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Ua hoʻomākaukau ʻia nā ʻāpana silica porous e kahi ʻano sol-gel me kekahi mau hoʻololi e loaʻa ai nā ʻāpana macroporous. .8 mm id) ua paʻi ʻia e ka slurry packing.Evaluated PMP kolamu hoʻokaʻawale ʻana o kahi hui peptide i loaʻa i ʻelima peptides (Gly-Tyr, Gly-Leu-Tyr, Gly-Gly-Tyr-Arg, Tyr-Ile-Gly-Ser-Arg, leucine enkephalin) a me ka hoʻokō ʻana o ka chromatorum graphic o ke kanaka. theoretical plate count of the peptide mixture is as high as 280,000 plates/m². Comparing the separation performance of the developed column with the commercial Ascentis Express RP-Amide column, it is observed that separation performance of the PMP column is superior to the commercial column in terms of separation efficiency and resolution.
I nā makahiki i hala iho nei, ua lilo ka ʻoihana biopharmaceutical i mea hoʻonui i ka mākeke honua me ka piʻi nui ʻana o ka māhele mākeke. ʻO ka wai, nā ʻiʻo a me nā cell he hana paʻakikī loa ma muli o ka nui o nā ʻano mea hiki ke ʻike ʻia i loko o kahi laʻana. mika i ka manawa i hāʻawi ʻia 4,5,6. Eia kekahi, i ka wā o ka hoʻokaʻawale ʻana o ka wai, hiki ke hoʻopaʻa ʻia nā analytes i nā ʻāpana liʻiliʻi, a laila e noʻonoʻo ana i kēia mau analytes a hoʻomaikaʻi i ka sensitivity ʻike MS.
Hoʻohana nui ʻia ka reversed-phase liquid chromatography (RP-LC) no ka hoʻomaʻemaʻe ʻana a me ka hoʻokaʻawale ʻana o nā hui peptide me ka hoʻohana ʻana i ka silica octadecyl-modified (ODS) ma ke ʻano he stationary phase11,12,13. Akā naʻe, ʻaʻole hāʻawi nā RP stationary phases i ka hoʻokaʻawale maikaʻi ʻana o nā peptides a me nā protein ma muli o ko lākou ʻano paʻakikī 14,12,13. e kālailai i nā peptides a me nā proteins me ka polar a me ka non-polar moieties e launa pū me a mālama i kēia mau analytes16.Mixed-mode chromatography, e hāʻawi ana i nā pilina multimodal, hiki ke lilo i mea ʻokoʻa i ka RP-LC no ka hoʻokaʻawale ʻana i nā peptides, proteins, a me nā hui paʻakikī ʻē aʻe. 20,21. He kūpono no ka peptide a me ka protein i hoʻokaʻawale ʻia ma muli o ka hiki ʻana mai o nā pūʻulu polar a me ka polar ʻole22,23,24,25,26,27,28 a me nā analytes non-polar, no ka mea, pili ka hoʻokaʻawale ʻana i ka pilina ma waena o ka analyte a me ka pae paʻa.ʻO nā pilina multimodal 29, 30, 31, 32. ʻO Zhang et al.30 hoʻomākaukau i ka dodecyl-terminated polyamine stationary phase a me ka hoʻokaʻawale maikaʻi hydrocarbons, antidepressants, flavonoids, nucleosides, estrogens, a me kekahi mau analytes. ʻO ka polar intercalator he polar a me ka ole polar pūʻulu, no laila, hiki ke hoʻohana 'ia e hoʻokaʻawale peptides a me nā proteins i loaʻa nā hydrophobic a me ka hydrophilic moieties. ma lalo o ka inoa kālepa ʻo Ascentis Express RP-Amide kolamu, akā hoʻohana ʻia kēia mau kolamu no ka nānā ʻana i ka amine 33 wale nō.
