Detection method of 3-[4-(2-oxiranylmethoxy) butoxy]-1, 2-propylene glycol

Abstract

The invention discloses a detection method of 3-[4-(2-oxiranyl methoxy) butoxy]-1, 2-propylene glycol. The detection method comprises the following steps: firstly, adding a reagent; the invention relates to a method for detecting 3-[4-(2-oxiranyl methoxy) butoxy]-1, 2-propylene glycol, which comprises the following step: carrying out qualitative or quantitative detection on 3-[4-(2-oxiranyl methoxy) butoxy]-1, 2-propylene glycol in a liquid to be detected by adopting a high performance liquid chromatography-mass spectrometry method. According to the present invention, the 3-[4-(2-oxiranylmethoxy) butoxy]-1, 2-propylene glycol in the to-be-detected sample is qualitatively and quantitatively detected by using the liquid chromatography-mass spectrometry method, such that the sensitivity and the accuracy are high, and the reproducibility is good. The method can be used for monitoring the quality of cross-linked sodium hyaluronate or a cross-linked product adopting BDDE as a cross-linking agent, and the safety risk of the materials is reduced.

Description

Technical Field

[0001] The present invention relates to the field of chemical detection, and particularly to a detection method and application of 3-[4-(2-oxiranylmethoxy)butoxy]-1,2-propanediol. Background Art

[0002] Hyaluronic acid (abbreviated as HA) is a naturally occurring biodegradable polysaccharide composed of alternating units of D-glucuronic acid and N-acetyl-D-glucosamine, and has ideal biocompatibility; hyaluronic acid is currently the most widely used soft tissue filling material, such as a corrective material in the aesthetic medicine field or a defect filling material in the therapeutic medicine field. However, hyaluronic acid is affected by factors such as hyaluronidase and free radicals in the body, has a fast degradation rate in the body, a short retention time, and poor mechanical strength in an aqueous system. Therefore, BDDE is often used to modify hyaluronic acid to prepare cross-linked sodium hyaluronate gel, and its physical and chemical properties are significantly improved compared with natural hyaluronic acid.

[0003] BDDE reacts with hydroxyl groups in HA through epoxy groups at both ends of the molecular chain to form stable ether bonds. Although it has lower toxicity compared with cross-linking agents such as divinyl sulfone, the residual BDDE in cross-linked sodium hyaluronate is often considered to have potential toxicity or carcinogenicity due to the presence of epoxy groups with relatively high reactivity, and is also a group of toxic impurities that need to be strictly quality controlled by the Food and Drug Administration. YY / T0962-2021 stipulates that the content of BDDE in cross-linked sodium hyaluronate shall not be higher than 2 ppm. In the cross-linking process, part of BDDE will undergo single-end ring opening, that is, it is very likely that the impurity 3-[4-(2-oxiranylmethoxy)butoxy]-1,2-propanediol exists in cross-linked sodium hyaluronate. This type of impurity also contains epoxy groups and belongs to genotoxic impurities. However, this substance has not received wide attention at present, and there is no method that can effectively detect this substance in cross-linked sodium hyaluronate.

[0004] Therefore, in order to control the quality of cross-linked sodium hyaluronate or other materials using BDDE as a cross-linking agent in the art and reduce the potential risks of the above materials, there is an urgent need to develop an analytical method for 3-[4-(2-oxiranylmethoxy)butoxy]-1,2-propanediol. Summary of the Invention

[0005] The technical problem to be solved by this application is to overcome the defects that the prior art pays little attention to 3-[4-(2-oxiranylmethoxy)butoxy]-1,2-propanediol remaining in cross-linked sodium hyaluronate and there is no suitable detection method, and to provide a detection method for 3-[4-(2-oxiranylmethoxy)butoxy]-1,2-propanediol. This application uses liquid chromatography-mass spectrometry to qualitatively and quantitatively detect 3-[4-(2-oxiranylmethoxy)butoxy]-1,2-propanediol in the sample to be tested, with high sensitivity and accuracy and good reproducibility. This method can be used to monitor the quality of cross-linked sodium hyaluronate or cross-linked products using BDDE as a cross-linking agent, and reduce the potential risks of the above materials.

[0006] This application solves the above technical problems through the following technical solutions.

[0007] This application provides a detection method for 3-[4-(2-oxiranylmethoxy)butoxy]-1,2-propanediol, including the following steps: using high performance liquid chromatography-mass spectrometry to qualitatively or quantitatively detect 3-[4-(2-oxiranylmethoxy)butoxy]-1,2-propanediol in the liquid to be tested.

[0008] In some embodiments, the liquid to be tested includes a sample to be tested containing 3-[4-(2-oxiranylmethoxy)butoxy]-1,2-propanediol and a solvent, and the solvent includes an alcohol solvent and / or water.

[0009] In a preferred embodiment, the alcohol solvent is selected from C1-C3 alcohol solvents, preferably methanol.

[0010] In some embodiments, the concentration of 3-[4-(2-oxiranylmethoxy)butoxy]-1,2-propanediol in the liquid to be tested is 4-85 ng / mL, such as 5 ng / mL, 10 ng / mL, 20 ng / mL, 30 ng / mL, 40 ng / mL, 50 ng / mL, 60 ng / mL, 70 ng / mL, 80 ng / mL or 85 ng / mL.

[0011] In a preferred embodiment, the sample of the liquid to be tested is a sample containing 3-[4-(2-oxiranylmethoxy)butoxy]-1,2-propanediol prepared by chemical synthesis.

[0012] In another preferred embodiment, the sample of the liquid to be tested is a cross-linked product prepared using BDDE as a cross-linking agent, preferably a cross-linked polysaccharide prepared using BDDE as a cross-linking agent, more preferably cross-linked sodium hyaluronate prepared using BDDE as a cross-linking agent.

[0013] Among them, the cross-linked product may further include a degradation treatment before detection. Preferably, the degradation method includes enzymatic hydrolysis or chemical degradation. The purpose of the degradation is to fully dissolve 3-[4-(2-epoxyethylmethoxy)butoxy]-1,2-propanediol in the cross-linked product in a solvent to avoid false negative phenomena.

[0014] When the sample to be tested is cross-linked sodium hyaluronate prepared using BDDE as a cross-linking agent, the degradation method includes the following steps: in water, enzymatic hydrolysis treatment is performed on the cross-linked sodium hyaluronate using hyaluronidase.

[0015] Preferably, the volume ratio of the water to the solvent in the sample to be tested is (5-20):1, more preferably (8-15):1, such as 9:1.

[0016] Preferably, based on the unit mass of the cross-linked sodium hyaluronate, the dosage of the hyaluronidase is 1000-3000 IU / g, such as 2500 IU / g.

