A method for detecting residual solvent in alpha-ribose methanesulfonate by gas chromatography

Abstract

The invention discloses a method for detecting residual solvents in alpha-ribose mesylate by gas chromatography, belongs to the technical field of chemical detection, and adopts gas chromatography to detect residual solvents in alpha-ribose mesylate. The instrument is a gas chromatograph, the detector is an FID detector, the chromatographic column adopts a DB-WAX quartz capillary chromatographic column, the carrier gas is N2, the flow rate is 3.0 mL / min, the split ratio is 20:1, the column temperature is maintained at 40 DEG C for 10 minutes, then the temperature is increased to 80 DEG C at 10 DEG C per minute, and then the temperature is increased to 250 DEG C at 40 DEG C per minute and maintained for 3 minutes, headspace injection, equilibrium temperature is 90 DEG C, quantitative loop temperature is 100 DEG C, transmission line temperature is 110 DEG C, and equilibrium time is 30 minutes; the injection volume is 1.0 ml, the residual solvents in alpha-ribose mesylate can be effectively separated and the content thereof can be quantitatively detected, the method is simple to operate, has strong specificity, high sensitivity and accuracy, good linearity and durability, and can accurately and quickly determine the residual amounts of 8 kinds of solvents in alpha-ribose mesylate.

Description

Technical Field

[0001] The invention belongs to the technical field of chemical detection, and particularly relates to a method for detecting residual solvent in alpha-ribose methanesulfonate by gas chromatography. Background Art

[0002] α-Ribose methanesulfonate is an important starting material in the synthesis of gemcitabine hydrochloride raw materials. Gemcitabine hydrochloride is an anticancer drug with wide clinical application. Its injection is marketed in many countries. It is particularly important to control the quality of the starting materials at the source. At present, there is no report on the detection method of residual solvents in α-ribose methanesulfonate. The residual organic solvents in α-ribose methanesulfonate have a certain influence on the product quality. Therefore, it is very necessary to develop a detection method that can effectively detect the residual solvents in α-ribose methanesulfonate. After a long period of research, we finally developed a method that can detect the residual solvents in α-ribose methanesulfonate. This method is easy to operate and has a good detection effect. Summary of the invention

[0003] In view of the deficiencies of the prior art, the object of the present invention is to provide a method for detecting residual solvents in α-ribose methanesulfonate by gas chromatography, comprising the following steps:

[0004] 1) Selection of instruments and chromatographic conditions: gas chromatograph; chromatographic column is DB-WAX (30m×0.53mm×1.0μm) capillary chromatographic column; FID detector is used, the initial temperature of the chromatographic column is 35-45°C, the holding time is 8-12min, then the temperature is increased to 80°C at a heating rate of 8-12°C / min, and then the temperature is increased to a final temperature of 250°C at a heating rate of 35-45°C / min, and the final temperature of the chromatographic column is held for 2-5min; the carrier gas is nitrogen, the carrier gas flow rate is 2.5-3.5ml / min, and the split ratio is 20:1; the injection port temperature is 190-210°C, and the detector temperature is 240-260°C; the injection mode 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 equilibrium time is 30min; the injection volume is 1.0mL.

[0005] 2) Prepare the following solutions:

[0006] a. Blank solution (diluent): dimethyl sulfoxide;

[0007] b. Reference solution: weigh 0.5g acetone, 0.06g dichloromethane, 0.072g tetrahydrofuran, 0.5g ethyl acetate, 0.089g toluene, 0.5g ether, 0.041g acetonitrile, 0.1g ethyl difluorobromoacetate, place in a 50ml volumetric flask, dilute to scale with diluent, shake well, as a stock solution; transfer 2.0ml of the stock solution to a 100ml volumetric flask, dilute to scale with diluent, shake well, and then accurately measure 5.0ml to a 20ml headspace bottle, cover and seal. C. Test solution: take 0.2g of α-ribose methanesulfonate, place in a 20ml headspace bottle, add 5.0ml diluent to dissolve, cover and seal, shake well, and prepare 2 copies in parallel.

[0008] Inject according to the following sequence table:

[0009] name Number of injection needles blank solution ≥1 needle Reference solution 6-pin Test solution 1 needle each

[0010] The peak areas of each solvent in the α-ribose methanesulfonate test solution were calculated using the external standard method.

