Analytical methods for multivesicular liposome compositions

Abstract

The application discloses an analysis method of a multivesicular liposome composition and belongs to the technical field of drug analysis. The analysis method is characterized by adopting a C8 chromatographic column, a buffer aqueous solution and an organic phase as a mobile phase, gradient elution, analysis and separation of composition components and hydrolysis products of the multivesicular liposome composition, and quantitative calculation of the composition components and the hydrolysis product content. The application is favorable for better monitoring of the quality of the multivesicular liposome composition product and provides a powerful guarantee for development of the multivesicular liposome composition.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of pharmaceutical analysis, and particularly relates to a method for determining the composition of a multivesicular liposome (MVL) composition and hydrolysis products. BACKGROUND

[0002] Liposomes are vesicles composed mainly of phospholipids and cholesterol, which are similar to the bilayer structure of biological membranes. The particle size of liposomes is in the nanometer and micrometer range, and they are a new type of drug carrier. Lipid components can be biodegraded in vivo and have good tolerance. According to whether the liposome is concentric, the liposome can be divided into single-chamber liposomes and multi-chamber liposomes with concentric structures, and multivesicular liposomes (MVL) with non-concentric structures. The concentric structure liposome is composed of one or more concentric lipid bilayers, and the structure is similar to a ping-pong ball or an onion. The multivesicular liposome composed of non-concentric multiple lipid layers has a honeycomb structure similar to a pomegranate. Among them, the multivesicular liposome (MVL) is a new type of liposome prepared by using a reservoir foam technology. Its characteristics are that it can accumulate in large quantities at the injection site or the drug delivery chamber, has good sustained-release effect and reservoir effect, can effectively reduce the number of drug administrations for patients, and improve the treatment compliance of patients.

[0003] The drug-encapsulating multivesicular liposome (MVL) composition usually contains a drug, which is often present in multiple internal aqueous chambers of the MVL separated by a lipid membrane, wherein the lipid membrane contains 1,2-dierucoylphosphatidylcholine (DEPC), 1,2-palmitoylphosphatidylglycerol (DPPG) and at least one neutral lipid, and an aqueous medium, the neutral lipid is often triolein (TO), tripalmitin, trimyristin, trilinolein, tributyrin, trihexanoin, tricaprylin and tricaprin. The drug-encapsulating multivesicular liposome (MVL) composition also often adds cholesterol and / or phytosterol to promote the accumulation of lipid chains and the formation of bilayers, reduce the fluidity of bilayers, and reduce the transmembrane transport of water-soluble drugs.

[0004] The prior art discloses simultaneous quantitative determination of 1,2-dierucoylphosphatidylcholine (DEPC), 1,2-palmitoylphosphatidylglycerol (DPPG) and fatty acid glyceride in a pharmaceutical preparation by using gas chromatography, which needs a pretreatment operation such as saponification on the sample, and the pretreatment means is complex; patent CN104931618A discloses a multiple detection method for six kinds of phospholipids, which uses chloroform / methanol / water as a mobile phase system, and needs to use a normal phase chromatographic column, and a large amount of organic solvents such as chloroform is consumed in the analysis process, which does not meet the requirements of green environmental protection. In view of the shortcomings of the prior art, a method for simultaneously analyzing multiple substances in a multivesicular liposome (MVL) composition is needed, which can better monitor the quality of the multivesicular liposome (MVL) composition product, and at the same time avoid controlling the multivesicular liposome (MVL) composition product by multiple sets of analysis methods, so as to meet the concept of green environmental protection and economic efficiency. SUMMARY

[0005] The application provides a multivesicular liposome (MVL) composition analysis method.

[0006] The names and structures of part of the detected substances are as follows:

[0007] LPEC: 1-erucoyl-2-hydroxyphosphatidylcholine, the structural formula is

[0008]

[0009] LPPG: 1-palmitoyl-2-hydroxy-phosphatidylglycerol, the structural formula is

[0010]

[0011] The application provides a multivesicular liposome (MVL) composition analysis method:

[0012] The chromatographic column is a high-performance liquid reversed-phase chromatographic column, preferably a C8 chromatographic column.

