CN115814772A - Silica gel stationary phase of bond and dipeptide and application of silica gel stationary phase in chitosan oligosaccharide chromatographic separation - Google Patents
Silica gel stationary phase of bond and dipeptide and application of silica gel stationary phase in chitosan oligosaccharide chromatographic separation Download PDFInfo
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- CN115814772A CN115814772A CN202211282631.8A CN202211282631A CN115814772A CN 115814772 A CN115814772 A CN 115814772A CN 202211282631 A CN202211282631 A CN 202211282631A CN 115814772 A CN115814772 A CN 115814772A
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- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 title claims abstract description 104
- 239000000741 silica gel Substances 0.000 title claims abstract description 100
- 229910002027 silica gel Inorganic materials 0.000 title claims abstract description 100
- 230000005526 G1 to G0 transition Effects 0.000 title claims abstract description 55
- 108010016626 Dipeptides Proteins 0.000 title claims abstract description 49
- RQFQJYYMBWVMQG-IXDPLRRUSA-N chitotriose Chemical compound O[C@@H]1[C@@H](N)[C@H](O)O[C@H](CO)[C@H]1O[C@H]1[C@H](N)[C@@H](O)[C@H](O[C@H]2[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O2)N)[C@@H](CO)O1 RQFQJYYMBWVMQG-IXDPLRRUSA-N 0.000 title claims abstract description 45
- 238000013375 chromatographic separation Methods 0.000 title claims abstract description 16
- 238000006243 chemical reaction Methods 0.000 claims abstract description 32
- 239000006087 Silane Coupling Agent Substances 0.000 claims abstract description 25
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 25
- 150000001875 compounds Chemical class 0.000 claims abstract description 17
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- 230000004048 modification Effects 0.000 claims abstract description 7
- 238000012986 modification Methods 0.000 claims abstract description 7
- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Chemical compound CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 claims description 42
- 238000000967 suction filtration Methods 0.000 claims description 16
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- 125000003368 amide group Chemical group 0.000 claims description 6
- 239000008367 deionised water Substances 0.000 claims description 6
- 229910021641 deionized water Inorganic materials 0.000 claims description 6
- JXUKBNICSRJFAP-UHFFFAOYSA-N triethoxy-[3-(oxiran-2-ylmethoxy)propyl]silane Chemical compound CCO[Si](OCC)(OCC)CCCOCC1CO1 JXUKBNICSRJFAP-UHFFFAOYSA-N 0.000 claims description 6
- -1 without limitation Chemical compound 0.000 claims description 6
- 239000007864 aqueous solution Substances 0.000 claims description 5
- 239000011159 matrix material Substances 0.000 claims description 5
- 229910004298 SiO 2 Inorganic materials 0.000 claims description 3
- 239000007822 coupling agent Substances 0.000 claims description 3
- 238000003760 magnetic stirring Methods 0.000 claims description 3
- 239000012046 mixed solvent Substances 0.000 claims description 3
- HJCMDXDYPOUFDY-WHFBIAKZSA-N Ala-Gln Chemical compound C[C@H](N)C(=O)N[C@H](C(O)=O)CCC(N)=O HJCMDXDYPOUFDY-WHFBIAKZSA-N 0.000 claims description 2
- XBGGUPMXALFZOT-VIFPVBQESA-N Gly-Tyr Chemical compound NCC(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 XBGGUPMXALFZOT-VIFPVBQESA-N 0.000 claims description 2
- BLRPTPMANUNPDV-UHFFFAOYSA-N Silane Chemical compound [SiH4] BLRPTPMANUNPDV-UHFFFAOYSA-N 0.000 claims description 2
- CCUAQNUWXLYFRA-UHFFFAOYSA-N alanyl-asparagine Chemical compound CC(N)C(=O)NC(C(O)=O)CC(N)=O CCUAQNUWXLYFRA-UHFFFAOYSA-N 0.000 claims description 2
- 108010044940 alanylglutamine Proteins 0.000 claims description 2
- 229960002648 alanylglutamine Drugs 0.000 claims description 2
- 238000004440 column chromatography Methods 0.000 claims description 2
- FWDBOZPQNFPOLF-UHFFFAOYSA-N ethenyl(triethoxy)silane Chemical compound CCO[Si](OCC)(OCC)C=C FWDBOZPQNFPOLF-UHFFFAOYSA-N 0.000 claims description 2
- NKSJNEHGWDZZQF-UHFFFAOYSA-N ethenyl(trimethoxy)silane Chemical compound CO[Si](OC)(OC)C=C NKSJNEHGWDZZQF-UHFFFAOYSA-N 0.000 claims description 2
- XBGGUPMXALFZOT-UHFFFAOYSA-N glycyl-L-tyrosine hemihydrate Natural products NCC(=O)NC(C(O)=O)CC1=CC=C(O)C=C1 XBGGUPMXALFZOT-UHFFFAOYSA-N 0.000 claims description 2
- 108010087823 glycyltyrosine Proteins 0.000 claims description 2
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- 238000012856 packing Methods 0.000 claims 1
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- 239000001257 hydrogen Substances 0.000 abstract description 2
- 239000000047 product Substances 0.000 description 15
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 13
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 12
- 239000000203 mixture Substances 0.000 description 11
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 10
- 239000012043 crude product Substances 0.000 description 10
- OWEGMIWEEQEYGQ-UHFFFAOYSA-N 100676-05-9 Natural products OC1C(O)C(O)C(CO)OC1OCC1C(O)C(O)C(O)C(OC2C(OC(O)C(O)C2O)CO)O1 OWEGMIWEEQEYGQ-UHFFFAOYSA-N 0.000 description 9
- GUBGYTABKSRVRQ-PICCSMPSSA-N Maltose Natural products O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@@H](CO)OC(O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-PICCSMPSSA-N 0.000 description 9
- 229930006000 Sucrose Natural products 0.000 description 9
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 9
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 9
- 239000012265 solid product Substances 0.000 description 9
- 239000005720 sucrose Substances 0.000 description 9
- 229920001661 Chitosan Polymers 0.000 description 8
- 238000010168 coupling process Methods 0.000 description 8
- 238000010521 absorption reaction Methods 0.000 description 7
- 230000000694 effects Effects 0.000 description 7
