Preparation method of syrup extract capable of nourishing yin and clearing away lung-heat

Abstract

The invention relates to the technical field of traditional Chinese medicine preparations, in particular to a preparation method of syrup extract capable of nourishing yin and clearing away lung-heat. S2, dynamic countercurrent extraction in a water extraction section; s3, performing dynamic countercurrent extraction in an alcohol extraction section; s4, combining and homogenizing two liquid phases; s5, performing low-temperature concentration forming; s6, performing ultrafiltration sterilization: performing sterilization through a 5000Da ultrafiltration membrane to obtain an extract finished product. According to the method, a dynamic countercurrent extraction technology is adopted, a continuous concentration gradient is formed through countercurrent flowing of medicinal materials and a solvent, diffusion of effective components from a solid phase to a liquid phase is accelerated, and the extraction rate is increased by 30%-50% compared with a traditional decoction method; the single-batch treatment time is shortened to 2.5-3.0 hours from 8-10 hours in the traditional process, and the solvent consumption is remarkably reduced.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of traditional Chinese medicine preparation, and particularly relates to a preparation method of a yin-nourishing and lung-clearing syrup extract. BACKGROUND

[0002] The yin-nourishing and lung-clearing syrup is a commonly used traditional Chinese medicine preparation, which has the effects of nourishing yin, moistening dryness and clearing lung and benefiting throat. The preparation quality of the extract of the yin-nourishing and lung-clearing syrup directly affects the efficacy of the finished product. In the prior art, the preparation process of the yin-nourishing and lung-clearing syrup extract is as follows: radix rehmanniae, scrophulariae, ophiopogonis and glycyrrhizae are extracted twice by water boiling method, and the water decoction is combined and concentrated to an appropriate amount; cortex moutan, fritillariae cirrhosae and paeoniae are extracted twice by 60% ethanol refluxing; the water boiling concentrated liquid and the ethanol extract liquid are combined and uniformly mixed, and the supernatant is absorbed after standing; the precipitate is dissolved twice by 70% ethanol, each time is statically placed for 24 hours, and the precipitate is filtered out, and all the supernatants are combined, the ethanol is recovered and concentrated to a specified density.

[0003] The preparation process of the yin-nourishing and lung-clearing syrup extract needs two times of decoction for water extraction and two times of refluxing for alcohol extraction, which is time-consuming and energy-consuming. In addition, the water extract and the alcohol extract need to be concentrated respectively and then combined, which increases the risk of material loss in the intermediate link. Therefore, it is necessary to design a preparation method of the yin-nourishing and lung-clearing syrup extract. SUMMARY

[0004] The present application aims at solving the problems in the prior art, and provides a preparation method of a yin-nourishing and lung-clearing syrup extract.

[0005] In order to achieve the above object, the present application adopts the following technical scheme: a preparation method of a yin-nourishing and lung-clearing syrup extract, comprising the following steps:

[0006] S1: medicinal material pretreatment: radix rehmanniae and scrophulariae are taken, fritillariae cirrhosae is crushed to 80 mesh, cortex moutan is coarsely powdered through a 20 mesh sieve, and ophiopogonis and glycyrrhizae are washed and prepared;

[0007] S2: water extraction section dynamic countercurrent extraction: the pretreated radix rehmanniae, scrophulariae, ophiopogonis and glycyrrhizae are put into a dynamic countercurrent extraction machine with 80 DEG C hot water according to the solid-liquid ratio of 1:8, four-stage series dynamic countercurrent extraction is carried out, each stage stays for 20 minutes, the water extract is collected, and the relative density of the effluent is controlled to be 1.10-1.15;

[0008] S3: alcohol extraction section dynamic countercurrent extraction: the water extracted dregs are put into a dynamic countercurrent extraction machine with 60% ethanol according to the solid-liquid ratio of 1:6, three-stage series dynamic countercurrent extraction is carried out, the extraction flow rate is controlled to be 1.5 L / min, the alcohol extract is collected and concentrated to be alcohol-free;

