Extracting solution and extraction method for extracting vitamin K2 from multivitamin mineral compound preparation

Abstract

The present application discloses an extracting solution and an extraction method for extracting vitamin K2 from a vitamin-mineral compound preparation. On the one hand, the present application provides an extracting solution for extracting vitamin K2 from a vitamin-mineral compound preparation, the extracting solution comprising a combination of a wall material lysing solution and an extraction solvent; the base liquid of the wall material lysing solution adopts an acidic buffer solution, the wall material lysing solution also comprises a surfactant added to the base liquid, the concentration of the surfactant in the base liquid is 0.05~1.5mg / mL, and the pH value of the wall material lysing solution is 2~6; the extraction solvent is an organic solvent. On the other hand, the present application also provides a method for extracting vitamin K2 from a vitamin-mineral compound preparation, using the above-mentioned extracting solution for extracting vitamin K2 from a vitamin-mineral compound preparation. The present application can effectively dissolve the wall material of vitamin K2 microcapsules, has excellent extraction effect on vitamin K2, and can effectively improve the quality control detection accuracy of vitamin K2 from vitamin-mineral compound preparations.

Description

Technical Field

[0001] The present application relates to the technical field of quality control and detection, and in particular to an extract and an extraction method for extracting vitamin K2 from a vitamin-mineral compound preparation. Background Art

[0002] Vitamin K2 is a fat-soluble vitamin and a derivative of the naphthoquinone group with biological activity of phylloquinone. It is one of the important vitamins indispensable to the human body and the only biologically active form of vitamin K. It promotes the deposition of calcium ions into bones, increases bone density, and prevents fractures. It can produce insulin, maintain blood sugar stability, and prevent diabetes. It can prevent cirrhosis from transforming into liver cancer, etc.

[0003] Vitamin K2 is unstable when exposed to light and alkali, and is easily decomposed. It is also easily affected by other components in vitamin-mineral compound preparations. The above characteristics determine that vitamin K2 needs to be processed by microencapsulation technology to increase its stability. The embedding process will increase the difficulty of extraction for content analysis, especially when the embedding material is more complex or multiple embedding treatments are used. It is difficult to completely extract the low-content vitamin components to be tested in the preparation using only traditional solvents, which greatly increases the difficulty of detection. There are pre-treatment methods for cracking wall materials in the prior art, which fully dissolve the composite wall materials. Although the cracking of the wall materials can be achieved and the above problems can be solved, it is difficult for such pre-treatment methods to ensure that vitamin K2 with poor stability is not destroyed. In addition, the content of vitamin K2 in vitamin-mineral compound preparations is low and is a trace component, which further increases the difficulty of detection.

[0004] At present, the detection method of vitamin K2 in raw materials usually adopts the method recommended in the Announcement No. 8 of 2016 of the National Health and Family Planning Commission, "Announcement on New Varieties of Food Additives Such as Calcium Alginate", which uses isopropyl alcohol for direct ultrasonic extraction and then uses high-performance liquid chromatography for detection. Isopropyl alcohol is an organic solvent, and the microencapsulation wall material is often a polymer material that is insoluble in organic solvents. Direct ultrasonic extraction with isopropyl alcohol alone cannot effectively cleave the microencapsulation wall material to fully release vitamin K2, and the extraction rate is low. Therefore, the above method is not suitable for the extraction of microencapsulated vitamin K2. If it is used to determine the content of microencapsulated vitamin K2 in vitamin-mineral compound preparations, the accuracy is difficult to meet the requirements, which is not conducive to product quality control. Summary of the invention

[0005] In order to solve at least one of the above technical problems, a microcapsule wall material that can effectively dissolve vitamin K2 is developed, which has excellent extraction effect on vitamin K2 and can effectively improve the accuracy of vitamin K2 content detection in vitamin-mineral compound preparations. The present application provides an extract and an extraction method for extracting vitamin K2 from a vitamin-mineral compound preparation.

[0006] On the one hand, the present application provides an extract for extracting vitamin K2 from a vitamin-mineral compound preparation, the extract comprising a wall material lysing solution and an extraction solvent; the base liquid of the wall material lysing solution adopts an acidic buffer solution, the wall material lysing solution also includes a surfactant added to the base liquid, the concentration of the surfactant in the base liquid is 0.05~1.5 mg / mL, and the pH value of the wall material lysing solution is 2~6; the surfactant adopts a non-ionic surfactant, or a sulfate ester surfactant or sodium dodecylbenzene sulfonate among anionic surfactants; the extraction solvent is an organic solvent.

[0007] By adopting the above technical scheme, the present application designs a solution scheme of a combination of a lysis solution and an extraction solvent. The lysis solution uses an acidic buffer as a base liquid, which can provide a stable acidic pH environment for the extraction of vitamin K2, and can effectively avoid the destruction or decomposition of vitamin K2 during the extraction process, thereby further ensuring the accuracy of detection; a surfactant is added to the acidic buffer base liquid to be configured into a wall material lysis solution, which can effectively cause swelling or dissolution damage to various existing wall materials when the sample composition is extremely complex and there are multiple microcapsule-encapsulated components, thereby enabling the encapsulated vitamin K2 to be fully released; and an organic solvent is used as an extraction solvent, which can effectively and completely extract the released vitamin K2, thereby effectively ensuring the completeness of the extraction and ensuring the accuracy of detection.

[0008] Optionally, the acidic buffer is selected from one of acetate buffer, phosphate buffer, borate buffer or citrate buffer.

[0009] Optionally, the concentration of the surfactant in the base liquid is 0.5-1.2 mg / mL

[0010] Optionally, the surfactant is selected from one or more of polyoxyethylene ether surfactants (such as octylphenyl polyoxyethylene ether, nonylphenol polyoxyethylene ether, lauryl alcohol polyoxyethylene ether), polyol surfactants (such as Tween-60, Tween-80), alkyl glycoside surfactants (such as octyl glucoside, decyl glucoside), sulfate surfactants (such as sodium dodecyl sulfate) or sodium dodecylbenzene sulfonate.

[0011] Further optionally, the surfactant is a single surfactant or a compound surfactant, and the compound surfactant is a compound of two surfactants.

[0012] Further optionally, the concentration ratio of the two surfactants in the compound surfactant is 1:1~5.

[0013] Optionally, the organic solvent is selected from one of 80% to 100% ethanol, isopropanol, tetrahydrofuran, n-hexane, ethyl acetate, ether, cyclohexane, isooctane or chloroform.

