A method for detecting polyols

Abstract

The present application discloses a method for detecting polyols, which comprises the following steps: preparing a reference solution: dissolving polyols to obtain a reference solution; preparing a test solution: dissolving a test sample containing polyols to obtain a test solution; and determining the content of the polyols by high performance liquid chromatography-mass spectrometry. The method has good repeatability and high accuracy in detecting polyols. When the test sample contains polyols glycerol and butanediol, the recovery rates of glycerol and butanediol are within the range of 80-120%, and the detection limit is low and the intermediate precision is high.

Description

Technical Field

[0001] The present application relates to the field of detection technology, and in particular to a method for detecting polyols. Background Art

[0002] Cross-linking reaction refers to the reaction of two or more molecules (generally linear molecules) bonding and cross-linking to form a relatively stable molecule (bulk molecule) with a network structure. This reaction transforms linear or slightly branched macromolecules into a three-dimensional network structure, thereby improving strength, heat resistance, wear resistance, solvent resistance and other properties. For example, in the field of soft tissue filling, cross-linking reactions are often used to modify easily degradable polymers to prolong their degradation time in the body.

[0003] Cross-linking reactions are divided into chemical cross-linking and physical cross-linking. 1,4-Butanediol diglycidyl ether (BDDE) is a commonly used chemical cross-linking agent, which is often used for the cross-linking of high-molecular-weight mucopolysaccharides such as hyaluronic acid and polyglutamic acid. BDDE will degrade under certain conditions, and its degradation products are mainly polyols, such as glycerol and butanediol. Detecting the content of polyols in cross-linked products is an important indicator for evaluating the quality of cross-linked products. Currently, the commonly used polyol detection methods are liquid chromatography detection and gas chromatography detection. However, these detection methods cannot detect glycerol and butanediol at the same time. They can only be detected separately by two methods. The detection process is complicated, time-consuming, inefficient, and costly. Summary of the invention

[0004] In order to solve the problem that conventional polyol detection methods in the prior art are difficult to detect the content of polyols such as glycerol and / or butylene glycol, the present application provides a polyol detection method, which can accurately detect the content of polyols, such as the content of glycerol and / or butylene glycol, and can detect the content of polyols in test samples such as cross-linked sodium hyaluronate, cross-linked sodium polyglutamate, cross-linked polysaccharide fibers, cross-linked agarose, cross-linked starch or cross-linked chitosan.

[0005] Furthermore, when the sample contains two polyols, such as glycerol and butanediol, the method described in the present application can completely separate glycerol from butanediol and has specificity, applicability and repeatability.

[0006] The specific technical solutions of this application are as follows:

[0007] 1. A method for detecting a polyol, comprising:

[0008] Dissolving the polyol to obtain a standard solution;

[0009] dissolving a test sample containing a polyol to obtain a test sample solution;

[0010] The content of the polyol was determined by high performance liquid chromatography-mass spectrometry.

[0011] 2. The detection method according to item 1, wherein the chromatographic column in the high performance liquid chromatography is an alkylsilane bonded silica gel column, preferably an octadecylsilane bonded silica gel column.

[0012] 3. The detection method according to item 1 or 2, wherein the mobile phase A is a formic acid solution, preferably a 0.05-0.25 wt % formic acid solution; and the mobile phase B is methanol;

[0013] 4. The detection method according to item 3, wherein the volume ratio of the mobile phase A to the mobile phase B is 35:65-50:50.

[0014] 5. The detection method according to any one of items 1 to 4, wherein the flow rate is 0.15-0.25 ml / min.

[0015] 6. The detection method according to any one of items 1 to 5, wherein the column temperature of the chromatographic column is 30-40°C and the injection volume is 1-5 μL.

[0016] 7. The detection method according to any one of items 1 to 6, wherein the ion source is an electrospray ion source, and preferably, the ion source temperature is 100-120° C.;

[0017] Preferably, the capillary voltage is 0.1-0.5KV;

[0018] Preferably, the desolvation temperature is 450-550°C;

[0019] Preferably, the desolvation gas flow rate is 800-1200 L / Hr.

[0020] 8. A detection method according to any one of items 1 to 7, wherein the cone voltage is 10-15V.

[0021] 9. The detection method according to any one of items 1 to 8, wherein the content of the polyol is determined by an external standard method.

[0022] 10. The detection method according to any one of items 1 to 9, wherein the polyol is glycerol and / or butanediol.

[0023] 11. The detection method according to item 10, wherein the detection limit of glycerol is greater than or equal to 10x10 -3 ppm, the detection limit of butanediol is greater than or equal to 22x10 -3 ppm.

