Method for detecting content of key auxiliary material sucrose octasulfate salt in liposome medicine and application of key auxiliary material sucrose octasulfate salt

Abstract

The invention discloses a method for detecting the content of sucrose octasulfate salt serving as a key auxiliary material in a liposome medicine. The method comprises the following steps: determining a reference substance solution by adopting a high performance liquid chromatography-evaporative light scattering detector, constructing a regression equation, and performing quantitative detection on a test solution according to the regression equation, the detection conditions of the high performance liquid chromatography are as follows: a YMC-Triart C18 chromatographic column is used, a mixed solution of ammonium acetate and acetonitrile is used as a mobile phase, and detection is performed after isocratic elution; the detection conditions of the evaporative light scattering detector are as follows: the temperature of a drift tube is 20-100 DEG C, the air velocity is 0.1-5L / min, and the gain is 1, 2, 4, 8 and 16; the test sample solution is a mixed solution of a test sample and a tetrahydrofuran solution. The method has the advantages of strong specificity, high sensitivity, high stability, good repeatability, good accuracy, low quantitation limit and low cost, and can realize quantitative detection of sucrose octasulfate in liposome drugs.

Description

Technical Field

[0001] The present invention belongs to the technical field of analysis and detection of pharmaceutical excipients, and more specifically, relates to a method for detecting the content of sucrose octasulfate, a key excipient in liposome drugs, and an application thereof. Background Art

[0002] Liposomes are artificial membranes formed primarily from amphiphilic molecules with hydrophilic heads and hydrophobic tails, typically composed of phospholipids, cholesterol, PEG, and other components. When amphiphilic molecules such as phospholipids and sphingolipids are dispersed in an aqueous phase, the hydrophobic tails of the molecules tend to aggregate, avoiding the aqueous phase, while the hydrophilic heads are exposed to the aqueous phase, forming closed vesicles with a bilayer structure called liposomes.

[0003] Liposomes are drugs that are encapsulated within an artificial biological membrane, forming a microscopic vesicle structure. When used as a pharmaceutical formulation, liposomes are able to bind to a variety of bioactive compounds due to their inherent amphiphilicity.

[0004] Sucrose octasulfate is a key excipient in liposome-derived pharmaceuticals. It is a salt-forming compound composed of sucrose octasulfate as an anion and metal ions such as potassium and sodium. During the preparation of some liposome-derived pharmaceuticals, a certain amount of sucrose octasulfate is added as an ion gradient builder to improve drug encapsulation efficiency. Therefore, during the preparation of liposome-derived pharmaceuticals, it is necessary to accurately measure the amount of sucrose octasulfate added to the raw material, track the amount lost during the process, and track the amount contained in the finished product. Detecting sucrose octasulfate ions can achieve these goals.

[0005] However, in liposome drugs, the detection method is often easily affected due to the complex drug composition, the small amount of target excipient sucrose octasulfate added, and the target existing in the form of ions inside the liposome.

[0006] Sucralfate (also known as aluminum sucrose sulfate), a variety listed in the United States Pharmacopoeia, was determined for sucrose octasulfate content using high-performance liquid chromatography-differential refractive index detection (HPLC-RID). However, experimental results showed that this method had poor reproducibility and sensitivity, with a tailing factor as high as 10 and a low number of theoretical plates.

[0007] "Comparison of Two Methods for the Quantification of Sucrose Octasulfate in Irinotecan Hydrochloride Liposomal Injection, Wang Huijia, School of Pharmacy, Shenyang Pharmaceutical University" describes a comparison of the determination of sucrose octasulfate in irinotecan hydrochloride liposomal injection using high-performance liquid chromatography with differential refractive index detection (HPLC-RID) and high-pressure ion chromatography with conductivity detection (HPIC-CD). The HPLC-RID method still suffers from poor sensitivity and a high limit of quantification (12.15 μg / mL). The HPIC-CD method, on the other hand, suffers from the high cost of ion chromatographs, high testing costs, high instrument maintenance costs, and demanding technical expertise, hindering widespread application.