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 6
- 230000008878 coupling Effects 0.000 description 6
- 238000005859 coupling reaction Methods 0.000 description 6
- 238000001514 detection method Methods 0.000 description 5
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- 239000011259 mixed solution Substances 0.000 description 4
- 230000007935 neutral effect Effects 0.000 description 4
- 239000000126 substance Substances 0.000 description 4
- 229920002101 Chitin Polymers 0.000 description 3
- 230000021736 acetylation Effects 0.000 description 3
- 238000006640 acetylation reaction Methods 0.000 description 3
- 125000003277 amino group Chemical group 0.000 description 3
- GUBGYTABKSRVRQ-QUYVBRFLSA-N beta-maltose Chemical compound OC[C@H]1O[C@H](O[C@H]2[C@H](O)[C@@H](O)[C@H](O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@@H]1O GUBGYTABKSRVRQ-QUYVBRFLSA-N 0.000 description 3
- 230000008859 change Effects 0.000 description 3
- 238000000921 elemental analysis Methods 0.000 description 3
- QQQSFSZALRVCSZ-UHFFFAOYSA-N triethoxysilane Chemical compound CCO[SiH](OCC)OCC QQQSFSZALRVCSZ-UHFFFAOYSA-N 0.000 description 3
- 238000001291 vacuum drying Methods 0.000 description 3
- 238000005303 weighing Methods 0.000 description 3
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 description 2
- 229910008051 Si-OH Inorganic materials 0.000 description 2
- 229910006358 Si—OH Inorganic materials 0.000 description 2
- 150000001413 amino acids Chemical class 0.000 description 2
- 235000011114 ammonium hydroxide Nutrition 0.000 description 2
- 239000000538 analytical sample Substances 0.000 description 2
- 239000007795 chemical reaction product Substances 0.000 description 2
- 125000004093 cyano group Chemical group *C#N 0.000 description 2
- GYZLOYUZLJXAJU-UHFFFAOYSA-N diglycidyl ether Chemical compound C1OC1COCC1CO1 GYZLOYUZLJXAJU-UHFFFAOYSA-N 0.000 description 2
- 239000000945 filler Substances 0.000 description 2
- 238000011049 filling Methods 0.000 description 2
- 125000000524 functional group Chemical group 0.000 description 2
- 125000003147 glycosyl group Chemical group 0.000 description 2
- 238000002329 infrared spectrum Methods 0.000 description 2
- 150000002500 ions Chemical class 0.000 description 2
- 125000005647 linker group Chemical group 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- 229920001542 oligosaccharide Polymers 0.000 description 2
- 150000002482 oligosaccharides Chemical class 0.000 description 2
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- 229910001220 stainless steel Inorganic materials 0.000 description 2
- 239000010935 stainless steel Substances 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- MSWZFWKMSRAUBD-IVMDWMLBSA-N 2-amino-2-deoxy-D-glucopyranose Chemical compound N[C@H]1C(O)O[C@H](CO)[C@@H](O)[C@@H]1O MSWZFWKMSRAUBD-IVMDWMLBSA-N 0.000 description 1
- NLXLAEXVIDQMFP-UHFFFAOYSA-N Ammonium chloride Substances [NH4+].[Cl-] NLXLAEXVIDQMFP-UHFFFAOYSA-N 0.000 description 1
- 229910002808 Si–O–Si Inorganic materials 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 125000004442 acylamino group Chemical group 0.000 description 1
- QGZKDVFQNNGYKY-UHFFFAOYSA-N ammonia Natural products N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 238000005452 bending Methods 0.000 description 1
- MSWZFWKMSRAUBD-UHFFFAOYSA-N beta-D-galactosamine Natural products NC1C(O)OC(CO)C(O)C1O MSWZFWKMSRAUBD-UHFFFAOYSA-N 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 125000002915 carbonyl group Chemical group [*:2]C([*:1])=O 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
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- 235000015872 dietary supplement Nutrition 0.000 description 1
- 150000002009 diols Chemical class 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 230000007515 enzymatic degradation Effects 0.000 description 1
- 125000001495 ethyl group Chemical group [H]C([H])([H])C([H])([H])* 0.000 description 1
- 239000003337 fertilizer Substances 0.000 description 1
- 229960002442 glucosamine Drugs 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- WGCNASOHLSPBMP-UHFFFAOYSA-N hydroxyacetaldehyde Natural products OCC=O WGCNASOHLSPBMP-UHFFFAOYSA-N 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
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- 231100000053 low toxicity Toxicity 0.000 description 1
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- 125000004433 nitrogen atom Chemical group N* 0.000 description 1
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/55—Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups
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- Solid-Sorbent Or Filter-Aiding Compositions (AREA)
Abstract
The invention relates to a silica gel stationary phase of a bond and dipeptide and application thereof in chitosan oligosaccharide chromatographic separation. Silica gel substrate is adopted, and a dipeptide compound is bridged by a silane coupling agent, so that the modification of silica gel bond and reaction is realized, and the silica gel stationary phase of bond and dipeptide is obtained. The bond and the dipeptide silica gel chromatographic column mainly provide hydrophilic action in the separation process of the chitosan oligosaccharide, and the retention on the surface of the stationary phase is completed by establishing a 'rich water layer' on the surface of the stationary phase and leading the analyte to enter the 'rich water layer' from the mobile phase. The amide and hydroxyl groups in the chromatographic column stationary phase have strong capability of forming hydrogen bonds, and can enhance the retention of analytes on the surface of the stationary phase. According to different degrees of polymerization of chitosan oligosaccharide in the 'water-rich layer' and the flowing phase distribution acting force, the effective separation of chitosan oligosaccharide can be realized.
