Rehabilitation new liquid and preparation method thereof

Abstract

The application provides a rehabilitation new liquid and a preparation method thereof. The application uses Periplaneta americana as a raw material, extracts, concentrates and defats the Periplaneta americana, directly uses defatted liquid of the Periplaneta americana and specified components to prepare the rehabilitation new liquid, omits multiple process steps such as filtering and concentrating and collecting cream in the preparation process, simplifies the production process, shortens the production cycle, improves the production efficiency, and reduces the production cost.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical technology, specifically to a new rehabilitation liquid and its preparation method. Background Technology

[0002] The American cockroach (Periplaneta Americana Linnaeus), commonly known as the "cockroach," is the largest insect in the Blattidae family, native to South America. It is a traditional Chinese medicine, first mentioned in the *Compendium of Materia Medica Supplement*, also called *Shijiang* or *Huachong*, with the adult insect used medicinally. It is cold in nature, salty in taste, and poisonous, with a pungent flavor. It is used for its effects of clearing heat and dampness, eliminating stagnation, detoxifying, promoting diuresis, and reducing swelling. Modern research has found that the active ingredients in the American cockroach have effects such as protecting and repairing the liver, anti-inflammatory and analgesic properties, anti-tumor effects, and enhancing immunity.

[0003] Kangfuxin Liquid is an extract of dried American cockroach body. It can promote blood circulation, nourish yin and promote tissue regeneration. For internal use: it is used for blood stasis, stomach pain and bleeding, gastric and duodenal ulcers; and as an adjunct treatment for yin deficiency pulmonary tuberculosis. For external use: it is used for wounds, injuries, ulcers, fistulas, burns, scalds, and bedsores.

[0004] Existing technologies, such as CN111728984A, disclose a method for preparing defatted American cockroach extract, including the following steps: (1) inactivation and pulverization; (2) defatting; (3) extraction; (4) vacuum concentration; (5) defatting filtration; (6) macroporous adsorption resin column chromatography; and (7) vacuum concentration. However, after defatting filtration, it also involves cumbersome chromatography and concentration steps. The existing technology for preparing Kangfuxin liquid suffers from problems such as cumbersome processes, low production efficiency, and high production costs, which are not conducive to industrial production. Summary of the Invention

[0005] The present invention provides a Kangfuxin liquid, characterized in that, calculated by weight percentage, the Kangfuxin liquid comprises: 5.0-12.0% American cockroach defatted liquid, 5.0-15.0% glycerin, 0-2.0‰ licorice extract, 0-0.8‰ vitamin C, 0-5.0‰ sodium benzoate, and 72.5-90.0% purified water.

[0006] In some embodiments, the Kangfuxin solution comprises, by weight percentage: 6.0%-10.0% American cockroach defatted solution, 8.0%-12.0% glycerin, 0%-2.0‰ licorice extract, 0%-0.3‰ vitamin C, 0%-2.5‰ sodium benzoate, and 78.0%-86.0% purified water.

[0007] In some embodiments, each 1 mL of the Kangfuxin solution contains 0.80-1.20 mg of total amino acids, calculated as alanine.

[0008] In some embodiments, each 1 mL of the Kangfuxin solution contains 0.82-1.18 mg of total amino acids, calculated as alanine.

[0009] In some embodiments, the American cockroach defatted liquid is a liquid obtained by defatting American cockroach extract.

[0010] In some embodiments, the American cockroach defatting liquid is a liquid obtained by defatting and filtering an extract of American cockroach alcohol (preferably ethanol; more preferably 75%-95% ethanol).

[0011] This invention also provides a method for preparing Kangfuxin Liquid, comprising:

[0012] Step 2: Prepare Kangfuxin Liquid directly using American cockroach degreasing solution.

[0013] In some embodiments, step 2 includes: adding the American cockroach defatting solution, glycerin, licorice extract, vitamin C and sodium benzoate to water, stirring, filtering, filling and sterilizing to obtain the Kangfuxin solution.

[0014] In some embodiments, the preparation method further includes: Step 1: preparing the American cockroach degreasing solution.