[0014] On the other hand, the present application also provides a method for extracting vitamin K2 from a vitamin-mineral compound preparation, using the above-mentioned extract for extracting vitamin K2 from the vitamin-mineral compound preparation, comprising the following steps:

[0015] S1. Pre-treat the vitamin-mineral compound preparation sample (crush the solid sample into powder), accurately weigh an appropriate amount of the pre-treated sample, and place it in a light-proof container;

[0016] S2, adding the wall material lysis solution to the light-proof container containing the sample in step S1, and treating for more than 10 minutes to obtain a sample mixture;

[0017] S3, adding an extraction solvent to the sample mixture obtained in step S2, mixing, and ultrasonically extracting for more than 10 minutes to obtain an extraction mixture;

[0018] S4. Filter the mixed extraction solution obtained in step S3 to remove insoluble matter, thereby obtaining a sample extract of vitamin K2.

[0019] By adopting the above-mentioned technical scheme, the extraction method of the present application uses the specific lysis solution of the present application to efficiently destroy the wall material in a static manner, and then extracts vitamin K2 in conjunction with the ultrasonic extraction step. The entire extraction process is simple and efficient, and the extraction of vitamin K2 is relatively complete, which can effectively ensure the accuracy of subsequent tests.

[0020] Optionally, in step S2, the processing temperature is controlled at 30-75°C.

[0021] Optionally, in step S3, the ultrasonic extraction parameters are: temperature 30~75°C, ultrasonic frequency 35~50kHz, and ultrasonic power 1kW±5W.

[0022] In summary, the present invention includes at least one of the following beneficial technical effects:

[0023] 1. This application designs an extraction solution, which uses an acidic buffer as a base solution, which can provide a stable acidic pH environment for the extraction of vitamin K2, and can effectively prevent the destruction or decomposition of vitamin K2 during the extraction process, further ensuring the accuracy of the detection;

[0024] 2. The extract of the present application is a wall material lysis solution prepared by adding a surfactant to an acidic buffer base liquid. When the sample composition is extremely complex and there are multiple microcapsule encapsulated components, it can effectively cause swelling or dissolution damage to various existing wall materials, thereby enabling the encapsulated vitamin K2 to be fully released;

[0025] 3. The extract of the present application uses an organic solvent as the extraction solvent, which can effectively extract the released vitamin K2 completely, thereby effectively ensuring the completeness of the extraction and the accuracy of the detection. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 Detection chromatogram of blank sample for this application;

[0027] Figure 2 The chromatogram of the vitamin K2 reference substance test for this application;

[0028] Figure 3 Chromatogram of the sample test for this application. DETAILED DESCRIPTION

[0029] The present application is further described in detail below in conjunction with the accompanying drawings and embodiments.

[0030] The present application provides an extract for extracting vitamin K2 from a vitamin-mineral compound preparation, wherein the extract comprises a combination of a wall material lysing solution and an extraction solvent; the base liquid of the wall material lysing solution adopts an acidic buffer solution, the wall material lysing solution also comprises a surfactant added to the base liquid, the concentration of the surfactant in the base liquid is 0.05-1.5 mg / mL, and the pH value of the wall material lysing solution is 2-6; the extraction solvent is an organic solvent.

[0031] The present application also provides a method for extracting vitamin K2 from a vitamin-mineral compound preparation, using the above-mentioned extract for extracting vitamin K2 from the vitamin-mineral compound preparation, comprising the following steps:

[0032] S1. Pre-treat the vitamin-mineral compound preparation sample (crush the solid sample into powder), accurately weigh an appropriate amount of the pre-treated sample, and place it in a light-proof container;

[0033] S2, adding the wall material lysis solution to the light-proof container containing the sample in step S1, and treating for more than 10 minutes to obtain a sample mixture;

[0034] S3, adding an extraction solvent to the sample mixture obtained in step S2, mixing, and ultrasonically extracting for more than 10 minutes to obtain an extraction mixture;

[0035] S4. Filter the mixed extraction solution obtained in step S3 to remove insoluble matter, thereby obtaining a sample extract of vitamin K2.

[0036] The extracting solution and extraction method of the present application can lyse various wall materials and effectively extract vitamin K2 from the wall materials without decomposing vitamin K2; even if the sample is a vitamin-mineral compound preparation with an extremely low content of vitamin K2, the present application can still achieve better extraction.

[0037] The following are experimental verification and examples of this application.

[0038] The experimental verification of this application adopts HPLC-UV method to detect the content of vitamin K2.

[0039] The specific method is basically the same as the detection method of vitamin K2 in the National Health Commission Announcement No. 8 of 2016, except that the chromatographic parameters were adjusted.

[0040] The specific chromatographic parameters are as follows:

[0041] Chromatographic column: Waters C 18 Column (150×4.6 mm, 5 μm);

[0042] Mobile phase: 97% methanol;

[0043] Column temperature: 50°C; flow rate: 1.0 mL / min;

[0044] Detection wavelength: 254 nm; injection volume: 50 μL.

[0045] The instruments used in the examples and experimental verification of this application are as follows:

[0046] CPA225D electronic analytical balance (accuracy: 1 / 100,000) and BSA224S-CW electronic analytical balance (accuracy: 1 / 100,000) were purchased from Sartorius (Beijing) Co., Ltd.

[0047] Model 1260 high performance liquid chromatograph, purchased from Agilent, USA;

[0048] IKA Multi Drive MI 250 pulverizer, purchased from IKA, Germany;

[0049] KQ-300DE ultrasonic oscillator, purchased from Kunshan Ultrasonic Instrument Co., Ltd.;

[0050] ST16R centrifuge, purchased from Thermo Corporation, USA;

[0051] C 18 The column (150×4.6 mm, 5 μm) was purchased from Waters Company, USA.

[0052] The reagents and samples used in the methodological validation and examples of this application were all from the laboratory and commercially available; among them, the vitamin K2 reference substance (MK-7), batch number R080L0, purity 97.80%, was purchased from USP Reference Standard Company, USA; methanol used chromatographically pure reagents purchased from Merck, USA; ethanol, isopropanol and other organic solvents used analytically pure reagents purchased from China National Pharmaceutical Group Co., Ltd.