[0024] 12. The detection method according to any one of items 1 to 11, wherein the test solution comprises a solution of cross-linked sodium hyaluronate, cross-linked sodium polyglutamate, cross-linked polysaccharide fiber, cross-linked agarose, cross-linked starch or cross-linked chitosan containing a polyol.

[0025] Effects of the Invention

[0026] The method described in the present application has good repeatability and high accuracy in detecting polyols such as glycerol and butanediol. The recovery rates of glycerol and butanediol are in the range of 80-120%, and the detection limit is low and the intermediate precision is high.

[0027] The method disclosed in the present application has specificity, applicability and repeatability, and can be used to detect the content of residual glycerol and / or butylene glycol in cross-linked sodium hyaluronate, cross-linked polyglutamate, cross-linked polysaccharide fiber, cross-linked agarose, cross-linked starch or cross-linked chitosan, and then the preparation process of cross-linked sodium hyaluronate, cross-linked sodium polyglutamate, cross-linked polysaccharide fiber, cross-linked agarose, cross-linked starch or cross-linked chitosan can be adjusted to reduce the residual glycerol and / or butylene glycol, and the residual amount can be controlled within 0.15%, thereby improving the quality of cross-linked sodium hyaluronate, cross-linked polyglutamate, cross-linked polysaccharide fiber, cross-linked agarose, cross-linked starch or cross-linked chitosan products.

[0028] The method described in the present application does not require special treatment of the test sample, and a single injection can simultaneously perform qualitative and quantitative analysis on the compounds in the sample, which is simple to operate and has wide applicability.

[0029] Instruction Manual

[0030] Figure 1 This is a schematic diagram of the spectrum of blank solvent.

[0031] Figure 2 This is a schematic diagram of the spectrum of the glycerol reference substance.

[0032] Figure 3 This is a schematic diagram of the spectrum of 1,4-butanediol reference substance.

[0033] Figure 4 It is a schematic diagram of the spectrum of the test sample which is a cross-linked sodium hyaluronate solution. DETAILED DESCRIPTION

[0034] The present application is described in detail below in conjunction with the embodiments described in the accompanying drawings, wherein the same numbers in all the accompanying drawings represent the same features. Although specific embodiments of the present application are shown in the accompanying drawings, it should be understood that the present application can be implemented in various forms and should not be limited by the embodiments described herein. On the contrary, these embodiments are provided in order to enable a more thorough understanding of the present application and to be able to fully convey the scope of the present application to those skilled in the art.

[0035] It should be noted that certain words are used in the specification and claims to refer to specific components. Those skilled in the art should understand that technicians may use different nouns to refer to the same component. This specification and claims do not use the difference in nouns as a way to distinguish components, but use the functional differences of components as the criterion for distinction. As mentioned throughout the specification and claims, "including" or "comprising" are open-ended terms and should be interpreted as "including but not limited to". The subsequent description of the specification is a preferred embodiment of the present application, but the description is based on the general principles of the specification and is not intended to limit the scope of the present application. The scope of protection of the present application shall be determined by the attached claims.

[0036] The present application provides a method for detecting polyols, which comprises:

[0037] Dissolving the polyol to obtain a standard solution;

[0038] dissolving a test sample containing a polyol to obtain a test sample solution;

[0039] The content of the polyol was determined by high performance liquid chromatography-mass spectrometry.

[0040] Wherein, the calculation of the content of polyol can adopt external standard method well known in the art, and the external standard method is one of the commonly used methods of instrumental analysis and is a kind of comparative method.Compared with internal standard method, external standard method is not to add standard substance to the sample to be tested, but to measure separately under the chromatographic conditions identical with the sample to be tested, and the chromatographic peak area obtained is compared with the chromatographic peak area of ​​the tested component to obtain the content of the tested component.External standard and tested component are the same substance but require it to have certain purity, and the concentration of external standard should be close to the measured substance concentration during analysis, so as to facilitate the accuracy of quantitative analysis.The application can configure reference solution and test solution respectively, and utilize the peak area or peak height of reference to quantify the content of polyol in test solution with its corresponding concentration.

[0041] In the present application, there is no limitation on the preparation of the reference solution, which can be prepared according to conventional methods in the art. For example, a polyol can be dissolved in water to obtain a reference solution.

[0042] In the present application, there is no limitation on the preparation of the test solution, which can be prepared according to conventional methods in the art. For example, the test solution can be obtained by swelling the test sample containing the polyol with water, and preferably filtering the solution after swelling with water to obtain the filtrate as the test solution.

