Purification method of dipeptide diaminobutyryl benzylamide diacetate
During the purification process of dipeptide diamine butyryl benzylamide diacetate, the combination technology of medium-pressure purification preparation machine and PDA detector was used to solve the problem of by-products affecting product purity, and efficient and accurate purification effect was achieved.
Patent Information
- Application Number
- CN202411943822.3
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-12-27
- Publication Date
- 2025-04-08
AI Technical Summary
The prior art is difficult to effectively remove by-products in dipeptide diaminobutyryl benzylamide diacetate, resulting in difficulty in improving product purity.
Using a medium-pressure purification preparation machine and PDA detector, the ratio of the mobile phase is accurately controlled to achieve efficient purification of the product through gradient elution program and chromatography separation technology.
It effectively improves the purification efficiency and purity of the product, simplifies the operating process, improves the purification yield, and has good stability and practical application value.
Smart Images
Figure CN119775343A_ABST
Abstract
Description
Technical Field
[0001] The invention relates to a method for purifying amide diacetate, in particular to a method for purifying dipeptide diaminobutyryl benzyl amide diacetate. Background Art
[0002] As the main intermediate of beauty cosmetics, dipeptide diaminobutyryl benzylamide diacetate has strict control over the by-products produced during production, and the product purity is ≥99%. The conventional impurity removal technology is to use the different solubility of the solvent to the product and impurities for pulping and purification. If the selected solvent can dissolve the product but it is difficult to dissolve the impurities, the product is dissolved in the solvent and filtered, and then the filtrate is concentrated and crystallized into the final product; if the solvent can dissolve the impurities but it is difficult to dissolve the product, the solvent is used to pulp and filter the product, and finally the filtered solid is dried to obtain the finished product; now the product and impurities have similarities, and different solvents have been tried for purification. Basically, either they can be dissolved or they cannot be dissolved. Conventional purification methods are difficult to effectively remove impurities and improve product purity. Summary of the invention
[0003] The present invention provides a method for purifying dipeptide diaminobutyryl benzylamide diacetate, which mainly solves the technical problem that by-products generated during production affect the purity of the main components of the product, thereby accurately controlling the residual amount of the by-products and the like.
[0004] The present invention adopts the following technical scheme: a method for purifying dipeptide diaminobutyryl benzylamide diacetate, comprising the following steps: Step 1, prepare the reagents: acetonitrile-preparative grade, deionized water, chromatography grade-formic acid;
[0005] Step 2: Sample pretreatment: Weigh a certain amount of sample and place it in a beaker, add acetonitrile, dissolve it by ultrasonic at room temperature, then add deionized water, and finally filter the sample with a polypropylene organic filter membrane before entering the preparation column; Step 3: separation by preparation machine: the preparation conditions are room temperature of the sample, manual sample loading, preparation column: Claricep Flash Spherical C18, flow rate: 50 mL / min, mobile phase A is 0.1% molar concentration of formic acid in water; mobile phase B is 0.1% mass percentage concentration of formic acid in acetonitrile, and gradient elution program is used.
[0006] The preparation machine used is the Fenome Astra medium pressure purification preparation machine, which uses a PDA detector. The preparation column model and specifications are Claricep Flash Spherical C18, 20-35µm 100 A, 330g / column. The gradient elution program is: 0-20min, the volume ratio of mobile phase B is from 25% to 85%, 20-20.1 min, the volume ratio of mobile phase B is from 85% to the initial ratio of 25%, 20.1-22min, the volume ratio of mobile phase B is from 25% to the equilibrium system.
[0007] The present invention has the following beneficial effects: the present invention mainly utilizes the precise flow rate of the medium-pressure purification preparation machine, and then monitors through the PDA detector, emulating the chromatographic separation technology, to solve the purification problem of impurities produced in conventional chemical reactions and products under the conditions of similar polarity and solubility, thereby effectively improving the effectiveness of the product in the purification process, and also improving the purification yield. This technology is simple, fast, accurate, and has good stability. BRIEF DESCRIPTION OF THE DRAWINGS
[0008] Figure 1 LCMSC profile of the product before preparation and purification.
