CN111135808B - Silicon compound, stationary phase and application thereof - Google Patents
Silicon compound, stationary phase and application thereof Download PDFInfo
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- B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
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- B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
- B01D15/08—Selective adsorption, e.g. chromatography
- B01D15/10—Selective adsorption, e.g. chromatography characterised by constructional or operational features
- B01D15/16—Selective adsorption, e.g. chromatography characterised by constructional or operational features relating to the conditioning of the fluid carrier
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- B01D15/08—Selective adsorption, e.g. chromatography
- B01D15/26—Selective adsorption, e.g. chromatography characterised by the separation mechanism
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- C07C403/00—Derivatives of cyclohexane or of a cyclohexene or of cyclohexadiene, having a side-chain containing an acyclic unsaturated part of at least four carbon atoms, this part being directly attached to the cyclohexane or cyclohexene or cyclohexadiene rings, e.g. vitamin A, beta-carotene, beta-ionone
- C07C403/24—Derivatives of cyclohexane or of a cyclohexene or of cyclohexadiene, having a side-chain containing an acyclic unsaturated part of at least four carbon atoms, this part being directly attached to the cyclohexane or cyclohexene or cyclohexadiene rings, e.g. vitamin A, beta-carotene, beta-ionone having side-chains substituted by six-membered non-aromatic rings, e.g. beta-carotene
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- C07F7/00—Compounds containing elements of Groups 4 or 14 of the Periodic System
- C07F7/02—Silicon compounds
- C07F7/08—Compounds having one or more C—Si linkages
- C07F7/18—Compounds having one or more C—Si linkages as well as one or more C—O—Si linkages
- C07F7/1804—Compounds having Si-O-C linkages
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- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11B—PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES
- C11B3/00—Refining fats or fatty oils
- C11B3/10—Refining fats or fatty oils by adsorption
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
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- C07C2601/00—Systems containing only non-condensed rings
- C07C2601/12—Systems containing only non-condensed rings with a six-membered ring
- C07C2601/16—Systems containing only non-condensed rings with a six-membered ring the ring being unsaturated
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- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C2602/00—Systems containing two condensed rings
- C07C2602/02—Systems containing two condensed rings the rings having only two atoms in common
- C07C2602/04—One of the condensed rings being a six-membered aromatic ring
- C07C2602/10—One of the condensed rings being a six-membered aromatic ring the other ring being six-membered, e.g. tetraline
Abstract
The invention relates to a silicon compound, a stationary phase and application thereof. The structural formula of the silicon compound is shown as the formula (I):
the silicon compound provided by the invention contains polar groups (amide groups, sulfonamide groups, carbamido groups or carbamate groups) and is bonded into the matrix to prepare the stationary phase, so that silicon hydroxyl on the surface of the stationary phase can be effectively shielded during chromatographic separation, the peak shape of the alkaline compound is improved, and the silicon compound can be used in a mobile phase environment with higher water content; and the silicon compound is bonded with C 20 ～C 29 The long-chain alkyl enables the prepared stationary phase to have higher stereoselectivity, and has good separation effect on compounds with long chains and high structural similarity.
Description
Technical Field
The invention relates to the technical field of liquid chromatography packing, in particular to a silicon compound, a stationary phase and application thereof.
Background
HPLC is an important analytical tool, and is suitable for analyzing various compounds with different charges, hydrophobicity, polarity, structures, sizes, and the like. Among these, the most commonly used stationary phase for HPLC is C 18 A stationary phase. However, in the specific application C 18 The stationary phase still has some defects, such as peak tailing caused by the interaction of the basic compound and underivatized silicon hydroxyl on the surface of the stationary phase. Based on this, researchers have reduced the residual of surface silicon hydroxyl groups by capping with high purity silica. However, the use of the prepared stationary phase in water causes "hydrophobic collapse" and thus in many cases remainsThere is a tail of the basic compound peak, thereby greatly reducing the retention of the analyte and affecting reproducibility, limiting C 18 The application of stationary phase.