[0043] In the present application, the high performance liquid chromatography (HPLC) is developed on the basis of gas chromatography and classical chromatography. There is no essential difference between modern liquid chromatography and classical liquid chromatography. The only difference is that modern liquid chromatography has higher efficiency and realizes automated operation than classical liquid chromatography. In classical liquid chromatography, the mobile phase is transported at normal pressure, the column efficiency of the stationary phase used is low, and the analysis cycle is long. Modern liquid chromatography cites the theory of gas chromatography, and the mobile phase is changed to high-pressure transportation (the maximum transportation pressure can reach 4.9×107Pa); the chromatographic column is filled with small-particle fillers by a special method, so that the column efficiency is much higher than that of classical liquid chromatography (the number of plates per meter can reach tens of thousands or hundreds of thousands); at the same time, a highly sensitive detector is connected to the back of the column, which can continuously detect the effluent. Therefore, high performance liquid chromatography has the characteristics of fast analysis speed, high separation efficiency, and automation. So people call it high pressure, high speed, high efficiency or modern liquid chromatography.

[0044] In some embodiments, the chromatographic column in the high performance liquid chromatography is an alkylsilane bonded silica gel column, preferably, the chromatographic column is an octadecylsilane bonded silica gel column, such as a C18 column.

[0045] In some embodiments, mobile phase A is formic acid solution, preferably 0.05-0.20wt% formic acid solution, further preferably 0.05-0.15wt% formic acid solution; mobile phase B is methanol; preferably, the volume ratio of mobile phase A to mobile phase B is 35:65-50:50, preferably 38:62-42:58.

[0046] For example, mobile phase A can be 0.05wt%, 0.1wt%, 0.15wt%, 0.20wt% formic acid solution;

[0047] The volume ratio of mobile phase A to mobile phase B (v 流动相A :v 流动相B ) can be 35:65, 36:64, 37:63, 38:62, 39:61, 40:60, 41:59, 42:58, 43:57, 44:56, 45:55, 46:54, 47:53, 48:52, 49:51, 50:50, etc.

[0048] In some embodiments, the flow rate is 0.15-0.21 ml / min, preferably 0.19-0.21 ml / min.

[0049] In some embodiments, the column temperature of the chromatographic column is 30-40° C.; preferably, the injection volume is 0.1-0.5 μL, preferably 0.3-0.5 μL.

[0050] For example, the flow rate may be 0.15 ml / min, 0.19 ml / min, 0.20 ml / min, 0.21 ml / min, etc.;

[0051] The column temperature of the chromatographic column can be 30°C, 31°C, 32°C, 33°C, 34°C, 35°C, 36°C, 37°C, 38°C, 39°C, 40°C, etc.

[0052] In some embodiments, the ion source is an electrospray ion source, and preferably, the ion source temperature is 100-120°C;

[0053] Preferably, the capillary voltage is 0.1-0.5 KV, preferably 0.3-0.5 KV;

[0054] Preferably, the desolvation temperature is 450-550°C;

[0055] Preferably, the desolvation gas flow rate is 800-1200 L / Hr.

[0056] For example, the ion source temperature may be 100°C, 105°C, 110°C, 115°C, 120°C, etc.;

[0057] Capillary voltage is 0.1KV, 0.2KV, 0.3KV, 0.4KV, 0.5KV, etc.;

[0058] The desolvation temperature is 450°C, 460°C, 470°C, 480°C, 490°C, 500°C, 510°C, 520°C, 530°C, 540°C, 550°C, etc.

[0059] The desolvation gas flow rate is 800L / Hr, 900L / Hr, 1000L / Hr, 1100L / Hr, 1200L / Hr, etc.

[0060] In this application, Hr means hours.

[0061] In some embodiments, the polyol is glycerol and / or butanediol. When the sample contains glycerol and butanediol, the data acquisition mode m / z of glycerol is 115.0; the data acquisition mode m / z of butanediol is 72.8;

[0062] Preferably, the cone voltage is 10-15V, preferably 13-15V. For example, the cone voltage can be 10V, 11V, 12V, 13V, 14V, 15V, etc.

[0063] In some embodiments, the detection limit of glycerol is greater than or equal to 10x10 -3 ppm, the detection limit of butanediol is greater than or equal to 22x10 -3 ppm.

[0064] When the sample contains a polyol, such as glycerol or butanediol, the method described in the present application can detect the content of glycerol or butanediol. When the sample contains glycerol and butanediol, the method described in the present application can completely separate glycerol and butanediol with good repeatability and high accuracy. The recovery rates of glycerol and butanediol are in the range of 80-120%. The operation is simple, the detection limit is low, and no sample pretreatment is required.

