Method for detecting 1,3-dichloro-2-propanol, 2,3-dichloro-1-propanol, 3-chloro-1,2-propanediol residues in dyphylline

Abstract

The present application belongs to the technical field of chemical component detection, and particularly relates to a detection method of 1,3-dichloro-2-propanol, 2,3-dichloro-1-propanol and 3-chloro-1,2-propanediol residues in dyphylline. The detection method comprises the following steps: (1) diluting dyphylline with a solvent, shaking and preparing a test sample solution; (2) detecting the test sample solution by using gas chromatography-tandem mass spectrometry, and calculating the contents of 1,3-dichloro-2-propanol, 2,3-dichloro-1-propanol and 3-chloro-1,2-propanediol in dyphylline by using an external standard method; and the solvent is a mixture of formic acid and DMF. The method provided by the present application has good specificity, high sensitivity, good repeatability, high accuracy and can simultaneously detect 1,3-dichloro-2-propanol, 2,3-dichloro-1-propanol and 3-chloro-1,2-propanediol in dyphylline.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of chemical component detection, and particularly relates to a detection method for residues of 1,3-dichloro-2-propanol, 2,3-dichloro-1-propanol and 3-chloro-1,2-propanediol in dyphylline. BACKGROUND

[0002] Dyphylline belongs to theophylline drugs, and is mainly used as a smooth muscle relaxant, has the effects of dilating bronchus and coronary artery, and has a diuretic effect, is stable in gastric juice, has a similar antiasthmatic effect to aminophylline, and is particularly suitable for tachycardia. 3-chloro-1,2-propanediol is a reactant of an intermediate of the drug, and has trace residues in dyphylline, and 1,3-dichloro-2-propanol and 2,3-dichloro-1-propanol impurities can be generated. 1,3-dichloro-2-propanol, 2,3-dichloro-1-propanol and 3-chloro-1,2-propanediol in dyphylline belong to genotoxic impurities, and have a genotoxic impurity warning structure. According to relevant research, 1,3-dichloro-2-propanol, 2,3-dichloro-1-propanol and 3-chloro-1,2-propanediol are considered to have potential genotoxicity, and can have adverse effects on human health, so it is necessary to monitor the content thereof in drugs.

[0003] There are few reports on the detection of chloropropanol content in dyphylline in the prior art. Li Runyan et al. published “Determination of Trace 3-chloro-1,2-propanediol in Dyphylline by Solid Phase Extraction-Gas Chromatography-Mass Spectrometry” in the Journal of Pharmaceutical Analysis. In the technical scheme, the sample is extracted by saturated sodium chloride, separated and purified by a Florisil chromatographic column, concentrated by rotary evaporation, derivatized by trifluoroacetic anhydride, and then detected by gas chromatography-mass spectrometry. However, the derivatization operation of the technical scheme is complex, and only 3-chloro-1,2-propanediol can be detected.

[0004] There is no method for simultaneously detecting residues of 1,3-dichloro-2-propanol, 2,3-dichloro-1-propanol and 3-chloro-1,2-propanediol in dyphylline in the prior art, so it is necessary to establish a method for simultaneously detecting residues of 1,3-dichloro-2-propanol, 2,3-dichloro-1-propanol and 3-chloro-1,2-propanediol in dyphylline with high sensitivity, good specificity, high accuracy and the like. SUMMARY

[0005] The present invention aims to solve one or more technical problems existing in the prior art, and at least provide a beneficial solution. Specifically, the present invention provides a method for detecting residues of 1,3-dichloro-2-propanol, 2,3-dichloro-1-propanol, and 3-chloro-1,2-propanediol in dihydroxypropyltheophylline. The method provided by the present invention has good specificity, high sensitivity, good repeatability, and high accuracy, and can simultaneously detect 1,3-dichloro-2-propanol, 2,3-dichloro-1-propanol, and 3-chloro-1,2-propanediol in dihydroxypropyltheophylline.