[0008] Therefore, how to provide a detection method with strong specificity, high sensitivity, high stability, good repeatability, good accuracy, low limit of quantification and low cost has become a technical problem that needs to be solved urgently. Summary of the Invention

[0009] In response to the above-mentioned existing technical problems, the primary purpose of the present invention is to provide a method for detecting the content of sucrose octasulfate, a key excipient in liposome drugs. The method has the advantages of strong specificity, high sensitivity, high stability, good repeatability, good accuracy, low limit of quantification and low cost.

[0010] The second object of the present invention is to provide the application of the above method in the quality monitoring of liposome drugs or sucrose octasulfate.

[0011] In order to achieve the above object, the present invention is implemented through the following technical solutions:

[0012] The present invention claims protection for a method for detecting the content of sucrose octasulfate, a key excipient in liposome drugs, comprising the following steps: measuring a reference solution using a high performance liquid chromatography-evaporative light scattering detector, constructing a regression equation, and then quantitatively detecting a test solution based on the regression equation;

[0013] The detection conditions of the high performance liquid chromatography are as follows: using a YMC-Triart C18 chromatographic column, a mixed solution of ammonium acetate and acetonitrile as the mobile phase, and detecting after isocratic elution; the detection conditions of the evaporative light scattering detector are as follows: a drift tube temperature of 20 to 100° C., an air flow rate of 0.1 to 5 L / min, and a gain of 1, 2, 4, 8, or 16; and the test solution is a mixed solution of the test sample and a tetrahydrofuran solution.

[0014] The present invention uses a high-performance liquid chromatography-evaporative light scattering detector (HPLC-ELSD) to detect the content of sucrose octasulfate, a key excipient in liposome drugs. Based on the characteristics of the high-performance liquid chromatography-evaporative light scattering detector and the properties of sucrose octasulfate, the inventors optimized the HPLC method's chromatographic column, mobile phase, ELSD detector, and other conditions, thereby summarizing a method for detecting sucrose octasulfate in liposome drugs. The method has the advantages of strong specificity, high sensitivity, high stability, good repeatability, good accuracy, low limit of quantification, and low cost. Through research, the inventors found that when the YMC-Triart C18 chromatographic column was selected, the test spectrum had the advantages of good main peak shape and good separation. However, when other C18 chromatographic columns (such as Welchrom Ultimate XB-C18) were used, the test spectrum had poor main peak shape and poor separation.

[0015] Furthermore, because sucrose octasulfate is present within the liposomes of the liposome drug, it requires demulsification (destroying the liposome structure and exposing the substances stored within the liposomes) to dissolve the sucrose octasulfate salt before use in chromatographic analysis. The inventors discovered that when tetrahydrofuran was used to treat the sample, the sample solution became clear and transparent, demonstrating a better demulsification effect (e.g., the measured value in Comparative Example 2 was 0.9808 mg / ml). However, conventional methods using methanol for dissolution or demulsification resulted in an opaque, turbid solution, indicating inadequate demulsification, resulting in insufficient exposure of the sucrose octasulfate salt encapsulated by the liposomes (e.g., the measured value in Comparative Example 2 was 0.8450 mg / ml).

[0016] The method overcomes the shortcomings of the high performance liquid chromatography-differential refractive index detector (HPLC-RID) method, such as poor sensitivity and poor durability; and the shortcomings of the high pressure ion chromatography-conductivity detector (HPIC-CD) method, such as high cost and poor practicality. In the reverse engineering of liposome drug formulations, the method can accurately analyze the content of sucrose octasulfate in the original preparation, facilitating the development of generic liposome drugs. In the research of liposome drug formulation process, the method can accurately monitor the whereabouts of sucrose octasulfate, providing support for determining the prescription quantity, intermediate control, and the content limit of sucrose octasulfate in the finished product. The method can be used for the quality control of the excipient sucrose octasulfate in the establishment of quality standards for raw and excipient materials of liposome drugs.