[0065] In some embodiments, the test solution comprises a cross-linked mucopolysaccharide solution such as a solution of cross-linked sodium hyaluronate, cross-linked sodium polyglutamate, cross-linked polysaccharide fiber, cross-linked agarose, cross-linked starch or cross-linked chitosan containing a polyol.

[0066] For the above-mentioned cross-linked mucopolysaccharide, since it is a relatively viscous substance, the conventional method for detecting glycerol and butylene glycol is difficult to detect the residual amount of glycerol and butylene glycol in these test samples. The method described in the present application can detect the content of glycerol and / or butylene glycol in these test samples, and when glycerol and butylene glycol are contained, glycerol and butylene glycol can be completely separated with a high degree of separation.

[0067] Example

[0068] The present application provides a general and / or specific description of the materials and test methods used in the experiments. In the following examples, unless otherwise specified, % means wt%, i.e., weight percentage. The reagents or instruments used without indicating the manufacturer are all conventional reagent products that can be obtained commercially.

[0069] Example 1-1

[0070] 1. Preparation of reference solution:

[0071] Take glycerol and 1,4-butanediol reference substances respectively, weigh accurately, dissolve in water and quantitatively dilute to make solutions containing 0.75 μg of each per ml, shake well and set aside.

[0072] 2. Test solution:

[0073] Take about 10 mg of cross-linked sodium hyaluronate (Huaxi Biotechnology Co., Ltd.), accurately weigh it, put it in a 20 ml volumetric flask, add an appropriate amount of water to swell it, dilute it to the scale with water, let it stand, filter the supernatant, and take the filtrate; prepare two parallel portions.

[0074] Blank solution: Use solvent water as the blank solution.

[0075] The chromatographic conditions are:

[0076] Infinitylab Poroshell 120EC-C18 column, Agilent, 4.6x100nm, 2.7μm,

[0077] Flow rate: 0.2ml / min,

[0078] Column temperature 35°C,

[0079] Mobile phase: Mobile phase A (0.1% formic acid aqueous solution): Mobile phase B (methanol) = 40:60,

[0080] Injection volume: 5 μL.

[0081] Mass spectrometer detector conditions:

[0082] Ion source: ESI source, electrospray positive ion mode ESI+, single ion monitoring (SIR)

[0083] Ion source temperature: 110°C

[0084] Capillary voltage: 0.5KV

[0085] Desolvation temperature: 500℃

[0086] Desolvation gas flow rate: 1000L / Hr

[0087] Cone voltage: 15V

[0088] The data acquisition mode is: the mass-to-charge ratio of the glycerol detection ion is M / Z 115.0, and the mass-to-charge ratio of the 1,4-butanediol detection ion is M / Z 72.8

[0089] Take blank solution, reference solution and test solution for injection, and determine the content of glycerol and 1,4-butanediol in cross-linked sodium hyaluronate by external standard method, wherein the spectra of blank solvent, reference solution and test solution are as follows: Figure 1-4 shown.

[0090] The content was calculated using the following formula:

[0091] Calculation formula for glycerol and 1,4-butanediol content in the test sample:

[0092]

[0093] Note: i ---Content of glycerol or 1,4-butanediol in the test sample, %

[0094] C i ---Concentration of glycerol or 1,4-butanediol in the reference solution, ug / mL

[0095] A i---Peak area of ​​glycerol or 1,4-butanediol in the test solution

[0096] A r ---Peak area of ​​glycerol or 1,4-butanediol in the reference solution

[0097] W i ---Weighing amount of test sample, mg

[0098] After measurement, the contents of glycerol and 1,4-butanediol in the cross-linked sodium hyaluronate (batch number: 20201116) in Example 1-1 are as follows.

[0099] Table 1 Results of the content of glycerol and 1,4-butanediol in Example 1-1

[0100]

[0101]

[0102] Example 1-2

[0103] The ratio of the chromatographic mobile phase A to the mobile phase B in Example 1-1 was changed to 35:65. Other conditions were the same as those in Example 1-1. The measured contents of glycerol and 1,4-butanediol were shown in Table 2.

[0104] Table 2 Results of the content of glycerol and 1,4-butanediol in Example 1-2

[0105] Glycerol peak area 1,4-Butanediol peak area Average peak area of ​​reference substance 342473 24623 Test sample parallel 1 35422 3756 Test sample parallel 2 34744 3812 content(%) 0.015 0.030 Detection limit (ppm) <![CDATA[22x10 -3 ]]> <![CDATA[29x10 -3 ]]>

[0106] Examples 1-3

[0107] The volume ratio of the chromatographic mobile phase A to the mobile phase B in Example 1-1 was changed to 38:62, and the other conditions were the same as in Example 1-1. The measured contents of glycerol and 1,4-butanediol are shown in Table 3.