[0006] To achieve the above objectives, the present invention adopts the following technical solution:

[0007] A method for detecting residues of 1,3-dichloro-2-propanol, 2,3-dichloro-1-propanol, and 3-chloro-1,2-propanediol in dihydroxypropyltheophylline, comprising the following steps:

[0008] (1) Dilute dihydroxypropyltheophylline with solvent, shake well, and prepare a test solution;

[0009] (2) Gas chromatography-tandem mass spectrometry was used to detect the test solution, and the contents of 1,3-dichloro-2-propanol, 2,3-dichloro-1-propanol and 3-chloro-1,2-propanediol in dihydroxypropyltheophylline were calculated by external standard method.

[0010] The solvent is a mixture of formic acid and DMF (N,N-dimethylformamide).

[0011] Furthermore, the dihydroxypropyltheophylline includes dihydroxypropyltheophylline active pharmaceutical ingredient and / or preparation.

[0012] Furthermore, the volume ratio of formic acid to DMF is 0.5-1.5:1000.

[0013] Furthermore, the volume ratio of formic acid to DMF is 1:1000.

[0014] Furthermore, the concentration of dihydroxypropyltheophylline in the test solution is 5 mg / mL.

[0015] Furthermore, the gas chromatographic column uses polyethylene glycol as the stationary phase.

[0016] Furthermore, the column temperature of the gas chromatograph is as follows: the initial temperature is 50-70℃, held for 1-3 min, and then increased to 240-260℃ at a heating rate of 5-15℃ / min, held for 3-5 min.

[0017] Furthermore, the gas chromatography conditions are as follows: injection port temperature is 230-250℃; column flow rate is 0.5-1.5mL / min; injection volume is 0.8-1.2μL.

[0018] Further, the conditions of the gas chromatography are as follows:

[0019] Column temperature: initial temperature 60℃, hold for 2 min, increase to 250℃ at a rate of 10℃ / min, hold for 4 min;

[0020] Inlet temperature: 240℃;

[0021] Carrier gas: helium;

[0022] Column flow rate: 1.0 mL / min;

[0023] Injection volume: 1 μL;

[0024] Split ratio: 5:1.

[0025] Further, the chromatographic column is DB-WAX, 30 m x 0.25 mm x 0.15 μm.

[0026] Further, the conditions of the mass spectrometry are as follows:

[0027] Ion source: EI;

[0028] Ion source temperature: 230℃;

[0029] Quadrupole temperature: 150℃;

[0030] Auxiliary heating box temperature: 260℃;

[0031] Solvent delay: 9 min;

[0032] Dwell time: 100 ms;

[0033] Sampling time: 15 min

[0034] MS acquisition time: 25 min;

[0035] Scan mode: SIM:

[0036] The quantitative ions of 1,3-dichloro-2-propanol, 2,3-dichloro-1-propanol and 3-chloro-1,2-propanediol are 49 (m / z), 62 (m / z) and 61 (m / z), respectively;

[0037] The qualitative ions of 1,3-dichloro-2-propanol, 2,3-dichloro-1-propanol and 3-chloro-1,2-propanediol are 81 (m / z), 64 (m / z) and 79 (m / z), respectively.

[0038] Further, step (1) further comprises preparation of the control solution: 1,3-dichloro-2-propanol control standard stock solution, 2,3-dichloro-1-propanol control standard stock solution, 3-chloro-1,2-propanediol control standard stock solution are mixed, diluted with a solvent, shaken to obtain the control solution.

[0039] Further, the concentrations of 1,3-dichloro-2-propanol, 2,3-dichloro-1-propanol and 3-chloro-1,2-propanediol in the control solution are all 65 ng / mL.

[0040] Further, the concentration of 1,3-dichloro-2-propanol in the 1,3-dichloro-2-propanol control standard stock solution is 2.0 mg / mL.

[0041] Further, the concentration of 2,3-dichloro-1-propanol in the 2,3-dichloro-1-propanol control standard stock solution is 2.0 mg / mL.

[0042] Further, the concentration of 3-chloro-1,2-propanediol in the 3-chloro-1,2-propanediol control standard stock solution is 2.0 mg / mL.