[0013] The mobile phase is mobile phase A: a buffer saline solution, and mobile phase B: an organic phase; the detector is an evaporative light scattering detector (ELSD) or a corona aerosol detector (CAD), preferably a corona aerosol detector (CAD).

[0014] The flow rate is 0.9-2 ml / min.

[0015] In some embodiments of the present application, the mobile phase A is an aqueous ammonium formate solution, an aqueous ammonium acetate solution, preferably an aqueous ammonium acetate solution, preferably the mobile phase A is an aqueous ammonium formate solution or an aqueous ammonium acetate solution with a concentration of 0.09-0.11 mol / L; more preferably the mobile phase A is an aqueous ammonium acetate solution with a concentration of 0.09-0.11 mol / L; the mobile phase B is an organic phase of methanol or acetonitrile.

[0016] In some embodiments of the present application, the pH of the mobile phase A is 7.4-7.6, and the pH of the mobile phase is adjusted by using ammonia water.

[0017] In some embodiments of the present application, the column temperature is 28-32℃.

[0018] In some embodiments of the present application, gradient elution is used, and the elution program is as follows: 0 min, 9%-11% mobile phase A, 91%-89% mobile phase B; 1 min, 0-2% mobile phase A, 100%-98% mobile phase B; 13 min, 0-2% mobile phase A, 100%-98% mobile phase B; 13.1 min, 9%-11% mobile phase A, 91%-89% mobile phase B; 23 min, 9%-11% mobile phase A, 91%-89% mobile phase B.

[0019] In some embodiments of the present application, the gradient elution program is as follows: 0 min, flow rate 0.9 ml / min, 9% mobile phase A, 91% mobile phase B; 1 min, flow rate 0.9 ml / min, 100% mobile phase B; 5 min, flow rate 0.9 ml / min, 100% mobile phase B; 6 min, flow rate 2 ml / min, 13 min, flow rate 2 ml / min, 100% mobile phase B; 13.1 min, flow rate 0.9 ml / min, 9% mobile phase A, 91% mobile phase B; 23 min, flow rate 0.9 ml / min, 9% mobile phase A, 91% mobile phase B.

[0020] In some embodiments of the present application, the gradient elution program is as follows: 0 min, flow rate 1.1 ml / min, 11% mobile phase A, 89% mobile phase B; 1 min, flow rate 1.1 ml / min, 100% mobile phase B; 5 min, flow rate 1.1 ml / min, 100% mobile phase B; 6 min, flow rate 2 ml / min, 13 min, flow rate 2 ml / min, 100% mobile phase B; 13.1 min, flow rate 1.1 ml / min, 11% mobile phase A, 89% mobile phase B; 23 min, flow rate 1.1 ml / min, 11% mobile phase A, 89% mobile phase B.

[0021] In some embodiments of the present application, the gradient elution procedure is: 0 min, 9%-11% mobile phase A, 91%-89% mobile phase B; 5 min, 0-2% mobile phase A, 100%-98% mobile phase B; 18 min, 0-2% mobile phase A, 100%-98% mobile phase B; 18.1 min, 9%-11% mobile phase A, 91%-89% mobile phase B; 23 min, 9%-11% mobile phase A, 91%-89% mobile phase B.

[0022] In some embodiments of the present application, the gradient elution procedure is: 0 min, 10% mobile phase A, 90% mobile phase B; 5 min, 100% mobile phase B; 18 min, 100% mobile phase B; 18.1 min, 10% mobile phase A, 90% mobile phase B; 23 min, 10% mobile phase A, 90% mobile phase B.

[0023] In some embodiments of the present application, the flow rate is 0.9 ml / min, and the gradient elution procedure is: 0 min, 9% mobile phase A, 91% mobile phase B; 5 min, 100% mobile phase B; 18 min, 100% mobile phase B; 18.1 min, 9% mobile phase A, 91% mobile phase B; 23 min, 9% mobile phase A, 91% mobile phase B.

[0024] In some embodiments of the present application, the flow rate is 1.1 ml / min, and the gradient elution procedure is: 0 min, 11% mobile phase A, 89% mobile phase B; 5 min, 100% mobile phase B; 18 min, 100% mobile phase B; 18.1 min, 11% mobile phase A, 89% mobile phase B; 23 min, 11% mobile phase A, 89% mobile phase B.