[0108] Table 3 Results of the content of glycerol and 1,4-butanediol in Example 1-2

[0109] Glycerol peak area 1,4-Butanediol peak area Average peak area of ​​reference substance 385785 26983 Test sample parallel 1 47695 5584 Test sample parallel 2 47146 5727 content(%) 0.018 0.041 Detection limit (ppm) <![CDATA[17x10 -3 ]]> <![CDATA[25x10 -3 ]]>

[0110] Examples 1-4

[0111] The volume ratio of the chromatographic mobile phase A to the mobile phase B in Example 1-1 was changed to 42:58, and the other conditions were the same as in Example 1-1. The measured contents of glycerol and 1,4-butanediol are shown in Table 4.

[0112] Table 4 Results of the content of glycerol and 1,4-butanediol in Examples 1-4

[0113]

[0114]

[0115] Examples 1-5

[0116] The ratio of the chromatographic mobile phase A to the mobile phase B in Example 1-1 was changed to 50:50, and the other conditions were the same as those in Example 1-1. The measured contents of glycerol and 1,4-butanediol are shown in Table 5.

[0117] Table 5 Glycerol and 1,4-Butanediol content results of Examples 1-5

[0118] Glycerol peak area 1,4-Butanediol peak area Average peak area of ​​reference substance 325753 23924 Test sample parallel 1 31856 3144 Test sample parallel 2 32865 3174 content(%) 0.015 0.026 Detection limit (ppm) <![CDATA[17x10 -3 ]]> <![CDATA[31x10 -3 ]]>

[0119] Examples 1-6

[0120] The chromatographic flow rate in Example 1-1 was changed to 0.19 ml / min, and other conditions were the same as in Example 1-1. The measured contents of glycerol and 1,4-butanediol are shown in Table 6.

[0121] Table 6 Glycerol and 1,4-Butanediol content results of Examples 1-6

[0122] Glycerol peak area 1,4-Butanediol peak area Average peak area of ​​reference substance 386534 29133 Test sample parallel 1 47356 4757 Test sample parallel 2 47785 4698 content(%) 0.018 0.035 Detection limit (ppm) <![CDATA[12x10 -3 ]]> <![CDATA[29x10 -3 ]]>

[0123] Examples 1-7

[0124] The chromatographic flow rate in Example 1-1 was changed to 0.15 ml / min, and the other conditions were the same as in Example 1-1. The measured contents of glycerol and 1,4-butanediol are shown in Table 26.

[0125] Table 7 Results of the content of glycerol and 1,4-butanediol in Examples 1-7

[0126]

[0127]

[0128] Examples 1-8

[0129] The chromatographic flow rate in Example 1-1 was changed to 0.21 ml / min, and other conditions were the same as in Example 1-1. The measured contents of glycerol and 1,4-butanediol are shown in Table 8.

[0130] Table 8 Glycerol and 1,4-Butanediol content results of Examples 1-8

[0131] Glycerol peak area 1,4-Butanediol peak area Average peak area of ​​reference substance 375744 28648 Test sample parallel 1 43854 5485 Test sample parallel 2 43380 5598 content(%) 0.017 0.037 Detection limit (ppm) <![CDATA[12x10 -3 ]]> <![CDATA[27x10 -3 ]]>

[0132] Examples 1-9

[0133] The column temperature of the chromatographic column in Example 1-1 was changed to 30° C., and other conditions were the same as those in Example 1-1. The measured contents of glycerol and 1,4-butanediol are shown in Table 9.

[0134] Table 9 Glycerol and 1,4-Butanediol Content Results of Examples 1-9

[0135] Glycerol peak area 1,4-Butanediol peak area Average peak area of ​​reference substance 384783 28239 Test sample parallel 1 47325 4645 Test sample parallel 2 47553 4633 content(%) 0.018 0.036 Detection limit (ppm) <![CDATA[14x10 -3 ]]> <![CDATA[24x10 -3 ]]>

[0136] Examples 1-10

[0137] The column temperature of the chromatographic column in Example 1-1 was changed to 40° C., and other conditions were the same as those in Example 1-1. The measured contents of glycerol and 1,4-butanediol are shown in Table 10.

