CN116465985A - Method for detecting residual solvent in alpha-ribose mesylate by gas chromatography - Google Patents

Abstract

The invention discloses a method for detecting residual solvent in alpha-ribose mesylate by using gas chromatography, which belongs to the technical field of chemical detection, wherein the residual solvent in the alpha-ribose mesylate is detected by using the gas chromatography, the instrument is a gas chromatograph, the detector is a FID detector, a chromatographic column adopts a DB-WAX quartz capillary chromatographic column, and carrier gas is N ₂ The flow rate is 3.0mL/min, the split ratio is 20:1, and the column temperature is as follows: maintaining at 40deg.C for 10 min, heating to 80deg.C at 10deg.C per minute, heating to 250deg.C per minute for 3 min, introducing sample into the headspace, balancing temperature at 90deg.C, quantifying ring temperature at 100deg.C, and transmitting line temperature at 110deg.C for 30min; sample injection volume: 1.0ml, can effectively separate the residual solvent in the alpha-ribose mesylate and quantitatively detect the content of the residual solvent, and the method has the advantages of simple operation, strong specificity, high sensitivity, high accuracy, good linearity and durability, and can accurately and rapidly determine the alpha-room ratioResidual amount of 8 solvents in ribomethane sulfonate.

Description

A method for detecting residual solvent in α-ribose mesylate by gas chromatography

technical field

The invention belongs to the technical field of chemical detection, in particular to a method for detecting residual solvent in α-ribose mesylate by gas chromatography.

Background technique

α-Ribose mesylate is an important starting material in the synthesis of gemcitabine hydrochloride raw materials. Gemcitabine hydrochloride is an anticancer drug widely used in clinical practice. Its injections are listed in many countries. It is particularly important to control the quality of starting materials at the source. At present, there is no report on the detection method of residual solvents in α-ribose mesylate. Residual organic solvents in α-ribose mesylate have a certain impact on product quality. Yes, after a long period of research, we have finally developed a method that can detect residual solvents in α-ribose mesylate, and this method is easy to operate and has better detection results.

Contents of the invention

For the deficiencies in the prior art, the object of the present invention is to provide a method for gas chromatography detection of residual solvent in α-ribose mesylate, comprising the following steps:

1) Selection of instrument and chromatographic conditions: gas chromatograph; the chromatographic column is DB-WAX (30m×0.53mm×1.0μm) capillary column; using FID detector, the initial temperature of the chromatographic column is 35-45°C, the holding time is 8-12min, and then the temperature is raised to 80°C at a rate of 8-12°C/min, and then the final temperature of the column is raised to 250°C at a rate of 35-45°C/min. The time is 2-5min; the carrier gas is nitrogen, the carrier gas flow rate is 2.5-3.5ml/min, the split ratio is 20:1; the inlet temperature is 190-210°C, the detector temperature is 240-260°C; the sampling method is headspace injection, the equilibrium temperature is 90°C, the loop temperature is 100°C, the transfer line temperature is 110°C, and the equilibrium time is 30min; the injection volume is 1.0mL.

2) Prepare the solution and prepare the following solutions:

a. blank solution (diluent): dimethyl sulfoxide;

b. Reference substance solution: Weigh 0.5g of acetone, 0.06g of dichloromethane, 0.072g of tetrahydrofuran, 0.5g of ethyl acetate, 0.089g of toluene, 0.5g of diethyl ether, 0.041g of acetonitrile, and 0.1g of ethyl difluorobromoacetate, put them in a 50ml measuring bottle, dilute to the mark with a diluent, shake well, and use it as a stock solution; pipette 2.0ml of the stock solution and put it in a 100ml measuring bottle , dilute to the mark with a diluent, shake well, then accurately measure 5.0ml, place it in a 20ml headspace bottle, and seal it with a cap. C. The test solution: take 0.2g of α-ribose mesylate, put it in a 20ml headspace bottle, add 5.0ml of diluent to dissolve, seal it with a cap, shake well, and prepare 2 parts in parallel.

Inject samples according to the following sequence table:

name Number of needles blank solution ≥1 needle Reference solution 6 pins Test solution 1 stitch each

Each solvent in the α-ribose mesylate solution of the test product is calculated by the peak area by adding an external standard method.

