocyanate (100 mg) li ile tsa qhibiliha ka toluene e omileng, 'me 0.1 mL ea 2,2'-azoisobutyronitrile (AIBN) e ile ea kenngoa ka har'a botlolo ea ho arabela ho lokisetsa phenylmaleimide-methyl vinyl isocyanate copolymer ka lihora tsa 3, C ° 6. e tlhotliloeng le ho omisoa ka ontong ea 40 ° C bakeng sa lihora tse 3.
Likaroloana tse omisitsoeng tsa silica (2 g) li ne li qhalakantsoeng ka toluene e omileng (100 mL), e tsositsoe mme e sonicated ka 500 mL e pota-potileng ka tlas'a botlolo bakeng sa 10 min.PMCP (10 mg) e ile ea qhibiliha ka toluene 'me ea eketsoa ka dropwise ho botlolo ea ho arabela ka funnel e theoha. 60 ° C bakeng sa lihora tsa 3. Joale, likaroloana tsa silica tse kopantsoeng le PMCP (100 g) li ne li qhibiliha ka toluene (200 ml) le 4-hydroxy-TEMPO (2 mL) e ile ea eketsoa ka pel'a 100 µL ea dibutyltin dilaurate e le catalyst.Motsoako o ile oa hlohlelletsoa bakeng sa 5 ° C le ho omisoa ka lihora tsa 50 ° C ka 50 ° C ka lihora tse 50. lihora.
Styrene (1 mL), benzoyl peroxide BPO (0.5 mL), le TEMPO-PMCP-attached silica particles (1.5 g) li ile tsa qhalakanngoa ka toluene le ho hloekisoa ka naetrojene.Polymerization ea styrene e ile ea etsoa ka 100 ° C bakeng sa lihora tse 12. Sehlahisoa se hlahisoang se ile sa hlatsuoa ka methanol ka kakaretso ea 60 ° C ka bosiu le karabelo e omisitsoeng. 1.
Li-samples li ile tsa tlosoa ka 393 K bakeng sa hora ea 1 ho fumana khatello e setseng e ka tlaase ho 10-3 Torr. Palo ea N2 e adsorbed ka khatello e lekanyelitsoeng ea P / P0 = 0.99 e ne e sebelisetsoa ho fumana palo eohle ea pore. The morphology of bare and ligand-bonded silica particles, e ile ea hlahlojoa ka microscopyelectron ea Tokyo High Technologies. Japane) . Li-sampuli tse omisitsoeng (li-silica tse se nang letho le likaroloana tsa silika tse kopantsoeng le ligand) li ne li behoa holim'a mohala oa aluminium ho sebelisa tepi ea carbon e khomarelang. Khauta e ne e pentiloe holim'a mehlala e sebelisang Q150T sputter coater, 'me 5 nm Au layer e ne e behiloe holim'a mehlala.Sena se ntlafatsa ts'ebetso ea ts'ebetso ka ho sebelisa li-voltages tse tlaase le ho fana ka lijo-thollo tse ntle tsa Amerika, USA. Flash EA1112 elemental analyzer was used for elemental analysis.A Malvern (Worcestershire, UK) Mastersizer 2000 particle size analyzer was used to find the particle size distribution.Naked silica particles and ligand-bonded silica particles (5 mg mong le e mong) ba ile ba qhalakantsoeng ka 5 mL ea isopropanol, beha ka 5 mins, sonicated bakeng sa op, sonicated, sonicated, 5 milligrams le silika. benche ea Mastersizer.Tlhahlobo ea Thermogravimetric e entsoe ka sekhahla sa 5 °C ka motsotso ho feta mocheso oa 30 ho isa ho 800 °C.
Litšepe tse sa hloekang tse entsoeng ka khalase tse nang le litšepe tse tšesaane tsa litekanyo (id ea 100 × 1.8 mm) li ne li pakiloe ho sebelisoa mokhoa oa ho paka seretse, ho sebelisoa mokhoa o tšoanang o sebelisitsoeng ho Ref. 31.Kolumo ea tšepe e sa hloekang (e kentsoeng ka khalase, 100 × 1.8 mm id) e nang le mochini o kenang o nang le 1 µm frit e ne e hokahantsoe le sephutheloana sa slurry packer (Alltech Deerfield, IL, USA). Lokisetsa slurry ea mokhahlelo o emeng ka ho emisa 150 mg ea mokhahlelo oa stationary mL le ho romela kholomo ea menotha ho menotha ho 1. e ne e sebelisoa e le solvent e slurry hammoho le propelling solvent.Tlatsa kholomo ka tatellano ka ho sebelisa likhatello tsa 100 MP bakeng sa metsotso e 10, 80 MP bakeng sa metsotso e 15, le 60 MP bakeng sa metsotso e 30. Nakong ea ho paka, ho thothomela ha mechine ho ne ho sebelisoa ka li-shaker tse peli tsa GC (Alltech, Deerfield) ho netefatsa hore ho na le li-columns IL, Deerfield, USA. 'me u lokolle khatello butle ho thibela tšenyo leha e le efe ka har'a kholomo.Hakolla kholomo ho tloha slurry packing unit' me u kopanye e 'ngoe e loketseng ho kena le ho LC system ho hlahloba tshebetso ea eona.