[0138] Table 10 Results of the content of glycerol and 1,4-butanediol in Examples 1-10

[0139]

[0140]

[0141] Embodiment 1-11

[0142] The injection volume in Example 1-1 was changed to 3 μL, and other conditions were the same as in Example 1-1. The measured contents of glycerol and 1,4-butanediol are shown in Table 11.

[0143] Table 11 Results of the content of glycerol and 1,4-butanediol in Examples 1-11

[0144] Glycerol peak area 1,4-Butanediol peak area Average peak area of ​​reference substance 236574 17548 Test sample parallel 1 26536 3364 Test sample parallel 2 27643 3542 content(%) 0.017 0.038 Detection limit (ppm) <![CDATA[12x10 -3 ]]> <![CDATA[23x10 -3 ]]>

[0145] Examples 1-12

[0146] The injection volume in Example 1-1 was changed to 4 μL, and the other conditions were the same as in Example 1-1. The measured contents of glycerol and 1,4-butanediol are shown in Table 12.

[0147] Table 12 Results of the content of glycerol and 1,4-butanediol in Examples 1-12

[0148] Glycerol peak area 1,4-Butanediol peak area Average peak area of ​​reference substance 326562 23175 Test sample parallel 1 38214 4686 Test sample parallel 2 36314 4716 content(%) 0.017 0.033 Detection limit (ppm) <![CDATA[10x10 -3 ]]> <![CDATA[27x10 -3 ]]>

[0149] Examples 1-13

[0150] The concentration of mobile phase A in Example 1-1 was changed to 0.05 wt %. Other conditions were the same as in Example 1-1. The measured contents of glycerol and 1,4-butanediol are shown in Table 13.

[0151] Table 13 Results of the content of glycerol and 1,4-butanediol in Examples 1-13

[0152]

[0153]

[0154] Examples 1-14

[0155] The concentration of mobile phase A in Example 1-1 was changed to 0.15 wt %. Other conditions were the same as in Example 1-1. The measured contents of glycerol and 1,4-butanediol are shown in Table 14.

[0156] Table 14 Results of the content of glycerol and 1,4-butanediol in Examples 1-14

[0157] Glycerol peak area 1,4-Butanediol peak area Average peak area of ​​reference substance 398432 30112 Test sample parallel 1 47543 5235 Test sample parallel 2 47255 5319 content(%) 0.017 0.034 Detection limit (ppm) <![CDATA[11x10 -3 ]]> <![CDATA[24x10 -3 ]]>

[0158] Examples 1-15

[0159] The concentration of the chromatographic mobile phase A in Example 1-1 was changed to 0.20 wt %. Other conditions were the same as in Example 1-1. The measured contents of glycerol and 1,4-butanediol are shown in Table 15.

[0160] Table 15 Glycerol and 1,4-Butanediol content results of Examples 1-15

[0161] Glycerol peak area 1,4-Butanediol peak area Average peak area of ​​reference substance 355439 26353 Test sample parallel 1 38464 4327 Test sample parallel 2 37161 4642 content(%) 0.016 0.033 Detection limit (ppm) <![CDATA[13x10 -3 ]]> <![CDATA[31x10 -3 ]]>

[0162] Examples 1-16

[0163] The ion source temperature of the mass spectrometer in Example 1-1 was changed to 100°C, the capillary voltage of the mass spectrometer was changed to 0.3KV, the desolvation gas temperature was changed to 450°C, the desolvation gas flow rate was changed to 800L / Hr, the cone voltage was changed to 13V, and the other conditions were the same as in Example 1-1. The measured contents of glycerol and 1,4-butanediol are shown in Table 16.

[0164] Table 16 Glycerol and 1,4-Butanediol content results of Examples 1-16

[0165] Glycerol peak area 1,4-Butanediol peak area Average peak area of ​​reference substance 389322 29568 Test sample parallel 1 47121 5742 Test sample parallel 2 47936 5419 content(%) 0.018 0.037 Detection limit (ppm) <![CDATA[12x10 -3 ]]> <![CDATA[23x10 -3 ]]>

[0166] Examples 1-17

[0167] The ion source temperature of the mass spectrometer in Example 1-1 was changed to 120°C, the capillary voltage of the mass spectrometer was changed to 0.1 KV, the desolvation gas temperature was changed to 500°C, the desolvation gas flow rate was changed to 1200 L / Hr, the cone voltage was changed to 14 V, and the other conditions were the same as in Example 1-1. The measured contents of glycerol and 1,4-butanediol are shown in Table 17.