Description
Technical Field
The invention belongs to the technical field of separation, and particularly relates to a silica gel stationary phase of a bond and a dipeptide and application thereof in chitosan oligosaccharide chromatographic separation.
Background
At the present day when the terrestrial resources are gradually deficient, the development and application of marine resources are paid more attention by researchers. Chitosan Oligosaccharide (COS) is an effective extension product, mainly comes from chitin in marine resources, and is the most important derivative of chitin. Based on the chemical structure similar to that of chitin, chitosan oligosaccharide has excellent biocompatibility, biodegradability, low toxicity and other physical and chemical characteristics. The unique good water solubility promotes the application of COS in the industrial fields of cosmetics, organic fertilizers, dietary supplements and the like. Since the 70 s of the 20 th century, the preparation and application of COS have been the focus of research by researchers. Currently, most of COS in industrial production is prepared by enzymatic degradation, the obtained product is usually in a wider molecular weight range, the impurity components are more, an effective separation technical means is lacked, and the COS with single polymerization degree cannot be obtained, so that the further application of the COS at the molecular level is limited.
The prior art on chromatographic separation of chitosan includes: chinese patent CN114544788A discloses a chromatographic separation method of chitosan oligosaccharide isomers at different acetylation sites. Particularly, under the conditions that the pH value is 8-12 and the column temperature is 40-90 ℃, a hydrophilic chromatographic column is adopted for separating the materials. The hydrophilic chromatographic column used in the patent is a silica gel chromatographic column or a silica gel chromatographic column bonded with a polar functional group; the polar bonding group is one or more of amide, amino acid, amino, cyano, glycol, carboxyl, glycosyl, zwitter ion and the like; in this patent, polar functional groups are added to a column mainly by amide groups, carboxyl groups, and the like, but the influence of polarity on chromatographic separation is not described. The patent also states that the hydrophilic chromatographic column has an inner diameter of 2.1 to 200mm; the length of the hydrophilic chromatographic column is 50-250 mm; the grain diameter of the filler is 1.7-30 mu m; the specific surface area of the filler is 180-350 m 2 (ii) in terms of/g. The bonding amount of the polar group on the silica gel substrate chromatographic column is 0.1-20 mu mol/m 2 . Chinese patent CN114544789A discloses a chromatographic separation method of chitosan oligosaccharide with different acetylation degrees. Separating the mixture by a hydrophilic chromatographic column under the condition that the pH value is 2-6. The hydrophilic chromatographic column used in the patent is one or more than two of silica gel chromatographic columns or silica gel chromatographic columns bonded with polar groups; the polar bonding group is amide, amino acid, amino group, cyano groupOne or more of diol, carboxyl, glycosyl, zwitter ion and the like; the method separates the chitosan oligosaccharide according to the number of the glucosamine, so that the chitosan oligosaccharide with different acetylation degrees can be separated, and the method still has the separation capability on samples with high polymerization degrees.
Disclosure of Invention
Different from the scheme of using a chromatographic column bonded with polar groups to realize chitosan chromatographic separation in the prior art, the application provides a silica gel stationary phase bonded with dipeptide and application thereof in chitosan oligosaccharide chromatographic separation.
Based on the characteristics of strong polarity, good hydrophilicity, more amino and hydroxyl contained in molecules, strong intermolecular or intramolecular action and the like, the application designs a mixed-mode chromatographic stationary phase taking dipeptide as a functional monomer and realizes the chromatographic separation of chitosan oligosaccharide by utilizing the mixed-mode chromatographic stationary phase.
In the application, the peptide bond is a chemical bond formed by dehydrating and condensing carboxyl and amino, and means that pi electrons on carbonyl in an amide group and lone pair electrons on nitrogen atoms in adjacent C-N bonds jointly form a three-center four-electron delocalized pi bond, so that other groups in the neighborhood display stronger polarity under the conjugation effect. Therefore, a dipeptide-structured molecule having a free carboxyl group, amino group or hydroxyl group can exhibit a stronger polar force than a single structure or a simple combination of amino group, amide group or carboxyl group.
The purpose of the invention can be realized by the following technical scheme:
the invention provides a silica gel stationary phase of bond and dipeptide, which adopts a silica gel substrate, and bridges a dipeptide compound through a silane coupling agent to realize the modification of silica gel bond and reaction, thereby obtaining the silica gel stationary phase of bond and dipeptide.
In one embodiment of the invention, the silica gel matrix is column chromatography white powder, and the main component is SiO 2 ·nH 2 O, insoluble in water, inorganic acid and other organic solvent, and has particle size of 8-12 micron, preferably 10 micron, 100-1300 mesh.
In one embodiment of the present invention, before the silica gel substrate is used, activated silicon hydroxyl groups are formed on the surface of the silica gel microspheres after acidification treatment.
In one embodiment of the present invention, the silica gel matrix is acidified before use by: refluxing and acid-washing 10-30 wt% hydrochloric acid solution at 100-120 deg.c, suction filtering after refluxing, washing with great amount of water to neutrality, suction filtering to obtain solid product, and drying to obtain activated silica gel with activated silica hydroxyl group formed on the surface.