[0015] In some implementations, step 1 includes:

[0016] Step 1.1: American cockroaches were impregnated with alcohol to obtain an American cockroach extract;

[0017] Step 1.2: Filter and concentrate the American cockroach extract to obtain an American cockroach alcohol concentrate;

[0018] Step 1.3: Add water to the concentrated extract of American cockroaches, let it stand, separate the water layer, filter the water layer, and obtain the defatted American cockroach solution.

[0019] In some embodiments, the alcohol in step 1.1 is selected from ethanol; preferably, the alcohol is 75%-95% ethanol (e.g., 80% ethanol, etc.).

[0020] In some embodiments, the immersion temperature in step 1.1 is 60±4℃.

[0021] In some implementations, the number of immersions in step 1.1 is at least once; preferably, the number of immersions is three times.

[0022] In some embodiments, the soaking time in step 1.1 is 1-30 hours per cycle; preferably, the soaking time is 12-24 hours per cycle; more preferably, the soaking time is 12 hours per cycle.

[0023] In some embodiments, step 1.1 includes immersing American cockroaches three times in 75%-95% ethanol at 60±4°C, each time for 12-24 hours, to obtain an American cockroach extract.

[0024] In some embodiments, American cockroaches are impregnated with ethanol (e.g., 75%-95% ethanol) to obtain an American cockroach ethanol extract.

[0025] In some embodiments, the concentration described in step 1.2 is a vacuum-concentrated filtrate.

[0026] In some implementations, the temperature for vacuum concentration is 40-60°C.

[0027] In some implementations, the pressure for vacuum concentration is ≤-0.6MPa.

[0028] In some embodiments, step 1.2 includes filtering the American cockroach extract and concentrating the filtrate under reduced pressure at 40-60°C and a vacuum degree ≤-0.6MPa to obtain an American cockroach alcohol extract concentrate.

[0029] In some embodiments, the volume ratio of the concentrated American cockroach extract to water in step 1.3 is 1:(1-10); preferably, the volume ratio is 1:5.

[0030] In some embodiments, the filtration described in step 1.3 is filtration using a 0.22 μm filter membrane.

[0031] In some embodiments, step 1.3 includes adding water to the concentrated American cockroach alcohol extract, allowing it to stand, separating and discarding the precipitate and oil layer to obtain an aqueous layer, which is then filtered through a 0.22 μm filter membrane to obtain the American cockroach defatted liquid.

[0032] In some implementations, step 1 includes:

[0033] Step 1.1: Soak American cockroaches three times with 75%-95% ethanol at 60±4℃, each time for 12-24 hours, to obtain American cockroach extract;

[0034] Step 1.2: Filter the American cockroach extract and concentrate the filtrate under reduced pressure at 40~60℃ and vacuum degree ≤-0.6MPa to obtain American cockroach alcohol extract concentrate;

[0035] Step 1.3: Add water to the concentrated extract of American cockroaches, let it stand, separate and discard the precipitate and oil layer to obtain an aqueous layer, filter the aqueous layer through a 0.22μm filter membrane to obtain the defatted extract of American cockroaches; the volume ratio of the concentrated extract of American cockroaches to water is 1:(1-10) (preferably the volume ratio is 1:5).

[0036] In some embodiments, the preparation method of Kangfuxin solution includes: adding 5.0-12.0% of American cockroach defatting solution, 5.0-15.0% of glycerin, 0-2.0‰ of licorice extract, 0-0.8‰ of vitamin C, and 0-5.0‰ of sodium benzoate to 71.5-90.0% of purified water by weight percentage, stirring, filtering, filling, and sterilizing to obtain the Kangfuxin solution. Optionally, each 1 mL of the Kangfuxin solution contains 0.80-1.20 mg of total amino acids, calculated as alanine.

[0037] In some embodiments, the preparation method of Kangfuxin solution includes: adding 6.0-10.0% of American cockroach defatting solution, 8.0-12.0% of glycerin, 0-2.0‰ of licorice extract, 0-0.3‰ of vitamin C, and 0-2.5‰ of sodium benzoate to 78.0-86.0% of purified water by weight percentage, stirring, filtering, filling, and sterilizing to obtain the Kangfuxin solution. Optionally, each 1 mL of the Kangfuxin solution contains 0.82-1.18 mg of total amino acids, calculated as alanine.