[0168] Table 17 Results of the content of glycerol and 1,4-butanediol in Examples 1-17

[0169] Glycerol peak area 1,4-Butanediol peak area Average peak area of ​​reference substance 369162 25753 Test sample parallel 1 41863 5433 Test sample parallel 2 41152 5242 content(%) 0.017 0.040 Detection limit (ppm) <![CDATA[21x10 -3 ]]> <![CDATA[34x10 -3 ]]>

[0170] Examples 1-18

[0171] The determination was carried out according to the chromatographic conditions of Example 1-1, except that the test sample used in Example 1-18 was cross-linked sodium polyglutamate. The determined contents of glycerol and 1,4-butanediol are shown in Table 18.

[0172] Table 18 Glycerol and 1,4-butanediol content results of Examples 1-18

[0173] Glycerol peak area 1,4-Butanediol peak area Average peak area of ​​reference substance 357749 26847 Test sample parallel 1 17463 6743 Test sample parallel 2 15278 6157 content(%) 0.007 0.047 Detection limit (ppm) <![CDATA[17x10 -3 ]]> <![CDATA[31x10 -3 ]]>

[0174] Examples 1-19

[0175] The determination was carried out according to the chromatographic conditions of Example 1-6, except that the test sample used in Example 1-19 was cross-linked sodium polyglutamate. The determined contents of glycerol and 1,4-butanediol are shown in Table 19.

[0176] Table 19 Glycerol and 1,4-Butanediol content results of Examples 1-19

[0177] Glycerol peak area 1,4-Butanediol peak area Average peak area of ​​reference substance 377553 27975 Test sample parallel 1 17595 6275 Test sample parallel 2 19371 6847 content(%) 0.007 0.045 Detection limit (ppm) <![CDATA[15x10 -3 ]]> <![CDATA[26x10 -3 ]]>

[0178] Examples 1-20

[0179] The determination was carried out according to the chromatographic conditions of Example 1-13, except that the test sample used in Example 1-20 was cross-linked sodium polyglutamate. The determined contents of glycerol and 1,4-butanediol are shown in Table 20.

[0180] Table 20 Glycerol and 1,4-Butanediol content results of Examples 1-20

[0181] Glycerol peak area 1,4-Butanediol peak area Average peak area of ​​reference substance 374462 29653 Test sample parallel 1 15245 6911 Test sample parallel 2 16932 6928 content(%) 0.006 0.046 Detection limit (ppm) <![CDATA[17x10 -3 ]]> <![CDATA[23x10 -3 ]]>

[0182] Table 21 Chromatographic conditions used in Examples 1-1 to 1-20

[0183]

[0184]

[0185] Example 2-1 Determination of Repeatability

[0186] According to the test sample processing method in Example 1-1, the same batch number sample as in Example 1-1 was taken and 6 parallel preparations were made. The reference solution and detection conditions were the same as in Example 1-1. The measurement results are shown in Tables 22 and 23.

[0187] Table 22 Glycerol content results of Example 2-1

[0188]

[0189] Table 23 Results of 1,4-butanediol content in Example 2-1

[0190]

[0191]

[0192] Example 2-2 Determination of inter-day precision

[0193] According to the test sample processing method in Example 1-1, the same batch number samples as in Example 1-1 were taken and 6 samples were prepared in parallel at different times. The reference solution and detection conditions were the same as in Example 1-1, but different instruments were used. The measurement results are shown in Tables 22 and 23.

[0194] Table 24 Glycerol content results of Example 2-2

[0195]

[0196] Table 25 1,4-Butanediol content results of Example 2-2

[0197]

[0198] Determination of Accuracy of Example 2-3

[0199] Take a blank solution without glycerol and 1,4-butanediol, treat the blank solution according to the test sample treatment method in Example 1-1, accurately weigh the reference substance in Example 1-1, simulate the prescription to prepare the test solution with glycerol and 1,4-butanediol content of 80%, 100%, and 120%, respectively, prepare 3 portions in parallel, a total of 9 portions. The reference solution and detection conditions are the same as those in Example 1-1, and the measurement results are shown in Tables 26 and 27.

[0200] Table 26 Glycerol accuracy verification results

[0201]

[0202] Table 27 1,4-Butanediol accuracy verification results

[0203]

[0204] Comparative Example 1

[0205] The mobile phase A in Example 1-1 was changed to 0.1% trifluoroacetic acid, and the other conditions were the same as in Example 1-1. Glycerol and 1,4-butanediol may not be completely detected. The detection results are shown in Table 28.