Ma ka haʻawina o kēia manawa, ua hoʻomākaukauʻia a loiloiʻia kahi papa hana polar-embedded (N-phenylmaleimide-embedded polystyrene) no ka hoʻokaʻawaleʻana i nā peptides a me nā trypsin digests o HSA. Ua hoʻomākaukauʻia ka māhele paʻa me ka hoʻohanaʻana i ka papahana. e hoʻomākaukau i nā ʻāpana silica me ka nui pore nui. ʻO ka lua, kahi ligand hou, phenylmaleimide-methyl vinyl isocyanate, ua synthesized a hoʻohana ʻia e derivatize silica particles e hoʻomākaukau ai i ka polar embedded stationary phase. ʻO ke kolamu. E loiloi i ka hoʻokaʻawale ʻana o ke kolamu i hoʻopaʻa ʻia o nā hui peptide me ʻelima peptides;(Gly-Tyr, Gly-Leu-Tyr, Gly-Gly-Tyr-Arg, Tyr-Ile-Gly-Ser-Arg, Leucine Enkephalin) a me Trypsin digest o ke kanaka serum albumin (HAS). .Uaʻikeʻia nā peptides a me nā protein e hoʻoholo maikaʻi a maikaʻi ma ka kolamu PMP, iʻoi aku ka maikaʻi ma mua o ke kolamu 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 (MCTEMP), Stydroxyl Chloride (MCTEMP), Stydroxyl Oxide (MCTEMP), Stydroxyl Oxide (MCTEMP), Benedroxyl Oxide (MCTEMP), Benedroxyl Oxide (MCTEMP) trile (ACN), Methanol, 2-Propanol, a me Acetone i kūʻai ʻia mai Sigma-Aldrich (St. Louis, MO, USA).
Ua hoʻoulu ʻia kahi hui ʻana o ka urea (8 g), polyethylene glycol (8 g), a me 8 mL o 0.01 N acetic acid no 10 mau minuke, a laila ua hoʻohui ʻia he 24 mL o TMOS ma lalo o nā kūlana hau. 70 ° C no nā hola 12. ʻO ka'ōpala maloʻo maloʻo he lepo maʻemaʻe i loko o ka umu a calcined ma 550 ° C no nā hola 12. Ua hoʻomākaukauʻiaʻekolu mau pahu a hoʻohālikelikeʻia no ka nānāʻana i ka hana houʻana i ka nui o ka pore, ka nui o ka pore a me ka ili.
Ma ka hoʻololi ʻana i ka ʻili o nā ʻāpana silica me ka ligand pre-synthesized phenylmaleimide-methylvinylisocyanate (PCMP) i ukali ʻia e ka polymerization radial me ka styrene, ua hoʻomākaukau ʻia kahi hui pūʻulu polar.ʻO ka pae paʻa no nā aggregates a me nā kaulahao polystyrene. Ua wehewehe ʻia ke kaʻina hana hoʻomākaukau ma lalo nei.
N-phenylmaleimide (200 mg) a me ka methyl vinyl isocyanate (100 mg) ua hoʻoheheʻeʻia i loko o ka toluene maloʻo, a me 0.1 mL o 2,2′-azoisobutyronitrile (AIBN) i hoʻohuiʻia i ka pahu hopena e hoʻomākaukau i ka phenylmaleimide-methyl isocyanate copolymer (PMC) i hoʻoheheʻeʻia i ka 3. 40 ° C no 3 mau hola.
Ua hoʻopuehu ʻia nā ʻāpana silica maloʻo (2 g) i loko o ka toluene maloʻo (100 mL), hoʻoulu ʻia a sonicated i loko o kahi pahu 500 mL puni lalo no 10 min. Ua hoʻoheheʻe ʻia ka PMCP (10 mg) i toluene a hoʻohui ʻia i ka dropwise i ka pahu hopena ma o ka hāʻule ʻana o ka funnel. hola. A laila, hoʻoheheʻe ʻia nā ʻāpana silica paʻa PMCP (100 g) i toluene (200 ml) a ua hoʻohui ʻia ka 4-hydroxy-TEMPO (2 mL) i mua o 100 µL o dibutyltin dilaurate ma ke ʻano he catalyst.
