CN116850231A - A method of preparing raw rehmannia glutinosa using lily - Google Patents

Abstract

The present invention relates to a method of using lily to prepare raw rehmannia glutinosa, which can effectively solve the problem of enhancing the yin-nourishing, heat-clearing and hemostatic effects of raw rehmannia glutinosa and increasing the soothing, cough-relieving and expectorant effects of raw rehmannia glutinosa. The method is to add lily slices to boil 3 times in water. , concentrate to a lily decoction with a mass concentration of 0.50~0.83g/mL, mix the raw rehmannia glutinosa and lily in a mass fraction ratio of 100:15~25, and moisten it until the medicinal juice is gone. The raw rehmannia glutinosa slices are processed and dried to obtain the finished product; for the first time, the present invention selects lily, a traditional Chinese medicine with the same origin as medicine and food, to process raw rehmannia glutinosa. It aims to innovatively develop "Xianzizwei" slices made from lily to make raw rehmannia glutinosa. The processed product can enhance the raw rehmannia glutinosa. It has the functions of nourishing yin, clearing away heat and stopping bleeding, and adds the soothing, cough-relieving and expectorant effects of raw rehmannia glutinosa, enhancing the efficacy of raw rehmannia glutinosa and the pertinence of medication, expanding the selection range and indication range of raw rehmannia glutinosa decoction pieces, and improving the clinical application of raw rehmannia glutinosa. accuracy and efficiency.

Description

A method for preparing raw Rehmannia root by using lily

Technical Field

The invention relates to the technical field of Chinese medicinal material processing, in particular to a method for preparing raw rehmannia root by using lily as an auxiliary material.

Background Art

Rehmannia glutinosa is the fresh or dried root of Rehmannia glutinosa Libosch., a plant of the Scrophulariaceae family. It is mainly produced in Jiaozuo, Henan Province. It is one of the four famous Huai medicines and has extremely high medicinal value. In particular, raw Rehmannia glutinosa, which has the effects of clearing heat and cooling blood, nourishing yin and promoting fluid production, is favored. Many classic prescriptions of the medical saint Zhang Zhongjing (such as Baihe Dihuang Decoction, Zhigancao Decoction, Fangji Dihuang Decoction, etc.) contain raw Rehmannia glutinosa and it is often used as an important drug or even the main drug in the prescription. Judging from the current 2020 edition of the "Chinese Pharmacopoeia", national and provincial processing specifications, and textbooks such as Chinese medicine and Chinese medicine processing, new pieces of raw Rehmannia glutinosa with special processing are extremely scarce. Although cooked rehmannia root was first recorded in "Compendium of Materia Medica" by Chen Cangqi in the Tang Dynasty, firstly, the medicinal properties, effects and main indications of cooked rehmannia root have undergone fundamental changes. The effects of raw rehmannia root are mainly sweet and cold, nourishing yin and clearing away heat, while the effects of cooked rehmannia root have changed to mainly sweet and warm, nourishing blood and benefiting essence. Cooked rehmannia root and raw rehmannia root are already two completely different Chinese medicines; secondly, even with cooked rehmannia root, the processed products of raw rehmannia root are still too limited, and the legal processed products are very scarce. Raw rehmannia root is an important drug component in many prescriptions (such as the aforementioned Baihe Dihuang Decoction and other famous prescriptions of Zhang Zhongjing). The clinical demand for raw rehmannia root is huge, and the diseases are often complex and changeable. In clinical practice, patients with different conditions, different syndromes and different symptoms have different targeted needs for raw rehmannia root. However, due to the lack of processed products, raw rehmannia root cannot provide more processed products suitable for different targeted medication needs of patients with different conditions and syndromes. This seriously limits the selectivity, accuracy and efficiency of raw rehmannia root in clinical application, and restricts the industrial application process of raw rehmannia root. Processing is one of the characteristics of Chinese medicine. Through methods such as "mutual assistance" to enhance efficacy and "mutual fear" to reduce toxicity, it can enhance efficacy and reduce toxicity of Chinese medicine. Lily is the dried fleshy scales of Lilium lancifolium Thunb., Lilium brownii F.E.Brownvar.viridulum Baker or Lilium pumilum DC. of the Liliaceae family. It is a Chinese medicine with both medicinal and edible properties. It has the effects of nourishing yin and moistening the lungs, clearing the heart and calming the mind. It is mainly used to treat yin deficiency and dry cough, cough with blood due to fatigue, restlessness and palpitations, insomnia and dreaminess, and mental confusion. Lily and Rehmannia glutinosa are a typical pair of drugs that are mutually beneficial and effective, which can be seen from the famous prescription of Lily and Rehmannia Decoction by Zhang Zhongjing. This laid a theoretical foundation for the "mutual assistance" of lily and Rehmannia glutinosa. However, to date, there has been no report on the "mutual assistance" of lily to process Rehmannia glutinosa at home and abroad.