[0017] Preferably, the hyaluronidase can be added in the form of an aqueous hyaluronidase solution according to the conventional method in the art, and the concentration of the hyaluronidase in the aqueous hyaluronidase solution is 8000-15000 IU / mL, preferably 10000 IU / mL.

[0018] Preferably, the temperature of the enzymatic hydrolysis is 30-45°C, such as 37°C.

[0019] Preferably, the time of the enzymatic hydrolysis is 0.5-5 h, such as 1 h.

[0020] Preferably, after the enzymatic hydrolysis, a filtration operation may further be included. The pore size of the filter membrane used for filtration is 0.2-0.5 μm, such as 0.22 μm.

[0021] In the high performance liquid chromatography of some embodiments, the mobile phase includes mobile phase A and mobile phase B; the mobile phase A is an aqueous solution of a volatile acid, and the mass percentage of the volatile acid in the aqueous solution of the volatile acid is 0.05%-0.2%; the mobile phase B is methanol.

[0022] Preferably, the aqueous solution of the volatile acid is selected from formic acid aqueous solution and / or acetic acid aqueous solution.

[0023] Preferably, the mass percentage of the volatile acid in the aqueous solution of the volatile acid is 0.08%-0.12%, such as 0.1%.

[0024] In the high performance liquid chromatography of some embodiments, the flow rate of the mobile phase is 0.15-0.5 mL / min, preferably 0.3 mL / min.

[0025] In the high performance liquid chromatography of some embodiments, the elution mode is isocratic elution or gradient elution.

[0026] When using the isocratic elution, the volume percentage of the mobile phase A in the mobile phase is more than 85%, preferably 85% - 95%, such as 85%. When the volume percentage of the mobile phase A in the mobile phase is less than 85%, such as 80%, the separation effect is poor and the detection accuracy is low.

[0027] When using the gradient elution, from 0 to 8.5 min, the volume percentage of the mobile phase A is 85% - 92%, and the volume percentage of the mobile phase B is 8% - 25%; from 8.6 to 11 min, the volume percentage of the mobile phase A is 10% - 30%, and the volume percentage of the mobile phase B is 70% - 90%; from 11.1 to 15 min, the volume percentage of the mobile phase A is 75% - 92%, and the volume percentage of the mobile phase B is 8% - 25%.

[0028] In the high performance liquid chromatography of some embodiments, the temperature of the injector is 5 - 30 °C, preferably 8 - 15 °C, such as 10 °C.

[0029] In the high performance liquid chromatography of some embodiments, the injection volume of the test solution is 0.5 - 5 μL, such as 1 μL.

[0030] In the high performance liquid chromatography of some embodiments, the chromatographic column is an octadecylsilane-bonded silica gel chromatographic column, preferably Agilent Infinitylab poroshell 120 EC-18 100 mm × 3 mm, 1.9 μm.

[0031] In the high performance liquid chromatography of a preferred embodiment, the particle size of the packing of the octadecylsilane-bonded silica gel chromatographic column is 1.5 - 5 μm, such as 1.9 μm.

[0032] In the high performance liquid chromatography of a preferred embodiment, the column length of the octadecylsilane-bonded silica gel chromatographic column is 50 - 250 mm, such as 100 mm.

[0033] In the high performance liquid chromatography of a preferred embodiment, the inner diameter of the octadecylsilane-bonded silica gel chromatographic column is 1.8 - 4.6 mm, such as 3 mm.

[0034] In the high performance liquid chromatography of some embodiments, the column temperature is 30 - 50 °C, such as 40 °C.

[0035] In the mass spectrometry detection of some embodiments, the effluent with a retention time of 5 - 8.5 min in the high performance liquid chromatography is detected.

[0036] In the mass spectrometry of some embodiments, the ion mode is the positive ion mode.

[0037] In the mass spectrometry of some embodiments, the scanning mode is the multiple reaction monitoring mode.

[0038] In the mass spectrometry of some embodiments, the ion source is an electrospray ion source.

[0039] In the mass spectrometry of some embodiments, the drying gas is nitrogen.

[0040] In the mass spectrometry of some embodiments, the temperature of the drying gas is 100 - 350 °C, preferably 300 - 340 °C, such as 325 °C.

[0041] In the mass spectrometry of some embodiments, the flow rate of the drying gas is 1 - 13 L / min, such as 10 L / min.

[0042] In the mass spectrometry of some embodiments, the nebulizing gas pressure is 10 - 40 psi, such as 40 psi.

[0043] In the mass spectrometry of some embodiments, the sheath gas is nitrogen.

[0044] In the mass spectrometry of some embodiments, the temperature of the sheath gas is 250 - 400 °C, preferably 380 - 400 °C, such as 400 °C.

[0045] In the mass spectrometry of some embodiments, the flow rate of the sheath gas is 10 - 14 L / min, such as 11 L / min.

[0046] In the mass spectrometry of some embodiments, the capillary voltage is 3500 - 6000 V, preferably 3500 - 4500 V, such as 4000 V.

[0047] In the mass spectrometry of some embodiments, the fragmentation voltage is 0 - 200 V, preferably 60 - 150 V, more preferably 70 - 100 V, such as 60 V, 70 V, 80 V, 90 V, 100 V, 110 V, 120 V, 130 V, 140 V, 150 V, 160 V, 170 V, 180 V, 190 V or 200 V.

[0048] In the mass spectrometry of some embodiments, the collision energy is 5 - 35 V, preferably 10 - 20 V, such as 5 V, 8 V, 10 V, 12 V, 14 V, 16 V, 18 V, 20 V, 22 V, 24 V, 26 V, 28 V, 30 V, 32 V, 34 V or 35 V.

[0049] In the mass spectrometry of some embodiments, the detection ion pair of 3-[4-(2 - epoxyethylmethoxy)butoxy]-1,2 - propanediol is at least one of 221 / 57.1, 221 / 73.2 and 221 / 129.

[0050] On the basis of conforming to the common knowledge in the art, the above preferred conditions can be combined arbitrarily to obtain various preferred examples of the present application.

[0051] The reagents and raw materials used in the present application are all commercially available.