[0025] In some embodiments of the present application, the components of the analysis composition are DPPG, tricaprylin, cholesterol, and DEPC.

[0026] In some embodiments of the present application, the hydrolyzed substances analyzed are erucic acid, LPEC, LPPG, and palmitic acid.

[0027] In some embodiments of the present application, the contents of DPPG, tricaprylin, cholesterol, and DEPC are calculated by the peak area method using an external standard.

[0028] In some embodiments of the present application, liquid chromatography is used to analyze linear solutions of erucic acid, LPPG and palmitic acid respectively, and the peak area is recorded according to the liquid chromatography conditions. The natural logarithm Y of the peak area of the linear series solution of erucic acid, LPPG and palmitic acid is linearly regressed with the natural logarithm of the concentration x (μg / mL), and the standard curve regression equation is obtained. The prepared liposome injection is determined by the same method, and the content of erucic acid, LPPG and palmitic acid is calculated according to the obtained standard curve regression equation.

[0029] Compared with the prior art, the beneficial effects of the present application include: without extraction of the sample, and avoiding the use of chloroform and other toxic organic solvents, avoiding the use of phosphate buffer salt, using simple organic acid ammonium aqueous phase and conventional methanol or acetonitrile mobile phase can effectively detect and separate DPPG, DEPC, triolein of neutral lipid, cholesteryl of acidic phospholipid, and can also analyze erucic acid, LPEC, LPPG, palmitic acid, the hydrolysis product of DPPG and DEPC, avoiding the consumption of a large amount of organic reagents in the control of multi-vesicular liposome (MVL) composition product quality by multiple sets of methods, and meeting the green environmental protection, economic and efficient concept of pharmaceutical preparation detection and analysis method. BRIEF DESCRIPTION OF DRAWINGS

[0030] Figure 1 The liquid chromatogram of the solution to be detected in Example 1 is shown in Figure 1.

[0031] Figure 2 The liquid chromatogram of the solution to be detected in Example 1 is shown in Figure 1.

[0032] Figure 3 The liquid chromatogram of the solution to be detected in Example 1 is shown in Figure 1.

[0033] Figure 4 The liquid chromatogram of the solution to be detected in Example 1 is shown in Figure 1.

[0034] Figure 5 The liquid chromatogram of the solution to be detected in Example 3 is shown in Figure 3.

[0035] Figure 6 The liquid chromatogram of the solution to be detected in Example 4 is shown in Figure 4.

[0036] Figure 7 The liquid chromatogram of the solution to be detected in Example 1 is shown in Figure 1.

[0037] Figure 8 The liquid chromatogram of the solution to be detected in Example 1 is shown in Figure 1. DETAILED DESCRIPTION

[0038] The application is further described by the following examples, which are only used to illustrate the technical solutions of the application and not used to limit the scope of the application. Those skilled in the art can make some non-essential improvements and adjustments, which still belong to the protection scope of the application.

[0039] Example 1 Quantitative detection method of DPPG, glyceryl trioctanate, cholesterol and DEPC

[0040] 1.1 Sample preparation

[0041] Take 1 ml of bupivacaine liposome injection, place it in a 50 ml volumetric flask, dissolve and dilute to the calibration mark with isopropyl alcohol aqueous solution to obtain the test solution 1.

[0042] Prepare a solution containing DPPG at a concentration of 0.018 mg / ml, 0.09 mg / ml of cholesterol, 0.04 mg / ml of glyceryl trioctanate, and 0.16 mg / ml of DEPC to obtain the test solution 2.

[0043] According to the prescription of bupivacaine liposome injection, prepare a solution containing no DPPG, glyceryl trioctanate, cholesterol and DEPC, take the solution, dissolve and dilute with isopropyl alcohol aqueous solution to obtain the excipient blank solution.

[0044] Solvent: isopropyl alcohol aqueous solution.

[0045] 1.2 Detection chromatographic conditions

[0046] Filler (C8) with octylsilane bonded silica gel; mobile phase A is 0.09 mol / L ammonium acetate solution, mobile phase B is methanol; column temperature: 28℃; detector: CAD; injection volume: 10 μl.

