阅读说明：本技术 一种多糖衍生物涂覆型手性固定相的制备工艺 (Preparation process of polysaccharide derivative coating type chiral stationary phase ) 是由 吴海波 周永正 梁新建 于 2018-07-10 设计创作，主要内容包括：本发明公开了一种多糖衍生物涂覆型手性固定相的制备工艺,该工艺以烷基化的大孔硅胶作基质,加入溶解多糖衍生物的四氢呋喃、二氯甲烷或氯仿溶液,再加入一定量添加剂,如N,N’-二甲基甲酰胺、苯酚、苯甲酸甲酯、苯乙酮或硝基苯,拌匀后加入玻璃珠,于旋转蒸发仪上将溶剂缓慢旋干,加乙醇分散,过滤,洗涤,乙醇回流,过滤,干燥,得到最终手性固定相。本工艺制备的固定相稳定性好,适合正、反相条件下的分离,多糖衍生物涂层均匀,填料分散性好,便于放大生产。通过调节涂覆时添加剂的种类或者量,甚至可以改善某些对映体在固定相上的分离,对于工业制备分离具有较大意义。(The invention discloses a process for preparing a polysaccharide derivative coating type chiral stationary phase, which comprises the steps of taking alkylated macroporous silica gel as a substrate, adding tetrahydrofuran, dichloromethane or chloroform solution for dissolving the polysaccharide derivative, adding a certain amount of additives such as N, N' -dimethylformamide, phenol, methyl benzoate, acetophenone or nitrobenzene, adding glass beads after uniformly stirring, slowly spin-drying the solvent on a rotary evaporator, adding ethanol for dispersion, filtering, washing, refluxing the ethanol, filtering and drying to obtain the final chiral stationary phase. The stationary phase prepared by the process has good stability, is suitable for separation under positive and reverse phase conditions, has uniform polysaccharide derivative coating and good filler dispersibility, and is convenient for large-scale production. By adjusting the type or amount of the additive during coating, the separation of certain enantiomers on the stationary phase can be even improved, which is of great significance for industrial preparative separations.)

1. A preparation process of a polysaccharide derivative coating type chiral stationary phase is characterized in that: taking alkylated macroporous silica gel as a matrix, adding tetrahydrofuran, dichloromethane or chloroform solution for dissolving polysaccharide derivatives, adding a certain amount of additive, stirring uniformly, adding a certain amount of glass beads, spin-drying the solvent on a rotary evaporator, adding ethanol for dispersion, filtering, washing, refluxing the ethanol, filtering, and drying to obtain the final chiral stationary phase.

2. The process according to claim 1, wherein the polysaccharide derivative is amylose-tris (3, 5-dimethylphenylcarbamate), cellulose-tris (4-methylbenzoate), amylose-tris [ (S) - α -methylbenzylcarbamate ], amylose-tris (3, 5-dichlorophenylcarbamate), cellulose-tris (4-chlorobenzoate), cellulose-tribenzoate, amylose-tris (5-chloro-2-methylphenylcarbamate), cellulose-tris (3-chloro-4-methylphenylcarbamate) or cellulose-tris (4-chloro-3-methylphenylcarbamate).

3. The process for preparing a polysaccharide derivative-coated chiral stationary phase according to claim 1, wherein: the stationary phase takes alkylated macroporous silica gel as a substrate, silane for modifying the silica gel comprises one or more alkyl groups, the length of an alkyl carbon chain is 1-10, and the silane is bonded with the silica gel through reaction of methoxyl, ethoxyl or chlorine on the silicon; the silica gel matrix is modified with one or more silanes.

4. A process for preparing a polysaccharide derivative-coated chiral stationary phase according to claim 3, wherein: the silane for modifying the silica gel is one or more of n-octyl trichlorosilane, n-octyl dimethylchlorosilane and trimethylchlorosilane.

5. The process for preparing a polysaccharide derivative-coated chiral stationary phase according to claim 1, wherein: the additive is N, N' -dimethylformamide, phenol, methyl benzoate, acetophenone or nitrobenzene.