[0043] Compared with the prior art, the present application has the following beneficial effects:

[0044] 1. The present application adopts gas chromatography-tandem mass spectrometry to detect 1,3-dichloro-2-propanol, 2,3-dichloro-1-propanol and 3-chloro-1,2-propanediol in dyphylline, uses a mixture of formic acid and DMF as a solvent, does not need to perform derivatization treatment, and can realize simultaneous detection of 1,3-dichloro-2-propanol, 2,3-dichloro-1-propanol and 3-chloro-1,2-propanediol in dyphylline by controlling the type and parameters of the chromatographic column in gas chromatography and the type and acquisition mode of the detector in mass spectrometry, which has an important role in formulating and improving the quality standards of dyphylline raw materials and preparations.

[0045] 2. The detection method provided by the present application has good specificity and high sensitivity, the detection limit of 1,3-dichloro-2-propanol is 1.3 ppm, the quantification limit is 3.9 ppm; the detection limit of 2,3-dichloro-1-propanol is 1.3 ppm, the quantification limit is 3.8 ppm; and the detection limit of 3-chloro-1,2-propanediol is 1.3 ppm, the quantification limit is 3.9 ppm.

[0046] 3、The detection method provided by the present application has good linear relationship in the range of 19.5195 ng / mL-130.1300 ng / mL for 1,3-dichloro-2-propanol, in the range of 19.0115 ng / mL-126.7436 ng / mL for 2,3-dichloro-1-propanol, and in the range of 19.4163 ng / mL-129.4419 ng / mL for 3-chloro-1,2-propanediol.

[0047] 4、The detection method provided by the present application has high accuracy, and the average spiked recovery rate of 1,3-dichloro-2-propanol at the levels of 50%, 100% and 150% is 93.8%-104.4%, and the RSD of the average spiked recovery rate is 4.2%; the average spiked recovery rate of 2,3-dichloro-1-propanol at the levels of 50%, 100% and 150% is 94.1%-98.3%, and the RSD of the average spiked recovery rate is 1.3%; the average spiked recovery rate of 3-chloro-1,2-propanediol at the levels of 50%, 100% and 150% is 98.7%-110.7%, and the RSD of the average spiked recovery rate is 3.1%.

[0048] 5、The detection method provided by the present application also has the advantages of good repeatability, high intermediate precision, good durability and good solution stability. BRIEF DESCRIPTION OF DRAWINGS

[0049] Figure 1 The mass spectrum of the blank solution is shown in Figure 1;

[0050] Figure 2 The mass spectrum of the reference solution is shown in Figure 2;

[0051] Figure 3 The mass spectrum of the sample solution is shown in Figure 3;

[0052] Figure 4 The mass spectrum of the spiked sample solution is shown in Figure 4;

[0053] Figure 5 The mass spectrum of the 1,3-dichloro-2-propanol positioning solution is shown in Figure 5;

[0054] Figure 6 The mass spectrum of the 2,3-dichloro-1-propanol positioning solution is shown in Figure 6;

[0055] Figure 7 The mass spectrum of the 3-chloro-1,2-propanediol positioning solution is shown in Figure 7;

[0056] Figure 8 The mass spectrum of the detection limit solution is shown in Figure 8;

[0057] Figure 9 The mass spectrum of the quantification limit solution is shown in Figure 9;

[0058] Figure 10Linear plot for 1,3-dichloro-2-propanol;

[0059] Figure 11 Linear plot for 2,3-dichloro-1-propanol;

[0060] Figure 12 Linear plot for 3-chloro-1,2-propanediol;

[0061] Figure 13 Mass spectrum of 100% limit spiked test solution for Comparative Example 1;

[0062] Figure 14 Mass spectrum of 100% limit spiked test solution for Comparative Example 2;

[0063] Figure 15 Mass spectrum of 100% limit spiked test solution for Comparative Example 3;

[0064] Figure 16 Mass spectrum of 100% limit spiked test solution for Comparative Example 4;

[0065] Figure 17 Mass spectrum of 100% limit spiked test solution for Comparative Example 5;

[0066] Figure 18 Mass spectrum of 100% limit spiked test solution for Comparative Example 6. DETAILED DESCRIPTION

[0067] In order to make the objects, technical solutions and advantages of the embodiments of the present application clearer, the technical solutions in the embodiments of the present application will be described clearly and completely below. If specific conditions are not indicated in the embodiments, conventional conditions or conditions suggested by manufacturers are adopted. If the reagents or instruments used are not indicated by manufacturers, they are all conventional products that can be purchased in the market.