[0038] In some embodiments, the preparation method of Kangfuxin liquid includes: Step 1.1: using alcohol to impregnate American cockroaches to obtain American cockroach extract;

[0039] Step 1.2: Filter and concentrate the American cockroach extract to obtain an American cockroach alcohol concentrate;

[0040] Step 1.3: Add water to the concentrated extract of American cockroach alcohol, let stand, separate the water layer, filter the water layer, and obtain the defatted American cockroach liquid;

[0041] Step 2: Calculate by weight percentage and add 5.0-12.0% of American cockroach defatting solution, 5.0-15.0% of glycerin, 0-2.0‰ of licorice extract, 0-0.8‰ of vitamin C, and 0-5.0‰ of sodium benzoate to 72.5-90.0% of purified water. Stir, filter, fill, and sterilize to obtain the Kangfuxin solution. Optionally, each 1 mL of the Kangfuxin solution contains 0.80-1.20 mg of total amino acids, calculated as alanine.

[0042] In some embodiments, the preparation method of Kangfuxin solution includes:

[0043] Step 1.1: Soak American cockroaches three times with 75%-95% ethanol at 60±4℃, each time for 12-24 hours, to obtain American cockroach extract;

[0044] Step 1.2: Filter the American cockroach extract and concentrate the filtrate under reduced pressure at 40~60℃ and vacuum degree ≤-0.6MPa to obtain American cockroach alcohol extract concentrate;

[0045] Step 1.3: Add water to the concentrated American cockroach alcohol extract, let stand, separate and discard the precipitate and oil layer to obtain an aqueous layer, filter the aqueous layer through a 0.22μm filter membrane to obtain the American cockroach defatted liquid; the volume ratio of the concentrated American cockroach alcohol extract to water is 1:(1-10) (preferably the volume ratio is 1:5);

[0046] Step 2: Calculate by weight percentage and add 6.0-10.0% of American cockroach defatting solution, 8.0-12.0% of glycerin, 0-2.0‰ of licorice extract, 0-0.3‰ of vitamin C, and 0-2.5‰ of sodium benzoate to 78.0-86.0% of purified water. Stir, filter, fill, and sterilize to obtain the Kangfuxin solution. Optionally, each 1 mL of the Kangfuxin solution contains 0.82-1.18 mg of total amino acids, calculated as alanine.

[0047] The beneficial effects of this invention include the fact that existing industrial processes use American cockroaches as raw materials, and obtain American cockroach extract through multiple steps such as extraction, concentration, defatting, filtration, and concentration, and then use the American cockroach extract to formulate Kangfuxin liquid, which is a complicated process.

[0048] This invention uses American cockroaches as raw material. After extraction, concentration, and degreasing, the degreased American cockroach solution and specified components are directly used to prepare Kangfuxin solution. While ensuring the quality of the finished product, multiple process steps such as filtration and concentration are omitted during the preparation process, simplifying the production process, shortening the product production cycle by more than 10 hours, and increasing the product production efficiency by more than 20%. For every 35,000L of Kangfuxin solution produced, the production cost is reduced by approximately 2,000 yuan. Attached Figure Description

[0049] Figure 1 The images show the thin-layer chromatography identification results of samples from Examples 1-10, where the shift value at point A is 0.7, the shift value at point B is 0.2, 1 represents the sample of Example 1, 2 represents the sample of Example 2, 3 represents the sample of Example 3, 4 represents the sample of Example 4, 5 represents the sample of Example 5, 6 represents the sample of Example 6, 7 represents the sample of Example 7, 8 represents the sample of Example 8, 9 represents the sample of Example 9, 10 represents the sample of Example 10, and the control represents the reference standard.

[0050] Figure 2 The images show the thin-layer chromatography identification results of samples from Examples 11-15, where the ratio shift value at point A is 0.7, the ratio shift value at point B is 0.2, 11 represents the sample of Example 11, 12 represents the sample of Example 12, 13 represents the sample of Example 13, 14 represents the sample of Example 14, 15 represents the sample of Example 15, and the control represents the reference standard.