6. The process for preparing a polysaccharide derivative-coated chiral stationary phase according to any one of claims 1 to 5, comprising the steps of:

① dissolving polysaccharide derivative in tetrahydrofuran, dichloromethane or chloroform 8-20 times (v/w), adding alkylated silica gel 3-5 times by mass, adding additive 0-2 times by mass, stirring, and adding glass beads 3-10mm 4-8 times by mass;

② evaporating the solvent of the mixture to dryness on a rotary evaporator, controlling the rotation speed at 10-60 rpm, and spin-drying the solvent within 2-8 hr by adjusting the temperature and vacuum degree;

③ adding 1-3 times volume (v/w) of ethanol to disperse the stationary phase, filtering, and washing with 1-2 times volume of ethanol;

④ dispersing the stationary phase with 2-5 times volume of ethanol, refluxing for 2-5 hr, filtering, and drying to obtain the final chiral stationary phase.

Technical Field

The invention relates to the technical field of chiral stationary phases, in particular to a preparation process of a polysaccharide derivative coating type chiral stationary phase.

Background

Biological macromolecules, such as proteins, polysaccharides, nucleic acids and enzymes, which are important bases of life activities, are almost all chiral, and thus many biochemical reactions closely related to organisms are associated with the chirality of substances. The enantiomers of chiral chemical drugs have significant differences in pharmacological activity, metabolic processes and toxicity in the human body. Chiral herbicides, insecticides and plant growth regulators which are widely used in agriculture also exhibit strong biological recognition effects. For this reason, the Food and Drug Administration (FDA) issued chiral drug guidelines in 1992, requiring all new racemic drugs to be marketed in the united states, the producers must provide reports that illustrate the pharmacological, toxic and clinical effects of the individual enantiomers contained in the drug. Therefore, the separation and analysis of chiral compounds are becoming more and more important in the development and production of pharmaceuticals.

Polysaccharide derivative-coated stationary phases are the most widely recognized class of stationary phases for chiral recognition, especially those coated with amylose-tris (3, 5-dimethylphenylcarbamate), cellulose-tris (4-methylbenzoate), and amylose-tris [ (S) - α -methylbenzylcarbamate ] are the most selective, and in total, can separate approximately 90% of the enantiomers.

The biggest problem in the preparation of the stationary phase is that the uniformity of the macromolecular polysaccharide coating is not easy to ensure, and particularly in the amplification preparation, when the solvent is nearly evaporated to dryness, the local solvent is evaporated too fast, so that the coating is not uniform and the caking phenomenon is caused. For this reason, many reports have used an aminated silica gel as a substrate, enhanced adsorption of the coating by dipolar, hydrogen bonding of the amino group, and improved uniformity of the coating by multiple coating. However, the aminated silica gel is easy to undergo self-corrosion, particularly has poorer stability under reverse phase conditions, and is easy to have stronger action with certain samples so as to influence the peak shape and separation; multiple coating increases the amount of work and solvent used in the preparation. Thus, the process for preparing the polysaccharide derivative-coated stationary phase remains to be improved.

Disclosure of Invention

The invention discloses a preparation process of an improved polysaccharide derivative coating type chiral stationary phase, aiming at improving the stability and coating uniformity of the polysaccharide derivative coating type stationary phase.

The technical scheme of the invention is as follows:

a preparation process of a polysaccharide derivative coating type chiral stationary phase is characterized in that: taking alkylated macroporous silica gel as a matrix, adding tetrahydrofuran, dichloromethane or chloroform solution for dissolving polysaccharide derivatives, adding a certain amount of additive, stirring uniformly, adding a certain amount of glass beads, slowly spin-drying the solvent on a rotary evaporator, adding ethanol for dispersion, filtering, washing, refluxing the ethanol, filtering, and drying to obtain the final chiral stationary phase.