[0068] In the embodiments and comparative examples of the present application:

[0069] 1,3-dichloro-2-propanol reference substance source: Macklin, batch number: C10563059;

[0070] 2,3-dichloro-1-propanol reference substance source: Macklin, batch number: C10801332;

[0071] 3-chloro-1,2-propanediol reference substance source: Macklin, batch number: C10660315.

[0072] Example 1: Method for detecting 1,3-dichloro-2-propanol, 2,3-dichloro-1-propanol and 3-chloro-1,2-propanediol residues in dyphylline:

[0073] (1) Preparation of solvent: 0.5 mL of formic acid and 500 mL of DMF were mixed evenly to obtain;

[0074] Preparation of test solution: 50.13 mg of dyphylline was placed in a 10 mL volumetric flask, diluted to the mark with solvent, and shaken to obtain;

[0075] (2) The test solution was detected by gas chromatography-tandem mass spectrometry, and the contents of 1,3-dichloro-2-propanol, 2,3-dichloro-1-propanol and 3-chloro-1,2-propanediol in dyphylline were calculated by external standard method; the detection conditions are shown in Table 1:

[0076] Table 1

[0077] .

[0078] Dyphylline, from Shanghai Wanyang Pharmaceutical Co., Ltd., three batches, batch numbers are 2407101, 2407102, 2407103, the detection results of the above three batches are shown in Table 2.

[0079] Table 2

[0080] .

[0081] Example 2 specificity:

[0082] (1) Preparation of blank solution: 0.5 mL of formic acid and 500 mL of DMF were mixed evenly to obtain;

[0083] (2) Preparation of test solution: same as Example 1;

[0084] (3) Preparation of control solution:

[0085] Control solution A: 10.14 mg of 1,3-dichloro-2-propanol control was accurately weighed and placed in a 5 mL volumetric flask, and the control solution A was prepared to 2.0280 mg / mL by solvent;

[0086] Control solution B: 10.05 mg of 2,3-dichloro-1-propanol control was accurately weighed and placed in a 5 mL volumetric flask, and the standard stock solution was prepared to 1.9499 mg / mL by solvent;

[0087] Control solution C: 10.10 mg of 3-chloro-1,2-propanediol control was accurately weighed and placed in a 5 mL volumetric flask, and the standard stock solution was prepared to about 1.9875 mg / mL by solvent;

[0088] Stock solution of reference substance: precisely pipette 32.5 μL of solution A, 32.5 μL of solution B and 32.5 μL of solution C1 of the stock solution of reference substance into the same 10 mL flask, dilute to the mark with solvent, shake well (1,3-DCP: 6.5910 μg / mL; 2,3-DCP: 6.3372 μg / mL; 3-MCPD: 6.4593 μg / mL);

[0089] Stock solution of reference substance: precisely pipette 32.5 μL of solution A, 32.5 μL of solution B and 32.5 μL of solution C1 of the stock solution of reference substance into the same 10 mL flask, dilute to the mark with solvent, shake well (1,3-DCP: 6.5910 μg / mL; 2,3-DCP: 6.3372 μg / mL; 3-MCPD: 6.4593 μg / mL);

[0090] Stock solution of reference substance: precisely pipette 32.5 μL of solution A, 32.5 μL of solution B and 32.5 μL of solution C1 of the stock solution of reference substance into the same 10 mL flask, dilute to the mark with solvent, shake well (1,3-DCP: 6.5910 μg / mL; 2,3-DCP: 6.3372 μg / mL; 3-MCPD: 6.4593 μg / mL);

[0091] 1,3-Dichloro-2-propanol positioning solution: precisely pipette 32.5 μL of solution A of the stock solution of reference substance into a 10 mL flask, dilute to the mark with solvent, shake well; precisely pipette 100 μL of the above solution into a 10 mL flask, dilute to the mark with solvent, shake well;

[0092] 2,3-Dichloro-1-propanol positioning solution: precisely pipette 32.5 μL of solution B of the stock solution of reference substance into a 10 mL flask, dilute to the mark with solvent, shake well; precisely pipette 100 μL of the above solution into a 10 mL flask, dilute to the mark with solvent, shake well;

[0093] 3-Chloro-1,2-propanediol positioning solution: precisely pipette 32.5 μL of solution C of the stock solution of reference substance into a 10 mL flask, dilute to the mark with solvent, shake well; precisely pipette 100 μL of the above solution into a 10 mL flask, dilute to the mark with solvent, shake well.