[0051] Figure 3-17 These are the characteristic spectra of the samples from Examples 1-15. Detailed Implementation

[0052] The present invention will be further described in detail below through specific embodiments. These embodiments are for illustrative purposes only and do not limit the scope of protection of the present invention. The American cockroaches used in the embodiments of the present invention were purchased from Yunnan Nanjing Xin Biotechnology Co., Ltd., batch number 2503002, and can also be purchased from other commercially available products sold by other manufacturers. The licorice extract powder used in the embodiments of the present invention was purchased from Xinjiang Alar Xinong Licorice Industry Co., Ltd., and can also be purchased from other commercially available products sold by other manufacturers. The American cockroach defatted solution used in the preparation of Kangfuxin Liquid in the present invention can also be prepared by referring to existing techniques. Other reagents or materials of the present invention can all be obtained commercially.

[0053] In this embodiment of the invention, the licorice extract can also be pre-prepared as a licorice extract pretreatment solution and then used to prepare Kangfuxin liquid. The specific operation is as follows: dissolve licorice extract (e.g., licorice extract powder) in five times its weight of water, stir for 20 minutes, heat to a gentle boil, cover and let stand at 0°C or below for more than 24 hours, then pump the supernatant of the solution to be centrifuged into a separator for centrifugation, weigh and discard the lower precipitate, control the discharge rate, collect the centrifuged liquid, and obtain the licorice extract pretreatment solution.

[0054] Example 1

[0055] Add 8.04% of the defatted American cockroach extract ①, 10% glycerin, 1.5‰ of licorice extract, 0.2‰ of vitamin C, and 2.5‰ of sodium benzoate sequentially to a 1000mL measuring cup, add water to make up to 1000mL, stir for 10 minutes, filter to obtain 1030g of Kangfuxin liquid preparation solution, fill and seal in 100mL oral liquid pharmaceutical polypropylene bottles, and sterilize in a water bath at 115℃ for 30 minutes.

[0056] The preparation process of the American cockroach degreasing solution ① is as follows:

[0057] Extraction: American cockroaches were soaked three times in 80% ethanol for 12 hours each time to obtain an ethanol extract of American cockroaches;

[0058] Concentration: The obtained American cockroach ethanol extract was filtered and concentrated under reduced pressure at 40~60℃ and vacuum degree ≤-0.6MPa to obtain the concentrated American cockroach ethanol extract.

[0059] Degreasing: Add water to the obtained concentrated American cockroach alcohol extract (the volume ratio of American cockroach alcohol extract to water is 1:5), let it stand, and then degrease it. Separate and discard the precipitate and oil layer. Filter the water layer through a 0.22 μm filter membrane to obtain American cockroach degreased solution ①.

[0060] The Kangfuxin liquid prepared in this embodiment is a light brown liquid with a slightly fishy odor and a sweet taste.

[0061] Two batches of Kangfuxin Liquid were prepared in accordance with Example 1 and named Example 2 and Example 3, respectively.

[0062] Example 4

[0063] Add 8.26% of the defatted American cockroach extract ②, 10% glycerin, 1.5‰ of licorice extract, 0.2‰ of vitamin C, and 2.5‰ of sodium benzoate to a 1000mL measuring cup, add water to make up to 1000mL, stir for 10 minutes, filter to obtain 1030g of Kangfuxin liquid preparation solution, fill and seal in 100mL oral liquid pharmaceutical polypropylene bottles, and sterilize in a 115℃ water bath for 30 minutes.

[0064] The preparation process of the American cockroach degreasing solution ② is as follows:

[0065] Extraction: American cockroaches were soaked three times in 80% ethanol for 12 hours each time to obtain an ethanol extract of American cockroaches;

[0066] Concentration: The obtained American cockroach ethanol extract was filtered and concentrated under reduced pressure at 40~60℃ and vacuum degree ≤-0.6MPa to obtain the concentrated American cockroach ethanol extract.

[0067] Degreasing: Add water to the obtained concentrated American cockroach alcohol extract (the volume ratio of American cockroach alcohol extract to water is 1:5), let it stand, and then degrease it. Separate and discard the precipitate and oil layer. Filter the water layer through a 0.22μm filter membrane to obtain American cockroach degreased solution ②.