[0052] The positive and progressive effects of the present application are as follows: The liquid chromatography - mass spectrometry method provided by the present application can qualitatively and quantitatively detect 3 - [4 - (2 - epoxyethanylmethoxy)butoxy] - 1,2 - propanediol in the sample to be tested, with high detection sensitivity and accuracy and good reproducibility. Using the method of the present application, the quality of cross - linked sodium hyaluronate or cross - linked products using BDDE as a cross - linker can be monitored, reducing the potential risks of the above - mentioned materials. Description of the Drawings

[0053] Figure 1 It is the gas chromatogram of 3 - [4 - (2 - epoxyethanylmethoxy)butoxy] - 1,2 - propanediol under the detection conditions of Comparative Example 1;

[0054] Figure 2 It is the gas chromatogram of BDDE under the detection conditions of Comparative Example 1;

[0055] Figure 3 It is the gas chromatogram of 3 - [4 - (2 - epoxyethanylmethoxy)butoxy] - 1,2 - propanediol under the detection conditions of Comparative Example 2;

[0056] Figure 4 It is the gas chromatogram of BDDE under the detection conditions of Comparative Example 2;

[0057] Figure 5 It is the gas chromatogram of the mixture of 3 - [4 - (2 - epoxyethanylmethoxy)butoxy] - 1,2 - propanediol and BDDE under the detection conditions of Comparative Example 2;

[0058] Figure 6 It is the detection graph of the reference solution when the collision energy is 15V in Effect Example 3;

[0059] Figure 7 It is the detection graph of the reference solution when the collision energy is 5V in Effect Example 3;

[0060] Figure 8 It is the detection graph of the reference solution when the collision energy is 10V in Effect Example 3;

[0061] Figure 9 It is the detection graph of the reference solution when the collision energy is 20V in Effect Example 3;

[0062] Figure 10 It is the characteristic spectrogram of the blank solution in Effect Example 4;

[0063] Figure 11 It is the characteristic spectrogram of the reference solution in Effect Example 4;

[0064] Figure 12 It is the characteristic spectrogram of the test solution in Effect Example 4;

[0065] Figure 13 It is the characteristic spectrogram of the spiked test solution in Effect Example 4;

[0066] Figure 14 It is the standard curve graph drawn in Effect Example 9. Detailed implementation manners

[0067] The present application will be further described below by way of examples, but the present application is not limited to the scope of the described examples. For the experimental methods without specific conditions noted in the following examples, they are carried out according to conventional methods and conditions, or selected according to the product specifications.

[0068] In the following examples and comparative examples, hyaluronidase was purchased from Anhui Zesheng Technology Co., Ltd., and the product model was 005XR5FH;

[0069] In the following examples and comparative examples, the purities of formic acid and methanol were both chromatographically pure;

[0070] In the following examples and comparative examples, the water was ultrapure water;

[0071] In the following examples and comparative examples, the purity of the 3-[4-(2-oxiranylmethoxy)butoxy]-1,2-propanediol standard was 96.46%.

[0072] Example 1

[0073] (1) Preparation of solutions

[0074] Preparation of the test solution: Weigh 200 mg of crosslinked sodium hyaluronate, place it in a 10 mL volumetric flask, add 1 mL of water and 50 μL of an aqueous solution of hyaluronidase with a hyaluronidase concentration of 10000 IU / mL, incubate in a water bath at 37 °C for 1 hour to obtain an enzymatically hydrolyzed test sample, dilute to 10 mL with methanol, shake well, and filter through a 0.22 μm filter membrane.

[0075] Preparation of the reference solution: Take 3-[4-(2-oxiranylmethoxy)butoxy]-1,2-propanediol, dissolve it in water to make a solution containing 400 ng of 3-[4-(2-oxiranylmethoxy)butoxy]-1,2-propanediol per 1 mL. Accurately measure 1 mL and place it in a 10 mL volumetric flask, dilute to the mark with methanol, and shake well;

[0076] (2) High performance liquid chromatography - mass spectrometry conditions

[0077] Equipment: Agilent liquid chromatography - mass spectrometry instrument, model 1290 HPLC / G6470BMSD;

[0078] Chromatographic column: Octadecylsilyl - bonded silica gel chromatographic column, model Agilent Infinitylab poroshell120 EC - 18, column length 100 mm, inner diameter 3 mm, packing particle size 1.9 μm.

[0079] Chromatographic conditions: Injector temperature is 10 °C; Column temperature is 40 °C; Injection volume is 1 μL; 0.1% formic acid aqueous solution is used as mobile phase A, methanol is used as mobile phase B, the flow rate of the mobile phase is 0.3 mL / min; The elution gradient of the mobile phase is shown in Table 1 below.

[0080] Table 1

[0081]

[0082] Mass spectrometry conditions:

[0083] Ion source is ESI source (electrospray ionization source); Drying gas is nitrogen, drying gas temperature is 325 °C, drying gas flow rate is 10 L / min; Nebulizing gas pressure is 40 psi; Sheath gas is nitrogen, sheath gas temperature is 400 °C, sheath gas flow rate is 11 L / min; Capillary voltage is 4000 V; Fragmentor voltage is 70 V; Collision energy (CE) is 15 V; Ionization mode is positive ion mode; Scanning mode is multiple reaction monitoring mode (MRM); Quantitative ion pair is 221 / 73.2, qualitative ion pair is 221 / 57.1.

[0084] Valve switching mode is: to waste from 0 min, to mass spectrometry from 5 min, to waste from 8.5 min.

[0085] (3) Detection and calculation method: Detect the test solution and reference solution under the high - performance liquid chromatography - mass spectrometry conditions of step (2), calculate the concentration of 3 - [4 - (2 - oxiranylmethoxy) butoxy] - 1,2 - propanediol in the test solution by the external standard method, and further calculate the concentration of 3 - [4 - (2 - oxiranylmethoxy) butoxy] - 1,2 - propanediol in cross - linked sodium hyaluronate.

[0086] Comparative Example 1 uses gas chromatography

[0087] Gas chromatographic column: DB - 624 chromatographic column (30 m × 0.53 mm × 3.0 μm), stationary phase is 6% cyanopropyl - 94% dimethyl polysiloxane;

[0088] Gas chromatography conditions: initial temperature 150 °C, ramp to 240 °C at a rate of 30 °C / min, hold for 9 min, then ramp to 250 °C at a rate of 10 °C / min, hold for 12 min;

[0089] Detector: FID;

[0090] Carrier gas: nitrogen;

[0091] Carrier gas flow rate: 2 mL / min;

[0092] Injector temperature: 250 °C;

[0093] Detector temperature: 280 °C;

[0094] Split ratio: 1:1;

[0095] Injection volume: 2 μL;

[0096] Detector: FID;

[0097] Using the above gas chromatography conditions, 3-[4-(2-epoxyethanylmethoxy)butoxy]-1,2-propanediol and BDDE were tested respectively, and the results are shown in Figure 1 and Figure 2 .

[0098] It can be seen that the peak positions of 3-[4-(2-epoxyethanylmethoxy)butoxy]-1,2-propanediol and BDDE are both in the range of retention time 8 - 10 min and cannot be separated. At the same time, the peak area of BDDE is 1,511,836; the peak area of 3-[4-(2-epoxyethanylmethoxy)butoxy]-1,2-propanediol is only 106,229, which is much lower than the peak area of BDDE. When both substances are present in the system, the detection sensitivity of the analyte 3-[4-(2-epoxyethanylmethoxy)butoxy]-1,2-propanediol is low and the method is not applicable.