[0011] Preferably, the chromatographic conditions selected for the gas chromatography analysis of the present invention are as follows:

[0012] Adopt FID detector;

[0013] Chromatographic column: DB-WAX (30m×0.53mm×1.0μm) capillary column;

[0014] Temperature program: start at 40°C, maintain for 10 minutes, increase to 80°C at 10°C per minute, then increase to 250°C at 40°C per minute and maintain for 3 minutes.

[0015] Carrier gas: nitrogen, flow rate: 3.0 mL / min, split ratio: 20:1;

[0016] Inlet temperature: 200°C;

[0017] Detector (FID) temperature: 250°C;

[0018] Injection method: headspace injection, equilibrium temperature 90°C, quantitative loop temperature 100°C, transfer line temperature 110°C, equilibrium time 30 min;

[0019] Injection volume: 1.0ml.

[0020] The beneficial effects of the present invention are as follows:

[0021] The invention can effectively separate the residual solvent in alpha-ribose methanesulfonate and can quantitatively detect the content thereof. The residual solvent is one or more of ether, acetone, tetrahydrofuran, ethyl acetate, dichloromethane, acetonitrile, toluene and ethyl difluorobromoacetate. The method is simple to operate, has strong specificity, and is high in sensitivity and accuracy, thereby ensuring that the quality of alpha-ribose methanesulfonate is controllable. Currently, there is no report on the detection method of multiple residual solvents in alpha-ribose methanesulfonate. The invention can provide a reliable detection method for residual solvent detection in alpha-ribose methanesulfonate, and is suitable for comprehensive promotion and application. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 is the blank solution chromatogram;

[0023] Figure 2 is the chromatogram of the reference solution;

[0024] Figure 3 is the chromatogram of the test solution;

[0025] Figure 4 The linear test result diagram of each solvent;

[0026] Figure 5 The figure is the solution chromatogram for accuracy. DETAILED DESCRIPTION

[0027] The present invention will further supplement the above contents with reference to the embodiments (attached drawings), but it should not be understood that the scope of the present invention is limited to the following embodiments.

[0028] The instruments and chromatographic conditions used in the following examples are as follows:

[0029] The gas chromatograph is Agilent 7890A, and an FID detector is used. The initial column temperature is 35-45°C, the holding time is 8-12min, and then the temperature is increased to 80°C at a heating rate of 8-12°C / min, and then the temperature is increased to a final column temperature of 250°C at a heating rate of 35-45°C / min, and the final column temperature is held for 2-5min; the carrier gas is nitrogen, and preferably, the carrier gas flow rate is 2.5-3.5ml / min, and the split ratio is 20:1; the injection port temperature is 190-210°C, and the detector temperature is 240-260°C; the injection mode is headspace injection, the equilibrium temperature is 90°C, the quantitative loop temperature is 100°C, the transmission line temperature is 110°C, and the equilibrium time is 30min; the injection volume is 1.0mL.

[0030] Example 1

[0031] Specificity inspection, such as Figure 1-2 , Figure 5 As shown:

[0032] Accurately measure 1.0 mL of blank solution, reference solution, and each impurity location solution, inject them into the gas chromatograph, and record the chromatogram. The results of the impurity location and separation experiments show that the blank has no interference, and other solvents have no interference with the sample determination. The minimum separation between adjacent solvents is 2.60. The specificity of this analysis condition is good, and the test results are as follows:

[0033]

[0034]

[0035] Example 2

[0036] Sensitivity test results:

[0037] The limit of quantitation (LOQ) results are as follows:

[0038]

[0039] The limit of detection (LOD) results are as follows:

[0040]

[0041] The results of the quantitative limit and detection limit determination of each solvent showed that the detection sensitivity of each solvent was high (far higher than the limit standard value) and the method sensitivity met the requirements.