[0068] The Kangfuxin liquid prepared in this embodiment is a light brown liquid with a slightly fishy odor and a sweet taste.

[0069] Two batches of Kangfuxin liquid were prepared in accordance with Example 4, namely Example 5 and Example 6.

[0070] Example 7

[0071] Add 8.06% of the defatted American cockroach extract ③, 10% glycerin, 1.5‰ of licorice extract, 0.2‰ of vitamin C, and 2.5‰ of sodium benzoate to a 1000mL measuring cup, add water to make up to 1000mL, stir for 10 minutes, filter to obtain 1030g of Kangfuxin liquid preparation solution, fill and seal in 100mL oral liquid pharmaceutical polypropylene bottles, and sterilize in a water bath at 115℃ for 30 minutes.

[0072] The preparation process of the American cockroach degreasing solution ③ is as follows:

[0073] Extraction: American cockroaches were warm-soaked three times with 80% ethanol, each time for 12 hours, to obtain an ethanol extract of American cockroaches;

[0074] Concentration: The obtained American cockroach ethanol extract was filtered and concentrated under reduced pressure at 40~60℃ and vacuum degree ≤-0.6MPa to obtain the concentrated American cockroach ethanol extract.

[0075] Degreasing: Add water to the obtained concentrated American cockroach alcohol extract (the volume ratio of American cockroach alcohol extract to water is 1:5), let it stand, and then degrease it. Separate and discard the precipitate and oil layer. Filter the water layer through a 0.22μm filter membrane to obtain the degreased American cockroach solution ③.

[0076] The Kangfuxin liquid prepared in this embodiment is a light brown liquid with a slightly fishy odor and a sweet taste.

[0077] Two batches of Kangfuxin liquid were prepared in accordance with Example 7, namely Example 8 and Example 9.

[0078] Example 10

[0079] Add 7.63% of the defatted American cockroach extract, 10% glycerin, 1.5‰ of licorice extract, 0.2‰ of vitamin C, and 2.5‰ of sodium benzoate sequentially to a 1000mL measuring cup, add water to make up to 1000mL, stir for 10 minutes, filter to obtain 1030g of Kangfuxin liquid preparation solution, fill and seal in 100mL oral liquid pharmaceutical polypropylene bottles, and sterilize in a 115℃ water bath for 30 minutes.

[0080] The preparation process of the American cockroach degreasing solution ④ is as follows:

[0081] Extraction: American cockroaches were soaked three times in 80% ethanol for 12 hours each time to obtain an ethanol extract of American cockroaches;

[0082] Concentration: The obtained American cockroach ethanol extract was filtered and concentrated under reduced pressure at 40~60℃ and vacuum degree ≤-0.6MPa to obtain the concentrated American cockroach ethanol extract.

[0083] Degreasing: Add water to the obtained concentrated American cockroach alcohol extract (the volume ratio of American cockroach alcohol extract to water is 1:5), let it stand, and then degrease it. Separate and discard the precipitate and oil layer. Filter the water layer through a 0.22μm filter membrane to obtain the degreased American cockroach solution ④.

[0084] The Kangfuxin liquid prepared in this embodiment is a light brown liquid with a slightly fishy odor and a sweet taste.

[0085] Two batches of Kangfuxin liquid were prepared in accordance with Example 10, namely Example 11 and Example 12.

[0086] Example 13

[0087] Add 7.94% of the defatted American cockroach extract (⑤), 10% glycerin, 1.5‰ of licorice extract, 0.2‰ of vitamin C, and 2.5‰ of sodium benzoate to a 1000mL measuring cup, add water to make up to 1000mL, stir for 10 minutes, filter to obtain 1030g of Kangfuxin liquid preparation solution, fill and seal in 100mL oral liquid pharmaceutical polypropylene bottles, and sterilize in a water bath at 115℃ for 30 minutes.

[0088] The preparation process of the American cockroach degreasing solution ⑤ is as follows:

[0089] Extraction: American cockroaches were soaked three times in 80% ethanol for 12 hours each time to obtain an ethanol extract of American cockroaches;

[0090] Concentration: The obtained American cockroach ethanol extract was filtered and concentrated under reduced pressure at 40~60℃ and vacuum degree ≤-0.6MPa to obtain the concentrated American cockroach ethanol extract.