Ua hoʻopuehu ʻia ʻo Styrene (1 mL), benzoyl peroxide BPO (0.5 mL), a me TEMPO-PMCP-attached silica particles (1.5 g) i toluene a hoʻomaʻemaʻe ʻia me ka nitrogen. Ua lawe ʻia ka polymerization o styrene ma 100 ° C no nā hola 12. Ua holoi ʻia ka huahana me ka methanol a maloʻo i ka pō.
Ua hoʻokuʻuʻia nā mea hoʻohālike ma 393 K no ka hola 1 e loaʻa ai ke koena o ke kaomi ma lalo o 10-3 Torr.ʻO ka nui o N2 adsorbed ma kahi pili pili o P / P0 = 0.99 i hoʻohanaʻia no ka hoʻoholoʻana i ka nui o ka pore volume.The morphology of bare and ligand-bonded silica particles i nānāʻia me ka scanning electron micros, Japan-ligad samples (Hōʻikeʻikeʻikeʻoniʻoni kiʻekiʻe). ded silica particles) ua kau 'ia ma luna o ke kolamu alumini me ka ho'ohana 'ana i ke kalapona lipine adhesive. Ua uhi 'ia ke gula ma luna o nā la'ana me ka Q150T sputter coater, a he 5 nm Au i waiho 'ia ma na samples. This improves process efficiency using low voltages and provides fine grain, cold sputtering. Ua hoʻohana 'ia ka Mastersizer 2000 particle size analyzer no ka loaʻaʻana o ka mahele nui. Naked silica particles a me nā ligand-bonded silica particles (5 mg kēlā me kēia) ua hoʻopuehuʻia i loko o 5 mL o isopropanol, sonicated no 10 min, vortexed no 5 min, a kauʻia ma luna o kaʻeleʻele o ka Mastersizer.
Ua hoʻopili ʻia nā kolamu ololi o nā ana (100 × 1.8 mm id) me ka hoʻohana ʻana i ke ʻano slurry packing, e pili ana i ke kaʻina hana like i hoʻohana ʻia ma Ref.31. Ua hoʻohui ʻia kahi kolamu kila kuhiliʻole (ke aniani-laina, 100 × 1.8 mm id) me kahi puka puka i loaʻa kahi frit 1 µm i hoʻopili ʻia i kahi packer slurry (Alltech Deerfield, IL, USA). a me ka propelling solvent. E hoʻopiha i ke kolamu ma ka hoʻopili ʻana i nā kaomi o 100 MP no nā minuke 10, 80 MP no 15 mau minuke, a me 60 MP no 30 mau minuke. I ka wā o ka hoʻopili ʻana, ua hoʻopili ʻia ka haʻalulu mechanical me ʻelua GC column shakers (Alltech, Deerfield, IL, USA) e hōʻoia i ka paʻi paʻi ʻana o ke kolamu. kolamu mai ka ʻāpana slurry packing a hoʻohui i kekahi mea kūpono i ka puka komo a me ka ʻōnaehana LC e nānā i kāna hana.
He pamu LC (10AD Shimadzu, Iapana), injector (Valco (USA) C14 W.05) me 50nL injection loop, membrane degasser (Shimadzu DGU-14A), UV-VIS capillary window i kukuluia. ua hoʻokomo ʻia ka pahu pahu, nā capillaries (50 μm id 365 a me ka hoʻohaʻahaʻa ʻana i nā capillaries union (50 μm) ma ka 1/16 ″ puka o ka union hoʻemi. Ua hana ʻia ka hōʻiliʻili ʻikepili a me ka hana chromatographic me ka hoʻohana ʻana i ka polokalamu Multichro 2000. Ka nānā ʻana ma 254 nm Analytes ua hoʻāʻo ʻia no ka ʻikepili O UV.