[0047] Gradient elution condition program: 0 min, flow rate 0.9 ml / min, 9% mobile phase A, 91% mobile phase B; 1 min, flow rate 0.9 ml / min, 100% mobile phase B; 5 min, flow rate 0.9 ml / min, 100% mobile phase B; 6 min, flow rate 2 ml / min, 13 min, flow rate 2 ml / min, 100% mobile phase B; 13.1 min, flow rate 0.9 ml / min, 9% mobile phase A, 91% mobile phase B; 23 min, flow rate 0.9 ml / min, 9% mobile phase A, 91% mobile phase B.

[0048] 1.3 Detection results

[0049] 1.3.1 Detection separation

[0050] Take 10 μl of the test solution 2, the test solution 1, the excipient blank solution and the solvent respectively, and detect according to the above liquid chromatographic conditions, and the results are as follows Figures 1-4As shown, the results show that the solvent and excipient blank has no interference, DPPG, tricaprylin, cholesterol, DEPC are sequentially eluted, and the separation degree is greater than 2.0, indicating that DPPG, tricaprylin, cholesterol, DEPC can be effectively separated.

[0051] 1.3.2 Quantitative calculation

[0052] Take 10 μl of the above to-be-detected solution 1 and to-be-detected solution 2, respectively, and inject them into the liquid chromatograph, and detect them according to the above chromatographic conditions, and record the chromatogram. According to the external standard method, the content of DPPG, tricaprylin, cholesterol, DEPC in the to-be-detected solution 1 can be calculated by peak area.

[0053] Example 2 DPPG, tricaprylin, cholesterol, DEPC quantitative detection method

[0054] 2.1. Chromatographic conditions: octylsilane bonded silica gel filler (C8); mobile phase A is 1.1 mol / L ammonium acetate solution, mobile phase B is methanol; column temperature: 32°C; detector: CAD; injection volume 10 μl. Gradient elution program is: 0 min, flow rate 1.1 ml / min, 11% mobile phase A, 89% mobile phase B; 1 min, flow rate 1.1 ml / min, 100% mobile phase B; 5 min, flow rate 1.1 ml / min, 100% mobile phase B; 6 min, flow rate 2 ml / min, 13 min, flow rate 2 ml / min, 100% mobile phase B; 13.1 min flow rate 1.1 ml / min, 11% mobile phase A, 89% mobile phase B; 23 min, flow rate 1.1 ml / min, 11% mobile phase A, 89% mobile phase B.

[0055] The same as in Example 1, the results show that the detection results of DPPG, tricaprylin, cholesterol, DEPC are basically the same as in Example 1.

[0056] Example 3 Quantitative detection and analysis method of erucic acid, palmitic acid, LPPG

[0057] 3.1. Preparation of detection sample

[0058] Prepare a solution with a concentration of 20 μg / ml of palmitic acid, LPPG, and erucic acid as to-be-detected solution 3.

[0059] Take 1 ml of bupivacaine liposome injection, place it in a 5 ml volumetric flask, dilute it to the mark with isopropyl alcohol solution, and obtain it as to-be-detected solution 4.

[0060] 3.2. Detection chromatographic conditions

[0061] Chromatographic column: C8; mobile phase A is 0.1 mol / L ammonium acetate solution, PH = 7.5, mobile phase B is methanol, column temperature is 30 DEG C, sample volume is 50ul, CAD detector, flow rate is 1ml / min, gradient elution procedure is: 0min, 10% mobile phase A, 90% mobile phase B; 5min, 100% mobile phase B; 18min, 100% mobile phase B; 18.1min, 10% mobile phase A, 90% mobile phase B; 23min, 10% mobile phase A, 90% mobile phase B.

[0062] 3.3. Detection separation

[0063] Respectively take 50ul of the to-be-detected liquid 3 and the to-be-detected liquid 4, inject and detect according to the liquid chromatography condition of 3.2, and the results are respectively as Figure 5 、 Figure 6 LPPG, palmitic acid and erucic acid in the to-be-detected liquid 3 are sequentially eluted, and LPEC and erucic acid in the to-be-detected liquid 4 can be effectively separated. The results show that the liquid chromatography detection of the application can quickly and effectively detect and separate palmitic acid, LPPG, erucic acid and LPEC in the bupivacaine liposome injection.