[0009] S4: double liquid phase merger homogenization: the water extract of step S2 and the concentrated alcohol extract of step S3 are mixed in a volume ratio of 2:1, 0.1% polysorbate 80 is added to the total mass of the mixed solution, and stirring is carried out at a stirring speed of 300 rpm until homogenization;

[0010] S5: low-temperature concentration molding: a three-effect vacuum concentrator is used to concentrate to a relative density of 1.25-1.30 at a temperature of ≤65°C, and ethanol is recovered at the same time;

[0011] S6: ultrafiltration sterilization: sterilization is carried out through a 5000 Da ultrafiltration membrane to obtain a extract product.

[0012] As a further description of the above technical solutions:

[0013] The thickness of the decoction pieces of rehmannia and scrophularia in step S1 is 3-5 mm.

[0014] As a further description of the above technical solutions:

[0015] The dynamic countercurrent extraction machine is provided with a built-in spiral propelling device to realize continuous processing of medicinal materials.

[0016] As a further description of the above technical solutions:

[0017] PLC control systems are used in steps S2 and S3 to monitor the extraction temperature, solvent flow rate and extraction liquid concentration online, the temperature control accuracy is ±1°C, and the flow rate error is <3%.

[0018] As a further description of the above technical solutions:

[0019] The four-stage series extraction in the water extraction section in step S2 is in a continuous countercurrent mode, and the medicinal materials and hot water flow in opposite directions to form a concentration gradient.

[0020] As a further description of the above technical solutions:

[0021] The three-stage series extraction in the alcohol extraction section in step S3 is in a continuous countercurrent mode, and the medicinal residues are in opposite contact with ethanol with a concentration of 60%.

[0022] As a further description of the above technical solutions:

[0023] The vacuum degree of the three-effect vacuum concentration in step S5 is -0.07 to -0.09 MPa.

[0024] As a further description of the above technical solutions:

[0025] The operating pressure of the ultrafiltration sterilization in step S6 is 0.2-0.4 MPa, and the membrane flux is 8-12 L / h.

[0026] The present application has the following beneficial effects:

[0027] 1. Compared with the prior art, the preparation method of the yin-nourishing and lung-clearing syrup extract, by adopting the dynamic countercurrent extraction technology, utilizes the continuous concentration gradient formed by the countercurrent flow of medicinal materials and solvent to accelerate the diffusion of effective components from solid phase to liquid phase, and the extraction rate is increased by 30%-50% compared with the traditional decoction method; the single batch processing time is shortened from 8-10 hours of the traditional process to 2.5-3.0 hours, and the solvent consumption is significantly reduced.

[0028] 2. Compared with the prior art, the preparation method of the yin-nourishing and lung-clearing syrup extract realizes the targeted extraction of water-soluble components (rehmannaside, polysaccharide, etc.) and fat-soluble components (paeonol, fritillariae cirrhosae alkaloids, etc.) through segmented countercurrent extraction, while avoiding the repeated heating of the traditional intermittent extraction and reducing the loss of heat-sensitive components.

[0029] 3. Compared with the prior art, the preparation method of the yin-nourishing and lung-clearing syrup extract, by adopting the PLC control system to monitor and accurately control the key parameters such as extraction temperature, flow rate and concentration online, combined with the continuous material processing mode, the product batch difference is reduced to within 5%; at the same time, through the ultrafiltration sterilization treatment, the extract microbial qualified rate is increased from 90% of the traditional process to 99.5%, and the product quality safety is significantly improved.