In one embodiment of the present invention, the specific method of acidifying the silica gel matrix before use is: and (2) refluxing for 8 hours at 115 ℃ by using a hydrochloric acid solution with the mass fraction of 20% for acid washing, performing suction filtration after the reflux is finished, washing to be neutral by using a large amount of water, performing suction filtration to obtain a solid product, and drying the solid product in a 120 ℃ drying oven for 2 hours to obtain the activated silica gel.
In one embodiment of the present invention, the silane coupling agent is an epoxy-based silane including, but not limited to, vinyltrimethoxysilane, vinyltriethoxysilane, and 3-glycidoxypropyltriethoxysilane or other modified silane derivatives. Preferably, the silane coupling agent is 3- (glycidoxypropyl) triethoxysilane.
In one embodiment of the present invention, the dipeptide compound is a hydrophilic dipeptide molecule having a free amino, hydroxyl or amide group, including, without limitation, glycyltyrosine, alanylglutamine, alanylasparagine, and the like. Preferably, the dipeptide compound is alanyl-L-glutamine.
In one embodiment of the invention, the silica gel stationary phase of the linkage and dipeptide is: silica gel substrate is adopted, and the modification of silica gel bond and reaction is realized through bridging dipeptide compound alanyl-L-glutamine by 3- (glycidyl ether oxygen propyl) triethoxysilane, so as to obtain the silica gel stationary phase of bond and dipeptide.
In one embodiment of the present invention, when the dipeptide compound is bridged by a silica gel substrate via a silane coupling agent, the bridging reaction comprises the steps of:
mixing a silica gel substrate, anhydrous toluene and a silane coupling agent, placing the mixture into a round-bottom flask, carrying out reflux reaction at 120-150 ℃ for 16-30 h, carrying out suction filtration after the reaction is finished, washing the mixture by using an ethanol/toluene solution, standing the mixture overnight, and drying the mixture for 1-3 h at 100-120 ℃ to obtain bridged silica gel microspheres;
placing the bridged silica gel microspheres in a round bottom flask, adding a dipeptide compound aqueous solution into the round bottom flask, wherein the mass ratio of silica gel to dipeptide compound is 1:1-1:3, magnetically stirring the mixture at 50-100 ℃ for reaction for 36-60 h, performing suction filtration after the reaction is finished, washing the reaction product by using a large amount of deionized water, and drying the reaction product at 100-120 ℃ for 1-3 h to obtain the silica gel stationary phase of the bond and the dipeptide.
Preferably, the silica gel substrate, when the dipeptide compound is bridged by the silane coupling agent, the concentration of the coupling agent in the mixed solvent of the silane coupling agent and the anhydrous toluene is 0.10% -0.30%, preferably 0.20%.
The reaction time of the bridged silica gel microspheres with the dipeptide compound is preferably 36 to 48 hours, and more preferably 48 hours.
The invention also provides the application of the silica gel stationary phase of the bond and the dipeptide, and the silica gel stationary phase of the bond and the dipeptide is used for chromatographic separation of chitosan oligosaccharide after being filled into a column by a wet filling device.
In one embodiment of the invention, the silica gel stationary phase of the bond and the dipeptide can be used for chromatographic separation of chitosan oligosaccharide molecules with single polymerization degree after being filled into a column by a wet filling device.
Compared with the prior art, the invention has the advantages that:
the bond and dipeptide silica gel chromatographic column mainly provide a hydrophilic effect in the separation process of the chitosan oligosaccharide, the polarities of free acylamino, hydroxyl and carboxyl under the action of peptide bonds are more obvious, a 'water-rich layer' is established on the surface of the stationary phase, the space effect of the water-rich layer is larger, and an analyte enters the 'water-rich layer' from a mobile phase to finish the retention on the surface of the stationary phase. The amide and hydroxyl groups in the chromatographic column stationary phase have strong capability of forming hydrogen bonds, and can enhance the retention of analytes on the surface of the stationary phase. According to different degrees of polymerization of chitosan oligosaccharide in the 'water-rich layer' and the flowing phase distribution acting force, the effective separation of chitosan oligosaccharide can be realized.
In addition, the charged carboxyl in the stationary phase of the chromatographic column can be combined with free amino in the chitosan oligosaccharide, so that an ionic effect is provided for the retention of the chitosan oligosaccharide on the stationary phase, and the retention of the chitosan oligosaccharide on the surface of the stationary phase is further enhanced.
The preparation method has the advantages of simple reaction process, controllable reaction conditions and realization of large-scale production. The novel stationary phase is filled by a wet method, and the chitosan oligosaccharide with single polymerization degree can be effectively separated under the conventional conditions of high performance liquid chromatography.
Drawings
FIG. 1: the infrared spectra of the silica gel, epoxy-coupled silica gel, and alanyl-L-glutamine grafted silica gel of example 1;
FIG. 2: the results of the influence of the silane coupling agent concentration on the coupling process in example 2;
FIG. 3: a schematic diagram of the change of the mass fraction of the N element with the reaction time in example 3;
FIG. 4: the separation results of chitosan oligosaccharide standard products and crude products by different chromatographic columns in the embodiment 4;
FIG. 5: the results of the separation of sucrose and maltose in the different columns of example 5.
Detailed Description
The invention is described in detail below with reference to the figures and specific embodiments.
Example 1
This example provides a method for preparing alanyl-L-glutamine grafted stationary phase
Weighing 15g of 10 μm silica gel, placing the silica gel into a 250mL single-neck flat-bottom flask, adding 100mL of 10% hydrochloric acid solution by mass fraction, and refluxing for 6h at 110 ℃. And after the reflux is finished, carrying out suction filtration, washing the product to be neutral by using a large amount of water, obtaining a solid product through suction filtration, and drying the solid product in a vacuum drying oven at 110 ℃ for 2h to obtain the activated silica gel.