LC pump (10AD Shimadzu, Japan), injector (Valco (USA) C14 W.05) e nang le 50nL injection loop, membrane degasser (Shimadzu DGU-14A), UV-VIS capillary window e ile ea hahoa Special µLC device detector (UV-2075) le glass-lined micolum e nyenyane haholo e kopanyang likhalase tse nyenyane haholo. Extra column band broadening e hlahlojoa ke OriginPro8 (Northampton, MA).
Albumin e tsoang serum ea batho, phofo ea lyophilized, ≥ 96% (agarose gel electrophoresis) 3 mg e kopantsoeng le trypsin (1.5 mg), 4.0 M urea (1 mL), le 0.2 M ammonium bicarbonate (1 mL). Tharollo e ile ea tsosoa ka metsotso e 10 'me e bolokoa ka bateng ea metsi ka lihora tse 67 ka inched 1 mL, ° C e ntan'o ba 67. 0.1% TFA.Sefa tharollo 'me u boloke ka tlase ho 4 °C.
Karohano ea metsoako ea peptide le tšilo ea li-trypsin ea HSA e ile ea hlahlojoa ka thoko ho likholomo tsa PMP. Hlahloba karohano ea motsoako oa peptide le trypsin digest ea HSA ka kholomo ea PMP 'me u bapise liphetho ho kholomo ea Ascentis Express RP-Amide.Nomoro ea poleiti ea theory e baloa ka tsela e latelang:
Litšoantšo tsa SEM tsa likaroloana tsa silika tse se nang letho le likaroloana tsa silica tse kopantsoeng le ligand li bontšoa ho FIG. 2 .SEM litšoantšo tsa likaroloana tsa silika tse se nang letho (A, B) li bontša hore, ho fapana le lithuto tsa rona tse fetileng, likaroloana tsena ke spherical moo likaroloana li lelefalitsoeng kapa li nang le li-symmetry tse sa tloaelehang. likaroloana tsa silika.
Ho hlahloba litšoantšo tsa electron microscope ea likaroloana tse se nang letho tsa silica (A, B) le likaroloana tsa silica tse kopantsoeng le ligand (C, D).
Likarolo tsa boholo ba likaroloana tsa silika tse se nang letho le likaroloana tsa silika tsa ligand-bonded li bontšoa ho Setšoantšo sa 3 (A) . Li-curve tsa boholo ba likaroloana tsa boholo-holo li bontšitse hore boholo ba likaroloana tsa silika bo ile ba eketseha ka mor'a phetoho ea lik'hemik'hale (setšoantšo sa 3A) . d (0.5), ea PMP ke 3.36 μm, ha e bapisoa le thuto ea rona e fetileng le boleng ba papatso (0.5) ea 3.05 μm (likotoana tsa silika tse entsoeng ka polystyrene) 34. Sehlopha sena se ne se e-na le kabo ea boholo bo fokolang ha bo bapisoa le thuto ea rona e fetileng ka lebaka la likarolo tse fapaneng tsa PEG, urea, TMOS ka karoloana e kholo ea motsoako oa acid e nyenyane ea PMP. ea karolo ea polystyrene-bound silica particle eo re ithutileng eona pele.Sena se bolela hore tšebetso ea holim'a silika e nang le styrene e ne e kenya feela polystyrene layer (0.97 µm) holim'a silika, athe karolong ea PMP botenya ba lera e ne e le 1.38 µm.
Kabo ea boholo ba likaroloana (A) le kabo ea boholo ba pore (B) ea likaroloana tsa silika tse se nang letho le likaroloana tsa silika tse tlamang ligand.