[0009] Figure 2 LCMSC profile after product preparation and purification. DETAILED DESCRIPTION
[0010] Main instruments: Feinomie Astra medium-pressure purification preparation machine, equipped with PDA detector, conventional preparation column Claricep FlashSpherical C18; ultrasonic extractor (KQ5200E, Zhengzhou Ketai Experimental Equipment Co., Ltd.), 0.45 μm polypropylene organic filter membrane (Shanghai Anpu Scientific Instrument Co., Ltd.), electronic analytical balance (CPA225D, Sartorius Scientific Instrument (Shanghai) Co., Ltd.).
[0011] Main reagents: The reagents used for the measurement included deionized water; acetonitrile (preparative grade, Titan Chemicals), formic acid (chromatographic grade, Shanghai Sinopharm); and conventional deionized water.
[0012] Sample pretreatment: Weigh the dipeptide diaminobutyryl benzylamide diacetate sample and place it in a 100 ml beaker, add 70 ml of acetonitrile, dissolve it by ultrasonic at room temperature, then add deionized water to dissolve it, and finally filter the sample with a polypropylene organic filter membrane and load it into the column, 10 ml each time.
[0013] Preparation conditions: The preparation room is at room temperature, and the preparation machine uses a PDA detector; the preparation column model and specifications are: Claricep FlashSpherical C18, 20-35µm 100 A, 330g / column. The gradient preparation procedure is: 0-20min, the volume ratio of mobile phase B is from 25% to 85%, 20-20.1 min, the volume ratio of mobile phase B is from 85% to the initial ratio of 25%, 20.1-22min, the volume ratio of mobile phase B is from 25% to the equilibrium system. Currently, dipeptide diaminobutyryl benzylamide diacetate is a powdered solid on the market, and the sample measured in the present invention is a product produced by Jier Biochemical (Shanghai) Co., Ltd.
[0014] The temperature of the preparation chamber of the present invention is set to room temperature, which is beneficial to the preservation of the acetonitrile solvent in the preparation chamber.
[0015] LCMSC spectrum of the product before purification Figure 1 As shown, the method was used to purify the dipeptide diaminobutyryl benzylamide diacetate product, and the LCMSC spectrum was shown in Figure 2 ,The results show that this method is fast, accurate, highly sensitive, ,stable, and has practical application value.
Claims
1. A method for purifying dipeptide diaminobutyryl benzylamide diacetate; characterized in that: The following steps are involved: Step 1: Prepare the reagents used in the preparation: acetonitrile-preparative grade, deionized water, chromatographic grade-formic acid; Step 2, sample processing: weigh a certain amount of sample and place it in a beaker, add acetonitrile, dissolve it by ultrasonic at room temperature, then add deionized water, and finally filter the sample with a polypropylene organic filter membrane and enter the preparation column; Step 3: separation by preparation machine: the preparation conditions are that the sample chamber temperature is room temperature, manual sample loading is adopted, the preparation column is: preparation column Claricep Flash Spherical C18, flow rate: 50 mL / min, mobile phase A is 0.1% molar concentration of formic acid aqueous solution; mobile phase B is 0.1% mass percentage concentration of formic acid acetonitrile solution, and a gradient elution program is adopted.
2. The method for purifying dipeptide diaminobutyryl benzylamide diacetate according to claim 1, characterized in that: The preparation machine is a Fenome Astra medium-pressure purification preparation machine, which uses a PDA detector.
3. The method for purifying dipeptide diaminobutyryl benzylamide diacetate according to claim 1, characterized in that: The specifications of the preparation column are Spherical, 20-35µm 100 A, 330g / column.
4. The method for purifying dipeptide diaminobutyryl benzylamide diacetate according to claim 1, characterized in that: The gradient elution program is as follows: 0-20 min, the volume ratio of mobile phase B increases from 20% to 85%, 20-20.1 min, the volume ratio of mobile phase B decreases from 85% to an initial ratio of 25%, and 20.1-22 min, the volume ratio of mobile phase B decreases from 25% to a balanced system.