[0091] Degreasing: Add water to the obtained concentrated American cockroach alcohol extract (the volume ratio of American cockroach alcohol extract to water is 1:5), let it stand, and then degrease it. Separate and discard the precipitate and oil layer. Filter the water layer through a 0.22μm filter membrane to obtain the degreased American cockroach solution ⑤.

[0092] The Kangfuxin liquid prepared in this embodiment is a light brown liquid with a slightly fishy odor and a sweet taste.

[0093] Three batches of Kangfuxin Liquid were prepared in accordance with Example 13, namely Example 14 and Example 15.

[0094] Example 16 Quality Testing of Kangfuxin Liquid

[0095] The quality of the Kangfuxin liquid prepared in Examples 1-15 above was tested.

[0096] (1) The quality inspection items are shown in Table 1 below:

[0097] Table 1. Test Items for Kangfuxin Liquid

[0098]

[0099] (2) The detection method is as follows:

[0100] [Characteristics]

[0101] Procedure: Visually inspect the color of the liquid, smell its odor, and taste it.

[0102] Result: This product is a light brown liquid; it has a slightly fishy odor and a sweet taste.

[0103] [Appearance]

[0104] This product should be clarified, and a small amount of easily dispersed precipitate is permissible when shaken.

[0105] [Identification]

[0106] Physicochemical identification:

[0107] Procedure: Take 1 mL of this product, add a few drops of ninhydrin test solution, and heat. Result interpretation: A purple-blue color will appear.

[0108] Thin-layer chromatography identification:

[0109] Procedure: Take this product as the test solution; take alanine reference standard, add ethanol to prepare a solution with a concentration of 1 mg / mL, as the reference solution. Perform thin-layer chromatography (Pharmacopoeia of the People's Republic of China 2025 Edition, Part IV, General Chapter 0502). Apply 2 µL of each of the above two solutions separately to the same silica gel G thin-layer plate. Develop with n-butanol-glacial acetic acid-water (4:1:1) as the developing solvent. Remove, air dry, spray with 0.5% ninhydrin-acetone solution, and bake at 65℃ for about 20 minutes. Result interpretation: The chromatogram of the test sample should show 5 or more spots, with spots of the same color at the corresponding positions as those of the reference standard.

[0110]

Feature Map

[0111] Determined by high performance liquid chromatography (General Chapter 0512, Part IV, Pharmacopoeia of the People's Republic of China, 2025 edition).

[0112] Chromatographic conditions and system suitability test: Octadecylsilane-bonded silica gel was used as the stationary phase (Agela VenusilMP C18, 4.6 mm × 250 mm, 5 μm or equivalent column); 0.1% phosphoric acid solution was used as mobile phase A, and methanol as mobile phase B, with gradient elution as specified in the table below; column temperature was 25°C; detection wavelength was 254 nm; flow rate was 0.5 mL / min. The theoretical plate number, calculated based on the inosine peak, should not be less than 5000. Gradient elution conditions are shown in Table 2 below.

[0113] Table 2. Gradient elution conditions

[0114]

[0115] Preparation of reference solution: Take an appropriate amount of inosine reference standard, accurately weigh it, add water, and prepare a solution with a concentration of 30 μg / mL.

[0116] Preparation of the test solution: Measure 2 mL of this product into a 10 mL volumetric flask, dilute with water to the mark, shake well, filter, and take the filtrate to obtain the test solution.

[0117] Assay: Measure 10 µL of the reference solution and the test solution respectively, inject them into the liquid chromatograph, and record the chromatograms.

[0118] The reference chromatogram showed five characteristic peaks: 0.47 (peak 1), 0.56 (peak 2), 0.73 (peak 3), 1.00 [peak 4 (S)], and 1.71 (peak 5). Peak 1: uracil; Peak 2: hypoxanthine; Peak 3: xanthine; Peak 4 (S): inosine; Peak 5: protocatechuic acid.