Boholo ba pore, bophahamo ba pore le sebaka sa holim'a likaroloana tsa silika tsa thuto ea morao-rao li fanoe ho Lethathamo la 1 (B) . Litlhaloso tsa PSD tsa likaroloana tsa silika tse se nang letho le likaroloana tsa silika tsa ligand-bonded li bontšoa ho Setšoantšo sa 3 (B) . Liphetho li bapisoa le thuto ea rona e fetileng. e bontša hore boholo ba pore bo fokotseha ka 69 ka mor'a phetoho ea lik'hemik'hale, joalokaha ho bontšitsoe Lethathamong la 1 (B), 'me phetoho ea sekhahla e bontšoa ho Setšoantšo sa 3 (B) .Ka mokhoa o ts'oanang, palo ea pore ea likaroloana tsa silika e fokotsehile ho tloha 0.67 ho 0.58 cm3 / g ka mor'a phetoho ea lik'hemik'hale. thuto ea rona e fetileng (124 m2 / g) . Joalokaha ho bontšitsoe Lethathamong la 1 (B), sebaka se ka holimo (m2 / g) sa likaroloana tsa silika le sona se fokotsehile ho tloha 116 m2 / g ho ea ho 105 m2 / g ka mor'a phetoho ea lik'hemik'hale.
Liphetho tsa tlhahlobo ea mantlha ea mohato o emeng li bonts'itsoe ho Lethathamo la 2. The carbon loading ea sebaka sa hona joale sa stationary ke 6.35%, e leng tlase ho feta carbon loading ea thuto ea rona e fetileng (polystyrene bonded silica particles, 7.93% 35 le 10.21%, ka ho latellana) 42. The carbon loading of the current phase, SP, the current phase, SP. li-ligands tse ling tsa polar tse kang phenylmaleimide-methylvinylisocyanate (PCMP) le 4-hydroxy-TEMPO li ne li sebelisoa. Boima ba naetrojene lekholong ea karolo ea hona joale ea boemo bo emeng ke 2.21%, ha e bapisoa le 0.1735 le 0.85% ka boima ba nitrogen lithutong tse fetileng, ka ho latellana.Sena se bolela hore karolo ea naetrojene e phahame ka lebaka la boemo ba hona joale ba nitrogen wt% phenylmaleimide.Ka mokhoa o ts'oanang, likhase tsa carbon loadings (4) le (5) e ne e le 2.7% le 2.9%, ka ho latellana, ha carbon loading ea sehlahisoa sa ho qetela (6) e ne e le 6.35%, joalokaha ho bontšitsoe Lethathamong la 2. Ho lahleheloa ke boima ba 'mele ho ile ha hlahlojoa le PMP mohato o emisang,' me lekhalo la TGA le bontšoa ho 4 ea TGA curve. e lumellana hantle le khabone (6.35%) hobane li-ligand ha li na C feela empa hape le N, O, le H.
Phenylmaleimide-methylvinylisocyanate ligand e khethiloe bakeng sa phetoho ea holim'a likaroloana tsa silika hobane e na le lihlopha tsa polar phenylmaleimide le lihlopha tsa vinylisocyanate.Lihlopha tsa vinylisocyanate li ka tsoela pele ho itšoara ka styrene ka ho phela radical polymerization. phenylmaleimide moiety ha e na tefiso ea sebele ho pH e tloaelehileng. Polarity ea mohato o tsitsitseng o ka laoloa ke tekanyo e nepahetseng ea styrene le nako ea ho itšoara ka polymerization ea mahala. the carbon loading of the stationary phase and vice versa.SPs e lokiselitsoeng ka mekhahlelo e fapaneng ea styrene e na le mefuta e sa tšoaneng ea carbon loadings. Hape, kenya mekhahlelo ena e emeng ka har'a litšiea tsa tšepe e sa hloekang 'me u hlahlobe ts'ebetso ea tsona ea chromatographic (selectivity, resolution, N value, joalo-joalo) .Ho itšetlehile ka liteko tsena, ho khethiloe ho hlophisoa ho optimized ho lokisetsa PMP e hlophisitsoeng hantle ea polate le ho etsa bonnete ba hore ho na le ts'ebetso e ntle ea "PMP".