10g of activated silica gel, 100mL of anhydrous toluene and 20mL of 3- (glycidyloxypropyl) triethoxysilane (GPTES) were placed in a 250mL single-neck flat-bottomed flask, and subjected to reflux reaction at 125 ℃ for 24 hours, followed by suction filtration after the reaction. Washing the product with ethanol/toluene solution, standingAfter overnight, the mixture was dried at 120 ℃ for 2 hours to give 3- (glycidyloxypropyl) triethoxysilicane (SiO) 2 -GPTES)。
Weighing 2g of SiO 2 GPTES, put in 50mL round-bottomed flask, add 10mL aqueous solution containing 1g alanyl-L-glutamine solid into it, drop 0.5mL acetic acid, keep airtight reaction, react 48h under magnetic stirring at 65 ℃, suction filter after the reaction, wash with a large amount of deionized water, put in oven 110 ℃ and dry for 2h, get alanyl-L-glutamine grafted stationary phase (SiO) 2 -GPTES-Ala-Gln)。
Wherein, the results of the infrared spectrum and the elemental analysis of the alanyl-L-glutamine grafted stationary phase are shown in figure 1.
In the infrared curve of bare silica gel in FIG. 1 (c), 800cm -1 And 1100cm -1 The nearby absorption peak should be antisymmetric telescopic vibration absorption peak of Si-O-Si, 3500-3000cm -1 The nearby strong broad peak is the characteristic absorption peak of Si-OH, 1640cm -1 The nearby weak single peak is an H-O-H bending vibration peak of the silica gel without water after being dried at high temperature; siO in comparison (b) 2 The infrared curve of GPTES can be found to be 3500-3000cm -1 The intensity of the nearby absorption peak is obviously reduced, probably caused by consuming a large amount of Si-OH, 3000-2800cm -1 A new absorption peak appears nearby, probably caused by C-H carried by ethyl in the silane coupling agent, thereby proving that the 3- (glycidyl ether oxygen radical propyl) triethoxysilane is successfully bonded to the naked silica gel.
Comparing the two curves (a) and (b) in FIG. 1, siO 2 3500-3000cm in-GPTES-Ala-Gln -1 The absorption peak is obviously enhanced, and is probably caused by the stretching vibration of groups such as-OH, NH and the like brought by alanyl-L-glutamine; 1640cm -1 The water peak nearby is 1700-1650cm -1 Is covered by absorption peak and is 1700-1650cm -1 The inner stronger single peak should be caused by the increase of C = O in carboxyl group or amide group brought by alanyl-L-glutamine, which proves that alanyl-L-glutamine is successfully fixed on the surface of silica gel.
TABLE 1 elemental analysis Table for silica gel, epoxy-coupled silica gel, and alanyl-L-glutamine grafted silica gel
According to the data in the table 1, the mass fractions of C, H elements are greatly increased from silica gel to epoxy coupling silica gel, which proves that 3- (glycidyl ether oxypropyl) triethoxysilane is successfully coupled; from epoxy coupling silica gel to alanyl-L-glutamine grafting silica gel, the mass fraction of the N element is obviously increased, and the success of alanyl-L-glutamine grafting is proved. C. The decrease in the mass fraction of the H element is probably due to the fact that the mass fraction of the C element in alanyl-L-glutamine accounts for 44% of the total mass, the mass fraction of the H element accounts for 6% of the total mass, and the mass fraction of the C element is reduced as a whole due to successful grafting.
Example 2
This example investigated the optimum silane coupling agent concentration
20g of 10 μm silica gel was weighed and placed in a 250mL single-neck flat-bottom flask, and 100mL of a 10% hydrochloric acid solution by mass fraction was added, and the mixture was refluxed at 110 ℃ for 6 hours. And after the reflux is finished, carrying out suction filtration, washing the product to be neutral by using a large amount of water, obtaining a solid product through suction filtration, and drying the solid product in a vacuum drying oven at 110 ℃ for 2h to obtain the activated silica gel.
Take 3g of 10 μm activated silica as an example. Respectively adding 50mL of 3- (glycidyl ether oxypropyl) triethoxysilane (GPTES)/toluene solution with the concentration of 0.10%, 0.15%, 0.20%, 0.25% and 0.30% into a 250mL single-mouth bottle containing different 3g of 10-micron activated silica gel, carrying out reflux reaction for 24 hours under the protection of nitrogen, carrying out suction filtration after the reaction is finished, washing with ethanol/toluene solution, standing overnight, drying at 120 ℃ for 2 hours to obtain 10-micron epoxy coupling silica gel, and carrying out elemental analysis and characterization on a product.
In the coupling process of the silica gel, the main method for judging the bonding quantity of the silane coupling agent is to compare the variable quantity of the mass fraction of the C element of the synthesized epoxy coupling silica gel. And (3) drawing by taking the content of the coupling agent in the silane coupling agent/anhydrous toluene mixed solvent as an abscissa and the mass fraction of the element C as an ordinate. As a result, as shown in FIG. 2, the amount of change in the mass fraction of the element C in the coupling silica gel increased as the content of the silane coupling agent increased. In the range of 0-0.10% of silane coupling agent content, the variation of the C element mass fraction is increased by 3.87%, and in the range of 0.25-0.30% of silane coupling agent content, the variation of the C element mass fraction is increased by 0.18%, and the increase is obviously reduced. This means that the amount of the silane coupling agent bonded to the silica gel surface gradually becomes saturated with further increase in the content of the silane coupling agent, and the optimum content of the silane coupling agent is selected to be 0.20% in view of the increase in the mass fraction of the element C and the increase in the variation.