[0053] The methodological verification and examples of the present invention used homemade multivitamin mineral tablets (batch number: S04N001). Its nutrient composition: calcium 100.00~143.75 mg / g, magnesium 32.50~43.75 mg / g, manganese 0.5~0.81 mg / g, iron 2.5~3.94 mg / g, zinc 1.5~7.5 mg / g, selenium 22.5~37.5 mg / g, copper 0.1~0.19 mg / g, vitamin A 157.2~353.7 μg / g, vitamin D31.5~7.5μg / g, vitamin E 2.52~5.67mg / g, vitamin K2 8.0~18.0 μg / g, vitamin B10.4~3.6 mg / g, vitamin B2 0.4~3.78 mg / g, vitamin B6 0.4~3.78 mg / g, vitamin B12 0.5~5.0μg / g, niacinamide 2.0~13.5 mg / g, folic acid 80.0~234.0 μg / g, biotin 7.1~37.8 μg / g, vitamin C 25.2~90.3 mg / g, pantothenic acid 1.1~7.5 mg / g, choline 39.51~45.25 mg / g. Vitamin K2 uses commercially available microencapsulated vitamin K2 powder (batch number: GF04001), and the theoretical content of vitamin K2 is 13.0μg / g.

[0054] Methodological verification experiment of the detection method in the embodiment of the present application

[0055] This experiment is used to verify the linearity and range, specificity, detection limit, quantitative limit, precision, stability and accuracy of the detection method of the present application. The blank sample of this experiment is prepared with 10 mL of wall material lysis solution and 90 mL of extraction solvent.

[0056] The wall material lysis solution is an acetate buffer containing a surfactant. The preparation method of the acetate buffer is as follows: take 5.1 g of sodium acetate, add 20 mL of glacial acetic acid, dilute to 250 mL with water, add the surfactant octyl glucoside (APG), the concentration of which in the base solution is 1.0 mg / mL, and the remainder is deionized water. The extraction solvent is 95% ethanol.

[0057] The vitamin K2 reference solution in this experiment uses 95% ethanol as solvent, adds vitamin K2 reference, and prepares a series of reference solutions with concentrations of 0, 0.2, 0.4, 0.6, 0.8, and 1.0 μg / mL, respectively.

[0058] The sample extraction in this experiment was carried out using the following steps:

[0059] 1. Accurately weigh 4 g of sample and place it in a 100 mL brown volumetric flask;

[0060] 2. Add 10 mL of the wall material lysis solution of this experiment to the volumetric flask and heat in a 40°C water bath for 15 min;

[0061] 3. Add 60 mL of the extraction solvent of this experiment to the volumetric flask, vortex to mix, and perform ultrasonic extraction for 15 min (ultrasonic frequency 45 kHz, ultrasonic power 1 kW); the ultrasonic extraction parameters are: temperature 40°C. After ultrasonic extraction, use the extraction solvent of this experiment to make the volume to 100 mL to obtain the sample extract.

[0062] The vitamin K2 content in the experimental samples was calculated according to the formula c = (c0×v) / m.

[0063] In the formula, c is the content of vitamin K2 in the sample, in μg / g; c0 is the concentration of vitamin K2 in the sample calculated by the standard curve, in μg / mL; v is the fixed volume of the sample, in mL; m is the mass of the sample, in g.

[0064] Specific experiments

[0065] 1) Blank experiment

[0066] Use 10 mL of wall material lysis solution and 90 mL of extraction solvent, and follow the operations of steps S2 and S3 of the embodiment of the present application to prepare a blank sample solution without adding a sample.

[0067] According to the operation of step S3 of the embodiment of the present application, a vitamin K2 reference solution with a concentration of 0.8 μg / mL was prepared.

[0068] According to the operations of steps S2 and S3 of the embodiment of the present application, the sample extract is prepared.

[0069] According to the operation of step S4 of the embodiment of the present application, the blank sample solution, the vitamin K2 reference solution and the sample solution are tested respectively, and the test results obtained are as follows: Figure 1~Figure 3 shown.

[0070] pass Figure 1~Figure 3It can be seen that the vitamin K2 reference solution and the sample solution have absorption peaks at 20.5-22.5 min, while the blank sample solution has no absorption peak at this location, indicating that the blank sample has basically no interference with the measurement results and meets the requirements of detection accuracy.

[0071] 2) Linear relationship verification

[0072] Prepare vitamin K2 reference solutions with concentrations of 0, 0.2, 0.4, 0.6, 0.8, and 1.0 μg / mL, respectively. Perform the sample injection test according to the operation of step S4 of the present embodiment, plot the standard working curve with the concentration of vitamin K2 reference as the abscissa (X) and the peak area as the ordinate (Y), and obtain the linear regression equation Y=65.432X-0.278, and the correlation coefficient R 2 =0.9999, which proves that the detection method of the present application has a good linear relationship in the range of 0.2-1.0 μg / mL and meets the requirements of detection accuracy.

[0073] 3) Verification of detection limit and quantification limit

[0074] Prepare a vitamin K2 reference solution with a concentration of 0.001 to 0.030 µg / mL. Perform the sample injection test according to the operation of step S4 of the embodiment of the present application. The results show that when the signal-to-noise ratio (S / N) is 3, the corresponding reference concentration is the limit of detection (LOD); when the signal-to-noise ratio (S / N) is 10, the corresponding reference concentration is the limit of quantification (LOQ). Using the noise report to derive the results, the detection limit (LOD) of the detection method of the present application is 0.13 μg / g, and the limit of quantification (LOD) is 0.42 μg / g, which fully meets the requirements of detection accuracy.

[0075] 4) Precision verification test

[0076] Six samples were accurately weighed and the content of vitamin K2 was determined by the method of the embodiment of the present application. The results are shown in Table 1.

[0077] Table 1 Precision test results

[0078]

[0079] Note: * The test was repeated twice and the average value was taken. ** The theoretical value refers to the dosage of vitamin K2.

[0080] It can be seen from the data in Table 1 that the RSD of the vitamin K2 content of the six samples is 1.48%, and the deviation between the mean and the theoretical value is 1.42%, which has extremely high detection accuracy.

[0081] 5) Recovery rate verification test

[0082] Accurately weigh 9 portions of 2.0g sample, place them in 100mL brown volumetric flasks, and divide them into 3 groups, 3 portions in each group, and add 0.2, 0.25, and 0.3mL of 100μg / mL vitamin K2 standard solution according to low (80.00%), medium (100.00%), and high (120.00%) as test samples.