For analytes with long chains and high similarity of molecular structure, such as lipids, fat-soluble vitamins and carotenes, the separation capacity of the C30 stationary phase is superior to that of the C18 stationary phase. However, commercially available C30 silanes are of low purity and typically consist of C20-C32 homologues. In addition, the bonding process is also challenging.
The reversed phase separating medium with embedded polarity is known in the chromatographic field, and the stationary phase improves the peak shape of the alkaline compound, improves the stereoselectivity and enables the stationary phase to be used in the mobile phase environment with higher water content. However, most of the existing polar intercalation type stationary phases are C 18 Silanes, which are less effective in separation than long chain and structurally similar compounds. To our knowledge, polar intercalated reverse phases with alkyl moieties greater than 20 carbon atoms have not been synthesized and studied.
Disclosure of Invention
Therefore, it is necessary to provide a silicon compound, a stationary phase and applications thereof to solve the problem of poor separation effect of the conventional polar intercalation-type immobilization relative to a compound with a long chain and high structural similarity.
A silicon compound having a structural formula represented by formula (I):
wherein R is 1 、R 2 、R 3 Each independently selected from substituted or unsubstituted C 1 -C 4 Alkyl, and R 1 、R 2 And R 3 At least one of them is alkoxy, halogen, alkylamino or alkanoyl;
Y 1 is selected from C 2 -C 18 An alkylene group; y is 2 Is selected from C 20 -C 29 An alkyl group;
l is selected from
The silicon compound provided by the invention contains polar groups (amide groups, sulfonamide groups, carbamido groups or carbamate groups) and is bonded into the matrix to prepare the stationary phase, so that silicon hydroxyl on the surface of the stationary phase can be effectively shielded during chromatographic separation, the peak shape of the alkaline compound is improved, and the silicon compound can be used in a mobile phase environment with higher water content; and the silicon compound is bonded with C 20 ～C 29 The long-chain alkyl group ensures that the prepared stationary phase has higher stereoselectivity, and has especially prominent separation effect on compounds with long chains and high structural similarity, especially on substances of vitamins and lipids.
In one embodiment, R 1 、R 2 、R 3 Each independently selected from alkoxy;
Y 1 is selected from C 2 -C 10 An alkylene group; y is 2 Is selected from C 25 -C 28 An alkyl group; l is selected from
In one embodiment, the silicon compound is selected from:
the invention provides the use of any of the silicon compounds described herein for the preparation of a stationary phase.
The invention provides a stationary phase, and the preparation raw materials of the stationary phase comprise any one of the silicon compounds and a matrix.
In one embodiment, the stationary phase has a structure represented by formula (II):
wherein the content of the first and second substances,
is a silica gel matrix, R 4 、R 5 Each independently selected from a hydroxyl group or an oxygen atom covalently attached to the silica gel matrix.
In one embodiment, the stationary phase is selected from:
the invention provides the use of any of the stationary phases described herein for the detection and/or separation of compounds having a linear length greater than 16 atoms.
In one embodiment, the compounds include lipids, vitamins, and carotenes.
The invention also provides the application of the stationary phase in detecting and/or separating isomers.
Drawings
FIG. 1 is a graph of the results of the hydrophobicity retention test for example 3 and comparative examples 3-4;
FIG. 2 is a graph showing results of stereoselectivity tests of example 4 and comparative examples 5 to 6;
FIG. 3 is a separation spectrum of vitamin K1 and its isomers of example 5 and comparative example 7;
FIG. 4 is a separation spectrum of vitamin K2 and its isomers of example 6 and comparative example 8;
FIG. 5 is a spectrum of the separation of lipids in the edible oil of example 7;
FIGS. 1-4 have time (time) in min on the abscissa and Absorbance (Absorbance) in mAU on the ordinate; the abscissa of fig. 5 is time (time) in min and the ordinate is current (current) in pA.
Detailed Description
The present invention will be described in detail with reference to the accompanying drawings, which illustrate embodiments of the present invention. In the following description, numerous specific details are set forth in order to provide a thorough understanding of the present invention. This invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein, as those skilled in the art will recognize without departing from the spirit and scope of the present invention.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The terminology used in the description of the invention herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. As used herein, the term "and/or" includes any and all combinations of one or more of the associated listed items.