Preferably, the selected chromatographic conditions for gas chromatography analysis of the present invention are as follows:

Using FID detector;

Chromatographic column: DB-WAX (30m×0.53mm×1.0μm) capillary column;

Programmed temperature rise: the initial temperature is 40°C, maintained for 10 minutes, raised to 80°C at a rate of 10°C per minute, and then raised to 250°C at a rate of 40°C per minute and maintained for 3 minutes.

Carrier gas: nitrogen, flow rate 3.0mL/min, split ratio: 20:1;

Injection port temperature: 200°C;

Detector (FID) temperature: 250°C;

Sampling method: headspace sampling, equilibration temperature 90°C, quantitative loop temperature 100°C, transfer line temperature 110°C, equilibration time 30min;

Injection volume: 1.0ml.

The beneficial effects of the present invention are as follows:

The present invention can effectively separate the residual solvent in α-ribose mesylate and quantitatively detect its content. The residual solvent is ether, acetone, tetrahydrofuran, ethyl acetate, dichloromethane, acetonitrile, toluene, and ethyl difluorobromoacetate. The detection of residual solvents in ribose mesylate provides a reliable detection method, which is suitable for general promotion and application.

Description of drawings

Fig. 1 is blank solution chromatogram;

Fig. 2 is reference substance solution chromatogram;

Fig. 3 is need testing solution chromatogram;

Fig. 4 is each solvent linearity test result figure;

Fig. 5 is the accuracy solution chromatogram.

Detailed ways

The present invention will further supplement the above content with reference to the embodiments (accompanying drawings), but it should not be understood that the scope of the present invention is limited to the following examples.

The instrument and chromatographic conditions adopted in the following examples are as follows:

The gas chromatograph is Agilent7890A, using FID detector, the initial temperature of the column is 35-45°C, the holding time is 8-12min, then it is raised to 80°C at a heating rate of 8-12°C/min, and then the final temperature of the column is raised to 250°C at a heating rate of 35-45°C/min, and the holding time of the final temperature of the column is 2-5min; the carrier gas is nitrogen. The flow ratio is 20:1; the inlet temperature is 190-210°C, the detector temperature is 240-260°C; the injection method is headspace injection, the equilibrium temperature is 90°C, the quantitative loop temperature is 100°C, the transfer line temperature is 110°C, and the equilibration time is 30 minutes; the injection volume is 1.0mL.

Example 1

Specificity investigation, as shown in Figure 1-2 and Figure 5:

Precisely measure 1.0 mL of the blank solution, the reference solution, and each impurity positioning solution, inject them into the gas chromatograph, and record the chromatograms. The results of the impurity location and resolution experiments show that the blank has no interference, and other solvents have no interference to the sample determination, and the minimum resolution between adjacent solvents is 2.60. The specificity of the analysis conditions is good, and the test results are as follows:

Example 2

Sensitivity test investigation results:

The limit of quantitation (LOQ) results are as follows:

The limit of detection (LOD) results are as follows:

The determination results of quantification limit and detection limit of each solvent show that each solvent has high detection sensitivity (far higher than the limit standard value), and the sensitivity of the method meets the requirements.

Example 3

Linear investigation, as shown in Figure 4:

The peak area is plotted against the concentration, the concentration of ether is in the range of 40.664μg/mL-304.980μg/mL, the concentration of acetone is in the range of 40.120μg/mL-300.900μg/mL, the concentration of tetrahydrofuran is in the range of 5.928μg/mL-44.460μg/mL, and the concentration of ethyl acetate is in the range of 40.160μg/mL-301.200μg In the range of 5.128μg/mL-38.460μg/mL for dichloromethane, in the range of 3.232μg/mL-24.240μg/mL for acetonitrile, in the range of 7.016μg/mL-52.620μg/mL for toluene, in the range of 8.792μg/mL-65.940μg/mL for ethyl difluorobromoacetate, the sample concentration and The peak area has a linear relationship, and the linearity is good. The specific results are as follows:

name linear equation R ² Ether y=0.2849x-1.0310 0.9993 acetone y=0.0526x-0.2046 0.9996 Tetrahydrofuran y=0.0698x-0.0407 0.9996 ethyl acetate y=0.0393x-0.1650 0.9996 Dichloromethane y=0.0116x-0.0067 0.9995 Acetonitrile y=0.0154x-0.0052 0.9987 toluene y=0.0532x-0.0527 0.9994 Ethyl difluorobromoacetate y=0.0046x-0.0058 0.9993

