Method for determining content of sucrose octasulfate in sucrose octasulfate triethylamine

Abstract

The invention relates to a method for measuring the content of sucrose octasulfate in sucrose octasulfate triethylamine, which comprises the following steps: step 1, preparing a linear solution; 2, preparing a quality control solution; step 3, preparing a test solution; and step 4, determination: taking the linear solution, the quality control solution and the test solution, injecting into a high performance liquid chromatograph to obtain a chromatogram, performing linear regression on the logarithm of the concentration of the sucrose octasulfate reference substance solution and the logarithm of the peak area of the corresponding main peak according to the peak area of the chromatogram, and calculating the content of sucrose octasulfate in the test solution.

Description

Technical Field

[0001] The invention relates to a drug detection method, in particular to a method for determining the content of sucrose octasulfate in sucrose octasulfate triethylamine. Background Art

[0002] Sucrose Octasulfate Triethylamine is a chemical substance, English name: Sucrose OctasulfateOctatriethylamine Salt, CAS number: 869561-07-9, molecular weight: 1185.19, molecular formula: C 18 H 37 NO 35 S 8， Its structural formula is as follows:

[0003]

[0004] Sucrose octasulfate is a drug used in the treatment of ulcers. It is then used as a non-polymer highly charged multivalent anion capture agent to increase the delivery of anticancer drugs. As reported in the literature, sucrose octasulfate triethylamine was used as a liposome excipient and the gradient method was used to prepare vinorelbine bitartrate liposomes. The results showed that the vinorelbine bitartrate liposome encapsulation rate was about 95% and the particle size was 129.5nm. Conclusion: The vinorelbine bitartrate liposomes prepared by the sucrose octasulfate triethylamine gradient method have a high encapsulation rate and are feasible.

[0005] Regarding the detection method, the relevant document "Comparison of Two Quantitative Methods for Determining Sucrose Octasulfate in Irinotecan Hydrochloride Liposome Injection" discloses a method for determining sucrose octasulfate by high performance liquid chromatography. However, the inventors found that the method in the article has the following defects:

[0006] 1. Using high performance liquid chromatography-differential refractive index detector (HPLC-RID) method: using Kromasil

[0007] 100-5-NH2 column (250mm×4.6mm, 5μm), 0.8mol·L-1 ammonium sulfate solution (pH 3.5)-acetonitrile (83:17) as the mobile phase, column temperature 30℃, detector temperature 30℃, flow rate 1.0mL·min-1. The sample pretreatment method of this method is relatively complicated and time-consuming.

[0008] 2. High-pressure ion chromatography-conductivity detector (HPIC-CD) method: Dionex InPacTMAS11-HC anion exchange column (250mm×4mm, 9μm), flow rate 1.5mL·min-1, column temperature 30℃, detector temperature 35℃, eluent 30mmol·L-1 sodium hydroxide solution. The conductivity detector used in this method has high maintenance costs and complex detection methods.

[0009] The evaporative light scattering detector used in the present invention has low maintenance cost and simple detection method. In addition, the document "HPLC method for determination of sucrose octasulfate content in sucralfate tablets" discloses the use of Agela VenusilXBP NH2 column, 0.8mol·L-1 ammonium sulfate solution (pH adjusted to 3.5 with phosphoric acid)-acetonitrile (83:17) as mobile phase, flow rate of 1.0mL·min-1, column temperature of 35℃, and detector as differential refractive index detector. Results The linear range of sucrose octasulfate is 3.13~97.95mg·mL-1, r=1.0000. There are differences in the content of sucrose octasulfate in sucralfate tablets from different manufacturers. Conclusion The content of sucrose octasulfate in sucralfate tablets was determined by high performance liquid chromatography for the first time in China, and the method has good repeatability.

[0010] However, the inventors found that the method has the following defects:

[0011] The differential refractive index detector used in this method has lower sensitivity than the evaporative light scattering detector used in the present invention, and the differential refractive index detector is more complicated to use, while the evaporative light scattering detector is simple and convenient to use.

[0012] There are also documents such as "A Review of Determination Methods of Sucrose Octasulfate in Irinotecan Hydrochloride Liposome Injection" and "Analysis of Potassium Sucrose Octasulfate by Pheromone Mixed Column HPLC". These documents more or less disclose the HPLC method for determining sucrose octasulfate, but all methods have similar defects, such as:

[0013] 1. The conductivity detector has high maintenance cost and the eluent is expensive, while the evaporative light scattering detector used in the present invention has low maintenance cost and the price of preparing the mobile phase is also relatively low.

[0014] 2. The sensitivity of the charged aerosol detector is lower than that of the evaporative light scattering detector used in the present invention.