[0099] Comparative Example 2

[0100] Gas chromatography column: RTX-50 chromatographic column (30 m × 0.32 mm × 0.25 μm), the stationary phase is 50% phenyl + dimethylpolysiloxane;

[0101] Gas chromatography conditions: initial temperature 150 °C, ramp to 260 °C at a rate of 30 °C / min, hold for 10 min;

[0102] Carrier gas: nitrogen;

[0103] Carrier gas flow rate: 1.5 mL / min;

[0104] Injector temperature: 250 °C;

[0105] Detector temperature: 280 °C;

[0106] Split ratio: 1:1;

[0107] Injection volume: 2 μL;

[0108] Detector: FID;

[0109] Test solution of 3-[4-(2-oxiranylmethoxy)butoxy]-1,2-propanediol: Mix 3-[4-(2-oxiranylmethoxy)butoxy]-1,2-propanediol with methanol to prepare a test solution with a concentration of 1 mg / mL;

[0110] Test solution of BDDE: Mix BDDE with methanol to prepare a test solution with a concentration of 1 mg / mL;

[0111] Mixed test solution: Mix 3-[4-(2-oxiranylmethoxy)butoxy]-1,2-propanediol, BDDE and methanol to prepare a test solution with the concentration of each analyte being 100 μg / mL;

[0112] Test the above three prepared test solutions under the above gas chromatography conditions respectively, and the results are shown in Figures 3 - 5 .

[0113] It can be seen that according to Figure 3 and Figure 4 results, it can be known that impurities are generated for both 3-[4-(2-oxiranylmethoxy)butoxy]-1,2-propanediol and BDDE under the gas detection conditions. It is speculated that ring-opening reactions of the epoxy groups of the two analytes may occur during the detection process. In addition, according to Figure 5 and the data results, when the concentrations of BDDE and 3-[4-(2-oxiranylmethoxy)butoxy]-1,2-propanediol in the test solution are 1 mg / mL, the detection signal-to-noise ratios are 16.6 and 4.4 respectively. It can be seen that when the concentrations of the two substances are 1 mg / mL, the signal-to-noise ratios are both low, and this concentration is extremely significantly higher than the upper limit of the BDDE content control (2×10 -3 mg / mL) in the industry standard YY / T 0962-2021. When using this method to detect the contents of the two in cross-linked sodium hyaluronate, the signal-to-noise ratio will be smaller, the detection sensitivity is low, and the method is not applicable.

[0114] Example 2

[0115] Compared with Example 1, the difference is only that the solvent type in the preparation of the test solution in step (1) is different. Specifically, methanol is replaced with an equal amount of water, and the temperature of the injector in step (2) is adjusted to 25 °C. Other condition parameters are the same as those in Example 1. Test and calculate the concentration of 3-[4-(2-oxiranylmethoxy)butoxy]-1,2-propanediol in cross-linked sodium hyaluronate.

[0116] Examples 3 to 12

[0117] Compared with Example 1, the difference lies only in the breakdown voltage in step (2), and the breakdown voltage is adjusted to 80V, 90V, 100V, 110V, 120V, 130V, 140V, 150V, 180V or 200V respectively. Other condition parameters are the same as those in Example 1. Test and calculate the concentration of 3-[4-(2-oxiranylmethoxy)butoxy]-1,2-propanediol in cross-linked sodium hyaluronate.

[0118] Examples 13 to 15

[0119] Compared with Example 1, the difference lies only in the collision energy in step (2), and the collision energy is adjusted to 5V, 10V or 20V respectively. Other condition parameters are the same as those in Example 1. Test and calculate the concentration of 3-[4-(2-oxiranylmethoxy)butoxy]-1,2-propanediol in cross-linked sodium hyaluronate.

[0120] Recovery rate of Effect Example 1

[0121] Verify the recovery rates of the detection methods in Example 1 and Example 2 above. The specific experimental steps are as follows:

[0122] (1) Solution preparation

[0123] Test solution: The same as the preparation method of the test solution in Example 1 and Example 2 above;

[0124] Reference solution for Example 1: Take 3-[4-(2-oxiranylmethoxy)butoxy]-1,2-propanediol, dissolve it in water to prepare a solution containing 400 ng of 3-[4-(2-oxiranylmethoxy)butoxy]-1,2-propanediol per 1 mL as the stock solution; then take 1 mL of the stock solution into a 10 mL volumetric flask and make up the volume to 10 mL with methanol and shake well;

[0125] Spiked test solution for Example 1: Weigh 200 mg of cross-linked sodium hyaluronate, place it in a 10 mL volumetric flask, add 1 mL of the stock solution and 50 μL of hyaluronidase aqueous solution. The concentration of hyaluronidase in the hyaluronidase aqueous solution is 10000 IU / mL. Incubate in a water bath at 37 °C for 1 hour, make up the volume to 10 mL with methanol, shake well, and filter through a 0.22 μm filter membrane;

[0126] Reference solution for Example 2: Take 3-[4-(2-oxiranylmethoxy)butoxy]-1,2-propanediol, dissolve it in water to prepare a solution containing 400 ng of 3-[4-(2-oxiranylmethoxy)butoxy]-1,2-propanediol per 1 mL as the stock solution; then take 1 mL of the stock solution into a 10 mL volumetric flask and make up the volume to 10 mL with water and shake well;

[0127] Example 2 Test sample spiked solution: Weigh 200 mg of cross-linked sodium hyaluronate, place it in a 10 mL volumetric flask, add 1 mL of the stock solution and 50 μL of an aqueous solution of hyaluronidase. The concentration of hyaluronidase in the aqueous solution of hyaluronidase is 10000 IU / mL. Incubate in a water bath at 37 °C for 1 hour, make up the volume to 10 mL with water, shake well, and filter through a 0.22 μm filter membrane;

[0128] (2) Detect the corresponding reference solution, test solution and test sample spiked solution under the high performance liquid chromatography - mass spectrometry conditions in Example 1 or Example 2 above, and calculate the concentration of (3-[4-(2-oxiranylmethoxy)butoxy]-1,2-propanediol) in the test solution and the test sample spiked solution by the external standard method; then calculate the mass of 3-[4-(2-oxiranylmethoxy)butoxy]-1,2-propanediol in 200 mg of cross-linked sodium hyaluronate as the background amount; calculate the mass of 3-[4-(2-oxiranylmethoxy)butoxy]-1,2-propanediol in the test sample spiked solution prepared from 200 mg of cross-linked sodium hyaluronate, label it as the test amount, and calculate the recovery rate. Conduct parallel tests three times and take the average value. The results are shown in Table 2.

[0129] Recovery rate = (Test amount - Background amount) / Added amount × 100%, where the added amount is the mass of 3-[4-(2-oxiranylmethoxy)butoxy]-1,2-propanediol in 1 mL of the stock solution.