[0017] Preferably, the tetrahydrofuran solution is a mixture of tetrahydrofuran and water in a volume ratio of 1-10:9-0. Further preferably, the tetrahydrofuran solution is a mixture of tetrahydrofuran and water in a volume ratio of 3-7:7-3. More preferably, the tetrahydrofuran solution is a mixture of tetrahydrofuran and water in a volume ratio of 5:4.

[0018] Preferably, in the detection conditions of high performance liquid chromatography, the specifications of the C18 chromatographic column are 250 mm×4.6 mm, 5 μm.

[0019] Preferably, in the high performance liquid chromatography conditions, the column temperature is 10 to 50° C. More preferably, the column temperature is 20 to 40° C. More preferably, the column temperature is 30° C.

[0020] Preferably, in the HPLC conditions, the flow rate of the mobile phase is 0.2 to 2 mL / min. More preferably, the flow rate of the mobile phase is 0.5 to 1.2 mL / min. More preferably, the flow rate of the mobile phase is 0.8 mL / min.

[0021] Preferably, under HPLC conditions, the injection volume is 1 to 100 μL. More preferably, the injection volume is 10 to 30 μL. More preferably, the injection volume is 15 μL.

[0022] Preferably, in the HPLC conditions, the concentration of the mobile phase is 0.01 to 0.5 mol / L. More preferably, the concentration of the mobile phase is 0.02 to 0.1 mol / L. More preferably, the concentration of the mobile phase is 0.05 mol / L.

[0023] Optionally, the liposome drug is irinotecan hydrochloride liposome.

[0024] Preferably, the limit of quantification of the sucrose octasulfate is 0.37 μg / mL.

[0025] Preferably, in the detection conditions of the evaporative light scattering detector, the air flow rate is 1 to 3 L / min.

[0026] Preferably, in the detection conditions of the evaporative light scattering detector, the drift tube temperature is 80-100°C.

[0027] Furthermore, the present invention seeks to protect the application of the above method in the quality monitoring of liposome drugs or sucrose octasulfate.

[0028] Compared with the prior art, the present invention has the following beneficial effects:

[0029] The present invention uses high performance liquid chromatography-evaporative light scattering detector (HPLC-ELSD) to detect the content of sucrose octasulfate, a key excipient in liposome drugs. By optimizing the HPLC method's chromatographic column, mobile phase, ELSD detector and other conditions, the content of sucrose octasulfate can be accurately measured. The quantification limit of sucrose octasulfate in the method of the present invention is 0.37 μg / mL, which is much lower than the quantification limit of 12.15 μg / mL of high performance liquid chromatography-differential refractive index detector. The detection method based on high performance liquid chromatography-evaporative light scattering detector (HPLC-ELSD) provided by the present invention has the advantages of strong specificity, high sensitivity, high stability, good repeatability, good accuracy, low quantification limit and low cost. The method can be used to establish quality standards for raw materials and excipients of liposome drugs. In the research of liposome drug formulation process, the whereabouts of sucrose octasulfate can be accurately monitored, providing support for determining the prescription quantity, intermediate control, and the content limit of sucrose octasulfate in the finished product. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] Figure 1 is the linear regression equation constructed based on the reference solution in Example 1.

[0031] Figure 2 Chromatograms of water, blank solvent, sample matrix, reference solution, and test solution in the specificity investigation of Example 2.

[0032] Figure 3 This is the chromatogram of the sucrose octasulfate reference solution with a concentration of 0.37 μg / ml in the quantitative limit investigation of Example 2.

[0033] Figure 4 This is the chromatogram of the linear investigation of sucrose octasulfate reference solution at different concentrations.

[0034] Figure 5 The chromatogram is detected using the Welchrom Ultimate XB-C18 column.

[0035] Figure 6 The chromatogram is detected using the YMC-Triart C18 column.

[0036] Figure 7 The figure shows the sample prepared by dissolving in tetrahydrofuran and methanol. Figure 7 The left side shows the test product prepared using tetrahydrofuran; Figure 7 The right side shows the sample prepared using methanol. DETAILED DESCRIPTION

[0037] The present invention will be further described below with reference to the accompanying drawings and specific examples, but the examples do not limit the present invention in any way. Unless otherwise specified, the reagents, methods and equipment used in the present invention are conventional reagents, methods and equipment in the art.