Example 4

Accuracy inspection, the chromatogram is shown in Figure 5:

Respectively add samples to prepare recovery rate solutions according to the limit concentrations of ether, acetone, tetrahydrofuran, ethyl acetate, dichloromethane, acetonitrile, toluene, and ethyl difluorobromoacetate at 50%, 100%, and 150%. Prepare 3 parts in parallel for each group of concentrations, prepare a part of the test solution and a part of the reference solution at the same time, measure according to the above-mentioned chromatographic conditions, and calculate the recovery rate. Each solvent recovery rate is in the range of 90%-110%, the results are shown in the following table:

Example 5

To inspect the durability, replace the chromatographic column, change the initial column temperature (40±1°C), carrier gas flow rate (3.0±0.1ml/min), injection port temperature (200±2°C), and heating rate (±1°C/min), compare the RSD of the peak area of each solvent in the reference solution and the minimum resolution of each solvent’s peak separation, and investigate its durability. Changing the parameters of the chromatographic conditions, the maximum RSD of the 6-pin peak area of each solvent in the reference solution was 6.7%, which was less than 10%. The results are shown in the table below:

The above is only a preferred embodiment of the present invention. It should be pointed out that those skilled in the art should understand that some improvements can be made without departing from the principles of the present invention.

Claims (5)

1. A method for detecting residual solvent in alpha-ribose mesylate by gas chromatography, comprising the steps of:

1) Selection of instrument and chromatographic conditions: a gas chromatograph; the chromatographic column is DB-WAX (30 m×0.53mm×1.0 μm) capillary chromatographic column; using a FID detector;

1) Preparation of a control solution: weighing 0.5g of acetone, 0.06g of dichloromethane, 0.072g of tetrahydrofuran, 0.5g of ethyl acetate, 0.089g of toluene, 0.5g of diethyl ether, 0.041g of acetonitrile and 0.1g of difluorobromoacetic acid ethyl ester, placing into a 50ml measuring flask, diluting to a scale with a diluent, shaking uniformly to serve as a stock solution, transferring 2.0ml of the stock solution, placing into a 100ml measuring flask, diluting to the scale with the diluent, shaking uniformly, precisely weighing 5.0ml of the stock solution, placing into a 20ml overhead flask, and sealing by capping;

2) Preparation of test solution: taking 0.2g of alpha-ribose mesylate, placing the alpha-ribose mesylate into a 20ml headspace bottle, adding 5.0ml of diluent for dissolution, capping, sealing and shaking uniformly to obtain the finished product;

3) Determination of residual solvent: taking a reference substance solution and a sample solution, carrying out headspace sampling, carrying out gas chromatography analysis to obtain peak areas of all solvents, and calculating according to an external standard method by the peak areas to obtain the chromatographic column temperature change flow in gas phase detection, wherein the chromatographic column temperature change flow is as follows: the initial temperature of the chromatographic column is 35-45 ℃, the retention time is 8-12min, then the temperature is increased to 80 ℃ at the heating rate of 8-12 ℃/min, and the final temperature of the chromatographic column is 250 ℃ at the heating rate of 35-45 ℃/min, and the retention time of the final temperature of the chromatographic column is 2-5min.

2. The method for detecting residual solvent in α -ribose mesylate by gas chromatography according to claim 1, wherein the diluent is dimethyl sulfoxide solution.

3. The method for detecting residual solvent in α -ribosylate by gas chromatography according to claim 1, wherein: the carrier gas in the gas chromatography detection is nitrogen, the flow rate of the carrier gas is 2.5-3.5ml/min, and the split ratio is 20:1.

4. The method for detecting residual solvent in α -ribosylate by gas chromatography according to claim 1, wherein: the temperature of the sample inlet in the gas chromatography detection is 190-210 ℃, and the temperature of the detector is 240-260 ℃.

5. The method for detecting residual solvent in α -ribosylate by gas chromatography according to claim 1, wherein: the sample injection mode in the gas chromatography detection is headspace sample injection, the equilibrium temperature is 90 ℃, the quantitative loop temperature is 100 ℃, the transmission line temperature is 110 ℃, the equilibrium time is 30min, and the sample injection volume is 1.0mL.