[0094] (4) The blank solution, the stock solution of reference substance, the sample solution, the spiked sample solution, the 1,3-dichloro-2-propanol positioning solution, the 2,3-dichloro-1-propanol positioning solution and the 3-chloro-1,2-propanediol positioning solution were detected by the method of Example 1, and the detection results are shown in Table 3; the chromatogram is shown in Figures 1-7

[0095] Table 3

[0096] .​

[0097] From Table 3 and Figures 1-7 It can be seen that: blank solvent, test solution does not interfere with the determination of target peak, the method is specific.

[0098] Example 3 detection limit and quantification limit:

[0099] (1) Preparation of detection limit solution (LOD): take 10 μL of control product stock solution, place it in a 10 mL volumetric flask, add solvent to dilute to the mark, shake well (1,3-DCP: 6.5065 ng / mL; 2,3-DCP: 6.3372 ng / mL; 3-MCPD: 6.4721 ng / mL);

[0100] (2) Preparation of quantification limit solution (LOQ): take 30 μL of control product stock solution, place it in a 10 mL volumetric flask, add solvent to dilute to the mark, shake well (1,3-DCP: 19.5195 ng / mL; 2,3-DCP: 19.0115 ng / mL; 3-MCPD: 19.4163 ng / mL);

[0101] (3) Detection limit solution and quantification limit solution were detected according to the method of Example 1, the detection limit solution (sample 3 times) detection results as shown in Table 4, the spectrum as Figure 8 shown; the quantification limit solution (sample 6 times) detection results as shown in Table 5, the spectrum as Figure 9 shown.

[0102] Table 4

[0103] .

[0104] Table 5

[0105] .

[0106] From Table 4 and Table 5, it can be seen that: 1,3-DCP detection limit concentration is 6.5065 ng / mL (equivalent to the test sample concentration is 1.3 ppm), the quantification limit concentration is 19.5195 ng / mL (equivalent to the test sample concentration is 3.9 ppm); 2,3-DCP detection limit concentration is 6.3372 ng / mL (equivalent to the test sample concentration is 1.3 ppm), the quantification limit concentration is 19.0115 ng / mL (equivalent to the test sample concentration is 3.8 ppm); 3-MCPD detection limit concentration is 6.4721 ng / mL (equivalent to the test sample concentration is 1.3 ppm), the quantification limit concentration is 19.4163 ng / mL (equivalent to the test sample concentration is 3.9 ppm), the signal-to-noise ratio and peak area RSD meet the requirements.

[0107] Example 4 linear range:

[0108] Prepare a series of linear test solutions from LOQ to 200% limit concentration (30%, 50%, 100%, 150%, 200%):

[0109] 30% linear test solution: Pipet 30 μL of the control stock solution into a 10 mL volumetric flask, dilute to volume with solvent, and mix (1,3-DCP: 19.5195 ng / mL; 2,3-DCP: 19.0115 ng / mL; 3-MCPD: 19.4163 ng / mL);

[0110] 50% linear test solution: Pipet 50 μL of the control stock solution into a 10 mL volumetric flask, dilute to volume with solvent, and mix (1,3-DCP: 32.5325 ng / mL; 2,3-DCP: 31.6859 ng / mL; 3-MCPD: 32.3605 ng / mL);

[0111] 100% linear test solution: Pipet 100 μL of the control stock solution into a 10 mL volumetric flask, dilute to volume with solvent, and mix (1,3-DCP: 65.0650 ng / mL; 2,3-DCP: 63.3718 ng / mL; 3-MCPD: 64.7209 ng / mL);

[0112] 150% linear test solution: Pipet 150 μL of the control stock solution into a 10 mL volumetric flask, dilute to volume with solvent, and mix (1,3-DCP: 97.5975 ng / mL; 2,3-DCP: 95.0577 ng / mL; 3-MCPD: 97.0814 ng / mL);

[0113] 200% linear test solution: Pipet 200 μL of the control stock solution into a 10 mL volumetric flask, dilute to volume with solvent, and mix (1,3-DCP: 130.1300 ng / mL; 2,3-DCP: 126.7436 ng / mL; 3-MCPD: 129.4419 ng / mL).