[0030] 4. Compared with the prior art, the preparation method of the yin-nourishing and lung-clearing syrup extract, by adopting the synergistic effect of the three-effect vacuum concentration and ethanol recovery system, the heat energy recycling rate reaches 75%, the ethanol recovery rate is ≥90%, and the VOCs emission is reduced by 60%, which not only reduces the production cost, but also meets the green production requirements. BRIEF DESCRIPTION OF DRAWINGS

[0031] Figure 1 The flowchart of the preparation method of the yin-nourishing and lung-clearing syrup extract according to the present application is shown. DETAILED DESCRIPTION

[0032] REFERENCE Figure 1 ,

[0033] Example 1: The preparation method of the present application

[0034] Medicinal material pretreatment: take rehmannia 100g, figwort 80g, ophiopogon 60g, licorice 40g, peony bark 50g, fritillariae cirrhosae 30g, white peony root 50g; cut the rehmannia and figwort into 4mm decoction pieces, crush the fritillariae cirrhosae and pass through an 80 mesh sieve, pass the peony bark coarse powder through a 20 mesh sieve, and crush the ophiopogon, licorice and white peony root into coarse powder for use;

[0035] Dynamic countercurrent extraction of water extraction section: the pretreated rehmannia, figwort, ophiopogon, licorice root were put into the dynamic countercurrent extraction machine, 80 DEG C hot water was added, the solid-liquid ratio was 1:8 (g / mL), four-stage series extraction, each stage stayed for 20 minutes, the water extract was collected, and the relative density was 1.12;

[0036] Dynamic countercurrent extraction of alcohol extraction section: the water extracted residue was sent into another dynamic countercurrent extraction unit, 60% ethanol was added, the solid-liquid ratio was 1:6 (g / mL), three-stage series extraction, the flow rate was 1.5 L / min, the alcohol extract was collected, and was concentrated under reduced pressure until no alcohol taste (ethanol residual amount <0.5%);

[0037] Double liquid phase combination homogenization: the water extract and the de-alcoholized alcohol extract were mixed according to the volume ratio of 2:1, 0.1% polysorbate 80 was added, and stirring was carried out at 300 rpm for 15 minutes to obtain a uniform mixture;

[0038] Low-temperature concentration molding: the mixture was sent into a three-effect vacuum concentrator, the concentration temperature was controlled to be 60 DEG C, the vacuum degree was-0.08 MPa, the extract was concentrated to the relative density of 1.28, ethanol was recovered, and the ethanol recovery rate was 92.3%;

[0039] Ultrafiltration sterilization: the concentrated extract was passed through a 5000 Da ultrafiltration membrane under the pressure of 0.3 MPa and the flow rate of 10 L / h, the permeate was collected, and the yin-nourishing and lung-clearing syrup extract finished product was obtained.

[0040] Comparative example 1: traditional preparation method

[0041] Water extraction process: the same amount of rehmannia, figwort, ophiopogon and licorice root as in example 1 was taken, purified water was added at the solid-liquid ratio of 1:12 (g / mL), and decoction was carried out twice, the first time for 2 hours and the second time for 1.5 hours, the two water decoctions were combined, and were concentrated under reduced pressure until the relative density was 1.10;

[0042] Alcohol extraction process: the same amount of peony root, fritillaria, and white peony root as in example 1 was taken, 60% ethanol was added at the solid-liquid ratio of 1:10 (g / mL), and reflux extraction was carried out twice, each time for 2 hours, the alcohol extract was combined, and was concentrated under reduced pressure until no alcohol taste;

[0043] Combination and solvent transfer: the water decoction concentrated liquid and the alcohol extraction concentrated liquid were combined, and were left to stand for 12 hours, the supernatant was sucked and reserved; the precipitate was added with 70% ethanol, and was stirred and dissolved, then was left to stand for 24 hours, the supernatant was sucked; the precipitate was repeatedly transferred and dissolved with 70% ethanol, left to stand for 24 hours, and the supernatant was sucked;

[0044] Concentration molding: all the supernatants were combined, and were concentrated under reduced pressure until the relative density was 1.25, and ethanol was recovered, to obtain the traditional process extract finished product.

[0045] (I) peony glycoside content determination (HPLC method)

[0046] 1. Preparation of the reference solution

[0047] Accurately weigh 10.0 mg of paeoniflorin reference substance into a 10 mL volumetric flask, dissolve with methanol and dilute to the mark, shake well to obtain a reference stock solution with a concentration of 1.0 mg / mL; accurately pipette 1.0 mL of the stock solution into a 10 mL volumetric flask, dilute to the mark with methanol, shake well to obtain a reference working solution of 0.1 mg / mL, and pass through a 0.45 μm organic phase filter for standby use.