[0130] Table 2

[0131] Number Recovery Rate (%) Example 1 87.9 Example 2 78.2

[0132] Effect Example 2 Influence of breakdown voltage on response value

[0133] Replace the test solutions in Examples 1, 3 to 12 above with reference solutions, and detect the reference solutions using the detection methods corresponding to the above examples. Record the ion abundance at a mass-to-charge ratio of 221. The results are shown in Table 3.

[0134] Preparation of the reference solution: Take 3-[4-(2-oxiranylmethoxy)butoxy]-1,2-propanediol, dissolve it in water, and prepare a solution containing 50 μg of 3-[4-(2-oxiranylmethoxy)butoxy]-1,2-propanediol per 1 mL.

[0135] Table 3

[0136]

[0137]

[0138] According to the results in Table 3, the smaller the rupture voltage, the greater the ion abundance at the mass-to-charge ratio of 221, and the higher the detection sensitivity.

[0139] Effect Example 3 Effect of Collision Energy on Response Value

[0140] The test solution in the above-mentioned Examples 1 and 13 to 15 was replaced with the reference solution, and the reference solution was tested using the detection methods corresponding to the above-mentioned Examples. The test results are shown in FIG. Figures 6 - 9 , record the ion abundances at mass-to-charge ratios of 57.2 and 73.1. The results are shown in Table 4.

[0141] Preparation of reference solution: Take 3-[4-(2-oxiranylmethoxy)butoxy]-1,2-propylene glycol, dissolve it in water, and prepare a solution containing 50 μg of 3-[4-(2-oxiranylmethoxy)butoxy]-1,2-propylene glycol per 1 mL.

[0142] Table 4

[0143] Number Collision Energy Ion Abundance at m / z 57.2 Ion Abundance at m / z 73.1 Example 1 15V 23103 23012 Example 13 5V 3875 8282 Example 14 10V 12567 20226 Example 15 20V 27267 18635

[0144] According to the results in Table 4, when the detection method of the embodiment is used, both the qualitative ion pair (221 / 57.1) and the quantitative ion pair (221 / 73.2) have ideal response values, and when the collision energy is 15V, the detection sensitivity is the highest.

[0145] Effect Example 4 Specificity

[0146] The specificity of the detection method was verified with reference to the "Knowledge Principles for Registration Review of Qualitative and Quantitative Drug and In Vitro Release Studies in Drug-Device Combination Products with Medical Device Function as the Main Function".

[0147] (1) Solution preparation

[0148] Blank solution: Methanol and water were mixed in a volume ratio of 90:10 and labeled as Blank.

[0149] Reference solution: Take about 20 mg of 3-[4-(2-oxiranylmethoxy)butoxy]-1,2-propanediol standard, accurately weigh, place in a 10mL volumetric flask, dissolve and dilute to the scale with water, shake well, accurately measure 2 mL of the solution, place in a 100mL volumetric flask, dilute to the scale with water, shake well, accurately measure 1 mL of the solution, place in a 100mL volumetric flask, dilute to the scale with water, shake well, as the stock solution; accurately measure 2 mL of the stock solution, place in a 20mL volumetric flask, dilute to the scale with methanol, shake well, and obtain, labeled as STD.

[0150] Test solution: Accurately weigh about 200 mg of cross-linked sodium hyaluronate, place it in a 10 mL volumetric flask, add 1 mL of water and 50 μL of aqueous hyaluronidase solution (10000 IU / mL), seal, incubate in a water bath at 37 °C for about 1 hour, dilute to the mark with methanol, shake well, filter through a 0.22 μm filter membrane, and take the subsequent filtrate, labeled as SPL.

[0151] Spiked test solution: Accurately weigh about 200 mg of cross-linked sodium hyaluronate, place it in a 10 mL volumetric flask, accurately add 1 mL of the stock solution, add 1 mL of water and 50 μL of aqueous hyaluronidase solution (10000 IU / mL), seal, incubate in a water bath at 37 °C for about 1 hour, dilute to the mark with methanol, shake well, filter through a 0.22 μm filter membrane, and take the subsequent filtrate. Labeled as SPL-STD.

[0152] (2) Use the high performance liquid chromatography - mass spectrometry conditions in step (2) of Example 1 to test the blank solution, reference solution, test solution and spiked test solution respectively, and observe the retention time of the peaks. The results are shown in Table 5 and Figures 10 - 13 .

[0153] Table 5

[0154] Name Retention Time (min) Ion Abundance Blank Solution / 0 Reference Substance Solution 7.837 2854 Test Solution / 0 Spiked Test Solution 7.795 2546

[0155] (3) Verification conclusion

[0156] According to the results in Table 5, it can be seen that neither the blank solution nor the cross-linked sodium hyaluronate interferes with the determination of 3-[4-(2-oxiranylmethoxy)butoxy]-1,2-propanediol, and the detection method in Example 1 has specificity.

[0157] Effect Example 5 Quantitative limit and detection limit

[0158] Refer to the "Registration Review Guidelines for the Qualitative and Quantitative Determination of Drugs and In Vitro Release Studies in Drug - Device Combination Products with the Main Function of Medical Devices" to verify the quantitative limit and detection limit of the detection method.

[0159] (1) Solution preparation

[0160] Quantitation Limit Solution: Take about 20 mg of 3-[4-(2-epoxyethanylmethoxy)butoxy]-1,2-propanediol reference standard, weigh accurately, place it in a 10 mL volumetric flask, dissolve with water and dilute to the mark, shake well. Accurately measure 2 mL of this solution, place it in a 100 mL volumetric flask, dilute to the mark with water, shake well. Accurately measure 1 mL of this solution, place it in a 100 mL volumetric flask, dilute to the mark with water, shake well to obtain the stock solution; accurately measure 2 mL of the stock solution, place it in a 20 mL volumetric flask, dilute to the mark with methanol, shake well and label it as STD. Take 1 mL of the STD solution, place it in a 10 mL volumetric flask, dilute to the mark with methanol and shake well to obtain the solution. Prepare 6 portions in parallel and label them as LOQ-1 to LOQ-6 respectively.

[0161] Detection Limit Solution: Take 3 mL of the quantitation limit solution prepared above, place it in a 10 mL volumetric flask, dilute to the mark with methanol, shake well and label it as LOD.

[0162] (2) Test the quantitation limit solution and the detection limit solution respectively under the high performance liquid chromatography - mass spectrometry conditions in step (2) of Example 1, calculate the signal - to - noise ratio. The quantitation limit results are shown in Table 6 and the detection limit results are shown in Table 7.

[0163] Table 6

[0164]

[0165] Table 7

[0166]

[0167] (3) Verification Conclusion

[0168] The concentration of the LOQ solution is 4.01 ng / mL, which is equivalent to 10.0% of the limit concentration, not greater than 20% of the limit concentration. The signal - to - noise ratio of 3-[4-(2-epoxyethanylmethoxy)butoxy]-1,2-propanediol in 6 portions of the LOQ solution is between 10 and 14, all not less than 10. The quantitation limit verification result meets the standard.