[0114] Prepare a series of linear test solutions from LOQ to 200% limit concentration (30%, 50%, 100%, 150%, 200%):

[0115] Table 6

[0116] .

[0117] The linear plot for 1,3-dichloro-2-propanol is shown in Figure 1; the linear plot for 2,3-dichloro-1-propanol is shown in Figure 2; and the linear plot for 3-chloro-1,2-propanediol is shown in Figure 3. Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 Figure 11 Figure 12 Figure 10 <

[0118] It can be seen from Table 6 that 1,3-DCP has a good linear relationship in the range of 19.5195 ng / mL~130.1300 ng / mL, R2 is 1.000; 2,3-DCP has a good linear relationship in the range of 19.0115 ng / mL~126.7436 ng / mL, R2 is 1.000; 3-MCPD has a good linear relationship in the range of 19.4163 ng / mL~129.4419 ng / mL, R2 is 0.999, all meet the requirements.

[0119] Accuracy of Example 5:

[0120] Prepare three levels of sample standard solution of 50%, 100% and 150% respectively:

[0121] 50% spiked sample solution: accurately weigh the theophylline hydroxypropyl, place it in a 10 mL volumetric flask, add an appropriate amount of solvent to dissolve, accurately pipette 50 μL of the control substance stock solution into the same volumetric flask, add solvent to dilute to the mark, shake well (1,3-DCP: 32.9550 ng / mL; 2,3-DCP: 31.6859 ng / mL; 3-MCPD: 32.2965 ng / mL).

[0122] 100% spiked sample solution: accurately weigh the theophylline hydroxypropyl, place it in a 10 mL volumetric flask, add an appropriate amount of solvent to dissolve, accurately pipette 100 μL of the control substance stock solution into the same volumetric flask, add solvent to dilute to the mark, shake well (1,3-DCP: 65.9100 ng / mL; 2,3-DCP: 63.3718 ng / mL; 3-MCPD: 64.5930 ng / mL).

[0123] 150% spiked sample solution: accurately weigh the theophylline hydroxypropyl, place it in a 10 mL volumetric flask, add an appropriate amount of solvent to dissolve, accurately pipette 150 μL of the control substance stock solution into the same volumetric flask, add solvent to dilute to the mark, shake well (1,3-DCP: 98.8650 ng / mL; 2,3-DCP: 95.0577 ng / mL; 3-MCPD: 96.8896 ng / mL).

[0124] The detection was carried out according to the method of Example 1, and the results are shown in Tables 7-9.

[0125] Table 7

[0126] .

[0127] Table 8

[0128] .

[0129] Table 9

[0130] .

[0131] Conclusion: The recovery rates of 1,3-DCP at three concentration levels were 93.8%~104.4%, and the RSD of recovery rate was 4.2%; the recovery rates of 2,3-DCP at three concentration levels were 94.1%~98.3%, and the RSD of recovery rate was 1.3%; the recovery rates of 3-MCPD at three concentration levels were 98.7%~110.7%, and the RSD of recovery rate was 3.1%, all of which met the requirements and the method had good accuracy.

[0132] Example 6 repeatability:

[0133] Prepare 6 portions of 100% limit level of spiked sample solution:

[0134] 100% spiked sample solution: accurately weigh the dyphylline, place it in a 10 mL volumetric flask, add an appropriate amount of solvent to dissolve, accurately pipette 100 μL of the control substance stock solution into the same volumetric flask, add solvent to dilute to the calibration mark, and shake well (1,3-DCP: 65.9100 ng / mL; 2,3-DCP: 63.3718 ng / mL; 3-MCPD: 64.5930 ng / mL).

[0135] According to the method of Example 1, analyze one sample per solution, and the results are shown in Tables 10-12.

[0136] Table 10

[0137] .

[0138] Table 11

[0139] .

[0140] Table 12

[0141] .

[0142] Conclusion: The RSD of 1,3-DCP content in the 6 repeated solutions was 1.1%, the RSD of 2,3-DCP content was 0.9%, and the RSD of 3-MCPD content was 1.7%, all of which met the requirements and the method had good repeatability.