Summary of the invention

In view of the above situation, in order to overcome the defects of the prior art, the purpose of the present invention is to provide a method for preparing raw Rehmannia root with lily, which can effectively solve the problem of enhancing the nourishing yin, clearing heat and hemostatic effects of raw Rehmannia root, and increasing the calming, relieving cough and expectorant effects of raw Rehmannia root.

The technical solution provided by the present invention is a method for preparing raw Rehmannia root by using lily, comprising the following steps:

a. Take raw Rehmannia root slices, remove impurities and set aside;

b. Take lily slices, add 12 times the weight volume of lily slices of water each time and boil for 3 times, boil for 30 minutes each time, combine the filtrate, and concentrate to a lily water decoction with a mass concentration of 0.50-0.83 g/mL; the weight volume refers to the solid in g and the liquid in mL;

c. Take the Radix Rehmanniae slices in step a, add the decoction of lily in step b, the mass fraction ratio of Radix Rehmanniae to lily is 100:15-25, mix well, and stew until the medicine is soaked and the juice is exhausted (about 5h);

d. Processing and drying the raw Rehmannia root slices soaked in step c to obtain a finished product;

The processing and drying is stir-frying and drying at a low heat, or freeze-drying at -80°C.

The present invention selects lily, a Chinese medicinal material which is both a food and a medicine, to prepare raw rehmannia root for the first time, and aims to innovatively develop the "mutually-assisted" decoction pieces of raw rehmannia root prepared with lily. The prepared processed product can enhance the yin-nourishing, heat-clearing and hemostatic effects of raw rehmannia root, and also increases the tranquilizing, cough-relieving and expectorant effects of raw rehmannia root, thereby enhancing the efficacy of raw rehmannia root and the targeted use of the drug, expanding the selection range and indication range of raw rehmannia root decoction pieces to meet the needs of patients with different conditions and syndromes for targeted use of different drugs, improving the accuracy and efficiency of raw rehmannia root in clinical application, expanding the medicinal value and commercial value of raw rehmannia root, and having significant economic and social benefits.

DETAILED DESCRIPTION

The specific implementation methods and specific situations of the present invention are described in detail below in conjunction with embodiments.

The present invention can be specifically implemented by the following examples.

Example 1

The present invention discloses a method for preparing raw rehmannia root by using lily, comprising the following steps:

a. Take raw Rehmannia root slices, remove impurities and set aside;

b. Weigh 15.0 g of lily slices, add 12 times the weight volume of lily slices of water each time, decoct 3 times, each time for 30 minutes, combine the decoctions, and concentrate to a concentration of 0.50 g/mL lily water decoction (30 mL);

c. Take 100 g of the raw Rehmannia slices after removing impurities in step a, add the lily decoction in step b, mix well, and simmer for about 5 hours until the medicine is fully absorbed;

d. Place the soaked Rehmannia root slices in step c in a wok and stir-fry over low heat (4 min, 100° C.) until the cross section of the Rehmannia root is brownish yellow to black or jet black with slight burnt spots, then place in a 60° C. oven to dry, take out, and let cool.

Example 2

The present invention discloses a method for preparing raw rehmannia root by using lily, comprising the following steps:

a. Take raw Rehmannia root slices, remove impurities and set aside;

b. Weigh 20.0 g of lily slices, add 12 times the weight volume of the lily slices of water each time, decoct 3 times, each time for 30 minutes, combine the decoctions, and concentrate to a lily water decoction with a concentration of about 0.67 g/mL;

c. Take 100 g of the raw Rehmannia slices after removing impurities in step a, add the lily decoction in step b, mix well, and simmer for about 5 hours until the medicine is fully absorbed;

d. Place the soaked Radix Rehmanniae slices in step c in a wok, fry over low heat until the cross section of the Radix Rehmanniae is brownish yellow to black or jet black with slight burnt spots, dry in a 60°C oven, take out, and let cool.

Example 3

The present invention discloses a method for preparing raw rehmannia root by using lily, comprising the following steps:

a. Take raw Rehmannia root slices, remove impurities and set aside;

b. Weigh 25.0 g of lily slices, add 12 times the weight volume of the lily slices of water each time, decoct 3 times, each time for 30 minutes, combine the decoctions, and concentrate to a lily water decoction with a concentration of about 0.83 g/mL;

c. Take 100 g of the raw Rehmannia slices after removing impurities in step a, add the lily decoction in step b, mix well, and simmer for about 5 hours until the medicine is fully absorbed;

d. Place the soaked Radix Rehmanniae slices in step c in a wok, fry over low heat until the cross section of the Radix Rehmanniae is brownish yellow to black or jet black with slight burnt spots, dry in a 60°C oven, take out, and let cool.