Albumin mai ke kanaka serum, lyophilized pauka, ≥ 96% (agarose gel electrophoresis) 3 mg hui pu me trypsin (1.5 mg), 4.0 M urea (1 mL), a me 0.2 M ammonium bicarbonate (1 mL). Ua hoʻonāukiʻia ka hopena no 10 mau minuke a mālamaʻia i loko o ka waiʻauʻau wai ma 37,10°C me 67,1°C. E hoʻololi i ka hopena a mālama i lalo o 4 ° C.
Hoʻokaʻawale ʻia ka hoʻokaʻawale ʻana o ka hui peptide a me ka HSA trypsin digests ma nā kolamu PMP.
Hōʻike ʻia nā kiʻi SEM o nā ʻāpana silika ʻole a me nā ʻāpana silika i hoʻopaʻa ʻia i ka ligand ma FIG.2. Hōʻike nā kiʻi SEM o nā ʻāpana silica ʻole (A, B) i ka ʻokoʻa i kā mākou mau haʻawina mua, ʻo kēia mau ʻāpana he spherical kahi e hoʻolōʻihi ʻia ai nā ʻāpana a i ʻole like ʻole ke ʻano like ʻole.
Ke nānā nei i nā kiʻi microscope electron o nā ʻāpana silica ʻole (A, B) a me nā ʻāpana silica paʻa ligand (C, D).
Hōʻike ʻia nā puʻunaue nui o ka nui o nā ʻāpana silica a me nā ʻāpana liʻiliʻi i hoʻopaʻa ʻia ma ka Figure 3 (A). Hōʻike ʻia ka nui o ka nui o ka nui o nā silica ma hope o ka hoʻololi kemika (Fig. 3A). i kā mākou noiʻi mua me ka ad(0.5) waiwai o 3.05 μm (polystyrene-bound silica particles) 34. Ua ʻoi aku ka liʻiliʻi o ka puʻupuʻu o kēia pūʻulu ma mua o kā mākou noiʻi mua ʻana ma muli o nā ʻano like ʻole o ka PEG, urea, TMOS, a me ka acetic acid i ka hui ʻana. Ua waiho wale ka styrene i kahi papa polystyrene (0.97 µm) ma ka ʻili silica, akā ma ka pae PMP he 1.38 µm ka mānoanoa o ka papa.
Ka puunaue ana i ka nui o ka palaki (A) a me ka puunaue nui o ka pore (B) o na mea silika olohelohe a me na kinikini silica paa ligand.
ʻO ka nui o ka pore, ka nui o ka pore a me ka ʻili o nā ʻāpana silica o ka haʻawina o kēia manawa ua hāʻawi ʻia ma ka Papa 1 (B). ʻO nā kiʻi PSD o nā ʻāpana silica ʻole a me nā ʻāpana silica i hoʻopaʻa ʻia i ligand i hōʻike ʻia ma ke Kiʻi 3 (B). Ua hoʻohālikelike ʻia nā hopena i kā mākou noiʻi mua. ʻO ka nui o ka pore o ka silica a me ka ligand-bound silica a me nā mea liʻiliʻi he 291, e hōʻike ana i ka nui o ka pore ma hope o 4310. hoʻololi kemika, e like me ka mea i hōʻike ʻia ma ka Papa 1(B), a ua hōʻike ʻia ka hoʻololi ʻana o ka pihi ma Fig. 3 (B) .Pēlā hoʻi, ua emi ka nui o ka pore o nā silica particles mai 0.67 a i 0.58 cm3/g ma hope o ka hoʻololi kemika. ʻO ka ʻāpana kiko o nā silica i aʻo ʻia i kēia manawa he 116 m2/g i hōʻike ʻia ma mua o ka 116 m2/g. B), ua emi iho ka ili honua (m2/g) o na mea silica mai 116 m2/g a i 105 m2/g ma hope o ka hoololi kemika.