[0143] Example 7 intermediate precision:

[0144] Prepare 6 portions of 100% limit level of spiked sample solution by another researcher at different times, and detect according to the method of Example 1, and the results are shown in Tables 13-15.

[0145] Table 13

[0146] .

[0147] Table 14

[0148] .

[0149] Table 15

[0150] .

[0151] Conclusion: The RSD of 1,3-DCP content in 12 precision samples was 1.7%, the RSD of 2,3-DCP content was 2.6%, and the RSD of 3-MCPD content was 1.7% by different persons on different days, all meeting the requirements, and the precision of the method was good.

[0152] Example 8 Durability:

[0153] The injection port temperature and column flow rate were changed respectively, and two portions of 100% limit spiked sample solutions were prepared, 2 injections were performed for each condition, and detection was performed according to the method of Example 1, and the results are shown in Tables 16-18.

[0154] Table 16

[0155] .

[0156] Table 17

[0157] .

[0158] Table 18

[0159] .

[0160] Conclusion: The system suitability met the requirements when the injection port temperature and column flow rate were changed; the RSD of 1,3-DCP content in spiked samples determined under different conditions of changing the injection port temperature and column flow rate was 2.1%, the RSD of 2,3-DCP content was 1.5%, and the RSD of 3-MCPD content was 4.1%, all less than 10.0%, meeting the requirements, and the durability of the method was good.

[0161] Example 9 Solution Stability:

[0162] One portion of the control solution and one portion of the 100% limit spiked sample solution were prepared respectively, and the drift of the peak area of the target peak in the control solution and the spiked sample solution within 13 h relative to 0 h was investigated, and the results are shown in Tables 19-21.

[0163] Table 19

[0164] .

[0165] Table 20

[0166] .

[0167] Table 21

[0168] .

[0169] Conclusion: The peak area ratio of the target impurity at each time point relative to 0 h was within the range of 80.0% to 120.0% for the control solution and the 100% limit spiked sample solution, and the solution was stable.

[0170] Comparative Example 1: The only difference from Example 1 is that the solvent is DMF; the rest is the same; the spectrum of the 100% limit spiked sample solution is shown in Figure 13 , and 1,3-dichloro-2-propanol and 2,3-dichloro-1-propanol cannot be separated.

[0171] Comparative Example 2: The only difference from Example 1 is that the solvent is ethyl acetate; the rest is the same, and the spectrum of the 100% limit spiked sample solution is shown in Figure 14 , there are impurity peaks, and 1,3-dichloro-2-propanol and 2,3-dichloro-1-propanol cannot be separated.

[0172] Comparative Example 3: The only difference from Example 1 is that the solvent is 5 mL formic acid and 1 L DMF; the rest is the same, and the spectrum of the 100% limit spiked sample solution is shown in Figure 15 , 1,3-dichloro-2-propanol, 2,3-dichloro-1-propanol and 3-chloro-1,2-propanediol have impurity peaks.

[0173] Comparative Example 4: The only difference from Example 1 is that the gas chromatography conditions are as follows: adjust the split ratio to 3:1, and the column temperature program is as follows: 80°C for 1 min, 20°C / min to 250°C, and 5 min; the rest is the same, and the spectrum of the 100% limit spiked sample solution is shown in Figure 16 , the baseline is not flat, and the peak shape of 3-chloro-1,2-propanediol is poor.

[0174] Comparative Example 5: The only difference from Example 1 is that the gas chromatography conditions are as follows: adjust the split ratio to 4:1, and the column temperature program is as follows: 40°C for 3 min, 3°C / min to 250°C, and 4 min; the rest is the same, and the spectrum of the 100% limit spiked sample solution is shown in Figure 17 , the baseline is not flat, and the peak shape of 3-chloro-1,2-propanediol is poor.