[0042] Example 3

[0043] Linear investigation, such as Figure 4 As shown:

[0044] The peak area was plotted against the concentration. The concentration of ether was in the range of 40.664μg / mL-304.980μg / mL, the concentration of acetone was in the range of 40.120μg / mL-300.900μg / mL, the concentration of tetrahydrofuran was in the range of 5.928μg / mL-44.460μg / mL, the concentration of ethyl acetate was in the range of 40.160μg / mL-301.200μg / mL, the concentration of dichloromethane was in the range of 5.128μg / mL-38.460μg / mL, the concentration of acetonitrile was in the range of 3.232μg / mL-24.240μg / mL, the concentration of toluene was in the range of 7.016μg / mL-52.620μg / mL, and the concentration of difluoroethyl bromoacetate was in the range of 8.792μg / mL-65.940μg / mL. The sample concentration was linearly related to the peak area, and the linearity was good. The specific results are as follows:

[0045] name Linear equations <![CDATA[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

[0046] Example 4

[0047] Accuracy inspection, chromatogram such as Figure 5 As shown:

[0048] The recovery rate solution was prepared by adding samples at the limit concentration of 50%, 100%, and 150% of ether, acetone, tetrahydrofuran, ethyl acetate, dichloromethane, acetonitrile, toluene, and ethyl difluorobromoacetate, and 3 portions were prepared in parallel for each concentration group. At the same time, a test solution and a reference solution were prepared. The recovery rate was calculated according to the above chromatographic conditions. The recovery rates of each solvent were all within the range of 90%-110%. The results are shown in the following table:

[0049]

[0050] Example 5

[0051] Durability inspection, respectively replace the chromatographic column, change the initial column temperature (40±1℃), carrier gas flow rate (3.0±0.1ml / min), injection port temperature (200±2℃), heating rate (±1℃ / min), compare the RSD of the peak area of ​​each solvent in the reference solution and the minimum separation of each solvent peak, and inspect its durability. After changing the chromatographic condition parameters, the maximum RSD of the 6-needle peak area of ​​each solvent in the reference solution was 6.7%, which was less than 10%; the separation of the system suitability solution was greater than 1.5, indicating that the method has good durability. The results are shown in the table below:

[0052]

[0053] The above description is only a preferred embodiment of the present invention. It should be pointed out that those skilled in the art should understand that several improvements can be made without departing from the principles of the present invention. These improvements should also be regarded as the scope of protection of the present invention and should not be regarded as specific limitations of the present invention.

Claims

1. A method for detecting residual solvents in α-ribose methanesulfonate by gas chromatography, characterized in that: The following steps are involved: 1) Selection of instruments and chromatographic conditions: gas chromatograph; chromatographic column is DB-WAX capillary column, 30m×0.53mm×1.0μm; FID detector is used; 2) Preparation of reference solution: weigh 0.5g acetone, 0.06g dichloromethane, 0.072g tetrahydrofuran, 0.5g ethyl acetate, 0.089g toluene, 0.5g ether, 0.041g acetonitrile, 0.1g ethyl difluorobromoacetate, place in a 50ml volumetric flask, dilute to scale with diluent, shake well, as the stock solution, transfer 2.0ml of the stock solution to a 100ml volumetric flask, dilute to scale with diluent, shake well, and accurately measure 5.0ml to a 20ml headspace bottle, cover and seal; 3) Preparation of test solution: Take 0.2g of α-ribose methanesulfonate, place it in a 20ml headspace bottle, add 5.0ml of diluent to dissolve, cover and seal, shake well, and the solution is ready; 4) Determination of residual solvents: Take the reference solution and the test solution, inject the sample into the headspace, and perform gas chromatography analysis to obtain the peak area of ​​each solvent. The peak area is calculated according to the external standard method. The temperature change process of the chromatographic column in the gas phase detection is as follows: the initial temperature of the chromatographic column is 35-45℃, the holding time is 8-12min, then the temperature is increased to 80℃ at a heating rate of 8-12℃ / min, and then the temperature is increased to the final temperature of the chromatographic column at a heating rate of 35-45℃ / min. The final temperature of the chromatographic column is maintained for 2-5min; The diluent is dimethyl sulfoxide solution; The carrier gas in the gas chromatography detection is nitrogen, the carrier gas flow rate is 2.5-3.5 ml / min, and the split ratio is 20:1; In the gas chromatography detection, the injection port temperature is 190-210°C, and the detector temperature is 240-260°C.

2. The method for detecting residual solvent in α-ribose methanesulfonate by gas chromatography according to claim 1, characterized in that: The injection method in the gas chromatography detection is headspace injection, the equilibrium temperature is 90°C, the quantitative loop temperature is 100°C, the transmission line temperature is 110°C, the equilibrium time is 30min, and the injection volume is 1.0mL.

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