[0038] The irinotecan hydrochloride liposome injection used in the present invention (batch number: U22975, source: Les Laboratoires Servier, strength: 43 mg / 10 ml) comprises: irinotecan hydrochloride, distearoylphosphatidylcholine, cholesterol, phosphatidylethanolamine, potassium sucrose octasulfate, 4-hydroxyethylpiperazineethanesulfonic acid, sodium chloride, and water for injection. According to the EU Public Assessment Report on Irinotecan Hydrochloride Liposome Injection (onivyde-epar-public-assessment-report_en) (Assessment report EMA / CHMP / 589179 / 2016: pages 17-18), irinotecan hydrochloride liposome injection is a white to slightly yellowish opaque sterile liposome suspension. These are small, unilamellar lipid bilayer vesicles with a diameter of approximately 110 nm, enclosing an inner aqueous compartment. Irinotecan hydrochloride and sucrose octasulfate are present in the liposomes as a gel or precipitate. Therefore, its dissolution or demulsification is related to the formulation components of the liposome.

[0039] Example 1 A method for detecting the content of sucrose octasulfate, a key excipient in liposome drugs

[0040] (1) Solution preparation

[0041] Preparation of reference solution 1-3: Accurately weigh 12.395 mg of sucrose potassium octasulfate reference, dissolve in water, and quantitatively dilute to produce solutions containing 0.0751 mg, 0.0938 mg, and 0.1314 mg of sucrose potassium octasulfate per mL, respectively;

[0042] Preparation of test solution: Take 1 mL of irinotecan hydrochloride liposome injection (batch number: U22975, source: Les Laboratoires Servier, specification: 43 mg / 10 mL), which is a white opaque liquid, and place it in a 10 mL volumetric flask. After adding 5 mL of tetrahydrofuran, the solution becomes clear and transparent. Dilute with water to the scale, shake well, and prepare 2 parallel portions.

[0043] (2) Chromatographic conditions

[0044] Chromatographic column: Octadecylsilane bonded silica gel was used as the filler (specifically YMC-Triart C18, 250 mm × 4.6 mm, 5 μm);

[0045] Mobile phase: 0.05 mol / L ammonium acetate (3.854 g ammonium acetate was dissolved in 1000 mL of water)-acetonitrile (95:5) as the mobile phase;

[0046] Flow rate: 0.8 mL / min;

[0047] Run time: 7 minutes;

[0048] The detection conditions of the evaporative light scattering detector were Alltech 6100: drift tube temperature 100°C, air flow rate 2.0 L / min, and gain 1;

[0049] Column temperature: 30°C;

[0050] Injection volume: 15 μL.

[0051] (3) Detection: Accurately measure reference solution 1-3 and inject each into a liquid chromatograph. Perform detection using the above-mentioned chromatographic conditions and record the chromatogram. Calculate the linear regression equation using the logarithmic value of the reference solution concentration and the logarithmic value of the corresponding peak area. Separately, accurately measure the test solution and inject it into the liquid chromatograph. Record the chromatogram and calculate the sucrose octasulfate content in the test solution using the regression equation.

[0052] (4) Results

[0053] System suitability evaluation

[0054] System suitability was evaluated using a 0.0958 mg / mL sucrose octasulfate reference solution. The relative standard deviation (RSD) of the peak area for the five-point reference solution was 1.38%, less than 2.0%, indicating good system suitability. Specific data are shown in Table 1 below.

[0055] Table 1

[0056]

[0057] The linearity of the reference solution is shown in Table 2 and Figure 1 shown.

[0058] Table 2

[0059] Concentration of reference solution (mg / mL) 0.0766 0.0958 0.1150 Logarithmic value of the concentration of the reference solution -1.116 -1.019 -0.939 Sucrose octasulfate peak area 8202564 1146077 1500519 Logarithm of sucrose octasulfate peak area 5.914 6.059 6.176

[0060] Figure 1 is the regression equation constructed based on the reference solution of different concentrations in Example 1. Figure 1 It can be seen that the linear regression equation is y=1.4807x+7.5669, and the correlation coefficient is 0.9998, which is greater than 0.99, and the linearity is good.