[0015] In order to overcome the above-mentioned defects, the inventors have studied the existing technology and put forward improved ideas. Finally, through screening, a new chromatographic condition was obtained. The sucrose octasulfate content in sucrose octasulfate triethylamine was determined by high performance liquid chromatography. The precision, repeatability, sensitivity and simplicity of operation method are far superior to the existing technology. Summary of the invention

[0016] The present invention provides a method for determining the content of sucrose octasulfate in sucrose octasulfate triethylamine, wherein the method comprises the following steps:

[0017] Step 1, linear solution preparation

[0018] Step 2, quality control solution preparation

[0019] Step 3, preparation of test solution

[0020] Step 4, determination: take the linear solution, the quality control solution and the test solution, inject them into the high performance liquid chromatograph to obtain a chromatogram, and perform a linear regression on the logarithm of the concentration of the sucrose octasulfate reference solution and the logarithm of the corresponding main peak area according to the peak area of ​​the chromatogram to calculate the content of sucrose octasulfate in the test solution;

[0021] The chromatographic conditions are as follows:

[0022] High performance liquid chromatograph: evaporative light scattering detector Chromatographic column: Waters XBridge HILIC 4.6mm×250mm, 5μm or equivalent chromatographic column Mobile phase: Mobile phase A sodium hexane sulfonate solution (take 7.7g of 1-sodium hexane sulfonate, dissolve in water and dilute to 1000ml): mobile phase B acetonitrile = 15:85;

[0023] Flow rate: 1.0 ml / min; drift tube temperature: 80°C, carrier gas flow rate: 2.0 L / min, injection volume: 20 ul; column temperature: 35°C, run time: 20 min;

[0024] Isocratic elution

[0025] Preferably, the method of the present invention comprises the following steps:

[0026] Step 1, linear solution preparation: The method is as follows:

[0027] Take 35 mg of sucrose potassium octasulfate reference substance, accurately weigh it, put it in a 25 ml volumetric flask, dissolve it in water and dilute it to the scale, shake it well, and you will get the reference substance stock solution; prepare five linear test solutions of different concentrations according to the following table:

[0028] serial number Reference substance stock solution (ml) Volumetric flask (ml) STD-1 1 20 STD-2 2 20 STD-3 3 20 STD-4 4 20 STD-5 5 20

[0029] Step 2, quality control solution preparation: The method is as follows:

[0030] Take 35 mg of sucrose potassium octasulfate reference substance, accurately weigh it, put it in a 25 ml volumetric bottle, dissolve it with water and dilute it to the mark, shake it well; accurately measure 3 ml, put it in a 20 ml volumetric bottle, dilute it to the mark with water, shake it well, and you have it;

[0031] Step 3, preparation of test solution: The method is as follows:

[0032] Accurately weigh 1g of the test sample, place it in a 100ml volumetric flask, dilute it to the mark with water, and shake well; accurately measure 3ml, place it in a 10ml volumetric flask, dilute it to the mark with water, and shake well. Prepare two portions in parallel.

[0033] Step 4, determination, the method is as follows:

[0034] Take 5-50 μl of the linear solution, quality control solution, and test solution, inject them into the high performance liquid chromatograph to obtain a chromatogram, and perform a linear regression on the logarithm of the concentration of the sucrose octasulfate reference solution and the logarithm of the corresponding main peak area based on the peak area of ​​the chromatogram to calculate the content of sucrose octasulfate in the test solution.

[0035] Compared with the prior art, the present invention has the following differences:

[0036]

[0037] Compared with the prior art, the present invention has the following advantages:

[0038] The present invention Prior art Detector Low maintenance cost and easy to use High maintenance cost and complex to use Sampling method Convenient complex

[0039] The sample detection method of the present invention is simple and efficient. BRIEF DESCRIPTION OF THE DRAWINGS

[0040] Figure 1 blank solution

[0041] Figure 2 Blank excipient solution

[0042] Figure 3 Control solution

[0043] Figure 4 Test solution DETAILED DESCRIPTION

[0044] The present invention is further illustrated by the following examples, but are not intended to limit the present invention.

[0045] Example 1

[0046] The content of sucrose octasulfate in sucrose octasulfate triethylamine was determined by the following method.

[0047] 1. Chromatographic conditions:

[0048]

[0049] 2. Solution preparation:

[0050]

[0051]

[0052]

[0053] 3. Determination: Take 5-50μl of linear solution, quality control solution and test solution, inject into high performance liquid chromatograph, obtain chromatogram, and perform linear regression on the logarithm of the concentration of sucrose octasulfate reference solution and the logarithm of the corresponding main peak area according to the peak area of ​​the chromatogram to calculate the content of sucrose octasulfate in the test solution. The results are as follows:

[0054] The sucrose octasulfate content is 55 mmol / kg.