[0119] The chromatogram of the test sample should detect 5 characteristic peaks. The chromatographic peak corresponding to the S peak in the characteristic chromatogram of the reference sample is taken as the S peak. The relative retention times of the remaining characteristic peaks and the S peak are calculated. The relative retention times should all be within ±10% of the specified values. The specified values ​​are: 0.47 (peak 1), 0.56 (peak 2), 0.73 (peak 3), 1.00 [peak 4 (S) peak], and 1.71 (peak 5).

[0120]

examine

[0121] pH value:

[0122] Procedure: Determine the pH value according to the pH determination method (General Chapter 0631, Part IV, 2025 Edition of the Pharmacopoeia of the People's Republic of China). First, use phosphate standard buffer solution (pH=6.88, 20℃) for positioning, and then use phthalate standard buffer solution (pH=4.00, 20℃) for slope adjustment and verification. Then, determine the pH value of the test sample.

[0123] Result interpretation: pH value should be 4.5-7.5.

[0124] [Content Determination]

[0125] Preparation of reference solution: Take an appropriate amount of alanine reference standard, add water, and prepare a solution with an alanine concentration of 25 μg / mL.

[0126] Preparation of standard curve: Take 0.2 mL, 0.4 mL, 0.6 mL, 0.8 mL, and 1.0 mL of the reference solution respectively, and place them in stoppered test tubes. Add water to 1 mL, add 1.0 mL of 0.2 mol / L citrate buffer (pH 5.0), 0.1 mL of 1% ascorbic acid solution, and 3.0 mL of 10% ninhydrin ethanol solution. Shake well, heat in a 90℃ water bath for 25 minutes, remove, cool, add 3.0 mL of 80% ethanol, shake well, and use the corresponding reagents as blanks. Measure the absorbance at 570 nm using ultraviolet-visible spectrophotometry (General Chapter 0401, Part IV, Pharmacopoeia of the People's Republic of China, 2025 Edition). Plot the standard curve with absorbance as the ordinate and concentration as the abscissa.

[0127] Assay: Measure 2 mL of this product into a 100 mL volumetric flask, dilute to the mark with water, shake well, and measure 1 mL into a stoppered test tube. Following the method under the Preparation of Standard Curve section, starting from "Add 1.0 mL of 0.2 mol / L citrate buffer (pH 5.0)," determine the absorbance according to the method. Read the amino acid content in the test solution from the standard curve (i.e., C in the following calculation formula), and calculate it as follows:

[0128]

[0129] Where: C represents the sample detection concentration (mg / ml); D represents the dilution factor; v represents the sampling volume (ml); N represents the total amino acid content calculated as alanine (mg / ml).

[0130] Result judgment: The total amino acid content of this product per 1 mL, calculated as alanine, should be 0.8 to 1.2 mg.

[0131] (3) Test results

[0132] The quality test results of the Kangfuxin solution prepared in Examples 1-15 are shown in the table below.

[0133] Table 3. Quality inspection results of Example 1

[0134]

[0135] Table 4. Quality inspection results of Example 2

[0136]

[0137] Table 5. Quality inspection results of Example 3

[0138]

[0139] Table 6. Quality inspection results of Example 4

[0140]

[0141] Table 7. Quality inspection results of Example 5

[0142]

[0143] Table 8. Quality inspection results of Example 6

[0144]

[0145] Table 9. Quality Inspection Results of Example 7

[0146]

[0147] Table 10. Quality inspection results of Example 8

[0148]

[0149] Table 11. Quality inspection results of Example 9

[0150]

[0151] Table 12. Quality inspection results of Example 10

[0152]

[0153] Table 13. Quality inspection results of Example 11

[0154]

[0155] Table 14. Quality inspection results of Example 12

[0156]

[0157] Table 15. Quality inspection results of Example 13

[0158]

[0159] Table 16. Quality Inspection Results of Example 14

[0160]

[0161] Table 17. Quality Inspection Results of Example 15

[0162]

[0163] The test results in the table above show that the Kangfuxin solution prepared directly from the defatted American cockroach solution in Examples 1-15 of this invention meets the quality standards of Kangfuxin solution.