Metsoako e mehlano ea peptide (Gly-Tyr, Gly-Leu-Tyr, Gly-Gly-Tyr-Arg, Tyr-Ile-Gly-Ser-Arg, leucine enkephalin) le eona e ile ea hlahlojoa ho sebelisoa mohala oa PMP o sebelisang mohato oa selefouno; 60/40 (v/v) acetonitrile/metsi (0.1% TFA) ka sekhahla sa phallo ea 80 μL/min. Tlas'a maemo a ntlafetseng a hlabollo, nomoro ea poleiti ea theoretical (N) ka kholomo (100 × 1.8 mm id) ke 20,000 ± 100 (200,000 plates for N² e fana ka 0). Litšiea tsa PMP le li-chromatogram li bonts'itsoe ho Setšoantšo sa 5A. Tlhahlobo e potlakileng ho kholumo ea PMP ka sekhahla se phallo se phahameng (700 μL/min), li-peptide tse hlano li ile tsa hlakoloa ka nako ea motsotso o le mong, boleng ba N bo ne bo le ntle haholo, 13,500 ± 330 kholomong (100 × 1.8 mm id), 0 Corresponds/00m (0 Corresponds/0) 5B) Litšiea tse tharo tse lekanang ka boholo (ID ea 100 × 1.8 mm) li ne li tletse lotho tse tharo tse fapaneng tsa PMP e emeng ho hlahloba ho ikatisa.The analyte concentration bakeng sa kholomo e 'ngoe le e 'ngoe e tlalehiloe ho sebelisoa maemo a holimo a elution le palo ea lipoleiti tsa khopolo-taba N le nako ea ho boloka ho arola motsoako o tšoanang oa teko ka har'a kholomo e 'ngoe le e 'ngoe ea PMP. Ho ikatisa ha kholomo ea PMP ho tsamaisana hantle le boleng bo tlase ba %RSD, joalo ka ha ho bonts'itsoe ho Lethathamo la 3.
Karohano ea motsoako oa peptide holim'a kholomo ea PMP (B) le kholomo ea Ascentis Express RP-Amide (A); mobile phase 60/40 ACN/H2O (TFA 0.1%), PMP columna litekanyo (100 × 1.8 mm id); analytical Taelo ea elution ea metsoako: 1 (Gly-Tyr), 2 (Gly-Leu-Tyr), 3 (Gly-Gly-Tyr-Arg), 4 (Tyr-Ile-Gly-Ser-Arg) le 5 (leucine) acid enkephalin)).
Kholomo ea PMP (ID ea 100 × 1.8 mm) e ile ea hlahlojoa bakeng sa karohano ea tšilo ea tryptic ea albumin ea serum ea motho ka chromatography ea mokelikeli e phahameng. Chromatogram e setšoantšong sa 6 e bontša hore sampole e arohane hantle 'me qeto e ntle haholo.Litlhaloso tsa HSA li ile tsa hlahlojoa ho sebelisoa sekhahla sa phallo ea 100min/3 µ Acetonitrile/metsi le 0.1% TFA. Joalo ka ha ho bonts'itsoe ho chromatogram (setšoantšo sa 6), tšilo ea HSA e arotsoe ka litlhōrō tse 17 tse tsamaellanang le li-peptide tse 17. Karohano ea tlhoro e 'ngoe le e' ngoe ho HSA digest e baloa mme litekanyetso li fanoe ho Lethathamo la 5.
Tryptic digest ea HSA (100 × 1.8 mm id) e ile ea aroloa holim'a kholomo ea PMP; sekhahla sa phallo (100 µL/min), karolo ea selefouno 60/40 acetonitrile/metsi ka 0.1% TFA.
moo L e leng bolelele ba litšiea, η ke viscosity ea mohato oa selefouno, ΔP ke khatello ea mokokotlo oa mokokotlo, 'me u ke lebelo la mola oa mohato oa selefouno. The permeability ea column ea PMP e ne e le 2.5 × 10-14 m2, tekanyo ea phallo e ne e le 25 μL / min, le 60/40 v / v / v. 1.8 mm id) e ne e tšoana le ea thuto ea rona e fetileng Ref.34. The permeability ea kholomo e tletseng likaroloana tse nang le li-porous tse ka holimo ke: 1.7 × 10-15 bakeng sa likaroloana tsa 1.3 μm, 3.1 × 10-15 bakeng sa likaroloana tsa 1.7 μm, 5.0-1 × 2 × 2 × 2 × 2 5.2 × 1 × 2 5. 2.6 μm likaroloana Bakeng sa likaroloana tsa 5 μm 43. Ka hona, ho khoneha ha karolo ea PMP ho tšoana le ea 5 μm core-shell particles.