Claims

1. A method for determining the content of sucrose octasulfate in sucrose octasulfate triethylamine, the method comprising the following steps: Step 1, linear solution preparation Step 2, quality control solution preparation Step 3, preparation of test solution Step 4, determination: take the linear solution, the quality control solution and the test solution, inject them into the high performance liquid chromatograph to obtain a chromatogram, and perform a linear regression on the logarithm of the concentration of the sucrose octasulfate reference solution and the logarithm of the corresponding main peak area according to the peak area of ​​the chromatogram to calculate the content of sucrose octasulfate in the test solution; The chromatographic conditions are as follows: High Performance Liquid Chromatography: Evaporative Light Scattering Detector Chromatographic column: Waters XBridge HILIC 4.6mm×250mm, 5μm or equivalent chromatographic column Mobile phase: Mobile phase A sodium hexane sulfonate solution (take 7.7g of 1-sodium hexane sulfonate, dissolve in water and dilute to 1000ml): mobile phase B acetonitrile = 15:85; Flow rate: 1.0 ml / min; drift tube temperature: 80°C, carrier gas flow rate: 2.0 L / min, injection volume: 20 ul; column temperature: 35°C, run time: 20 min.

2. The method according to claim 1, characterized in that in Step 1, linear solution preparation: The method is as follows: Take 35 mg of sucrose potassium octasulfate reference substance, accurately weigh it, put it in a 25 ml volumetric flask, dissolve it in water and dilute it to the scale, shake it well, and you will get the reference substance stock solution; prepare five linear test solutions of different concentrations according to the following table:

3. The method according to claim 1, characterized in that in Step 2, quality control solution preparation: the method is as follows: Take 35 mg of sucrose potassium octasulfate reference substance, accurately weigh it, place it in a 25 ml volumetric flask, dissolve it with water and dilute it to the scale, shake it well; accurately measure 3 ml, place it in a 20 ml volumetric flask, dilute it to the scale with water, shake it well, and you have it.

4. The method according to claim 1, characterized in that in Step 3, preparation of test solution: The method is as follows: Accurately weigh 1 g of the test sample, place it in a 100 ml volumetric flask, dilute it to the mark with water, and shake well; accurately measure 3 ml, place it in a 10 ml volumetric flask, dilute it to the mark with water, shake well, and prepare two parallel portions.

5. The method according to claim 1, characterized in that in Step 4, determination, the method is as follows: Take 5-50 μl of the linear solution, quality control solution, and test solution, inject them into the high performance liquid chromatograph to obtain a chromatogram, and perform a linear regression on the logarithm of the concentration of the sucrose octasulfate reference solution and the logarithm of the corresponding main peak area based on the peak area of ​​the chromatogram to calculate the content of sucrose octasulfate in the test solution.

6. The method according to claim 1, characterized in that The method comprises the following steps: Step 1, linear solution preparation: the method is as follows: Take 35 mg of sucrose potassium octasulfate reference substance, accurately weigh it, put it in a 25 ml volumetric flask, dissolve it in water and dilute it to the scale, shake it well, and you will get the reference substance stock solution; prepare five linear test solutions of different concentrations according to the following table: Step 2, quality control solution preparation: the method is as follows: Take 35 mg of sucrose potassium octasulfate reference substance, accurately weigh it, put it in a 25 ml volumetric bottle, dissolve it with water and dilute it to the mark, shake it well; accurately measure 3 ml, put it in a 20 ml volumetric bottle, dilute it to the mark with water, shake it well, and you have it; Step 3, preparation of test solution: The method is as follows: Accurately weigh 1g of the test sample, place it in a 100ml volumetric flask, dilute it to the mark with water, and shake well; accurately measure 3ml, place it in a 10ml volumetric flask, dilute it to the mark with water, and shake well. Prepare two portions in parallel. Step 4, determination, the method is as follows: Take 5-50 μl of the linear solution, quality control solution, and test solution, inject them into a high performance liquid chromatograph to obtain a chromatogram, and perform a linear regression on the logarithm of the concentration of the sucrose octasulfate reference solution and the logarithm of the corresponding main peak area according to the peak area of ​​the chromatogram to calculate the content of sucrose octasulfate in the test solution; The chromatographic conditions are as follows: High Performance Liquid Chromatography: Evaporative Light Scattering Detector Chromatographic column: Waters XBridge HILIC 4.6mm×250mm, 5μm or equivalent chromatographic column Mobile phase: Mobile phase A sodium hexane sulfonate solution (take 7.7g of 1-sodium hexane sulfonate, dissolve in water and dilute to 1000ml): mobile phase B acetonitrile = 15:85; Flow rate: 1.0 ml / min; drift tube temperature: 80°C, carrier gas flow rate: 2.0 L / min, injection volume: 20 ul; column temperature: 35°C, run time: 20 min.

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