[0083] The content of vitamin K2 was determined by the method of the embodiment of the present application, and the recovery rate, average recovery rate, and RSD between recovery rates were calculated. The results are shown in Table 2.

[0084] Table 2 Recovery test results

[0085]

[0086] Note: * The test was repeated twice and the average value was taken.

[0087] From the test results in Table 2, it can be seen that the recovery rate of this method is 95.12% to 100.05%, the average recovery rate is 97.80%, the RSD between the recoveries is 1.76%, ≤2.00%, and it has a high recovery rate.

[0088] 6) Stability verification test

[0089] According to step S3 of the embodiment of the present application, a vitamin K2 reference solution with a concentration of 0.5 μg / mL was prepared. After being placed at room temperature for 0, 1, 2, 4, 8, and 12 hours, the content of vitamin K2 was determined by the method of the embodiment of the present application, and its RSD was calculated. The results are shown in Table 3.

[0090] Table 3 Stability test results

[0091]

[0092] Note: * The test was repeated twice and the average value was taken

[0093] It can be seen from the data in Table 3 that after the vitamin K2 standard solution was placed at room temperature for 0, 1, 2, 4, 8, and 12 hours, its RSD was 1.08%, indicating that the vitamin K2 standard solution had good stability within 12 hours at room temperature and the method had good stability.

[0094] Through the above-mentioned methodological verification experiment, it can be seen that the detection method used in the embodiment of the present application has high detection precision, detection accuracy, detection sensitivity and detection stability, and is minimally interfered by the extract of the present application, which fully meets the verification needs. The detection method also complies with the requirements of GB / T 27404 "Laboratory Quality Control Specification for Food Physical and Chemical Testing", GB / T27417 "Guidelines for Confirmation and Validation of Chemical Determination Methods for Conformity Assessment", and "AOAC Single Laboratory Test Method Validation Guide".

[0095] The following are Examples 1 to 14 of this application

[0096] Example 1

[0097] The extraction solution of this embodiment includes a combination of a wall material cracking solution and an extraction solvent.

[0098] The wall material lysis solution includes acetate buffer and octyl glucoside (APG) surfactant, and the concentration ratio of each component is as follows: sodium acetate concentration is 0.28 mol / L, acetic acid concentration is 0.12 mol / L, octyl glucoside (APG) concentration is 1.0 mg / mL, the balance is deionized water, and the pH value is 4.

[0099] The extraction solvent was 95% ethanol.

[0100] Example 2

[0101] The extraction solution of this embodiment includes a combination of a wall material cracking solution and an extraction solvent.

[0102] The wall material lysis solution includes acetate buffer and Triton X-100 surfactant, and the concentration ratio of each component is as follows: sodium acetate concentration is 0.28 mol / L, acetic acid concentration is 0.12 mol / L, Triton X-100 concentration is 0.80 mg / mL, the balance is deionized water, and the pH value is 4.

[0103] The extraction solvent was 95% ethanol.

[0104] Example 3

[0105] The extraction solution of this embodiment includes a combination of a wall material cracking solution and an extraction solvent.

[0106] The wall material lysis solution includes acetate buffer and Tween-60 surfactant, and the concentration ratio of each component is as follows: sodium acetate concentration is 0.28 mol / L, acetic acid concentration is 0.12 mol / L, Tween-60 concentration is 0.50 mg / mL, the balance is deionized water, and the pH value is 4.

[0107] The extraction solvent was 95% ethanol.

[0108] Example 4

[0109] The extraction solution of this embodiment includes a combination of a wall material cracking solution and an extraction solvent.

[0110] The wall material lysis solution includes acetate buffer and monolaurin surfactant, and the concentration of each component is as follows: sodium acetate concentration is 0.28 mol / L, acetic acid concentration is 0.12 mol / L, monolaurin concentration is 0.80 mg / mL, the balance is deionized water, and the pH value is 4.

[0111] The extraction solvent was 95% ethanol.

[0112] Example 5

[0113] The extraction solution of this embodiment includes a combination of a wall material cracking solution and an extraction solvent.

[0114] The wall material lysis solution includes acetate buffer, and a composite surfactant composed of octyl glucoside (APG) and Triton X-100, and the added mass ratio of the two is 3:1; the concentration ratio of each component is as follows: sodium acetate concentration is 0.28 mol / L, acetic acid concentration is 0.12 mol / L, the concentration of the composite surfactant is 0.90 mg / mL, the balance is deionized water, and the pH value is 4.

[0115] The extraction solvent was 95% ethanol.

[0116] Example 6

[0117] The extraction solution of this embodiment includes a combination of a wall material cracking solution and an extraction solvent.

[0118] The wall material lysis solution includes acetate buffer, and a composite surfactant composed of octyl glucoside (APG) and Triton X-100, and the added mass ratio of the two is 3:1; the concentration ratio of each component is as follows: sodium acetate concentration is 0.28 mol / L, acetic acid concentration is 0.12 mol / L, the concentration of the composite surfactant is 0.50 mg / mL, the balance is deionized water, and the pH value is 4.

[0119] The extraction solvent was 95% ethanol.

[0120] Example 7

[0121] The extraction solution of this embodiment includes a combination of a wall material cracking solution and an extraction solvent.

[0122] The wall material lysis solution includes acetate buffer, and a composite surfactant composed of octyl glucoside (APG) and Triton X-100, and the added mass ratio of the two is 3:1; the concentration ratio of each component is as follows: sodium acetate concentration is 0.28 mol / L, acetic acid concentration is 0.12 mol / L, the concentration of the composite surfactant is 1.2 mg / mL, and the pH value is 4.

[0123] The extraction solvent was 95% ethanol.

[0124] Example 8

[0125] The extraction solution of this embodiment includes a combination of a wall material cracking solution and an extraction solvent.

[0126] The wall material lysis solution includes phosphate buffer and a composite surfactant composed of octyl glucoside (APG) and Triton X-100, and the added mass ratio of the two is 3:1; the concentration of the composite surfactant is 1.0 mg / mL, and the pH value of the phosphate buffer is 4.

[0127] The extraction solvent was 95% ethanol.

[0128] Example 9

[0129] The extraction solution of this embodiment includes a combination of a wall material cracking solution and an extraction solvent.

[0130] The wall material lysis solution includes phosphate buffer and a composite surfactant composed of octyl glucoside (APG) and Triton X-100, and the added mass ratio of the two is 3:1; the concentration of the composite surfactant is 1.0 mg / mL, and the pH value of the phosphate buffer is 3.