[0169] The concentration of the LOD solution is 1.20 ng / mL, which is equivalent to 3.0% of the limit concentration, not greater than 10% of the limit concentration. The signal - to - noise ratio of 3-[4-(2-epoxyethanylmethoxy)butoxy]-1,2-propanediol in the LOD solution is 5, not less than 3. The detection limit verification result meets the standard.

[0170] Effect Example 6 Solution Stability

[0171] Verify the stability of the detection method with reference to the "Registration Review Guidelines for Qualitative and Quantitative Determination of Drugs and In Vitro Release Studies in Medicinal Device - Drug Combination Products with the Main Function of Medical Devices".

[0172] (1) Solution Preparation

[0173] Reference solution: Weigh accurately about 20 mg of the standard of 3-[4-(2-epoxyethanylmethoxy)butoxy]-1,2-propanediol, place it in a 10-mL volumetric flask, dissolve and dilute to the mark with water, shake well. Accurately pipette 2 mL of this solution into a 100-mL volumetric flask, dilute to the mark with water, shake well. Then accurately pipette 1 mL of this solution into a 100-mL volumetric flask, dilute to the mark with water, shake well to obtain the stock solution. Accurately pipette 2 mL of the stock solution into a 20-mL volumetric flask, dilute to the mark with methanol, shake well, and label it as STD.

[0174] Spiked test solution: Weigh accurately about 200 mg of cross-linked sodium hyaluronate, place it in a 10-mL volumetric flask, accurately add 1 mL of the stock solution, 1 mL of water, and 50 μL of hyaluronidase aqueous solution (10000 IU / mL), seal, incubate in a water bath at 37 °C for about 1 hour, dilute to the mark with methanol, shake well, filter through a 0.22-μm filter membrane, and take the subsequent filtrate, which is labeled as SPL-STD.

[0175] (2) Using the high performance liquid chromatography - mass spectrometry conditions in step (2) of Example 1, test the peak areas of the reference solution stored at room temperature for 0 h, 4 h, and 8.5 h respectively, and calculate the percentage change in peak area at 4 h and 8.5 h relative to that at 0 h. The results are shown in Table 8.

[0176] Using the high performance liquid chromatography - mass spectrometry conditions in step (2) of Example 1, test the peak areas of the spiked test solution stored at room temperature for 0 h, 4 h, and 8.5 h respectively, and calculate the percentage change in peak area at 4 h and 8.5 h relative to that at 0 h. The results are shown in Table 9.

[0177] Table 8

[0178] Storage Time at Room Temperature (h) Peak Area Percentage Change in Peak Area Relative to 0 h 0 2854 / 1 2777 97.3 8.5 2710 95.0

[0179] Table 9

[0180]

[0181]

[0182] (4) Verification conclusion

[0183] When the reference solution is placed at room temperature for 8.5 h, the percentage of the peak area at different time points compared with the peak area at 0 h is between 95.0% and 97.3%, and all are between 80% and 120%. The verification results meet the acceptance criteria.

[0184] When the spiked test solution is placed at room temperature for 8.5 h, the percentage of the peak area at different time points compared with the peak area at 0 h is between 92.1% and 95.3%, and all are between 80% and 120%. The verification results meet the acceptance criteria.

[0185] Effect Example 7 Accuracy and Precision

[0186] Verify the accuracy and precision of the detection method with reference to the "Registration Review Guidelines for Qualitative and Quantitative Determination of Drugs and In Vitro Release Studies in Drug-Device Combination Products with the Main Function of Medical Devices".

[0187] (1) Solution Preparation

[0188] Blank solution: Methanol and water are mixed in a volume ratio of 90:10 and labeled as Blank.

[0189] Reference solution: Take about 20 mg of 3-[4-(2-epoxyethylmethoxy)butoxy]-1,2-propanediol reference standard, accurately weighed, placed in a 10 mL volumetric flask, dissolved with water and diluted to the mark, shaken well. Accurately pipette 2 mL of this solution into a 100 mL volumetric flask, diluted to the mark with water, shaken well. Accurately pipette 1 mL of this solution into a 100 mL volumetric flask, diluted to the mark with water, shaken well to obtain the stock solution; accurately pipette 1 mL of the stock solution into a 10 mL volumetric flask, diluted to the mark with methanol, shaken well, and labeled as STD.

[0190] Test solution: Take about 200 mg of cross-linked sodium hyaluronate, accurately weighed, placed in a 10 mL volumetric flask, add 1 mL of water and 50 μL of hyaluronidase aqueous solution (10000 IU / mL), seal, incubate in a water bath at 37 °C for about 1 hour, diluted to the mark with methanol, shaken well, filtered through a 0.22 μm filter membrane, and take the subsequent filtrate to obtain the test solution, labeled as SPL.

[0191] LOQ concentration spiked solution: Take about 200 mg of cross-linked sodium hyaluronate, accurately weighed, placed in a 10 mL volumetric flask, accurately add 1 mL of the reference solution, add 1 mL of water and 50 μL of hyaluronidase aqueous solution (10000 IU / mL), seal, incubate in a water bath at 37 °C for about 1 hour, diluted to the mark with methanol, shaken well, filtered through a 0.22 μm filter membrane, and take the subsequent filtrate to obtain the test solution. Prepare 3 parallel portions, labeled as SPL-LOQ-1 to SPL-LOQ-3 respectively.

[0192] 100% concentration spiked solution: Take about 200 mg of cross-linked sodium hyaluronate, accurately weighed, placed in a 10 mL volumetric flask, accurately add 1 mL of the stock solution, add 1 mL of water and 50 μL of hyaluronidase aqueous solution (10000 IU / mL), seal, incubate in a water bath at 37 °C for about 1 hour, diluted to the mark with methanol, shaken well, filtered through a 0.22 μm filter membrane, and take the subsequent filtrate to obtain the test solution. Prepare 6 parallel portions, labeled as SPL-100STD-1 to SPL-100STD-6 respectively.

[0193] 150% spiked solution: Weigh accurately about 200 mg of cross-linked sodium hyaluronate, place it in a 10 mL volumetric flask, accurately add 1.5 mL of the stock solution, 1 mL of water, and 50 μL of hyaluronidase aqueous solution (10000 IU / mL), seal it, heat it in a water bath at 37 °C for about 1 hour, dilute it to the mark with methanol, shake well, filter it through a 0.22 μm filter membrane, and take the subsequent filtrate to obtain it. Prepare 3 portions in parallel, and label them as SPL-150STD-1 to SPL-150STD-3 respectively.