Hōʻike ʻia nā hopena o ka hoʻopili elemental o ka pae paʻa ma ka Papa 2. ʻO ka hoʻouka kalapona o ka manawa paʻa o kēia manawa he 6.35%, ʻoi aku ka haʻahaʻa ma mua o ka hoʻouka kalapona o kā mākou haʻawina mua (polystyrene bonded silica particles, 7.93%35 a me 10.21%, kēlā me kēia) 42. ʻO ka hoʻouka kalapona o ka māhele paʻa SP o kēia manawa he haʻahaʻa, no ka mea, haʻahaʻa ka hoʻouka ʻana o ke kalapona o ka SP stationary i kēia manawa, no ka mea, he haʻahaʻa ka hoʻouka ʻana o ke kalapona o ka SP i kēia manawa i ka hoʻomākaukau ʻana i ka popherene i kēia manawa. Ua hoʻohana ʻia ka nylmaleimide-methylvinylisocyanate (PCMP) a me 4-hydroxy-TEMPO. ʻO ka pākēneka o ke kaumaha o ka nitrogen i kēia manawa he 2.21%, ke hoʻohālikelike ʻia me 0.1735 a me 0.85% ma ke kaumaha o ka nitrogen i nā haʻawina mua. ʻO ia hoʻi, ʻoi aku ka kiʻekiʻe o ka wt% o ka nitrogen i ka manawa kū hoʻokahi o kēia manawa, ka phenylmading . he 2.7% a me 2.9%, oiai o ka hoouka kalapona o ka hua hope (6) he 6.35%, e like me ka hoike ana ma ka Papa 2. Ua nanaia ka poho kaumaha me ka PMP stationary phase, a ua hoikeia ka TGA curve ma ke Kii 4. Ua hoike mai ka TGA curve i ka poho kaumaha o 8.6%, aia i loko o ka aelike maikaʻi me ka hoʻouka kalapona, ʻaʻole ʻo 6.
Ua koho ʻia ka ligand phenylmaleimide-methylvinylisocyanate no ka hoʻololi ʻana i ka ʻili o nā ʻāpana silica no ka mea he mau pūʻulu phenylmaleimide polar a me nā hui vinylisocyanate. Hiki i nā hui isocyanate Vinyl ke hana hou aku me ka styrene e ka polymerization radical ola. ʻO ke kumu ʻelua e hoʻokomo i kahi hui i loaʻa ka pilina ma waena o ka analyte a me ka hoʻoheheʻe analyteimide ikaika ma waena o ka analytestatic a me ka hoʻoheheʻe ʻana o ka moʻololo. ʻAʻohe o ka uku maʻamau i ka pH maʻamau. Hiki ke hoʻomalu ʻia ka polarity o ka pae paʻa e ka nui o ka styrene a me ka manawa pane o ka polymerization radical manuahi. Ua hoʻomākaukau ʻia me nā manaʻo like ʻole o ka styrene i nā ukana kalapona ʻokoʻa. Eia hou, e hoʻouka i kēia mau mea paʻa i loko o nā kolamu kila kila a nānā i kā lākou hana chromatographic (selectivity, hoʻonā, N waiwai, a me nā mea ʻē aʻe).