[0131] The extraction solvent was 95% ethanol.

[0132] Example 10

[0133] The extracting solution of this embodiment includes wall material pyrolysis solution and extraction solvent.

[0134] The wall material lysis solution includes phosphate buffer and a composite surfactant composed of octyl glucoside (APG) and Triton X-100, and the added mass ratio of the two is 3:1; the concentration of the composite surfactant is 1.0 mg / mL, and the pH value of the phosphate buffer is 5.

[0135] The extraction solvent was 95% ethanol.

[0136] Embodiment 11

[0137] The extraction solution of this embodiment includes a combination of a wall material cracking solution and an extraction solvent.

[0138] The wall material lysis solution includes acetate buffer, and a composite surfactant composed of octyl glucoside (APG) and Triton X-100, and the added mass ratio of the two is 3:1; the concentration ratio of each component is as follows: sodium acetate concentration is 0.28 mol / L, acetic acid concentration is 0.12 mol / L, the concentration of the composite surfactant is 1.5 mg / mL, and the pH value is 4.

[0139] Example 12

[0140] The extraction solution of this embodiment includes a combination of a wall material cracking solution and an extraction solvent.

[0141] The wall material lysis solution includes acetate buffer, and a composite surfactant composed of octyl glucoside (APG) and Triton X-100, and the added mass ratio of the two is 3:1; the concentration ratio of each component is as follows: sodium acetate concentration is 0.28 mol / L, acetic acid concentration is 0.12 mol / L, the concentration of the composite surfactant is 0.40 mg / mL, and the pH value is 4.

[0142] Example 13

[0143] The extraction solution of this embodiment includes a combination of a wall material cracking solution and an extraction solvent.

[0144] The wall material lysis solution includes acetate buffer, and a composite surfactant composed of octyl glucoside (APG) and Triton X-100, and the added mass ratio of the two is 3:1; the concentration ratio of each component is as follows: sodium acetate concentration is 0.28 mol / L, acetic acid concentration is 0.12 mol / L, the concentration of the composite surfactant is 0.20 mg / mL, and the pH value is 4.

[0145] Embodiment 14

[0146] The extraction solution of this embodiment includes a combination of a wall material cracking solution and an extraction solvent.

[0147] The wall material lysis solution includes acetate buffer, and a composite surfactant composed of octyl glucoside (APG) and Triton X-100, and the added mass ratio of the two is 3:1; the concentration ratio of each component is as follows: sodium acetate concentration is 0.28 mol / L, acetic acid concentration is 0.12 mol / L, the concentration of the composite surfactant is 0.05 mg / mL, and the pH value is 4.

[0148] Comparative Example

[0149] The reagents and extraction method of Example 4 in the invention entitled "A method for extracting vitamin K2 from a vitamin K2 embedding agent and a method for detecting vitamin K2" with publication number CN105301148A are used as a comparative example of this application.

[0150] Vitamin K2 extraction and detection experiment

[0151] This experiment used 4 kinds of commercially available vitamin K2 microcapsule powder raw materials as test samples, as follows:

[0152] Sample 1

[0153] Commercially available vitamin K2 microcapsule powder 1 (batch number: GF04001). This sample uses maltodextrin, edible corn starch, soybean oil, sodium caseinate, and ascorbyl palmitate as wall materials, and the content of vitamin K2 is 2.5 mg / g.

[0154] Sample 2

[0155] Commercially available vitamin K2 microcapsule powder 2 (batch number: LC04002). This sample uses ethyl cellulose, edible corn starch, soybean oil, sodium caseinate, and ascorbyl palmitate as wall materials, and the content of vitamin K2 is 2.5 mg / g.

[0156] Sample 3

[0157] Commercially available vitamin K2 microcapsule powder 3 (batch number: SJ04003), this sample uses xanthan gum, edible corn starch, soybean oil, sodium caseinate, and ascorbyl palmitate as wall materials, and the content of vitamin K2 is 2.5 mg / g.

[0158] Sample 4

[0159] Commercially available vitamin K2 microcapsule powder 4 (batch number: YH04001). This sample uses beeswax, edible corn starch, soybean oil, sodium caseinate, and ascorbyl palmitate as wall materials, and the content of vitamin K2 is 2.5 mg / g.

[0160] The above four samples were extracted with the extracts of Examples 1 to 14 of the present application, and then the content of vitamin K2 in the extracts was detected; the above four samples were extracted with the comparative example, and then the content of vitamin K2 in the extracts was detected. Examples 1 to 14 of the present application adopt the experimental method verified by methodology of the present application; the comparative example adopts the detection method corresponding to patent CN105301148A.

[0161] The extracts of Examples 1 to 14 of the present application all use the following steps to extract samples:

[0162] 1. Accurately weigh 4 g of sample and place it in a 100 mL brown volumetric flask;

[0163] 2. Add 10 mL of wall material lysis solution to the volumetric flask and heat in a 40°C water bath for 15 min.

[0164] 3. Add 60 mL of extraction solvent to the volumetric flask, vortex for 1 min to mix, and then perform ultrasonic extraction for 15 min; the ultrasonic extraction parameters are: temperature 40°C, ultrasonic frequency 45kHz±5Hz, ultrasonic power 1kW±5W; after ultrasonic extraction, dilute to 100 mL with extraction solvent;

[0165] 4. Filter the fixed volume of extraction solution through a 0.45µm microporous filter membrane to obtain the sample extract.

[0166] The specific experimental results of this experiment are shown in Table 4 below.

[0167] Table 4 Experimental results of vitamin K2 microcapsule powder extraction in Examples 1 to 14

[0168] Sample 1 (mg / g) Sample 2 (mg / g) Sample 3 (mg / g) Sample 4 (mg / g) Example 1 2.4546 2.4584 2.4534 2.4529 Example 2 2.4541 2.4577 2.4531 2.4524 Example 3 2.4536 2.4562 2.4526 2.4521 Example 4 2.4543 2.4571 2.4540 2.4527 Example 5 2.4878 2.4874 2.4871 2.4868 Example 6 2.4785 2.4812 2.4778 2.4774 Example 7 2.4949 2.4961 2.4941 2.4939 Example 8 2.4858 2.4873 2.4849 2.4846 Example 9 2.4792 2.4809 2.4784 2.4781 Example 10 2.4758 2.4771 2.4748 2.4745 Embodiment 11 2.4942 2.4957 2.4934 2.4932 Example 12 2.4769 2.4791 2.4762 2.4758 Embodiment 13 2.4752 2.4776 2.4738 2.4731 Embodiment 14 2.4602 2.4631 2.4593 2.4588 Comparative Example 2.3614 2.3539 2.2638 2.2456

[0169] It can be seen from the data in Table 4 that the extracts of Examples 1 to 14 of the present application can achieve good lysis of various wall materials and can achieve better extraction of vitamin K2. The deviation rate between the detection value of vitamin K2 detected by the extract and the actual value is controlled within 2%.