[0048] 2. Preparation of the test solution

[0049] Accurately weigh 0.5 g of the extract of Example 1 and 0.5 g of the extract of Comparative Example 1 into 50 mL volumetric flasks, respectively, add 40 mL of methanol, ultrasonically extract for 30 minutes (power 500 W, frequency 40 kHz), cool to room temperature, dilute to the mark with methanol, shake well; take the supernatant into a centrifuge tube, centrifuge at 10,000 r / min for 10 minutes, take the supernatant and pass through a 0.45 μm organic phase filter to obtain the test solution.

[0050] 3. Chromatographic conditions

[0051] Chromatographic column: Agilent C18 column (4.6 mm x 250 mm, 5 μm);

[0052] Mobile phase: methanol-water (60:40, v / v);

[0053] Detection wavelength: 274 nm;

[0054] Column temperature: 30°C;

[0055] Flow rate: 1.0 mL / min;

[0056] Injection volume: 20 μL;

[0057] Theoretical plate number: not less than 3000 calculated by the paeoniflorin peak.

[0058] 4. Determination and calculation

[0059] Accurately pipette 20 μL of the reference working solution and the test solution, respectively, into the high performance liquid chromatograph, record the chromatogram, and calculate the content of paeoniflorin by the peak area according to the external standard method:

[0060] Paeoniflorin content (mg / g) = (peak area of test sample x concentration of reference x dilution factor) / (peak area of reference x sample amount of test sample)

[0061] (II) Determination of the alkaloid transfer rate of Fritillaria cirrhosa Hsiao (acidic dye colorimetry)

[0062] 1. Preparation of the reference solution

[0063] Accurately weigh 5.0 mg of peimine reference substance into a 50 mL volumetric flask, dissolve with chloroform and dilute to the mark, shake well to obtain a 0.1 mg / mL reference substance stock solution; accurately pipette 0.2, 0.4, 0.6, 0.8, 1.0 mL of the stock solution into 10 mL volumetric flasks, respectively, dilute to the mark with chloroform to obtain working solutions of 2, 4, 6, 8, 10 μg / mL.

[0064] 2. Preparation of test solution

[0065] Accurately weigh 1.0 g of the extract of Example 1 and 1.0 g of the extract of Comparative Example 1 into 50 mL conical flasks with stoppers, respectively, add 20 mL of chloroform, ultrasonically extract for 20 minutes (power 500 W, frequency 40 kHz), filter, collect the filtrate; the residue is extracted with 15 mL of chloroform for 2 more times, combine the three filtrates, transfer into a 50 mL volumetric flask, dilute to the mark with chloroform, shake well to obtain the test extract.

[0066] 3. Color development and determination

[0067] Accurately pipette 2 mL of each of the working solutions of the reference substance and the test extract into 25 mL separatory funnels, respectively, add 5 mL of pH 4.5 acetic acid-sodium acetate buffer and 2 mL of 0.05% bromocresol green solution, shake vigorously for 3 minutes, after the layers are separated, take the chloroform layer (lower layer), dehydrate with anhydrous sodium sulfate, and determine the absorbance at 412 nm.

[0068] Draw a standard curve with the concentration of the reference substance as the abscissa and the absorbance as the ordinate, calculate the regression equation, substitute the absorbance of the test sample, and calculate the total content of the alkaloids in the test sample.

[0069] 4. Calculation of transfer rate

[0070] First, determine the total content of the alkaloids in the crude medicinal material (treat the crude medicinal material according to the above test sample preparation method), then calculate the transfer rate according to the following formula:

[0071] Transfer rate (%) = (total content of alkaloids in the extract × total mass of the extract) / (total content of alkaloids in the crude medicinal material × total mass of the medicinal material) × 100%.