The invention provides a silicon compound, the structural formula of which is shown as the formula (I):
wherein R is 1 、R 2 、R 3 Each independently selected from substituted or unsubstituted C 1 -C 4 Alkyl, and R 1 、R 2 And R 3 At least one of them is alkoxy, halogen, alkylamino or alkanoyl;
Y 1 is selected from C 2 -C 18 An alkylene group; y is 2 Is selected from C 20 -C 29 An alkyl group;
l is selected from
(amido group),. Or>
(sulfonamido),. Or>
(ureido) or->
(carbamate group).
Further, the alkoxy group may be selected from methoxy, ethoxy, propoxy, and the like; halogen may be selected from fluorine, chlorine, bromine, iodine, and the like; the alkylamino group can be selected from dimethylamino group, diethylamino group, etc.
In one embodiment, R 1 (or R) 2 Or R is 3 ) Selected from alkoxy, halogen, alkylamino or alkanoyl, R 2 、R 3 (or R) 1 、R 3 Or R is 1 、R 2 ) Each independently selected from alkoxy, halogen, alkylamino, alkanoyl, or substituted or unsubstituted C 1 -C 4 An alkyl group.
In one embodiment, R 1 、R 2 (or R) 2 、R 3 Or R is 1 、R 3 ) Each independently selected from alkoxy, halogen, alkylamino or alkanoyl; r is 3 (or R) 1 Or R is 2 ) Selected from alkoxy, halogen, alkylamino, alkanoyl, or substituted or unsubstituted C 1 -C 4 An alkyl group.
In one embodiment, R 1 、R 2 、R 3 Are respectively selected from alkoxy, halogen, alkylamino or alkanoyl.
Preferably, R 1 、R 2 、R 3 Each independently selected from alkoxy; y is 1 Is selected from C 2 -C 10 An alkylene group; y is 2 Is selected from C 25 -C 28 An alkyl group; l is selected from
In one embodiment, the silicon compound is selected from:
the preparation of the silicon compounds (L is selected from urea groups or urethane groups) provided by the present invention can be achieved by: in an anhydrous solvent (e.g., toluene), an isocyanatosilane (e.g., (3-isocyanopropyl) triethoxysilane) is reacted with a long chain alkyl alcohol (e.g., 1-octacosanol) or a long chain alkyl amine. Alternatively, the same objective is achieved by reacting a primary aminosilane, a secondary aminosilane or a silyl alcohol with a long chain alkyl isocyanate.
The preparation method provided by the invention is also suitable for Y 2 Is selected from C 30 -C 40 Preparation of the silicon compound of the alkyl group. The mixture containing long chains (C) is treated as described above 20 -C 29 Alkyl) compounds with long chains (C) 30 -C 40 Alkyl) compounds.
Further, the preparation method of the silicon compound (L is selected from urea group or urethane group) comprises the following steps:
the compound
Mixing the compound D-E with a solvent, and reacting at 60-80 ℃ for 20-24 h to obtain the compound D-E;
wherein R is 6 、R 7 、R 8 Are each independently selected from substituted or unsubstituted C 1 -C 4 Alkyl, and R 6 、R 7 And R 8 At least one of them is alkoxy, halogen, alkylamino or alkanoyl; a is selected from C 2 -C 18 An alkylene group;
d is selected from hydroxyl or amine (primary or secondary); e is selected from C 20 -C 29 An alkyl group.
In some embodiments, the solvent may be toluene, xylene, or like organic solvent.
The preparation method of the silicon compound provided by the invention has the advantages of easily obtained raw materials and simple and convenient route.
The invention also provides the use of the silicon compounds described above for the preparation of a stationary phase.
The silicon compound provided by the invention contains polar groups (amide groups, sulfonamide groups, carbamide groups or carbamate groups), and is bonded in a matrix to prepare a stationary phase, and chromatographic separation is carried out on the stationary phaseWhen the method is used, the silicon hydroxyl on the surface of the stationary phase can be effectively shielded, the peak shape of an alkaline compound is improved, and the method can be used in a mobile phase environment with higher water content; and the silicon compound is bonded with C 20 ～C 29 The long-chain alkyl group ensures that the prepared stationary phase has higher stereoselectivity, and has especially prominent separation effect on compounds with long chains and high structural similarity, especially on substances of vitamins and lipids.