[0061] The sucrose octasulfate content in the test solution was calculated using the regression equation. The measured values of two replicate samples were 0.9630 mg / mL and 0.9681 mg / mL, respectively. The average value was 0.9656 mg / mL.

[0062] Example 2 Methodological Investigation

[0063] (1) Specificity inspection

[0064] Referring to the method in Example 1, the specificity was investigated by chromatograms of water, blank solvent (tetrahydrofuran aqueous solution), sample matrix (the sample matrix was the liposome components of irinotecan hydrochloride liposome injection except sucrose octasulfate, mainly including active drug molecules, cholesterol, phospholipids, buffer salts, osmotic pressure regulators, etc.), reference solution and test solution.

[0065] Figure 2 The chromatograms of water, blank solvent (tetrahydrofuran aqueous solution), sample matrix, reference solution and test solution are used to investigate the specificity. Figure 2 It can be seen that both the blank solvent (tetrahydrofuran aqueous solution) and the sample matrix had no interference with the determination of sucrose octasulfate, and the specificity was good.

[0066] Figure 2 In the chromatogram of the test solution, the main peak is sucrose octasulfate peak; Figure 2 In the chromatogram of the reference solution, peak X is the sucrose octasulfate peak and peak Y is the potassium ion peak.

[0067] The inventors speculate that the appearance of the potassium ion peak here is the result of the combined effects of the ion pair effect of ammonium acetate in the mobile phase, the electrostatic adsorption of residual silanol groups on the C18 column, and the detection of non-volatile salts by the evaporative light scattering detector (ELSD). Even after the C18 column has been end-capped, a small amount of silanol groups (-SiOH) may still remain. Under the neutral or weakly alkaline conditions of the ammonium salt mobile phase of the present invention, the silanol groups partially dissociate into -SiO-, which adsorbs K through electrostatic interaction. + , resulting in weak retention of potassium ions in the C18 column. In addition, NH4 + and K + They are all cationic and may be competitively adsorbed on the residual silanol groups (-SiOH) on the surface of the silica matrix. + It is likely weakly retained by electrostatic interactions and finally detected as a peak by an evaporative light scattering detector (ELSD).

[0068] Figure 2 In the chromatogram of the reference solution, the peak areas of sucrose octasulfate peak and potassium ion peak are 1154447.45 and 1243391.95 respectively.

[0069] (2) Instrument precision inspection

[0070] System suitability was assessed using a 0.0949 mg / mL reference solution. The relative standard deviation (RSD) of the peak area for the five-point reference solution was 1.96%, less than 2.0%, indicating good instrument precision. Specific data are shown in Table 3 below.

[0071] Table 3

[0072]

[0073] (3) Quantitative limit investigation

[0074] Referring to the preparation and testing method of the reference solution in Example 1, a sucrose octasulfate reference solution with a concentration of 0.37 μg / mL was prepared for quantitative limit investigation.

[0075] Figure 3 The chromatogram of the sucrose octasulfate reference solution with a concentration of 0.37 μg / mL is shown in Figure 2. Figure 3 As can be seen, the signal-to-noise ratio of the chromatogram is 19, which is greater than 10, indicating that this concentration can be used as the limit of quantification. This shows that when the content of sucrose octasulfate in the sample is 0.37 μg / mL, this method can accurately detect its content.

[0076] (4) Linear investigation

[0077] Referring to the preparation and testing method of the reference solution in Example 1, the concentrations of the reference solutions were prepared to be 0.0380 mg / mL, 0.0664 mg / mL, 0.0949 mg / mL, 0.1235 mg / mL, and 0.1519 mg / mL, respectively, and the linearity of the reference solutions was investigated.