[0064] 3.4. Quantitative detection

[0065] Respectively prepare palmitic acid, LPPG and erucic acid solutions with concentrations of 5ug / ml, 20ug / ml, 50ug / ml and 80ug / ml, take 50ul of each, inject into the liquid chromatograph, and detect according to the above detection chromatography condition, record the chromatogram, calculate the linear regression equation with the logarithmic value of the concentration and the logarithmic value of the corresponding peak area, obtain the linear regression equation with a correlation coefficient (r) not less than 0.99, and calculate the content of palmitic acid, LPPG and erucic acid in the to-be-detected sample 4 by using the obtained linear regression equation.

[0066] The results show that the liquid chromatography condition can accurately quantitatively detect the content of palmitic acid, LPPG and erucic acid in the to-be-detected sample 4.

[0067] Example 4 DPPG, trioctanoin, cholesterol, DEPC methodological verification

[0068] 4.1 Linear range

[0069] Prepare a series of linear solutions containing DPPG, trioctanoin, cholesterol and DEPC with different concentrations.

[0070] Respectively take 10ul of each of the above series of linear solutions and inject into the liquid chromatograph, detect according to the liquid chromatography condition of example 1, and record the chromatogram.

[0071] The results show that: with the peak area to linear regression, DPPG, trioctanoin, cholesterol, DEPC in turn in 9 ~ 27 μg / ml, 20 ~ 60 μg / ml, 47 ~ 141 μg / ml, 82 ~ 246 μg / ml concentration range linear good.

[0072] 4.2 repeatability, precision

[0073] Example 1 to be tested solution 1, parallel 6 (2 groups).

[0074] Example 1 to be tested solution 2, parallel 2 (2 groups).

[0075] Take the solution each 10 μl, respectively, into the liquid chromatograph, according to the liquid chromatography conditions of example 1 detection, record chromatogram.

[0076] The results show that: 6 of the test solution 1 DPPG, trioctanoin, cholesterol, DEPC content of RSD are not greater than 2%, the method is good reproducibility, 12 of the test solution 1 DPPG, trioctanoin, cholesterol, DEPC content of RSD are not greater than 2%, the method of intermediate precision is good.

[0077] 4.3 accuracy

[0078] Example 1 to be tested solution 2, parallel 2

[0079] Take example 1 auxiliary material blank solution 0.5 ml, 1 ml, 0.5 ml respectively in 25, 50, 25 ml flask, respectively, add 2 ml, 5 ml, 3 ml containing DPPG 180 μg / ml, trioctanoin 400 μg / ml, cholesterol 940 μg / ml, DEPC 1640 μg / ml of the test solution 3, get recovery rate solution 1, recovery rate solution 2, recovery rate solution 3, each 3 groups.

[0080] Take recovery rate solution 1, recovery rate solution 2, recovery rate solution 3, test solution 3, each 10 μl, respectively, into the liquid chromatograph, according to the liquid chromatography conditions in example 1 detection, record chromatogram.

[0081] The results show that: according to the external standard method to calculate the recovery rate of DPPG, trioctanoin, cholesterol, DEPC, DPPG, trioctanoin, cholesterol, DEPC each recovery rate value is between 80% and 120%, the RSD value is not greater than 3%, the accuracy of the method is good.

[0082] Example 5 of erucic acid, palmitic acid, LPPG methodology validation

[0083] 5.1. sample preparation

[0084] System suitability solution: take 1 ml of bupivacaine liposome injection, put it in a 5 ml flask, add 1 ml of mixed solution of palmitic acid and LPPG with a concentration of 125 μg / ml, dissolve with 2 ml of isopropyl alcohol, and dilute with water to the mark.

[0085] 5.2. Specificity verification

[0086] Take 25 μg / ml of palmitic acid, LPPG, erucic acid solution, 80 μg / ml of palmitic acid positioning solution, 80 μg / ml of LPPG positioning solution, 100 μg / ml of erucic acid positioning solution and system suitability solution, 50 μl each into the liquid chromatograph, and detect using the liquid chromatography conditions of 3.2 in Example 3. The results show that the separation degree between each impurity in the system suitability solution is greater than 1.5, so the analysis method has good specificity.