moo Wx e leng boima ba kholomo e nang le chloroform, Wy ke boima ba kholomo e nang le methanol, le ρ ke boima ba mokelikeli.Densities of methanol (ρ = 0.7866) le chloroform (ρ = 1.484).Kakaretso ea porosity ea SILICA LIKAROLO 4 × 10 8 limilimithara (1.8 mm) le C18-Urea litšiea 31 tseo re ithutileng tsona pele e ne e le 0.63 le 0.55, ka ho latellana.Sena se bolela hore ho ba teng ha urea ligands ho fokotsa ho phunyeletsa ha karolo e emeng.Ka lehlakoreng le leng, kakaretso ea porosity ea kholomo ea PMP (100 × 1.8 mm id) ke 0.60 ea li-columns tse tlaase ho feta li-columns tse ka tlaase. Likaroloana tsa silika tsa C18 hobane ka mekhahlelo ea C18-mofuta oa li-stationary li-ligand tsa C18 li khomaretsoe ho likaroloana tsa silika e le liketane tse nang le mela, ha ka mekhahlelo ea mofuta oa polystyrene, ho thehoa A hoo e ka bang e teteaneng ea polymer layer e entsoeng ho e potoloha.
Setšoantšo sa 7A, B se bontša kholomo ea PMP (100 × 1.8 mm id) le kholomo ea Ascentis Express RP-Amide (100 × 1.8 mm id) ho sebelisoa maemo a tšoanang a elution (ke hore, 60/40 ACN/H2O le 0.1% TFA). ) ea morero oa van Deemter. Metsoako e khethiloeng ea peptide (Gly-Tyr, Gly-Leu-Tyr, Gly-Gly-Tyr-Arg, Tyr-Ile-Gly-Ser-Arg, Leucine Enkephalin) e lokisitsoe ka 20 µL/ Sekhahla sa phallo e tlase bakeng sa likholomo ka bobeli ke 800 µL/min. kholomo le kholomo ea Ascentis Express RP-Amide e ne e le 2.6 µm le 3.9 µm, ka ho latellana. Litekanyetso tsa HETP li bontša hore katleho ea karohano ea kholomo ea PMP (100 × 1.8 mm id) e molemo haholo ho feta kholomo ea Ascentis Express RP-Amide e fumanehang khoebong (100 × 8 mm Deem) 7. hore ho fokotseha ha boleng ba N ka phallo e ntseng e eketseha ha ho bohlokoa ha ho bapisoa le thuto ea rona e fetileng.Katleho e phahameng ea karohano ea kholomo ea PMP (100 × 1.8 mm id) ha e bapisoa le kholomo ea Ascentis Express RP-Amide e itšetlehile ka ntlafatso ea sebopeho sa likaroloana, boholo, le mekhoa e rarahaneng ea ho paka mela e sebelisoang mosebetsing oa hona joale34.
(A) van Deemter plot (HETP khahlano le mobile phase linear velocity) e fumanoe ka ho sebelisa mohala oa PMP (100 × 1.8 mm id) ka 60/40 ACN/H2O le 0.1% TFA. (B) van Deemter plot (HETP khahlanong le mobile phase linear velocity) e fumanoeng ka Ascentis × 1 kholomo ea Ascentis × 1 60/40 ACN/H2O e nang le 0.1% TFA.
Polar-embedded polystyrene stationary phase e ile ea lokisoa le ho hlahlojoa bakeng sa karohano ea metsoako ea maiketsetso ea peptide le trypsin digests ea human serum albumin (HAS) in high performance liquid chromatography.The chromatographic performance of PMP columns for peptide mixes is excellent in karohano katleho le qeto.The joalo ka karohano ea mefuta e fapaneng ea mabaka a karohano ke ka lebaka la ho ntlafala ha kholomo ea karohano ka lebaka la ts'ebetso ea karoloana ea PMP e joalo ka lebaka la ts'ebetso e ntlafetseng ea likaroloana tsa PMP, boholo ba li-silica particles, controlled synthesis of the stationary phase, and complex column packing.Ho phaella ho katleho e phahameng ea karohano, khatello e tlaase ea mokokotlo oa mokokotlo ka lebelo le phahameng la phallo ke molemo o mong oa karolo ena e tsitsitseng.PMP columns display good reproducibility and can be used for the analysis of peptide mixs and trypsin digestion of various proteins. lihlahisoa tsa fungal ho chromatography ea metsi. Nakong e tlang, litšiea tsa PMP li tla boela li hlahlojoe bakeng sa ho arohana ha liprotheine le li-antibodies tsa monoclonal.
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