The invention provides a stationary phase, and the preparation raw materials of the stationary phase comprise any one of the compounds and a matrix.
In the present invention, the above silicon compound may be covalently bonded to the substrate. Further, the above silicon compound may be covalently attached to the substrate in the form of a single layer or with multiple layers.
In one embodiment, the substrate is a solid substrate with silicon hydroxyl groups (Si-OH). In the present invention, the solid substrate may also be a non-porous material.
In the present invention, the solid matrix may be in various shapes such as granular shape, block shape, lamellar shape, etc. which are conventionally used in the prior art.
In the present invention, the solid substrate may be spherical, square, or irregular in shape.
Preferably, the solid matrix is selected from silica/organic hybrid microspheres and/or silica microspheres.
In one embodiment, the silica gel may be non-porous or of varying pore sizes. Preferably, the pore size is selected to be
Silica or silica/organic hybrid porous particles of (a).
Specifically, the preparation steps of the stationary phase are as follows: the above silicon compound is reacted with silica gel in an inert solvent such as toluene at a high temperature. Depending on the application, water, acid or base catalysts may be employed to enhance surface coverage; the terminal may be appropriately terminated as necessary.
In one embodiment, the stationary phase has a structural formula as shown in formula (II):
wherein the content of the first and second substances,
is a silica gel matrix, R 4 、R 5 Each independently selected from a hydroxyl group or an oxygen atom covalently attached to a silica gel matrix.
Preferably, the stationary phase is selected from:
the invention also provides the use of the above stationary phase in the detection and/or separation of compounds having a linear length greater than 16 atoms.
In one embodiment, compounds with linear lengths greater than 16 atoms include lipids, vitamins, and carotenes.
Vitamin K, also called thromboxane, is one of the vitamins, has phylloquinone bioactivity, which was first discovered and extracted in 1929 by danish chemist dam from animal livers and hemp oil. The vitamin K comprises K1, K2, K3, K4 and other forms, wherein the K1 and the K2 are naturally occurring and belong to fat-soluble vitamins; and K3 and K4 are artificially synthesized and are water-soluble vitamins. Vitamin K1 belongs to vitamin drugs, and is a substance necessary for liver to synthesize factors II, VII, IX and X. The vitamin K1 injection is a 2009 edition of national basic drug catalogue variety and is mainly used for treating hemorrhagic diseases caused by deficiency of various vitamin K. Due to the structural similarity, the separation of vitamins K1, K2 from their isomers is challenging and cannot be efficiently separated on C18.
Edible oils are lipids that are purified from plants and are usually liquids at room temperature. The main components are triglyceride, and small amount of free fatty acid and monoglyceride and diglyceride. The composition of edible oils is very complex due to the diversity of alkyl chain length, unsaturation, source, etc.
And the vitamin K1, the vitamin K2 and the lipid compound generally have strong hydrophobicity, so that the separation of the vitamin and the lipid compound in the prior art has great challenges. The inventor of the present invention has continuously studied to provide a silicon compound and prepare a stationary phase, the hydrophobic retention characteristic of which facilitates the rapid separation of the above-mentioned substances, and the stationary phase has a high stereoselectivity and a good separation effect for the separation of the above-mentioned substances. Similarly, the stationary phase provided by the invention is also suitable for separating other compounds with long chains and high structural similarity.
The invention also provides the use of any of the above stationary phases for the detection and/or separation of isomers.
The following are specific examples
Example 1
Synthesis of stationary phase P1
Under the protection of nitrogen, 12g of (3-isocyanopropyl) triethoxysilane is added to 100mL of a Toluene (Toluene) solution containing 24g of 1-octacosanol, then the reaction is stirred at 80 ℃ for 24 hours, and then the reaction solution is decompressed to remove the solvent, so that a silicon compound M1 is obtained.