[0087] 5.3. Linearity verification

[0088] Take 50 μl of palmitic acid, LPPG, erucic acid linear solution respectively into the liquid chromatograph, and detect according to the liquid chromatography conditions in Example 3. Record the chromatogram. Use the least square method for linear regression analysis of the logarithmic value of concentration-peak area logarithmic value. The results show that the logarithm of peak area and the logarithm of concentration of LPPG, palmitic acid and erucic acid within the limit of 1-100 μg / ml are linearly related, and the linear regression coefficients are all greater than 0.999. It shows that LPPG, palmitic acid and erucic acid have good linearity within the limit of 1-100 μg / ml.

[0089] 5.4. Sensitivity verification

[0090] Take 50 μl of 1 μg / ml LPPG, palmitic acid and erucic acid limit of quantitation solution into the liquid chromatograph, and detect according to the liquid chromatography method of Example 3. Record the chromatogram. The results show that the signal-to-noise ratio of LPPG, palmitic acid and erucic acid is greater than 10 at 1 μg / ml, indicating that the detection concentration of LPPG, palmitic acid and erucic acid does not exceed 1 μg / ml, and the method has good sensitivity.

[0091] 5.5. Accuracy verification

[0092] Prepare the recovery rate solution of erucic acid, LPPG and palmitic acid respectively, and detect according to the detection method of Example 3. Use the linear regression equation to calculate the recovery rate of erucic acid, LPPG and palmitic acid respectively. The calculation results show that the average recovery rate of erucic acid is between 90% and 110%, the average recovery rate of LPPG and palmitic acid is between 80% and 120%, and the RSD value is not greater than 3%, indicating that the method has good accuracy.

[0093] 5.6. Durability verification

[0094] The changes in column temperature (±2℃), flow rate (±0.1 ml / min), mobile phase ratio (organic phase ±2%), and pH (±0.1) of the liquid chromatography conditions in Example 3 were detected, as shown in Table 1.

[0095] Table 1. Durability of Chromatographic Conditions for Erucic Acid, LPPG, and Palmitic Acid

[0096]

[0097] Test results: The solvent did not interfere with the test under the various liquid chromatography conditions. The test results for erucic acid, palmitic acid and LPPG were basically the same as those under the conditions of Example 3. The method for detecting palmitic acid and LPPG has good robustness.

[0098] Comparative Example 1

[0099] The solutions of DPPG, octanoic acid glyceride, cholesterol, and DEPC at a concentration of 125 μg / ml were detected using the liquid chromatography conditions shown in Table 2. The results were as follows: Figure 7 .

[0100] Table 2 Chromatographic conditions

[0101]

[0102]

[0103] The results showed that, using the mobile phase described above, tricaprylic acid glyceride and cholesterol were not completely separated, DEPC was not detected, and DPPG, tricaprylic acid glyceride, and cholesterol had short retention times, making them unsuitable for the detection of multivesicular liposome (MVL) composition samples.

[0104] Comparative Example 2

[0105] Replace the mobile phase A:mobile phase B ratio of 50:50 in Comparative Example 1 with a ratio of 10:90. Analyze the solution from Comparative Example 1 and the DEPC targeting solution (containing 2.5 mg DEPC per ml). The HPLC chromatogram of the DEPC targeting solution is shown below. Figure 8 As shown, the results indicate that although DEPC can detect it, its peak shape is poor and it cannot be used for quantitative detection of its content.

[0106] The above results show that the method of the present invention can simultaneously and quantitatively detect DPPG, trioctyl glycerol, cholesterol, and DEPC, and can also be used to detect their hydrolysis products. It has accuracy and reliability and can achieve the purpose of controlling product quality.

[0107] The methods and products of the present application have been described by preferred embodiments, and it will be apparent to those skilled in the art that the methods and products described herein can be modified or adapted and combined appropriately without departing from the content, spirit and scope of the present application to implement the present technology. In particular, it is pointed out that all similar substitutions and modifications are obvious to those skilled in the art, and they are considered to be included in the spirit, scope and content of the present application.