[0194] (2) Detect the solutions prepared above under the high performance liquid chromatography-mass spectrometry conditions in step (2) of Example 1, and calculate the concentration of 3-[4-(2-oxiranylmethoxy)butoxy]-1,2-propanediol in each solution by the external standard method; then calculate the mass of 3-[4-(2-oxiranylmethoxy)butoxy]-1,2-propanediol in 200 mg of cross-linked sodium hyaluronate as the background amount; calculate the mass of 3-[4-(2-oxiranylmethoxy)butoxy]-1,2-propanediol in each spiked solution, label it as the test amount, and calculate the recovery rate. Test it three times in parallel and take the average value. The results are shown in Table 10.

[0195] Recovery rate = (test amount - background amount) / added amount × 100%, where the added amount is the mass of 3-[4-(2-oxiranylmethoxy)butoxy]-1,2-propanediol in 1 mL of the stock solution.

[0196] Table 10

[0197]

[0198]

[0199] (3) Verification conclusion

[0200] The recovery rates of 3-[4-(2-oxiranylmethoxy)butoxy]-1,2-propanediol at the LOQ%, 100% and 150% concentration levels are between 84.9% and 105.0%, the average recovery rate is 96.1%, all are between 75% and 120%, the RSD value is 7.6%, not greater than 8%, and the verification result of the accuracy test meets the standard.

[0201] The RSD of the content of 3-[4-(2-oxiranylmethoxy)butoxy]-1,2-propanediol in SPL-100STD-1 to SPL-100STD-6 is 7.2%, not greater than 8%, so the verification result of the precision test meets the standard.

[0202] Effect Example 8 Durability

[0203] Verify the durability of the detection method with reference to the "Registration Review Guidelines for Qualitative and Quantitative Determination of Drugs and In Vitro Release Studies of Drug-Device Combination Products with the Main Function of Medical Devices".

[0204] (1) Solution preparation

[0205] Reference solution: Take about 20 mg of 3-[4-(2-oxiranylmethoxy)butoxy]-1,2-propanediol standard, weigh accurately, place it in a 10 mL volumetric flask, dissolve and dilute to the mark with water, shake well. Pipette 2 mL of this solution accurately, place it in a 100 mL volumetric flask, dilute to the mark with water, shake well. Pipette 1 mL of this solution accurately, place it in a 100 mL volumetric flask, dilute to the mark with water, shake well to obtain the stock solution. Pipette 1 mL of the stock solution accurately, place it in a 10 mL volumetric flask, dilute to the mark with methanol, shake well.

[0206] Spiked test solution: Take about 200 mg of cross-linked sodium hyaluronate, weigh accurately, place it in a 10 mL volumetric flask, accurately add 1 mL of the stock solution, 1 mL of water and 50 μL of hyaluronidase aqueous solution (10000 IU / mL), seal, incubate in a water bath at 37 °C for about 1 hour, dilute to the mark with methanol, shake well, filter through a 0.22 μm filter membrane, and take the subsequent filtrate, labeled as SPL-STD.

[0207] (2) Detect the above reference solution and spiked test solution using the high performance liquid chromatography-mass spectrometry conditions in step (2) of Example 1, and calculate the concentration of 3-[4-(2-oxiranylmethoxy)butoxy]-1,2-propanediol in the spiked test solution according to the external standard method. The results are shown in Table 11.

[0208] Change the concentration of formic acid in mobile phase A by 0.1 ± 0.01%, column temperature by 40 ± 2 °C, and mobile phase flow rate by 0.3 ± 0.01 mL / min, and observe the durability of the method. The results are shown in Table 11.

[0209] Table 11

[0210]

[0211] (3) Verification conclusion

[0212] By changing the concentration of formic acid aqueous solution in mobile phase A, column temperature and mobile phase flow rate, compared with Example 1, the difference in the content of 3-[4-(2-oxiranylmethoxy)butoxy]-1,2-propanediol in the spiked test solution is between 0.00 ppm and 0.04 ppm, all not greater than 0.2 ppm. Therefore, the verification result of the durability test meets the standard.

[0213] Effect Example 9 Linearity

[0214] Verify the linearity of the detection method with reference to the "Registration Review Guidelines for the Qualitative and Quantitative Determination of Drugs and In Vitro Release Studies in Medicinal Device-Drug Combination Products with the Main Function of Medical Devices".

[0215] (1) Solution preparation

[0216] Take about 20 mg of 3-[4-(2-epoxyethanylmethoxy)butoxy]-1,2-propanediol standard, accurately weigh it, place it in a 10 mL volumetric flask, dissolve it with water and dilute to the mark, shake well. Accurately pipette 2 mL of this solution, place it in a 100 mL volumetric flask, and dilute to the mark with water, shake well; accurately pipette 1 mL of this solution, place it in a 100 mL volumetric flask, and dilute to the mark with water, shake well to obtain the stock solution.

[0217] L-200%: Accurately pipette 2 mL of the stock solution, place it in a 10 mL volumetric flask, and dilute to the mark with methanol, shake well;

[0218] L-150%: Accurately pipette 1.5 mL of the stock solution, place it in a 10 mL volumetric flask, and dilute to the mark with methanol, shake well;

[0219] L-100%: Accurately pipette 1.0 mL of the stock solution, place it in a 10 mL volumetric flask, and dilute to the mark with methanol, shake well;

[0220] L-50%: Accurately pipette 0.5 mL of the stock solution, place it in a 10 mL volumetric flask, and dilute to the mark with methanol, shake well;

[0221] L-LOQ: Accurately pipette 2 mL of the stock solution, place it in a 20 mL volumetric flask, and dilute to the mark with methanol, shake well. Take 1 mL of this solution, place it in a 10 mL volumetric flask, and dilute to the mark with methanol, shake well.

[0222] Detect the test solutions containing standard products (i.e., analytes) with different concentrations above using the high performance liquid chromatography-mass spectrometry conditions in step (2) of Example 1. In the range of the LOQ - 200% limit solution, use the concentration of the analyte as the abscissa and the peak area (ion abundance ratio) as the ordinate to plot a graph to obtain the linear equation and correlation coefficient R of the analyte. The results are shown in Table 12 and Figure 14 , According to the results, it can be seen that the peak area is positively correlated with the concentration of 3-[4-(2-epoxyethanylmethoxy)butoxy]-1,2-propanediol, the correlation coefficient R = 0.9994, and the intercept / Y100% = 2.1%; among them, R is not less than 0.990, and the intercept / Y100% is not greater than 25%, and the linear result meets the acceptance criteria.

[0223] Table 12

[0224]

[0225] Finally, it should also be noted that in this application, the terms "comprising", "including" or any other variants thereof are intended to cover non-exclusive inclusion, so that a process, method, article or device comprising a series of elements not only includes those elements, but also includes other elements not expressly listed, or further includes elements inherent to such process, method, article or device.