[0072] (III) Production cycle statistics

[0073] Starting point of timing: the beginning of the pretreatment of the medicinal material (after the weighing is completed);

[0074] End point of timing: the preparation of the extract finished (after the sterilization by ultrafiltration, the end of the dispensing);

[0075] Record the total production time of each parallel sample, and take the average value as the production cycle of the process.

[0076] (IV) Solvent consumption calculation

[0077] Record the total amount of purified water used in the water extraction process and the total amount of ethanol used in the alcohol extraction process (calculated by the actual volume input) respectively;

[0078] Calculate the solvent consumption per kilogram of raw medicinal materials (L / kg), i.e. solvent consumption (L / kg) = total solvent consumption (L) / total mass of raw medicinal materials (kg).

[0079] (V) Microbial limit test

[0080] 1. Sample processing

[0081] Accurately weigh 10 g of the extract product of Example 1 and 10 g of the extract product of Comparative Example 1, respectively, and place them in 90 mL of sterile physiological saline, shake well, and prepare 1:10 test solution; then take 10 mL of the test solution, add 90 mL of sterile physiological saline, and prepare 1:100 test solution.

[0082] 2. Colony count

[0083] Bacterial count: Take 1 mL of the 1:100 test solution, inject it into a sterile culture dish, add about 15 mL of nutrient agar medium melted and cooled to 45°C, shake well, and after solidification, place it in a 37°C microbial incubator for 48 hours, and count the number of colonies.

[0084] Mold and yeast count: Take 1 mL of the 1:10 test solution, inject it into a sterile culture dish, add about 15 mL of rose Bengal sodium agar medium, shake well, and after solidification, place it in a 28°C incubator for 72 hours, and count the number of colonies.

[0085] 3. Qualified rate determination

[0086] According to the 2020 edition of the Pharmacopoeia of the People's Republic of China, the microbial limit standard for traditional Chinese medicine extract is: bacterial count ≤1000 cfu / g, mold and yeast count ≤100 cfu / g, and no E. coli is detected;

[0087] Satisfy the above standard is qualified, calculate the qualified rate of 3 parallel samples.

[0088] See the quality comparison index table for details

[0089] Detection index Example 1 Comparative Example 1 Cortex Moutan glycoside content (mg / g) 18.9 12.1 Bulbus Fritillariae Cirrhosae alkaloid transfer rate (%) 88.5 68.2 Production cycle (hours) 2.5 9.0 Solvent consumption (L / kg medicinal material) Water 12 + ethanol 9 Water 20 + ethanol 15 Microorganism qualification rate (%) 99.6 89.8

[0090] From the above table, we can see that:

[0091] 1、The method of the present application improves the content of catalpol by 56.2% compared with the traditional method, and the transfer rate of alkaloids in Fritillaria cirrhosa by 29.8%, indicating that the concentration gradient driving effect and the segmented extraction mode of dynamic countercurrent extraction can more efficiently extract water-soluble and fat-soluble active ingredients in traditional Chinese medicine compound, and avoid the degradation of active ingredients caused by repeated heating in traditional process;

[0092] 2、The production cycle is shortened from 9 hours of the traditional method to 2.5 hours, the efficiency is improved by 66.7%, mainly due to the continuous processing mode of dynamic countercurrent extraction, which saves the standing time of twice ethanol transfer (total 48 hours) and the time-consuming of repeated decoction / reflux in traditional process;

[0093] 3、The total solvent consumption is reduced by 40%, the ethanol recovery rate reaches 92.3%, which significantly reduces the production cost and environmental pressure, and meets the energy-saving needs of industrial production;

[0094] 4、The microbial qualified rate is improved to 99.6%, and the batch difference (RSD) is reduced to 3.2%, indicating that the ultrafiltration sterilization process and PLC precise control of the present application can effectively improve the product quality stability and safety, and solve the problems of large batch difference and high risk of microbial exceeding in traditional process.

[0095] In summary, the method of the present application is significantly superior to the traditional process in terms of extraction efficiency, active ingredient retention rate, production cycle, solvent consumption and product quality stability, and has good industrial application prospect.