Example 3
This example investigates the optimum reaction duration
20g of 10 μm silica gel was weighed and placed in a 250mL single-neck flat-bottom flask, and 100mL of a 10% hydrochloric acid solution by mass fraction was added, and the mixture was refluxed at 110 ℃ for 6 hours. And after the reflux is finished, carrying out suction filtration, washing the mixture to be neutral by using a large amount of water, obtaining a solid product through suction filtration, and drying the solid product in a vacuum drying oven at 110 ℃ for 2 hours to obtain the activated silica gel.
15g of activated silica gel, 100mL of anhydrous toluene and 20mL of 3- (glycidyloxypropyl) triethoxysilane (GPTES) were placed in a 250mL single-neck flat-bottomed flask, and subjected to reflux reaction at 125 ℃ for 24 hours, followed by suction filtration after the reaction was completed. The product was washed with ethanol/toluene solution, left to stand overnight and dried at 120 ℃ for 2h to give 3- (glycidyloxypropyl) triethoxysilicane (SiO 2-GPTES).
Keeping the adding amount of each reactant to be 2g of epoxy coupling silica gel, 10mL of alanyl-L-glutamine aqueous solution at 1g/mL, 0.5mL of acetic acid, keeping the reaction under magnetic stirring at 65 ℃ for closed reaction, setting the reaction time to be 12h, 24h, 36h, 48h and 60h, carrying out suction filtration after the reaction is finished, washing by using a large amount of deionized water, drying in an oven at 110 ℃ for 2h, and characterizing the dried product by element analysis, wherein the result is shown in figure 3.
The change in the mass fraction of the N element was observed at the reaction temperature of 65 ℃ at 12-hour intervals. In fig. 3, the mass fraction of N element continuously increases as the reaction time length increases, indicating that the reaction is also continuously proceeding. In the initial 0-12h, the reaction speed is slow, the growth trend of the N element mass fraction is stable between 12h and 36h along with the increase of the reaction time, the reaction speed is fast and basically kept unchanged, after the reaction is carried out for 48h, the growth of the N element mass fraction tends to be stopped, the N element mass fraction only increases by 0.01% in the reaction time of 48h-60h, and the optimal reaction time is determined to be 48h by integrating the reaction time and the increase condition of the N element mass fraction.
Example 4
This example provides the use of a silica gel alanyl-L-glutamine grafted stationary phase (alanyl-L-glutamine grafted silica gel obtained in example 1) for the chromatographic separation of chitooligosaccharides
To further study the separation effect of 10 μm silica gel alanyl-L-glutamine grafted stationary phase on chitosan oligosaccharide, the stationary phase (alanyl-L-glutamine grafted silica gel obtained in example 1) was loaded on a 4.6X 150mm stainless steel analytical chromatographic column, connected to an Agilent model 1260 high performance liquid chromatograph, washed for 4h with pure acetonitrile solution as the mobile phase, and detected with a differential detector with oligochitosan oligosaccharide as the analytical sample, to investigate the separation ability of the chromatographic column. The separation was carried out under conditions of mobile phase composition acetonitrile/aqueous ammonia solution (70/30), mobile phase pH 10.24, flow rate of 1.0mL/min, column temperature 35 ℃, detection wavelength 254 nm.
Separating with chitosan oligosaccharide standard product and crude product as analytes. Weighing 250mg of chitosan oligosaccharide standard product and crude product, respectively placing into two 10mL test tubes, respectively adding 5mL deionized water, and ultrasonically dissolving at 35 deg.C for 5min. Filtering with 0.22 μm water system filter membrane after completely dissolving to obtain chitosan oligosaccharide standard mixed solution and chitosan oligosaccharide standard mixed solution with concentration of 50 mg/mL. The results of the separation are shown in FIG. 4, using a 10 μm preparative column and a commercially available Shodex 5 μm amino hydrophilic column as a control. The numbers in the figure correspond to the degree of polymerization of the corresponding chitosan oligosaccharide.
In the separation of chitosan oligosaccharide standard products and crude products, partial miscellaneous peaks exist in the separation of chitosan oligosaccharide crude products by the amino column, the miscellaneous peaks are less in the preparation column, but the retention trends of all retention curves are basically the same, the prepared 10-micron silica gel chromatographic column shows the separation capacity similar to that of a 5-micron amino chromatographic column while the reaction cost is greatly reduced, and the rapid detection of the chitosan oligosaccharide can be realized within 15 min.
In the separation of crude products, the retention time of partial chromatographic peak shifts on an amino column and a preparation column, the retention time of crude product chitosan 2-4 sugar in the amino column is advanced relative to the retention time of a standard product, and the retention time of crude product chitosan 3-5 sugar in the preparation column is delayed relative to the retention time of the standard product, which may be caused by different varieties of crude products of chitosan oligosaccharide and standard products, and the aqueous solution of the dissolved crude products has lower pH, so that the electrostatic repulsion acting force between the amino column and the chitosan oligosaccharide is enhanced, and the electrostatic adsorption acting force between the preparation column and the chitosan oligosaccharide is enhanced.
Example 5
This example provides the use of a silica gel alanyl-L-glutamine grafted stationary phase (alanyl-L-glutamine grafted silica gel obtained in example 1) for the chromatographic separation of chitooligosaccharides
To further study the separation effect of 10 μm silica gel alanyl-L-glutamine grafted stationary phase on chitosan oligosaccharide, the stationary phase (alanyl-L-glutamine grafted silica gel obtained in example 1) was loaded on a 4.6X 150mm stainless steel analytical chromatographic column, connected to an Agilent model 1260 high performance liquid chromatograph, washed for 4h with pure acetonitrile solution as the mobile phase, and detected with a differential detector with oligochitosan oligosaccharide as the analytical sample, to investigate the separation ability of the chromatographic column. The separation is carried out under the conditions that the mobile phase consists of acetonitrile/ammonia water solution (70/30), the pH value of the mobile phase is 10.24, the flow rate is 1.0mL/min, the column temperature is 35 ℃, and the detection wavelength is 254 nm.