[0164] The above are merely preferred embodiments of the present invention. It should be noted that the above preferred embodiments should not be considered as limitations on the present invention, and the scope of protection of the present invention should be determined by the scope defined in the claims. For those skilled in the art, several improvements and modifications can be made without departing from the spirit and scope of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A new rehabilitation liquid, characterized in that, The Kangfuxin solution comprises, by weight percentage: 5.0-12.0% American cockroach defatted solution, 5.0-15.0% glycerin, 0-2.0‰ licorice extract, 0-0.8‰ vitamin C, 0-5.0‰ sodium benzoate, and 72.5-90.0% purified water.

2. The Kangfuxin liquid according to claim 1, characterized in that, The Kangfuxin solution comprises, by weight percentage: 6.0%-10.0% American cockroach defatting solution, 8.0%-12.0% glycerin, 0%-2.0‰ licorice extract, 0%-0.3‰ vitamin C, 0%-2.5‰ sodium benzoate, and 78.0%-86.0% purified water.

3. The Kangfuxin liquid according to claim 1 or 2, characterized in that, Each 1 mL of the Kangfuxin liquid contains 0.80-1.20 mg of total amino acids, calculated as alanine.

4. The Kangfuxin Liquid according to claim 1, 2, or 3, characterized in that, The American cockroach defatted liquid is a liquid obtained by defatting American cockroach extract.

5. A method for preparing Kangfuxin Liquid as described in any one of claims 1-4, characterized in that, include: Step 2: Prepare Kangfuxin Liquid directly using American cockroach degreasing solution.

6. The preparation method according to claim 5, characterized in that, Step 2 includes: adding the American cockroach defatting solution, glycerin, licorice extract, vitamin C and sodium benzoate to water, stirring, filtering, filling and sterilizing to obtain the Kangfuxin solution.

7. The preparation method according to claim 5 or 6, characterized in that, The preparation method further includes: Step 1: preparing the American cockroach degreasing solution; The preparation of the American cockroach defatting solution includes: Step 1.1: American cockroaches were impregnated with alcohol to obtain an American cockroach extract; Step 1.2: Filter and concentrate the American cockroach extract to obtain an American cockroach alcohol concentrate; Step 1.3: Add water to the concentrated extract of American cockroaches, let it stand, separate the water layer, filter the water layer, and obtain the defatted American cockroach solution.

8. The preparation method according to claim 7, characterized in that, The alcohol mentioned in step 1.1 is selected from ethanol; preferably, the alcohol is selected from 75%-95% ethanol; The immersion temperature in step 1.1 is 60±4℃; The soaking process described in step 1.1 is at least once; preferably, it is three times. And / or, the soaking time in step 1.1 is 1-30 hours / time; preferably, the soaking time is 12-24 hours / time; more preferably, the soaking time is 12 hours / time.

9. The preparation method according to any one of claims 5-8, characterized in that, The preparation method includes: adding 5.0-12.0% (by weight) of defatted American cockroach solution, 5.0-15.0% (by weight) of glycerin, 0-2.0‰ (by weight) of licorice extract, 0-0.8‰ (by weight) of vitamin C, and 0-5.0‰ (by weight) of sodium benzoate to 72.5-90.0% purified water, stirring, filtering, filling, and sterilizing to obtain the Kangfuxin solution. Optionally, each 1 mL of the Kangfuxin solution contains 0.80-1.20 mg of total amino acids, calculated as alanine.

10. The preparation method according to any one of claims 5-9, characterized in that, The preparation method includes: Step 1.1: American cockroaches were impregnated with alcohol to obtain an American cockroach extract; Step 1.2: Filter and concentrate the American cockroach extract to obtain an American cockroach alcohol concentrate; Step 1.3: Add water to the concentrated extract of American cockroach alcohol, let stand, separate the water layer, filter the water layer, and obtain the defatted American cockroach liquid; Step 2: Calculate by weight percentage and add 5.0-12.0% of American cockroach defatting solution, 5.0-15.0% of glycerin, 0-2.0‰ of licorice extract, 0-0.8‰ of vitamin C, and 0-5.0‰ of sodium benzoate to 72.5-90.0% of purified water. Stir, filter, fill, and sterilize to obtain the Kangfuxin solution. Optionally, each 1 mL of the Kangfuxin solution contains 0.80-1.20 mg of total amino acids, calculated as alanine.

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