The preparation process of the polysaccharide derivative coating type chiral stationary phase is characterized by comprising the following steps:

the polysaccharide derivative is amylose-tris (3, 5-dimethylphenylcarbamate), cellulose-tris (4-methylbenzoate), amylose-tris [ (S) - α -methylbenzylcarbamate ], amylose-tris (3, 5-dichlorophenylcarbamate), cellulose-tris (4-chlorobenzoate), cellulose-tribenzoate, amylose-tris (5-chloro-2-methylphenylcarbamate), cellulose-tris (3-chloro-4-methylphenylcarbamate) or cellulose-tris (4-chloro-3-methylphenylcarbamate).

The stationary phase takes alkylated macroporous silica gel as a substrate, silane for modifying the silica gel comprises one or more alkyl groups, the length of an alkyl carbon chain is 1-10, and the silane is bonded with the silica gel through reaction of methoxyl, ethoxyl or chlorine on the silicon; the silica gel matrix is modified with one or more silanes. The silane for modifying the silica gel is preferably one or more of n-octyltrichlorosilane, n-octyldimethylchlorosilane and trimethylchlorosilane.

The additive is N, N' -dimethylformamide, phenol, methyl benzoate, acetophenone or nitrobenzene.

The preparation process of the polysaccharide derivative coating type chiral stationary phase is characterized by comprising the following steps:

① dissolving polysaccharide derivative in tetrahydrofuran, dichloromethane or chloroform 8-20 times (v/w), adding alkylated silica gel 3-5 times by mass, adding additive 0-2 times by mass, stirring, and adding glass beads 3-10mm 4-8 times by mass;

② evaporating the solvent of the mixture to dryness on a rotary evaporator, controlling the rotation speed at 10-60 rpm, and spin-drying the solvent within 2-8 hr by adjusting the temperature and vacuum degree;

③ adding 1-3 times volume (v/w) of ethanol to disperse the stationary phase, filtering, and washing with 1-2 times volume (v/w) of ethanol;

④ dispersing the stationary phase with 2-5 times volume (v/w) of ethanol, refluxing for 2-5 hr, filtering, and drying to obtain the final chiral stationary phase.

The substrate for coating in the process is alkylated silica gel, has good stability and is suitable for separation under the positive and reverse phase conditions. Certain additive is added during coating, so that the evaporation speed of the solvent can be slowed down in the later stage of rotary evaporation, the mass transfer of the polysaccharide derivative on the surface of the silica gel is improved, and the uniformity of the coating is ensured. In addition, glass beads are added during coating, so that the flowability during stationary phase coating is enhanced, and the uniformity of the coating can be improved. The separation of certain enantiomers on a stationary phase can be even improved by adjusting the type or the dosage of the additive, and the method has great significance for industrial preparation and separation.

The polysaccharide derivative stationary phase coated by the process has uniform coating, good filler dispersibility and convenient amplification production.

Drawings

FIG. 1 is a scanning electron micrograph of the stationary phase prepared in example 1 at 1 kXmagnification;

FIG. 2 is a scanning electron micrograph of the stationary phase prepared in example 1 magnified 3 ten thousand times;

FIG. 3 is a chromatogram of the separation of N- (2, 3-epoxypropyl) phthalimide after the stationary phase prepared in example 1 was packed in a column (column 4.6 mm. times.150 mm, acetonitrile/water. times. 40/60, flow rate 0.5mL/min, wavelength 254nm, column temperature 25 ℃);

FIG. 4 is a chromatogram showing the separation of zopiclone racemate after column packing in the stationary phase prepared in example 2 (column 4.6 mm. times.150 mm, acetonitrile/0.01M Na₂HPO₄(pH 6.5) 43/57, flow rate 0.5mL/min, wavelength 305nm, column temperature 25 ℃);

FIG. 5 is a simulated preparative spectrum of the ketorolac racemate after the stationary phase prepared in example 6 was packed in a column (column 4.6 mm. times.250 mm, methanol + 0.1% formic acid, flow rate 1mL/min, wavelength 254nm, single needle loading 10 mg).

Detailed Description

The following examples are provided to illustrate the improvement of the preparation process of a polysaccharide derivative-coated chiral stationary phase of the present invention.