10g of silica gel (particle size 5 μm, pore size
Surface area of 200m 2 /g) and 10g of silicon compound M1 are mixed with 50mL of toluene, ultrasonic treatment is carried out for 10min, then reflux reaction is carried out for 72h at 110 ℃, the mixed solution after reaction is filtered and washed by toluene, 1,4-dioxane and acetone in sequence, and then vacuum drying is carried out for 3h at 50 ℃, thus obtaining the stationary Phase P1 which is marked as Phase 1. The silica gel surface bonding density was calculated to be 2.0. Mu. Mol/m based on the elemental analysis data (13.3% C,0.55% N) of the bonding phase 2 。
Example 2
Synthesis of stationary phase P2
Under the protection of nitrogen, 1mol of (3-isocyanopropyl) triethoxysilane is added into 100mL of a toluene solution containing 1.1mol of 1-octacosylamine, then the mixture is stirred and reacted for 24 hours at 80 ℃, and then the reaction solution is decompressed and the solvent is removed to obtain a silicon compound M2.
10g of silica gel (particle size 5 μm, pore diameter: 10 g)
Surface area of 200m 2 And/g), 10g of silicon compound M2 and 50mL of toluene are mixed, ultrasonic treatment is carried out for 10min, then reflux reaction is carried out for 72h at 110 ℃, the mixed solution after reaction is filtered and washed by toluene, 1,4-dioxane and acetone in sequence, and then vacuum drying is carried out for 3h at 50 ℃, thus obtaining the stationary phase P2. The silica gel surface bonding density was calculated to be 2.1. Mu. Mol/m based on the elemental analysis data (13.0% C,1.2% N) of the bonding phase 2 。
Comparative example 1
Under nitrogen protection, 25g of n-triacontyldimethylchlorosilane and 10g of imidazole were added to 300mL of a solution containing 25g of silica gel (particle size 5 μm, pore diameter: 25 g)
Surface area of 200m 2 /g) at 100 ℃ for 24h, the reaction mixture was filtered, washed successively with toluene, 1,4-dioxane, water and acetone, and then dried in vacuo at 50 ℃ for 8h to give a C30 stationary phase.
Under the protection of nitrogen, 25g of trimethylchlorosilane and 10g of imidazole are added into 300mL of toluene dispersion liquid containing the intermediate, the mixture is stirred for 24 hours at 100 ℃, the reaction mixture is filtered, the mixture is washed by toluene, 1,4-dioxane, water and acetone in sequence, and then the mixture is dried for 8 hours in vacuum at 50 ℃ to obtain stationary phase P3, wherein the structure of the stationary phase is that
Is denoted as C30. The surface bonding density calculated from the elemental analysis data (13.01%; C) of the bonding phase was 2.0. Mu. Mol/m 2 。
Comparative example 2
25g of octadecyldimethylmethoxysilane and 1g of imidazole were added to 300mL of a solution containing 50g of silica gel (particle size 5 μm, pore diameter: 50 g) under nitrogen protection
Surface area of 200m 2 /g) at 100 ℃ for 24h, filtering the reaction mixture, washing with toluene, 1,4-dioxane, water and acetone in that order, and drying in a vacuum at 50 ℃ for 8h to give the stationary phase P4 which has the formula->
Denoted Polar C18. The surface bonding density calculated from the elemental analysis data (12.24% C,0.63% N) of the bonding phase was 2.9. Mu. Mol/m 2 。
Performance evaluation
The following tests of examples 3-7 employed the stationary Phase (Phase 1) prepared in example 1; testing of the following comparative examples the stationary phases prepared in comparative examples 1, 2 (C30, polar C18) were used; filling the stationary phase into a stainless steel column with the diameter of 4.6 multiplied by 150mm by adopting the traditional high-pressure slurry technology for testing;
the test samples in the following examples and comparative examples were formulated using the mobile phase used in the test method, except that the test sample of example 7 was formulated using isopropyl alcohol.
Hydrophobic retention test: the retention of the neutral hydrophobic compound by the stationary phase was evaluated, and the retention time of the neutral hydrophobic compound naphthalene was used as an index of the hydrophobic retention capacity, and a longer retention time indicates a stronger hydrophobic retention capacity.