Maltose and sucrose were used as analytes for separation. Specifically, 250mg of each of sucrose and maltose is weighed and placed into two 10mL test tubes, 5mL of deionized water is respectively added, and ultrasonic dissolution is carried out for 5min at 35 ℃. After completely dissolving, filtering with a 0.22 μm water system filter membrane to obtain a maltose mixed solution and sucrose mixed solution with the concentration of 50 mg/mL. The results of the separation are shown in FIG. 5, using a 10 μm preparative column and a commercially available Shodex 5 μm amino hydrophilic column as a control.
The separation effect of the sucrose and the maltose on the amino column and the preparation column is good, clear peaks are formed, the symmetry is good, the retention time of the sucrose and the maltose in the preparation column is short, and the rapid detection of the sucrose and the maltose can be completed within about 4 min. The successful separation of the maltose and the sucrose on the preparation column when the retention time of the sucrose and the maltose in the amino column is about 7min and 8min represents that the preparation column has good application potential and is to be applied to the separation and detection of other substances.
The embodiments described above are described to facilitate an understanding and use of the invention by those skilled in the art. It will be readily apparent to those skilled in the art that various modifications to these embodiments may be made, and the generic principles described herein may be applied to other embodiments without the use of the inventive faculty. Therefore, the present invention is not limited to the above embodiments, and those skilled in the art should make improvements and modifications within the scope of the present invention based on the disclosure of the present invention.
Claims (10)
1. A silica gel stationary phase of bond and dipeptide is characterized in that a silica gel substrate is adopted, a dipeptide compound is bridged by a silane coupling agent, and the silica gel stationary phase of bond and dipeptide obtained after modification of silica gel bond and reaction is realized.
2. The silica gel stationary phase of a bond and dipeptide according to claim 1, wherein the silica gel matrix is a column chromatography white powder with a major component of SiO 2 ·nH 2 O, insoluble in water, inorganic acid and other organic solvent, and has particle size of 8-12 microns, preferably 10 microns, and 100-1300 mesh.
3. The silica gel stationary phase of a bond and a dipeptide according to claim 1, wherein the silica gel matrix forms activated silica hydroxyl groups on the surface of the silica gel microspheres after acidification treatment before use.
4. The silica gel stationary phase of a bond and dipeptide according to claim 1, wherein the silane coupling agent is an epoxy based silane including, without limitation, vinyltrimethoxysilane, vinyltriethoxysilane, and 3-glycidoxypropyltriethoxysilane or other modified silane derivatives;
preferably, the silane coupling agent is 3-glycidoxypropyltriethoxysilane.
5. The silica gel stationary phase of a bond and a dipeptide according to claim 1, wherein the dipeptide compound is a hydrophilic dipeptide molecule having free amino, hydroxyl or amide groups, including and not limited to glycyl tyrosine, alanyl glutamine, alanyl asparagine;
preferably, the dipeptide compound is alanyl-L-glutamine.
6. The silica gel stationary phase of a bond and a dipeptide according to claim 1,
the silica gel stationary phase of the bond and dipeptide is: silica gel substrate is adopted, and the modification of silica gel bond and reaction is realized by bridging dipeptide compound alanyl-L-glutamine through 3- (glycidyl ether oxygen propyl) triethoxysilane, so that the obtained bond and dipeptide silica gel stationary phase are obtained.
7. A process for the preparation of a silica gel stationary phase of a bond and a dipeptide according to any of claims 1 to 6 comprising the steps of:
mixing a silica gel substrate, anhydrous toluene and a silane coupling agent, carrying out reflux reaction at 120-150 ℃ for 16-30 h, carrying out suction filtration after the reaction is finished, washing with an ethanol/toluene solution, standing overnight, and drying at 100-120 ℃ for 1-3 h to obtain bridged silica gel microspheres;
adding a dipeptide compound aqueous solution into the bridged silica gel microspheres, wherein the mass ratio of silica gel to the dipeptide compound is 1:1-1:3, carrying out magnetic stirring reaction for 36-60 h at 50-100 ℃, carrying out suction filtration after the reaction is finished, washing with deionized water, and drying at 100-120 ℃ for 1-3 h to obtain the silica gel stationary phase of the bond and the dipeptide.
8. The method according to claim 7, wherein when the silica gel substrate, the anhydrous toluene and the silane coupling agent are mixed, the concentration of the coupling agent in the mixed solvent of the silane coupling agent and the anhydrous toluene is 0.10% to 0.30%, preferably 0.20%.
9. The process according to claim 7, wherein the reaction time of the bridged silica gel microspheres with the dipeptide compound is 36 to 54 hours, more preferably 48 hours.
10. Use of the silica gel stationary phase of a bond and a dipeptide according to any of claims 1 to 6, wherein the silica gel stationary phase of a bond and a dipeptide is used for the chromatographic separation of chitosan oligosaccharide after being packed into a column by a wet packing apparatus.