ʻElima mau huila peptide (Gly-Tyr, Gly-Leu-Tyr, Gly-Gly-Tyr-Arg, Tyr-Ile-Gly-Ser-Arg, leucine enkephalin) ua loiloi pū ʻia me ka hoʻohana ʻana i kahi kolamu PMP me ka hoʻohana ʻana i kahi pae kelepona;60/40 (v/v) acetonitrile/wai ​​(0.1% TFA) ma kahi kahe kahe o 80 μL/min. Ma lalo o nā kūlana elution maikaʻi loa, ʻo ka helu papa helu (N) no kēlā me kēia kolamu (100 × 1.8 mm id) ʻo 20,000 ± 100 (200,000 mau papa chromato ​​m²). Hōʻike ʻia nā grams ma ka Figure 5A. ʻO ka loiloi wikiwiki ma kahi kolamu PMP ma ke kahe kahe kiʻekiʻe (700 μL / min), ʻelima mau peptides i hoʻokaʻawale ʻia i loko o hoʻokahi minuke, maikaʻi loa nā helu N, 13,500 ± 330 i kēlā me kēia kolamu (100 × 1.8 mm id), E like me ka 135,000 135,000 mm id (T 135,000). × 1.8 mm id) ua hoʻopiha ʻia me ʻekolu mau ʻokoʻa like ʻole o ka PMP stationary phase e nānā i ka reproducibility. The analyte concentration for each column was recorded using the optimal elution condition and the number of theoretical plates N and retention time to separate the same test mixture on each column.
ʻO ka hoʻokaʻawale ʻana o ka hui peptide ma ke kolamu PMP (B) a me ka kolamu Ascentis Express RP-Amide (A);ka pae kelepona 60/40 ACN/H2O (TFA 0.1%), PMP kolamu ana (100 × 1.8 mm id);analytical ʻO ke kauoha elution o nā pūhui: 1 (Gly-Tyr), 2 (Gly-Leu-Tyr), 3 (Gly-Gly-Tyr-Arg), 4 (Tyr-Ile-Gly-Ser-Arg) a me 5 (leucine) acid enkephalin)).
Ua loiloi ʻia kahi kolamu PMP (100 × 1.8 mm id) no ka hoʻokaʻawale ʻana o ka tryptic digests o ke kanaka serum albumin i ka hana kiʻekiʻe wai chromatography.The chromatogram ma Figure 6 hōʻike ʻia ua hoʻokaʻawale maikaʻi ʻia ka hāpana a maikaʻi loa ka hoʻonā. Ua kālai ʻia nā digest HSA me ka hoʻohana ʻana i kahi kahe kahe o 100 µL/3. i hōʻike ʻia ma ka chromatogram (Figure 6), ua māhele ʻia ka HSA digestion i 17 peaks e pili ana i nā peptides 17. Ua helu ʻia ka hoʻokaʻawale ʻana o kēlā me kēia peak i ka HSA digest a hāʻawi ʻia nā waiwai ma ka Papa 5.
Ua hoʻokaʻawale ʻia kahi digest tryptic o HSA (100 × 1.8 mm id) ma kahi kolamu PMP;kahe kahe (100 µL / min), ka pae kelepona 60/40 acetonitrile / wai me 0.1% TFA.
kahi L ka lōʻihi o ke kolamu, ʻo η ka viscosity o ka pae kelepona, ΔP ke kolamu hope kaomi, a ʻo u ka linear velocity o ka mobile phase.The permeability o ke kolamu PMP he 2.5 × 10-14 m2, ʻo ka kahe kahe ʻana he 25 μL / min, a ua hoʻohana ʻia ka 60/40 v / v / v PMP i kēlā me kēia wai. like me kā mākou noiʻi mua Ref.34. ʻO ka permeability o ke kolamu i hoʻopili ʻia me nā ʻāpana superficially porous: 1.7 × 10-15 no nā ʻāpana 1.3 μm, 3.1 × 10-15 no nā ʻāpana 1.7 μm, 5.2 × 10-15 a me 2.5 × 10-15 a me 2.5 × 10-15 m. refore, ua like ka permeability o ka PMP me ka 5 μm core-shell particles.