Claims

1. A method of analyzing a multivesicular liposome composition, characterized by: The analysis method is used for analyzing the multivesicular liposome composition 1,2-palmitoyl phosphatidylglycerol DPPG, tricaprylin, cholesterol, 1,2-dierucoylphosphatidylcholine DEPC component, multivesicular liposome hydrolysate erucic acid, 1-erucyl-2-hydroxyphosphatidylcholine LPEC, 1-palmitoyl-2-hydroxy-phosphatidylglycerol LPPG and palmitic acid; The high performance liquid phase reverse phase chromatographic column adopts a C8 chromatographic column; The mobile phase A is an aqueous solution of ammonium acetate with a concentration of 0.09-0.11 mol / L, and the mobile phase B is methanol; the pH of the mobile phase A is 7.4-7.6, the detector is a charged aerosol detector CAD, the flow rate is 0.9-2 ml / min, and the detection column temperature is 28-32 DEG C. When the analytical method is used to analyze the multivesicular liposome composition 1,2-palmitoyl phosphatidylglycerol DPPG, caprylic acid triglyceride, cholesterol and 1,2-dielaidoyl phosphatidylcholine DEPC components, a gradient elution is used with an elution program of 0 min, 9% mobile phase A, 91% mobile phase B 11% mobile phase A, 89% mobile phase B; 1-13 min, 0 2% mobile phase A, 100%-98% mobile phase B; 2% mobile phase A, 100%-98% mobile phase B; 13.1 min, 9% 11% mobile phase A, 91% mobile phase B 89% mobile phase B; 23 min, 9% 11% mobile phase A, 91% mobile phase B 89% mobile phase B; When the analytical method is used to analyze the polycystic liposome hydrolysate erucic acid, 1-erucyloyl-2-hydroxyphosphatidylcholine LPEC, 1-palmitoyl-2-hydroxy-phosphatidylglycerol LPPG and palmitic acid, gradient elution is used, and the elution program is: 0 min, 9% mobile phase A, 91% mobile phase B; 5 min, 0 mobile phase A, 100% mobile phase B; 18 min, 0 mobile phase A, 100% mobile phase B; 18.1 min, 9% mobile phase A, 91% mobile phase B; 23 min, 9% mobile phase A, 91% mobile phase B; 30 min, 9% mobile phase A, 91% mobile phase B. 11% mobile phase A, 91% 89% mobile phase B; 23 min, 9% 11% mobile phase A, 91% 89% mobile phase B. 11% mobile phase A, 91% 89% mobile phase B; 23 min, 9% 11% mobile phase A, 91% 89% mobile phase B.

2. The analysis method according to claim 1, characterized in that The pH of the mobile phase A is adjusted by using ammonia water.

3. The analysis method according to claim 1 or 2, characterized in that The contents of 1,2-palmitoyl phosphatidylglycerol DPPG, tricaprylin, cholesterol and 1,2-dierucoylphosphatidylcholine DEPC are calculated by using the peak area calculation method of the external standard method.

4. The analysis method according to claim 1 or 2, characterized in that The natural logarithm Y of the peak area of the linear series solution of erucic acid, 1-palmitoyl-2-hydroxy-phosphatidylglycerol LPPG and palmitic acid is linearly regressed to the natural logarithm of the concentration x, the unit of the concentration x is μg / mL, a standard curve regression equation is obtained, the prepared liposome injection is determined by the same method, and the contents of erucic acid, 1-palmitoyl-2-hydroxy-phosphatidylglycerol LPPG and palmitic acid are calculated according to the obtained standard curve regression equation.

5. The analysis method of claim 3, wherein The natural logarithm Y of the peak area of the linear series solution of erucic acid, 1-palmitoyl-2-hydroxy-phosphatidylglycerol LPPG and palmitic acid is linearly regressed to the natural logarithm of the concentration x, the unit of the concentration x is μg / mL, a standard curve regression equation is obtained, the prepared liposome injection is determined by the same method, and the contents of erucic acid, 1-palmitoyl-2-hydroxy-phosphatidylglycerol LPPG and palmitic acid are calculated according to the obtained standard curve regression equation.

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