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Citations (14)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4604207A (en) * | 1982-05-19 | 1986-08-05 | Sumitomo Chemical Co., Ltd. | Packing materials for chromatographic use and a method for analysis of an enantiomer mixture using the same |
| SU1478112A1 (en) * | 1987-05-25 | 1989-05-07 | МГУ им.М.В.Ломоносова | Method of obtaining sorbent for separation of biological fluids by liquid chromatography |
| EP0358758A1 (en) * | 1987-04-09 | 1990-03-21 | Terumo Kabushiki Kaisha | B lymphocyte separating material and body fluid clarifying material |
| CN1864853A (en) * | 2006-04-19 | 2006-11-22 | 劳文剑 | Biological micromolecule or its analogue bonded chiral stationery phase |
| CN101690887A (en) * | 2009-10-16 | 2010-04-07 | 华东理工大学 | Oligopeptide phase reversal chromatography stationary phase and preparation method thereof |
| CN103638913A (en) * | 2013-11-27 | 2014-03-19 | 华东理工大学 | Bonding polysaccharide type hydrophilic chromatographic stationary phase as well as preparation method and application thereof |
| US20140116932A1 (en) * | 2012-10-25 | 2014-05-01 | Guangqing Li | Novel Liquid Chromatographic Media and Methods of Synthesizing the Same |
| CN105199101A (en) * | 2015-10-21 | 2015-12-30 | 武汉理工大学 | Dipeptide polyethyleneimine polymer as well as preparation method and application thereof |
| JP5914731B1 (en) * | 2015-06-15 | 2016-05-11 | 富士シリシア化学株式会社 | Hydrophilic interaction chromatography filler and method for separating hydrophilic compound |
| CN105771943A (en) * | 2016-03-10 | 2016-07-20 | 中国药科大学 | Preparation method and application of bacitracin bonded type chromatographic packing |
| CN108114707A (en) * | 2016-11-29 | 2018-06-05 | 中国科学院大连化学物理研究所 | A kind of silica matrix hydrophobic amino acid bonded stationary phase and its preparation and application |
| CN108114705A (en) * | 2016-11-29 | 2018-06-05 | 中国科学院大连化学物理研究所 | A kind of silica matrix basic amino acid bonded stationary phase and its preparation and application |
| CN109078628A (en) * | 2018-08-27 | 2018-12-25 | 西北大学 | It is a kind of using Bian amine as the efficient hydrophobic interaction chromatograph medium of aglucon, preparation method and its application in protein renaturation and purifying |
| CN115245816A (en) * | 2022-07-08 | 2022-10-28 | 河北工业大学 | Amino nicotinic acid mixed-mode chromatographic stationary phase and preparation method and application thereof |
-
2022
- 2022-10-19 CN CN202211282631.8A patent/CN115814772B/en active Active
Patent Citations (14)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4604207A (en) * | 1982-05-19 | 1986-08-05 | Sumitomo Chemical Co., Ltd. | Packing materials for chromatographic use and a method for analysis of an enantiomer mixture using the same |
| EP0358758A1 (en) * | 1987-04-09 | 1990-03-21 | Terumo Kabushiki Kaisha | B lymphocyte separating material and body fluid clarifying material |
| SU1478112A1 (en) * | 1987-05-25 | 1989-05-07 | МГУ им.М.В.Ломоносова | Method of obtaining sorbent for separation of biological fluids by liquid chromatography |
| CN1864853A (en) * | 2006-04-19 | 2006-11-22 | 劳文剑 | Biological micromolecule or its analogue bonded chiral stationery phase |
| CN101690887A (en) * | 2009-10-16 | 2010-04-07 | 华东理工大学 | Oligopeptide phase reversal chromatography stationary phase and preparation method thereof |
| US20140116932A1 (en) * | 2012-10-25 | 2014-05-01 | Guangqing Li | Novel Liquid Chromatographic Media and Methods of Synthesizing the Same |
| CN103638913A (en) * | 2013-11-27 | 2014-03-19 | 华东理工大学 | Bonding polysaccharide type hydrophilic chromatographic stationary phase as well as preparation method and application thereof |
| JP5914731B1 (en) * | 2015-06-15 | 2016-05-11 | 富士シリシア化学株式会社 | Hydrophilic interaction chromatography filler and method for separating hydrophilic compound |
| CN105199101A (en) * | 2015-10-21 | 2015-12-30 | 武汉理工大学 | Dipeptide polyethyleneimine polymer as well as preparation method and application thereof |
| CN105771943A (en) * | 2016-03-10 | 2016-07-20 | 中国药科大学 | Preparation method and application of bacitracin bonded type chromatographic packing |
| CN108114707A (en) * | 2016-11-29 | 2018-06-05 | 中国科学院大连化学物理研究所 | A kind of silica matrix hydrophobic amino acid bonded stationary phase and its preparation and application |
| CN108114705A (en) * | 2016-11-29 | 2018-06-05 | 中国科学院大连化学物理研究所 | A kind of silica matrix basic amino acid bonded stationary phase and its preparation and application |
| CN109078628A (en) * | 2018-08-27 | 2018-12-25 | 西北大学 | It is a kind of using Bian amine as the efficient hydrophobic interaction chromatograph medium of aglucon, preparation method and its application in protein renaturation and purifying |
| CN115245816A (en) * | 2022-07-08 | 2022-10-28 | 河北工业大学 | Amino nicotinic acid mixed-mode chromatographic stationary phase and preparation method and application thereof |
Non-Patent Citations (4)
| Title |
|---|
| MAGDALENA SKOCZYLAS ET AL: ""Dipeptide-bonded stationary phases for hydrophilic interaction liquid chromatography"", 《RSC ADV.》, vol. 6 * |
| SZYMON BOCIAN ET AL: ""Amino acids, peptides, and proteins as chemically bonded stationary phases – A review"", 《JOURNAL OF SEPARATION SCIENCE》, vol. 39 * |
| 张景宇等: ""苯丙氨酸二肽手性固定相的合成"", 《第十七届全国色谱学术报告会及仪器展览会会议论文集》, pages 1 * |
| 李莹莹等: ""环二肽手性固定相的合成"", 《全国生物医药色谱学术交流会（2008）论文集》, pages 2 * |
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