ʻo Wx ke kaumaha o ke kolamu i hoʻopili ʻia me ka chloroform, ʻo Wy ke kaumaha o ke kolamu i hoʻopiha ʻia me ka methanol, a ʻo ρ ka mānoanoa o ka mea hoʻoheheʻe. ʻO nā ʻāpana o ka methanol (ρ = 0.7866) a me ka chloroform (ρ = 1.484). ʻO ka huina o ka SILICA PARTICLES-C18 × kolamu (103 mmU i nā kolamu) s 31 a mākou i aʻo mua ai he 0.63 a me 0.55. ʻO ia hoʻi, ʻo ka hiki ʻana mai o nā ligands urea e hoʻemi i ka permeability o ka pae paʻa. Ma kekahi ʻaoʻao, ʻo ka nui o ka porosity o ke kolamu PMP (100 × 1.8 mm id) ʻo 0.60. ʻO ka permeability o nā kolamu C18 i hoʻopaʻa ʻia ma lalo o ka C1 kolamu. nā ʻāpana, ua hoʻopili ʻia nā ligands C18 i nā ʻāpana silica ma ke ʻano he kaulahao laina, ʻoiai i loko o ka polystyrene-type stationary phases, ua hoʻokumu ʻia ka papa polymer mānoanoa a puni ia.
Hōʻike ka Figure 7A,B i ke kolamu PMP (100 × 1.8 mm id) a me ka kolamu Ascentis Express RP-Amide (100 × 1.8 mm id) me ka hoʻohana ʻana i nā kūlana elution like (ie, 60/40 ACN/H2O a me 0.1% TFA).) o ka mokuahi van Deemter.ʻO nā hui peptide i koho ʻia (Gly-Tyr, Gly-Leu-Tyr, Gly-Gly-Tyr-Arg, Tyr-Ile-Gly-Ser-Arg, Leucine Enkephalin) i hoʻomākaukau ʻia i 20 µL/ ʻO ka liʻiliʻi liʻiliʻi no nā kolamu ʻelua he 800 µL/min. ʻO ke kolamu Express RP-Amide he 2.6 µm a me 3.9 µm. ʻO nā helu HETP e hōʻike ana i ka hoʻokaʻawale ʻana o ke kolamu PMP (100 × 1.8 mm id) ʻoi aku ka maikaʻi ma mua o ka mea kūʻai aku ʻo Ascentis Express RP-Amide kolamu (100 × 1.8 mm id7). ʻoi aku ka nui ma mua o kā mākou noiʻi mua ʻana. ʻO ke kiʻekiʻe o ka hoʻokaʻawale ʻana o ke kolamu PMP (100 × 1.8 mm id) i hoʻohālikelike ʻia me ka kolamu Ascentis Express RP-Amide ma muli o ka hoʻomaikaʻi ʻana i ke ʻano ʻāpana, ka nui, a me nā kaʻina hana hoʻopili kolamu paʻakikī i hoʻohana ʻia i ka hana o kēia manawa34.
(A) van Deemter plot (HETP versus mobile phase linear velocity) i loaʻa me ka PMP column (100 × 1.8 mm id) ma 60/40 ACN/H2O me 0.1% TFA.(B) van Deemter plot (HETP versus mobile phase linear velocity) loaʻa me ka Ascentis Express RP-Amide × 1 column (10 mm0 × i 4) O me 0.1% TFA.
Ua hoʻomākaukau a loiloi ʻia kahi papa hana polystyrene i hoʻopaʻa ʻia no ka hoʻokaʻawale ʻana i nā hui peptide synthetic a me nā trypsin digests o ke kanaka serum albumin (HAS) i ka hana kiʻekiʻe wai chromatography. nthesis o ke kahua paʻa, a me ka paʻakikī kolamu packing.In addition to high separation efficiency, low column back pressure at high flow rate is another advantage of this stationary phase.PMP columns exhibit good reproducibility and can be used for the analysis of peptide mixtures and trypsin digestion of various proteins.We intend to use this column for the separation of natural products, biomedical PMP kolamu e hiki mai ana i ka wai puna i loko o ka chrominal mea kanu. e loiloi ʻia no ka hoʻokaʻawale ʻana o nā protein a me nā antibodies monoclonal.
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