Example 3
Test samples: a mixture of uracil (10. Mu.g/mL), dimethyl phthalate (0.1. Mu.L/mL), and naphthalene (300. Mu.g/mL);
and (3) testing conditions: mobile phase: CH (CH) 3 CN/H 2 O (60; flow rate: 1mL/min(ii) a Sample introduction amount: 5 mu L of the solution; temperature: 30 ℃; detection wavelength: 254nm;
comparative example 3
Essentially the same as in example 3, except that the stationary phase is C 30 。
Comparative example 4
Essentially the same as example 3, except that the stationary phase was Polar C18.
The results of the hydrophobicity retention test of example 3 and comparative examples 3 to 4 are shown in FIG. 1, and 1, 2, and 3 in FIG. 1 represent the peaks of uracil, dimethyl phthalate, and naphthalene, respectively.
As shown in FIG. 1, the naphthalene retention factor for the Phase1 test is 2.4, while the naphthalene retention factors for the C30 and Polar C18 tests are 3.0 and 3.2, respectively, under the same test conditions. It can be seen that under the conditions of comparable bonding density, due to the introduction of polar intercalating groups, phase1 has a weaker hydrophobic retention than C30, which does not contain polar intercalating groups; due to steric effects, phase1 has a lower bonding density than C18 with the same polar insertion group, resulting in a decrease in hydrophobic retention.
Since vitamin K1, vitamin K2 and lipid compounds have strong hydrophobicity, the hydrophobic retention property of Phase1 contributes to the rapid separation of such compounds.
And (3) stereoselectivity testing: stereoselectivity is represented by alpha 3/2 ＝(t 3 -t 0 )/(t 2 -t 0 )。
Example 4
Test samples: a mixture of uracil (40. Mu.g/mL), ortho-terphenyl (140. Mu.g/mL) and triphenylene (40. Mu.g/mL).
And (3) testing conditions are as follows: mobile phase: meOH/H 2 O (90; flow rate: 1mL/min; sample introduction amount: 5 mu L of the solution; column temperature: 30 ℃; detection wavelength: 254nm.
Comparative example 5
Essentially the same as example 4, except that the stationary phase is C 30 。
Comparative example 6
Essentially the same as example 4, except that the stationary phase is Polar C18.
The results of the stereoselectivity test of example 4 and comparative examples 5 to 6 are shown in FIG. 2, in which 1, 2 and 3 represent the peaks of uracil, ortho-terphenyl and triphenylene, respectively.
Alpha of stationary Phase1 is obtained by spectrogram analysis 3/2 α of =3.94, stationary phase C30 3/2 α of stationary phase Polar C18 =1.73 3/2 ＝2.36。
Although the surface bonding density of the stationary Phase1 is equivalent to that of the stationary Phase C30, the stereoselectivity of the stationary Phase1 is superior to that of the stationary Phase C30 due to the introduction of polar insertion groups and the length (35 atoms) of the bonded Phase.
In general, stereoselectivity increases with increasing surface bonding density. Although the surface bonding density of the stationary Phase1 is much lower than that of the stationary Phase Polar C18, the stereoselectivity is significantly higher due to the length (28 atoms) of the hydrophobic group of its bonded Phase. Therefore, the stationary phase provided by the invention is more beneficial to separating the substances to be separated with higher structural similarity.
Separation of vitamin K1 and its isomers
Example 5
Test samples: vitamin K1;
and (3) testing conditions are as follows: mobile phase: meOH/H 2 O (95; flow rate: 1mL/min; sample introduction amount: 10 mu L of the solution; column temperature: 25 ℃; detection wavelength: 254nm.
Comparative example 7
Essentially the same as in example 5, except that the stationary phase was C 30 。
The separation spectrum of vitamin K1 and its isomers of example 5 and comparative example 7 is shown in fig. 3, and the data of the spectrum analysis is shown in table 1.
TABLE 1
As can be seen from fig. 3 and table 1, the stationary phase C30 has a long detection time and a poor separation effect, and the separation degree is only 1.46; under the same test conditions, the separation degree of the stationary Phase1 is obviously improved (3.62), and the separation speed is doubled.