[0170] The comparative extraction method has different cleavage effects on different wall materials. The cleavage effect on starch and cellulose wall materials is better, while the cleavage effect on colloid and lipid wall materials is relatively poor; the detection value of vitamin K2 in the extract has a large deviation rate from the actual value.

[0171] It can also be seen from the data in Table 1 that the extraction rates of Examples 5 to 14 of the present application using a composite surfactant are significantly better than those of Examples 1 to 4 using a single surfactant; from the data results of Examples 5 to 7 and Examples 11 to 14, it can be seen that when the surfactant concentration of the present application is controlled at 0.50 to 1.20 mg / mL, the extraction effect is relatively better; from the results of Examples 8 to 10, it can be seen that when the pH value of the wall material lysate of the present application is controlled at about 4, the extraction effect is relatively better.

[0172] The applicant conducted experimental research on different acidic buffer solutions. Within the range of acidic buffer solutions disclosed in the present application, any acidic buffer solution with a pH value controlled in the range of 2 to 6 can effectively destroy the wall material. The deviation between the detected value and the theoretical value of the extracted vitamin K2 is within 2%, which meets the requirements of detection accuracy.

[0173] The following are Examples 15 to 20 of the present application, which verify different surfactant types and different surfactant concentrations.

[0174] Embodiment 15

[0175] The extract of this example is substantially the same as that of Example 1, except that an equal amount of sodium dodecyl sulfate (SDS) is used to replace octyl glucoside (APG).

[0176] Example 16

[0177] The extract of this example is substantially the same as that of Example 15, except that the concentration of sodium dodecyl sulfate in the extract is 0.05 mg / mL.

[0178] Embodiment 17

[0179] The extract of this example is substantially the same as that of Example 15, except that the concentration of sodium dodecyl sulfate in the extract is 0.50 mg / mL.

[0180] Embodiment 18

[0181] The extract of this example is substantially the same as that of Example 15, except that the concentration of sodium dodecyl sulfate in the extract is 1.2 mg / mL.

[0182] Embodiment 19

[0183] The extract of this example is substantially the same as that of Example 7, except that an equal amount of sodium dodecyl sulfate (SDS) is used to replace octyl glucoside (APG).

[0184] Embodiment 20

[0185] The extract of this example is substantially the same as that of Example 7, except that an equal amount of sodium dodecylbenzenesulfonate (SDBS) is used to replace octyl glucoside (APG).

[0186] The same extraction steps as above were used to extract samples 1 to 4, and then the obtained sample extracts were tested using the methodologically verified experimental method of this application. The specific results are shown in Table 5 below.

[0187] Table 5 Experimental results of vitamin K2 microcapsule powder extraction in Examples 15 to 20

[0188] Sample 1 (mg / g) Sample 2 (mg / g) Sample 3 (mg / g) Sample 4 (mg / g) Embodiment 15 2.4532 2.4541 2.4511 2.4504 Example 16 2.4322 2.4336 2.4297 2.4289 Embodiment 17 2.4418 2.4427 2.4409 2.4402 Embodiment 18 2.4541 2.4549 2.4508 2.4497 Embodiment 19 2.4921 2.4929 2.4901 2.4891 Embodiment 20 2.4928 2.4933 2.4908 2.4902

[0189] It can be seen from the data in Table 5 that the extracts of Examples 15 to 20 of the present application can achieve good destruction of various wall materials and can also achieve better extraction of vitamin K2. The deviation rate between the detection value of vitamin K2 detected by the extract and the actual value is also controlled within 2%.

[0190] It can also be seen from the data in Table 5 that in the extract of the present application, when the surfactant concentration is lower than 1.2 mg / mL, the extraction effect decreases as the surfactant concentration decreases. In addition, when anionic surfactants are used, the extraction effect of the composite surfactant is also better than that of a single surfactant.

[0191] The following are Examples 21 to 25 of the present application, which are basically the same as Example 7.

[0192] Embodiment 21

[0193] The difference between this embodiment and embodiment 7 is that tetrahydrofuran is used as the extraction solvent.

[0194] Embodiment 22

[0195] The difference between this embodiment and embodiment 7 is that ethyl acetate is used as the extraction solvent.

[0196] Embodiment 23

[0197] The difference between this embodiment and embodiment 7 is that a composite surfactant consisting of octyl glucoside (APG) and Triton X-100 is used, and the added mass ratio of the two is 2:1.

[0198] Embodiment 24

[0199] The difference between this embodiment and embodiment 7 is that a composite surfactant consisting of octyl glucoside (APG) and Triton X-100 is used, and the added mass ratio of the two is 4:1.

[0200] Embodiment 25

[0201] The difference between this embodiment and embodiment 7 is that a composite surfactant consisting of octyl glucoside (APG) and Triton X-100 is used, and the added mass ratio of the two is 5:1.

[0202] The vitamin K2 extraction and detection experiments were conducted again on the extracts of Examples 21 to 25 of the present application, and the results are shown in Table 6.

[0203] Table 6 Experimental results of vitamin K2 microcapsule powder extraction in Examples 21 to 25

[0204] Sample 1 (mg / g) Sample 2 (mg / g) Sample 3 (mg / g) Sample 4 (mg / g) Embodiment 21 2.4944 2.4953 2.4936 2.4934 Embodiment 22 2.4946 2.4956 2.4938 2.4935 Embodiment 23 2.4935 2.4943 2.4929 2.4928 Embodiment 24 2.4939 2.4948 2.4933 2.4932 Embodiment 25 2.4928 2.4934 2.4924 2.4925

[0205] It can be seen from the data in Table 6 that the use of different organic solvents as extraction solvents in this application has little effect on the test results. It can be seen that different organic solvents have no obvious effect on the extraction effect of vitamin K2 in the present application scheme, and the detection method used in this application also has no obvious interference.