[0078] From Table 4 and Figure 4 As shown, the linear regression equation is y=1.4832x+7.5796, and the correlation coefficient is 0.9993, which is greater than 0.99, indicating that the sucrose octasulfate concentration is in the range of 0.038 mg / mL to 0.1519 mg / mL, with good linearity.

[0079] Table 4

[0080]

[0081] (5) Accuracy assessment

[0082] Referring to the preparation and testing method of the reference solution in Example 1, a sucrose octasulfate reference stock solution with a concentration of 0.8361 mg / mL was prepared for accuracy investigation.

[0083] Accurately measure 0.3 ml of the test sample, place it in a 10 ml volumetric flask, accurately add 1.0 ml of the reference substance stock solution, add 5 ml of tetrahydrofuran to dissolve, quantitatively dilute with water to the scale, shake well, that is, 50% concentration level, prepare three parallel copies; accurately measure 0.3 ml of the test sample, accurately weigh, place it in a 10 ml volumetric flask, accurately add 2.0 ml of the reference substance stock solution, add 5 ml of tetrahydrofuran to dissolve, quantitatively dilute with water to the scale, shake well, that is, 100% concentration level, prepare three parallel copies; accurately measure 0.5 ml of the test sample, place it in a 10 ml volumetric flask, accurately add 3.0 ml of the reference substance stock solution, add 5 ml of tetrahydrofuran to dissolve, quantitatively dilute with water to the scale, shake well, that is, 150% concentration level, prepare three parallel copies.

[0084] As shown in Table 5 below, in 9 accuracy test solutions, the recovery of the added amount of sucrose octasulfate ranged from 95.916% to 101.974%, and the average recovery was 98.63%, ranging from 95.0% to 105.0%. The RSD of the recovery (n=9) was 2.2%, less than 5.0%, indicating good accuracy of the method.

[0085] Table 5

[0086]

[0087]

[0088] (6) Repeatability study

[0089] Referring to the preparation and testing methods of the reference solution and the test solution in Example 1, sucrose octasulfate reference solutions with concentrations of 0.0575 mg / mL, 0.0766 mg / mL, 0.0958 mg / mL, 0.1150 mg / mL, 0.1342 mg / mL, and 0.1533 mg / mL were prepared for repeatability investigation.

[0090] Accurately pipette 1 ml of this product (irinotecan hydrochloride liposome injection), accurately weigh it, and place it in a 10 ml volumetric flask. Add 5 ml of tetrahydrofuran to dissolve it, dilute it to the mark with water, and shake it well. Prepare 6 parallel portions. Experimental results:

[0091] As shown in Table 6 below, the RSD of the determination results of the 6 test solutions was 0.8%, which was less than 2%, indicating that the method had good repeatability.

[0092] Table 6

[0093]

[0094] (7) Solution stability study

[0095] Referring to the preparation and testing methods for the reference solution in Example 1, a sucrose octasulfate reference solution with a concentration of 0.0958 mg / mL was prepared for stability testing. Accurately pipette 1 ml of the product, accurately weigh it, and place it into a 10 ml volumetric flask. Dissolve it in 5 ml of tetrahydrofuran, dilute to the mark with water, shake well, and perform a stability test.

[0096] As shown in Table 7 below, the RSDs of the main peak areas of the test solution and the reference solution were 1.8% and 0.9%, respectively, indicating that the test solution and the reference solution were stable within 48 hours.

[0097] Table 7

[0098]

[0099]

[0100] Analytical methodology validation has shown that this method can be applied to the determination of sucrose octasulfate in liposomes, given the complex and high-content matrix (multiple phospholipid components, the active ingredient, and other excipients) and the low content of the target compound, sucrose octasulfate (only 10% of the phospholipid + cholesterol components). The method is sufficiently sensitive to accurately detect sucrose octasulfate at levels as low as 0.37 μg / mL.

[0101] Comparative Example 1

[0102] The difference between this comparative example and Example 1 is that a chromatographic column (Welchrom Ultimate XB-C18 4.6 mm×150 mm, 5 μm) was used.

[0103] Figure 5 The chromatogram is detected using the Welchrom Ultimate XB-C18 column. Figure 6 The chromatogram is detected using the YMC-Triart C18 column.