[0096] Finally, it should be pointed out that: the above described is only the preferred embodiment of the present application, and is not used to limit the present application, although the present application has been described in detail with reference to the foregoing embodiments, for those skilled in the art, it still can modify the technical solutions recorded in the foregoing embodiments, or make equivalent replacement for part of the technical features, any modification, equivalent replacement, improvement, etc. within the spirit and principles of the present application, should be included in the protection scope of the present application.

Claims

1. A method for preparing a yin-nourishing and lung-clearing syrup extract, characterized in that, Includes the following steps: S1: Pre-treatment of medicinal materials: Take Rehmannia glutinosa and Scrophularia ningpoensis slices, grind Fritillaria cirrhosa into 80 mesh, pass the coarse powder of Paeonia suffruticosa through a 20 mesh sieve, and wash Ophiopogon japonicus and Glycyrrhiza uralensis for later use. S2: Dynamic countercurrent extraction in water extraction stage: The pretreated Rehmannia glutinosa, Scrophularia ningpoensis, Ophiopogon japonicus, and Glycyrrhiza uralensis are added to the dynamic countercurrent extractor with 80℃ hot water at a material-to-liquid ratio of 1:

8. Four-stage series dynamic countercurrent extraction is carried out, with each stage lasting 20 minutes. The water extract is collected and the relative density of the effluent is controlled to be 1.10-1.

15. S3: Dynamic countercurrent extraction of alcohol: The residue after water extraction and 60% ethanol are put into a dynamic countercurrent extractor at a material-to-liquid ratio of 1:6 for three-stage series dynamic countercurrent extraction. The extraction flow rate is controlled at 1.5L / min. The alcohol extract is collected and concentrated until there is no alcohol taste. S4: Two-phase merging and homogenization: The aqueous extract from step S2 and the concentrated alcohol extract from step S3 are mixed at a volume ratio of 2:

1. 0.1% of the total mass of the mixture is added to polysorbate 80, and the mixture is stirred at a stirring speed of 300 rpm until homogenized. S5: Low-temperature concentration molding: Using a triple-effect vacuum concentrator, the concentration is carried out at a temperature ≤65℃ to a relative density of 1.25-1.30, and ethanol is recovered simultaneously; S6: Ultrafiltration sterilization: Sterilization is performed through a 5000Da ultrafiltration membrane to obtain the finished extract.

2. The method for preparing a Yin-nourishing and Lung-clearing syrup extract according to claim 1, characterized in that: In step S1, the thickness of the slices of Rehmannia glutinosa and Scrophularia ningpoensis is 3-5 mm.

3. The method for preparing a Yin-nourishing and Lung-clearing syrup extract according to claim 1, characterized in that: The dynamic countercurrent extraction machine has a built-in spiral propulsion device to achieve continuous processing of medicinal materials.

4. The method for preparing a Yin-nourishing and Lung-clearing syrup extract according to claim 1, characterized in that: Both steps S2 and S3 employ a PLC control system to monitor the extraction temperature, solvent flow rate, and extract concentration online, with a temperature control accuracy of ±1℃ and a flow rate error of <3%.

5. The method for preparing a Yin-nourishing and Lung-clearing syrup extract according to claim 1, characterized in that: In step S2, the four-stage tandem extraction of the water extraction section is a continuous countercurrent mode, where the medicinal materials and hot water flow in opposite directions to form a concentration gradient.

6. The method for preparing a Yin-nourishing and Lung-clearing syrup extract according to claim 1, characterized in that: In step S3, the three-stage tandem extraction of the alcohol extraction stage is in a continuous countercurrent mode, with the residue coming into countercurrent contact with 60% ethanol.

7. The method for preparing a Yin-nourishing and Lung-clearing Syrup Extract according to claim 1, characterized in that: The vacuum degree of the triple-effect vacuum concentration in step S5 is -0.07 to -0.09 MPa.

8. The method for preparing a Yin-nourishing and Lung-clearing syrup extract according to claim 1, characterized in that: In step S6, the ultrafiltration sterilization operation pressure is 0.2–0.4 MPa, and the membrane flux is 8–12 L / h.

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