Separation of vitamin K2 and isomers thereof
Example 6
Test samples: vitamin K2;
and (3) testing conditions are as follows: mobile phase, meOH/H 2 O (95; flow rate: 1mL/min; sample introduction amount: 10 mu L of the solution; column temperature: 25 ℃; detection wavelength: 254nm.
Comparative example 8
Essentially the same as example 6, except that the stationary phase was C 30 。
The separation profile of vitamin K2 and its isomers of example 6 and comparative example 8 is shown in fig. 4, and the profile analysis data is shown in table 2.
TABLE 2
As can be seen from table 2, the stationary Phase C30 achieved baseline separation (degree of separation of 2.41), while the stationary Phase1 achieved better degree of separation in a shorter time under the same test conditions (3.15).
Separation of lipids from edible oils
Example 7
Test samples: a mixture of olive oil (5 mg/mL), sesame oil (5 mg/mL), peanut oil (5 mg/mL) and soybean oil (5 mg/mL).
And (3) testing conditions: mobile phase: a: CH (CH) 3 CN, B:100mM ammonium acetate (NH) 4 OAc) buffer, ph5.0, C: and (3) isopropanol.
Gradient elution procedure: injecting the test sample after equilibrating for 10min with mobile phase 85% A, 5%B and 10% C0-10min, 85-65A, 5%B and 10-30% C10-60min, 65-20% A, 5%B and 30-75% C60-70min, 20-5%A, 5%B and 75-90% C, and then maintaining for 10min again at 5%A, 5%B and 90% C;
flow rate: 1mL/min; sample injection amount: 5 mu L of the solution; temperature: 30 ℃; the detection method comprises the following steps: the light is scattered by evaporation.
As can be seen from the analysis chart of fig. 5, the stationary Phase1 has high stereoselectivity and good separation degree for the components with similar structures in the edible oil.
The technical features of the embodiments described above may be arbitrarily combined, and for the sake of brevity, all possible combinations of the technical features in the embodiments described above are not described, but should be considered as being within the scope of the present specification as long as there is no contradiction between the combinations of the technical features.
The above-mentioned embodiments only express several embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present patent shall be subject to the appended claims.
Claims (10)
1. A stationary phase, wherein the stationary phase has a structural formula as shown in formula (II):
wherein the content of the first and second substances,
is a silica gel matrix, R 4 Selected from hydroxy, R 5 Selected from the group consisting of oxygen atoms covalently attached to the silica gel matrix;
Y 1 is selected from C 2 -C 18 An alkylene group; y is 2 Is selected from C 20 -C 29 An alkyl group;
l is selected from
2. A stationary phase according to claim 1, characterized in that Y is 1 Is selected from C 2 -C 10 An alkylene group; y is 2 Is selected from C 25 -C 28 An alkyl group; l is selected from
3. A stationary phase according to any of the claims 1 or 2, wherein the stationary phase is selected from the group consisting of:
4. the stationary phase according to any of claims 1 or 2, wherein the raw materials for preparing the stationary phase comprise a silicon compound and a matrix.
5. The stationary phase according to claim 4, wherein the silicon compound has a structural formula shown in formula (I):
wherein R is 1 、R 2 、R 3 Are each independently selected from substituted or unsubstituted C 1 -C 4 Alkyl, and R 1 、R 2 And R 3 At least one of them is alkoxy, halogen, alkylamino or alkanoyl.
6. A stationary phase according to claim 5, characterized in that in the silicon compound, R 1 、R 2 、R 3 Are respectively provided withIndependently selected from alkoxy groups.
7. A stationary phase according to any of the claims 5 to 6, characterized in that the silicon compound is selected from the group consisting of:
8. use of a stationary phase according to any of claims 1 to 7 for the detection and/or separation of compounds having a linear length of more than 16 atoms.
9. Use according to claim 8, wherein the compounds comprise lipids, vitamins and carotenes.
10. Use of a stationary phase according to any of claims 1 to 7 for the detection and/or separation of isomers.
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