[0206] Table 28 Results of the content of glycerol and 1,4-butanediol in Comparative Example 1

[0207] Glycerol peak area 1,4-Butanediol peak area Average peak area of ​​reference substance 314431 19754 Test sample parallel 1 16742 1652 Test sample parallel 2 18463 1811 content(%) 0.008 0.017 Detection limit (ppm) <![CDATA[81x10 -3 ]]> <![CDATA[76x10 -3 ]]>

[0208] Comparative Example 2

[0209] The ratio of mobile phase A to mobile phase B in Example 1-1 was changed to 30:70, and the other conditions were the same as in Example 1-1. Glycerol and 1,4-butanediol may not be completely detected. The detection results are shown in Table 29.

[0210] Table 29 Results of the content of glycerol and 1,4-butanediol in Comparative Example 2

[0211] Glycerol peak area 1,4-Butanediol peak area Average peak area of ​​reference substance 276311 15422 Test sample parallel 1 13743 1157 Test sample parallel 2 14223 1224 content(%) 0.007 0.015 Detection limit (ppm) <![CDATA[56x10 -3 ]]> <![CDATA[79x10 -3 ]]>

[0212] Comparative Example 3

[0213] The concentration of the chromatographic mobile phase A in Example 1-1 was changed to 0.01 wt %, and the other conditions were the same as in Example 1-1. Glycerol and 1,4-butanediol may not be completely detected. The detection results are shown in Table 30.

[0214] Table 30 Results of the content of glycerol and 1,4-butanediol in Comparative Example 3

[0215]

[0216]

[0217] Comparative Example 4

[0218] The chromatographic column in Example 1-1 was changed to an amino column, and other conditions were the same as in Example 1. The detection results showed that no glycerol and 1,4-butanediol were found in the test sample.

[0219] Comparative Example 5

[0220] The ratio of mobile phase A to mobile phase B in Example 1-18 was changed to 30:70, and other conditions were the same as in Example 1-18. Glycerol and 1,4-butanediol may not be completely detected. The detection results are shown in Table 31.

[0221] Table 31 Results of the content of glycerol and 1,4-butanediol in Comparative Example 5

[0222] Glycerol peak area 1,4-Butanediol peak area Average peak area of ​​reference substance 286547 14854 Test sample parallel 1 6475 755 Test sample parallel 2 7160 631 content(%) 0.003 0.009 Detection limit (ppm) <![CDATA[62x10 -3 ]]> <![CDATA[101x10 -3 ]]>

[0223] The above is only the preferred embodiment of the present application, and does not limit the present application in other forms. Any technician familiar with the profession may use the above disclosed technical content to change or modify it into an equivalent embodiment with equivalent changes. However, any simple modification, equivalent change and modification made to the above embodiment based on the technical essence of the present application without departing from the technical solution of the present application still falls within the protection scope of the technical solution of the present application.

Claims

1. A method for detecting a polyol, comprising: Dissolving the polyol to obtain a standard solution; dissolving a test sample containing a polyol to obtain a test sample solution; High performance liquid chromatography-mass spectrometry was used to measure and obtain the content of polyol; The chromatographic column in the high performance liquid chromatography is an alkylsilane bonded silica gel column; Mobile phase A is 0.05-0.25wt% formic acid solution; mobile phase B is methanol; The volume ratio of the mobile phase A to the mobile phase B is 35:65-50:50; The polyol comprises glycerol and butylene glycol.

2. The detection method according to claim 1, wherein: The chromatographic column in the high performance liquid chromatography is an octadecylsilane bonded silica gel column.

3. The detection method according to claim 1, wherein The flow rate is 0.15-0.25 ml / min.

4. The detection method according to claim 1, wherein: The column temperature of the chromatographic column is 30-40°C and the injection volume is 1-5 μL.

5. The detection method according to claim 1, wherein: The ion source is an electrospray ion source.

6. The detection method according to claim 5, wherein: The ion source temperature is 100-120°C.

7. The detection method according to claim 5, wherein: The capillary voltage is 0.1-0.5KV.

8. The detection method according to claim 5, wherein: The desolvation temperature is 450-550°C.

9. The detection method according to claim 5, wherein: The desolvation gas flow rate is 800-1200L / Hr.

10. The detection method according to claim 1, wherein: The cone voltage is 10-15V.

11. The detection method according to any one of claims 1 to 10, wherein: The content of polyols was determined by external standard method.

12. The detection method according to claim 1, wherein The detection limit of glycerol is greater than or equal to 10x10 -3 ppm, the detection limit of butanediol is greater than or equal to 22 x10 -3 ppm.

13. The detection method according to claim 1, wherein: The test solution comprises a solution of cross-linked sodium hyaluronate, cross-linked sodium polyglutamate, cross-linked polysaccharide fiber, cross-linked agarose, cross-linked starch or cross-linked chitosan containing a polyol.

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