[0206] It can also be seen from the data in Table 6 that the extraction effects of the extracts of Examples 19 to 21 of the present application are not as good as the extraction effect of the extract of Example 7 of the present application; thus, when the composite surfactant composed of octyl glucoside (APG) and Triton X-100 is used, the extraction effect is optimal when the mass ratio of the two is 3:1. It can also be seen from this that different ratios of composite surfactants have a certain influence on the extraction effect.

[0207] The following are Examples 26 to 29 of the present application, which use the extract of Example 7 of the present application, except that different extraction steps are used.

[0208] Embodiment 26

[0209] The extraction steps of this embodiment are as follows:

[0210] 1. Accurately weigh 4 g of sample and place it in a 100 mL brown volumetric flask;

[0211] 2. Add 10 mL of wall material lysis solution to the volumetric flask and heat in a 40°C water bath for 15 min.

[0212] 3. Add 60 mL of extraction solvent to the volumetric flask, vortex and shake for 1 min to mix, and then perform ultrasonic extraction for 15 min; the ultrasonic extraction parameters are: temperature 40°C, ultrasonic frequency 40kHz±5Hz, ultrasonic power 0.95kW±5W; after ultrasonic extraction, dilute to 100 mL with extraction solvent;

[0213] 4. Filter the fixed volume of extraction solution through a 0.45µm microporous filter membrane to obtain the sample extract.

[0214] Embodiment 27

[0215] The extraction steps of this embodiment are as follows:

[0216] 1. Accurately weigh 4 g of sample and place it in a 100 mL brown volumetric flask;

[0217] 2. Add 10 mL of wall material lysis solution to the volumetric flask and heat in a 40°C water bath for 15 min.

[0218] 3. Add 60 mL of extraction solvent to the volumetric flask, vortex and shake for 1 min to mix, and then perform ultrasonic extraction for 15 min; the ultrasonic extraction parameters are: temperature 40°C, ultrasonic frequency 50kHz±5Hz, ultrasonic power 1.05kW±5W; after ultrasonic extraction, dilute to 100 mL with extraction solvent;

[0219] 4. Filter the fixed volume of extraction solution through a 0.45µm microporous filter membrane to obtain the sample extract.

[0220] Embodiment 28

[0221] The extraction steps of this embodiment are as follows:

[0222] 1. Accurately weigh 4 g of sample and place it in a 100 mL brown volumetric flask;

[0223] 2. Add 10 mL of wall material lysis solution to the volumetric flask and heat in a 30°C water bath for 25 min.

[0224] 3. Add 60 mL of extraction solvent to the volumetric flask, vortex for 1 min to mix, and then perform ultrasonic extraction for 25 min; the ultrasonic extraction parameters are: temperature 30°C, ultrasonic frequency 45kHz±5Hz, ultrasonic power 1kW±5W; after ultrasonic extraction, dilute to 100 mL with extraction solvent;

[0225] 4. Filter the fixed volume of extraction solution through a 0.45µm microporous filter membrane to obtain the sample extract.

[0226] Embodiment 29

[0227] The extraction steps of this embodiment are as follows:

[0228] 1. Accurately weigh 4 g of sample and place it in a 100 mL brown volumetric flask;

[0229] 2. Add 10 mL of wall material lysis solution to the volumetric flask and heat in a 50°C water bath for 15 min.

[0230] 3. Add 60 mL of extraction solvent to the volumetric flask, vortex for 1 min to mix, and then ultrasonically extract for 15 min; the ultrasonic extraction parameters are: temperature 50°C, ultrasonic frequency 45kHz±5Hz, ultrasonic power 1kW±5W; after ultrasonic extraction, dilute to 100 mL with extraction solvent;

[0231] 4. Filter the fixed volume of extraction solution through a 0.45µm microporous filter membrane to obtain the sample extract.

[0232] For Examples 26 to 29 of the present application, vitamin K2 extraction and detection experiments were conducted again, and the results obtained are shown in Table 7.

[0233] Table 7 Experimental results of vitamin K2 microcapsule powder extraction in Examples 26 to 29

[0234] Sample 1 (mg / g) Sample 2 (mg / g) Sample 3 (mg / g) Sample 4 (mg / g) Embodiment 26 2.4941 2.4950 2.4935 2.4934 Embodiment 27 2.4949 2.4962 2.4942 2.4939 Embodiment 28 2.4938 2.4947 2.4932 2.4929 Embodiment 29 2.4945 2.4955 2.4938 2.4932

[0235] It can be seen from the data in Table 7 that the ultrasonic parameters of the present application are preferably at an ultrasonic frequency of 40kHz±5Hz and an ultrasonic power of 0.95kW±5W or above, and the temperature of the wall material destruction and ultrasonic extraction steps is preferably controlled at 40°C.

[0236] Quality control test experiment of compound vitamin preparation samples

[0237] This experiment is used to verify the extract and extraction method of this application and to verify the effect of quality control.

[0238] The extracts of Example 1 and Example 7 were used in this experiment.

[0239] The extracts of Examples 1 and 7 of the present application are extracted by the following steps:

[0240] 1. Crush the vitamin-mineral compound preparation (solid sample into powder), accurately measure 4 g of sample, and place it in a 100 mL brown volumetric flask;

[0241] 2. Add 10 mL of wall material lysis solution to the volumetric flask and heat in a 40°C water bath for 15 min.

[0242] 3. Add 60 mL of extraction solvent to the volumetric flask, vortex for 1 min to mix, and then perform ultrasonic extraction for 15 min; the ultrasonic extraction parameters are: temperature 40°C, ultrasonic frequency 45kHz±5Hz, ultrasonic power 1kW±5W; after ultrasonic extraction, dilute to 100 mL with extraction solvent;

[0243] 4. Filter the fixed volume of extraction solution through a 0.45µm microporous filter membrane to obtain the sample extract.

[0244] This experiment used the extraction method of the comparative example.

[0245] The samples in this experiment were two homemade vitamin and mineral compound preparations, as follows:

[0246] Sample a

[0247] 21 Gold Vita® Multivitamin Mineral Tablets (Batch No. S04N001), produced by Hangzhou Minsheng Health Pharmaceutical Co., Ltd.; using commercially available vitamin K2 microcapsule powder 1 (Batch No.: S04N001), the content of vitamin K2 in the microcapsule powder is 2.5 mg / g; the theoretical feed content of vitamin K2 in the sample is 13.0 μg / g.