[0226] Although the present application has been disclosed above through the description of specific embodiments of the present application, it should be understood that those skilled in the art can design various modifications, improvements or equivalents to the present application within the spirit and scope of the appended solutions. These modifications, improvements or equivalents should also be considered to be included within the scope claimed by the present application.

Claims

1. A method for detecting 3-[4-(2-oxiranylmethoxy)butoxy]-1,2-propylene glycol, characterized in that: The method comprises the following steps: using a high performance liquid chromatography-mass spectrometry method to conduct qualitative or quantitative detection of 3-[4-(2-oxiranylmethoxy)butoxy]-1,2-propylene glycol in a test liquid.

2. The detection method according to claim 1, characterized in that The test liquid satisfies the following conditions (1) and / or (2): (1) The test solution includes a test sample containing 3-[4-(2-oxiranylmethoxy)butoxy]-1,2-propylene glycol and a solvent, wherein the solvent includes an alcohol solvent and / or water; (2) The concentration of 3-[4-(2-oxiranylmethoxy)butoxy]-1,2-propanediol in the test solution is 4 to 85 ng / mL.

3. The detection method according to claim 2, characterized in that The test liquid satisfies the following conditions (1) and / or (2): (1) The alcohol solvent is selected from C1 to C3 alcohol solvents, preferably methanol; (2) The liquid sample to be tested is a sample containing 3-[4-(2-oxiranylmethoxy)butoxy]-1,2-propylene glycol obtained by chemical synthesis or a cross-linked product obtained using BDDE as a cross-linking agent.

4. The detection method according to claim 3, characterized in that The test liquid sample is a cross-linked polysaccharide prepared with BDDE as a cross-linking agent, preferably a cross-linked sodium hyaluronate prepared with BDDE as a cross-linking agent.

5. The detection method according to claim 3 or 4, characterized in that: The cross-linked product further includes a degradation treatment before detection; Preferably, the degradation method includes enzymatic degradation or chemical degradation.

6. The detection method according to claim 5, characterized in that The sample to be tested is cross-linked sodium hyaluronate prepared with BDDE as a cross-linking agent, and the degradation method comprises the following steps: enzymatically treating the cross-linked sodium hyaluronate with hyaluronidase in water; Preferably, the volume ratio of the water to the solvent in the test solution is (5-20):1; Preferably, the amount of the hyaluronidase is 1000 to 3000 IU / g based on the unit mass of the cross-linked sodium hyaluronate; Preferably, the hyaluronidase is added in the form of a hyaluronidase aqueous solution, and the concentration of the hyaluronidase in the hyaluronidase aqueous solution is 8000 to 15000 IU / mL; Preferably, the enzymatic hydrolysis temperature is 30-45°C; Preferably, the enzymatic hydrolysis time is 0.5 to 5 hours; Preferably, the enzymatic hydrolysis is followed by a filtration operation, wherein the filtration uses a filter membrane with a pore size of 0.2 to 0.5 μm.

7. The detection method according to any one of claims 1 to 6, characterized in that: The detection method satisfies at least one of the following conditions (1) to (7): (1) In high performance liquid chromatography, the mobile phase comprises a mobile phase A and a mobile phase B; the mobile phase A is an aqueous solution of a volatile acid, the mass percentage of the volatile acid in the aqueous solution of the volatile acid is 0.05% to 0.2%; the mobile phase B is methanol; (2) In HPLC, the flow rate of the mobile phase is 0.15 to 0.5 mL / min; (3) In high performance liquid chromatography, the elution mode is isocratic elution or gradient elution; (4) In high performance liquid chromatography, the temperature of the injector is 5 to 30°C; (5) In high performance liquid chromatography, the injection volume of the test solution is 0.5 to 5 μL; (6) In high performance liquid chromatography, the chromatographic column is an octadecylsilane bonded silica gel chromatographic column; (7) In high performance liquid chromatography, the column temperature is 30-50°C.

8. The detection method according to claim 7, characterized in that The detection method satisfies at least one of the following conditions (1) to (6): (1) The aqueous solution of the volatile acid is selected from an aqueous formic acid solution and / or an aqueous acetic acid solution; (2) The mass percentage of the volatile acid in the aqueous solution of the volatile acid is 0.08% to 0.12%; (3) The isocratic elution is adopted, and the volume percentage of the mobile phase A in the mobile phase is greater than 85%, preferably 85% to 95%; (4) using the gradient elution, 0-8.5 min, the volume percentage of the mobile phase A is 85%-92%, and the volume percentage of the mobile phase B is 8%-25%; 8.6-11 min, the volume percentage of the mobile phase A is 10%-30%, and the volume percentage of the mobile phase B is 70%-90%; 11.1-15min, the volume percentage of the mobile phase A is 75%-92%, and the volume percentage of the mobile phase B is 8%-25%; (5) In HPLC, the injector temperature is 8-15°C; (6) In the high performance liquid chromatography, the chromatographic column was Agilent Infinitylab poroshell 120EC-18 100 mm × 3 mm, 1.9 μm.

9. The detection method according to any one of claims 1 to 8, characterized in that: The detection method satisfies at least one of the following conditions (1) to (15): (1) In mass spectrometry detection, detecting the effluent with a retention time of 5 to 8.5 min by the high performance liquid chromatography; (2) In mass spectrometry, the ion mode is positive ion mode; (3) In mass spectrometry, the scanning mode is multiple reaction monitoring mode; (4) In mass spectrometry, the ion source is an electrospray ion source; (5) In mass spectrometry, the drying gas is nitrogen; (6) In mass spectrometry, the temperature of the drying gas is 100 to 350°C; (7) In mass spectrometry, the flow rate of the drying gas is 1 to 13 L / min; (8) In mass spectrometry, the nebulizer gas pressure is 10 to 40 psi; (9) In mass spectrometry, the sheath gas is nitrogen; (10) In mass spectrometry, the sheath gas temperature is 250–400 °C; (11) In mass spectrometry, the sheath gas flow rate is 10–14 L / min; (12) In mass spectrometry, the capillary voltage is 3500-6000 V; (13) In mass spectrometry, the fragmentation voltage is 0 to 200 V; (14) In mass spectrometry, the collision energy is 5–35 V; (15) In the mass spectrum, the detection ion pair of 3-[4-(2-oxiranylmethoxy)butoxy]-1,2-propanediol is at least one of 221 / 57.1, 221 / 73.2 and 221 / 129.

10. The detection method according to claim 9, characterized in that: The detection method satisfies at least one of the following conditions (1) to (5): (1) In mass spectrometry, the temperature of the drying gas is 300-340°C; (2) In mass spectrometry, the sheath gas temperature is 380-400°C; (3) In mass spectrometry, the capillary voltage is 3500-4500 V; (4) In mass spectrometry, the fragmentation voltage is 60 to 150 V; (5) In mass spectrometry, the collision energy is 10 to 20 V.

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