[0175] Comparative Example 6: The only difference from Example 1 is that the conditions of gas chromatography are as follows: no split, column temperature program: initial temperature 50°C, temperature rising at 20°C / min to 140°C, temperature rising at 5°C / min to 170°C, temperature rising at 20°C / min to 250°C for 3 min, and the rest are the same. The chromatogram of the 100% limit spiked sample solution measured using the above conditions is shown in Fig. 6. It can be seen that 1,3-dichloro-2-propanol and 2,3-dichloro-1-propanol are not separated and appear as a tailing peak. Figure 18

[0176] Finally, it should be noted that the above content is only used to illustrate the technical solutions of the present application, and is not a limitation on the protection scope of the present application. Simple modifications or equivalent replacements of the technical solutions of the present application made by those skilled in the art do not deviate from the essence and scope of the technical solutions of the present application.​

Claims

1. A method for detecting 1,3-dichloro-2-propanol, 2,3-dichloro-1-propanol, 3-chloro-1,2-propanediol residues in dyphylline, characterized in that, The method comprises the following steps: (1) dilute the dyphylline with a solvent, shake well, and prepare a test solution; (2) detect the test solution by gas chromatography-mass spectrometry, and calculate the content of 1,3-dichloro-2-propanol, 2,3-dichloro-1-propanol and 3-chloro-1,2-propanediol in the dyphylline by an external standard method; the solvent is a mixture of formic acid and DMF; the volume ratio of the formic acid to the DMF is 0.5-1.5:1000; the conditions of the gas chromatography are as follows: the column temperature is 50-70℃ at the beginning, and then is raised to 240-260℃ at a rate of 5-15℃ / min, and is kept for 3-5 min; the split ratio is 5:

1.

2. The method for detecting 1,3-dichloro-2-propanol, 2,3-dichloro-1-propanol, 3-chloro-1,2-propanediol residues in dyphylline according to claim 1, characterized in that, the concentration of the dyphylline in the test solution is 5 mg / mL.

3. The method for detecting 1,3-dichloro-2-propanol, 2,3-dichloro-1-propanol, 3-chloro-1,2-propanediol residues in dyphylline according to claim 2, characterized in that, the stationary liquid of the chromatographic column of the gas chromatography is polyethylene glycol.

4. The method for detecting 1,3-dichloro-2-propanol, 2,3-dichloro-1-propanol, 3-chloro-1,2-propanediol residues in dyphylline according to claim 3, characterized in that, the conditions of the gas chromatography are as follows: the injection port temperature is 230-250℃; the column flow rate is 0.5-1.5 mL / min; and the injection volume is 0.8-1.2 μL.

5. The method for detecting the residual amount of 1,3-dichloro-2-propanol, 2,3-dichloro-1-propanol, 3-chloro-1,2-propanediol in dyphylline according to claim 4, characterized in that, the conditions of the gas chromatography are as follows: the column temperature is 60℃ at the beginning, and is kept for 2 min, and then is raised to 250℃ at a rate of 10℃ / min, and is kept for 4 min; the injection port temperature is 240℃; the carrier gas is helium; the column flow rate is 1.0 mL / min; the injection volume is 1 μL.

6. The method for detecting 1,3-dichloro-2-propanol, 2,3-dichloro-1-propanol, 3-chloro-1,2-propanediol residues in dyphylline according to claim 5, characterized in that, the chromatographic column is DB-WAX, 30 m x 0.25 mm x 0.15 μm.

7. The method for detecting 1,3-dichloro-2-propanol, 2,3-dichloro-1-propanol, 3-chloro-1,2-propanediol residues in dyphylline according to claim 6, characterized in that, the conditions of the mass spectrometry include: the ion source is EI; the ion source temperature is 230℃; the quadrupole rod temperature is 150℃; the auxiliary heating box temperature is 260℃; the solvent delay is 9 min; the residence time is 100 ms; the sampling time is 15 min the MS acquisition time is 25 min.

8. The method for detecting 1,3-dichloro-2-propanol, 2,3-dichloro-1-propanol, 3-chloro-1,2-propanediol residues in dyphylline according to claim 7, characterized in that, the conditions of the mass spectrometry further include: the scanning mode is SIM; the quantitative ions of 1,3-dichloro-2-propanol, 2,3-dichloro-1-propanol and 3-chloro-1,2-propanediol are 49 m / z, 62 m / z and 61 m / z respectively; and the qualitative ions of 1,3-dichloro-2-propanol, 2,3-dichloro-1-propanol and 3-chloro-1,2-propanediol are 81 m / z, 64 m / z and 79 m / z respectively.

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