[0248] Sample b

[0249] 21 Gold Vita® Multivitamin Mineral Granules (Batch No. S04N002) were produced by Hangzhou Minsheng Health Pharmaceutical Co., Ltd.; commercially available vitamin K2 microcapsule powder 1 (Batch No.: GF04001) was used, and the content of vitamin K2 in the microcapsule powder was 2.5 mg / g; the theoretical feed content of vitamin K2 in the sample was 13.0 μg / g.

[0250] Sample c

[0251] 21 Gold Vita® Multivitamin Mineral Tablets (Batch No. S04N003), produced by Hangzhou Minsheng Health Pharmaceutical Co., Ltd.; commercially available vitamin K2 microcapsule powder 2 (Batch No.: GF04001) was used, the content of vitamin K2 in the microcapsule powder was 2.5 mg / g; the theoretical feed content of vitamin K2 in the sample was 13.0 μg / g.

[0252] Sample d

[0253] 21 Gold Vita® Multivitamin Mineral Tablets (Batch No. S04N004), produced by Hangzhou Minsheng Health Pharmaceutical Co., Ltd.; commercially available vitamin K2 microcapsule powder 3 (Batch No.: SJ04003) was used, the content of vitamin K2 in the microcapsule powder was 2.5 mg / g; the theoretical feed content of vitamin K2 in the sample was 13.0 μg / g.

[0254] Sample

[0255] 21 Jinvita® Multivitamin Mineral Complex Preparation (Batch No. S04N005), produced by Hangzhou Minsheng Health Pharmaceutical Co., Ltd.; commercially available vitamin K2 microcapsule powder 4 (Batch No.: YH04001) was used, and the content of vitamin K2 in the microcapsule powder was 2.5 mg / g; the theoretical feed content of vitamin K2 in the sample was 13.0 μg / g. After the sample extraction was completed, the five samples were tested using the same detection method as the vitamin K2 extraction test, and the results are shown in Table 8.

[0256] Table 8 Quality control test results

[0257] Sample a (μg / g) (mg / g) (mg / g) (μg / g) Sample b (μg / g) (μg / g) Sample c (μg / g) Sample d (μg / g) Sample e (μg / g) Example 1 12.8286 12.8343 12.8355 12.8199 12.8317 Example 7 12.9041 12.9133 12.8999 12.9341 12.9087 Comparative Example 12.1689 11.7028 11.9825 11.8976 12.2546

[0258] From the data in Table 8, it can be seen that the extracts of Examples 1 and 7 of the present application, using the specific extraction method of the present application, extract trace vitamin K2 from the vitamin-mineral composite preparation after embedding with different wall materials, and the extraction effect is better, and the deviation between the detection value and the theoretical value is within 2%; while after extraction using the comparative example, the deviation between the sample detection value and the theoretical value exceeds 2%. It can be seen that the extract and extraction method of the present application are applied to the K2 quality control inspection of the vitamin-mineral composite preparation, and the extraction effect of the microencapsulated vitamin K2 is significantly better than the existing method, which can greatly improve the accuracy of the detection and is beneficial to the quality control of the product.

[0259] The above are all preferred embodiments of the present application, and the protection scope of the present application is not limited thereto. Therefore, any equivalent changes made according to the structure, shape, and principle of the present application should be included in the protection scope of the present application.

Claims

1. An extract for extracting vitamin K2 from a vitamin-mineral compound preparation, It is characterized in that The extracting solution comprises a combination of a wall material lysing solution and an extraction solvent; the base liquid of the wall material lysing solution adopts an acidic buffer solution, and the wall material lysing solution also comprises a surfactant added to the base liquid, the concentration of the surfactant in the base liquid is 0.05-1.5 mg / mL, and the pH value of the wall material lysing solution is 2-6; the surfactant adopts a nonionic surfactant, or a sulfate surfactant or sodium dodecylbenzene sulfonate among anionic surfactants; the extraction solvent is an organic solvent; The acidic buffer is selected from one of acetate buffer, phosphate buffer, borate buffer or citrate buffer.

2. The extract for extracting vitamin K2 from a vitamin-mineral compound preparation according to claim 1, It is characterized in that The concentration of the surfactant in the base liquid is 0.5-1.2 mg / mL.

3. The extract for extracting vitamin K2 from a vitamin-mineral compound preparation according to claim 1, It is characterized in that The surfactant is selected from one or more of polyoxyethylene ether surfactants, polyol surfactants, alkyl glycoside surfactants, sulfate surfactants, and sodium dodecylbenzene sulfonate.

4. The extract for extracting vitamin K2 from a vitamin-mineral complex preparation according to claim 3, It is characterized in that The surfactant is a single surfactant or a compound surfactant, and the compound surfactant is a compound of two surfactants.

5. The extract for extracting vitamin K2 from a vitamin-mineral complex preparation according to claim 4, It is characterized in that The concentration ratio of the two surfactants in the compound surfactant is 1:1-5.

6. The extract for extracting vitamin K2 from a vitamin-mineral compound preparation according to claim 1, It is characterized in that The organic solvent is selected from one of 80% to 100% ethanol, isopropanol, tetrahydrofuran, n-hexane, ethyl acetate, ether, cyclohexane, isooctane or chloroform.

7. A method for extracting vitamin K2 from a vitamin-mineral compound preparation, It is characterized in that The method for extracting vitamin K2 from a vitamin-mineral composite preparation according to any one of claims 1 to 6 comprises the following steps: S1. Pre-treat the vitamin-mineral compound preparation sample, accurately weigh the pre-treated sample, and place it in a light-proof container; S2, adding the wall material lysis solution to the light-proof container containing the sample in step S1, and treating for more than 10 minutes to obtain a sample mixed solution; S3, adding an extraction solvent to the sample mixture obtained in step S2, mixing, and ultrasonically extracting for more than 10 minutes to obtain an extraction mixture; S4. Filter the mixed extraction solution obtained in step S3 to remove insoluble matter, thereby obtaining a sample extract of vitamin K2.

8. The method for extracting vitamin K2 from a vitamin-mineral compound preparation according to claim 7, It is characterized in that In step S2, the processing temperature is controlled at 30-75°C.

9. The method for extracting vitamin K2 from a vitamin-mineral compound preparation according to claim 7, It is characterized in that In step S3, the ultrasonic extraction parameters are: temperature 30-75°C, ultrasonic frequency 35-50kHz, and ultrasonic power 1kW±5W.

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