[0104] Depend on Figure 5 and Figure 6 As can be seen, using the Welchrom Ultimate XB-C18 column results in poor main peak shape and resolution. However, using the YMC-Triart C18 column provides excellent main peak shape and resolution. Therefore, the YMC-Triart C18 column is specifically recommended for use with high-performance liquid chromatography coupled with an evaporative light scattering detector (HPLC-ELSD).

[0105] Comparative Example 2

[0106] The difference between this comparative example and Example 1 is that in step (1), in the preparation of the test solution, irinotecan hydrochloride liposome injection (batch number: U22975, source: Les Laboratoires Servier, specification: 43 mg / 10 ml) was dissolved in 5 mL of methanol, diluted to the scale with water, and shaken well.

[0107] Figure 7 The figure shows the sample prepared by dissolving in tetrahydrofuran and methanol. Figure 7 The left side shows the test product prepared using tetrahydrofuran; Figure 7 The right side shows the sample prepared using methanol.

[0108] Depend on Figure 7 It can be seen that when the sample solution was prepared using methanol, the sample solution was turbid and flocculent, indicating that the sample was not fully dissolved and the emulsion was not fully broken, resulting in insufficient exposure of the sucrose octasulfate salt encapsulated by the liposomes. Therefore, the test result was low, with a measured value of 0.8450 mg / ml. On the other hand, the sample solution prepared using tetrahydrofuran was clear and transparent, indicating complete dissolution, so the test result was normal, with a measured value (average) of 0.9808 mg / ml.

[0109] The foregoing examples are merely illustrative, serving to illustrate some of the features of the method of the present invention. The appended claims are intended to claim the widest possible scope that can be envisioned, and the embodiments presented herein are demonstrated by the applicant's actual experimental results. Therefore, the applicant intends that the appended claims are not limited by the selection of examples illustrating the features of the present invention. Some numerical ranges used in the claims also include subranges therein, and variations in these ranges should also be interpreted as being covered by the appended claims where possible.

Claims

1. A method for detecting the content of sucrose octasulfate, a key excipient in liposome drugs, characterized in that: The following steps are involved: The reference solution was determined by high performance liquid chromatography-evaporative light scattering detection, and a regression equation was constructed. The test solution was then quantitatively detected based on the regression equation. The HPLC detection conditions are as follows: using a YMC-Triart C18 chromatographic column, using a mixed solution of ammonium acetate and acetonitrile as the mobile phase, and performing detection after isocratic elution; The detection conditions of the evaporative light scattering detector are as follows: drift tube temperature 20-100°C, air flow rate 0.1-5 L / min, gain 1, 2, 4, 8, 16; The test solution is a mixed solution of the test sample and tetrahydrofuran solution.

2. The method according to claim 1, characterized in that The tetrahydrofuran solution is a solution in which tetrahydrofuran and water are mixed in a volume ratio of 1-10:9-0.

3. The method according to claim 1, characterized in that Ammonium acetate and acetonitrile are mixed in a volume ratio of 95-50:5-50.

4. The method according to claim 1, characterized in that In the detection conditions of high performance liquid chromatography, the specifications of the C18 chromatographic column are 250 mm × 4.6 mm, 5 μm.

5. The method according to claim 1, characterized in that: The detection conditions of high performance liquid chromatography are as follows: the column temperature is 10 to 50°C.

6. The method according to claim 1, characterized in that In the detection conditions of high performance liquid chromatography, the flow rate of the mobile phase is 0.2 to 2 mL / min.

7. The method according to claim 1, characterized in that: In the detection conditions of high performance liquid chromatography, the concentration of the mobile phase is 0.01 to 0.5 mol / L.

8. The method according to claim 1, characterized in that: The liposome drug is irinotecan hydrochloride liposome.

9. The method according to claim 1, characterized in that: The limit of quantification of the sucrose octasulfate salt was 0.37 μg / mL.

10. Use of the method according to any one of claims 1 to 9 in quality monitoring of liposome drugs or sucrose octasulfate.

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