Example 4

The present invention discloses a method for preparing raw rehmannia root by using lily, comprising the following steps:

a. Take raw Rehmannia root slices, remove impurities and set aside;

b. Weigh 15.0 g of lily slices, add 12 times the weight volume of lily slices of water each time, decoct 3 times, each time for 30 minutes, combine the decoctions, and concentrate to a lily water decoction (30 mL) with a concentration of about 0.50 g/mL;

c. Take 100 g of the Radix Rehmanniae slices after removing impurities in step a, add the lily water decoction in step b, mix well, and simmer for about 5 hours until the medicine is fully absorbed;

d. Place the soaked Rehmannia root slices in step c in a freeze dryer and freeze-dry at -80°C, then take them out to obtain the product.

Example 5

a. Take raw Rehmannia root slices, remove impurities and set aside;

b. Weigh 20.0 g of lily slices, add 12 times the weight volume of the lily slices of water each time, decoct 3 times, each time for 30 minutes, combine the decoctions, and concentrate to a lily water decoction with a concentration of about 0.67 g/mL;

c. Take 100 g of the Radix Rehmanniae slices after removing impurities in step a, add the lily water decoction in step b, mix well, and simmer for about 5 hours until the medicine is fully absorbed;

d. Place the soaked Rehmannia root slices in step c in a freeze dryer and freeze-dry at -80°C, then take them out to obtain the product.

Example 6

The present invention discloses a method for preparing raw rehmannia root by using lily, comprising the following steps:

a. Take raw Rehmannia root slices, remove impurities and set aside;

b. Weigh 25.0 g of lily slices, add 12 times the weight volume of the lily slices of water each time, decoct 3 times, each time for 30 minutes, combine the decoctions, and concentrate to a lily water decoction with a concentration of about 0.83 g/mL;

c. Take 100 g of the raw Rehmannia slices after removing impurities in step a, add the lily decoction in step b, mix well, and simmer for about 5 hours until the medicine is fully absorbed;

d. Place the soaked Rehmannia root slices in step c in a freeze dryer and freeze-dry at -80°C, then take them out to obtain the product.

The present invention selects lily, a Chinese medicinal herb with both medicinal and edible properties, to prepare raw rehmannia for the first time. The prepared product enhances the yin-nourishing, heat-clearing and hemostatic effects of raw rehmannia, and increases the tranquilizing, cough-relieving and expectorant effects of raw rehmannia, enhances the efficacy of raw rehmannia and the pertinence of medication, changes the active ingredients of raw rehmannia slices, expands the selection range and indication range of raw rehmannia slices, so as to meet the needs of different medication pertinence of patients with different conditions and syndromes, and obtains good beneficial technical effects through experiments. The relevant experimental data are as follows:

1. Experimental Materials

1.1 Experimental animals

ICR mice, male, SPF grade, weight 16-18 g, provided by Jinan Pengyue Experimental Animal Breeding Co., Ltd. [SCXK(Lu)20220006].

1.2 Experimental drugs

According to Examples 1-6 of the present invention, the raw Rehmannia root slices prepared from lily are obtained.

2. Observation or measurement experiment on the nourishing yin, clearing heat, calming the nerves, relieving cough, removing phlegm and stopping bleeding effects of raw Rehmannia root slices prepared with lily.

2.1 Animal grouping and drug administration

After 90 male ICR mice were adaptively fed for one week, they were randomly divided into 9 groups according to body weight, namely, ① blank control group, ② raw rehmannia group, ③ lily decoction group, ④ embodiment 1, ⑤ embodiment 2, ⑥ embodiment 3, ⑦ embodiment 4, ⑧ embodiment 5, and ⑨ embodiment 6, with 10 mice in each group. Raw rehmannia, lily decoction and raw rehmannia slices prepared with different lilies were dissolved in 0.5% sodium carboxymethyl cellulose solution and all were intragastrically administered at a dose of 2g/kg. The blank control group was given an equal volume of 0.5% sodium carboxymethyl cellulose solution, administered once/d, and intragastrically for 14 days. One hour after the last administration, the effects of raw rehmannia characteristic processed products on nourishing yin and clearing heat, calming the nerves, relieving cough, removing phlegm and stopping bleeding were observed or measured.

2.2 Detection of related indicators of nourishing yin and clearing heat

2.2.1 Water intake

The water intake of mice was tested on the 14th day of administration. Each mouse was placed in a cage with a water bottle in each cage, and the water intake of the mice for 12 hours was tested and calculated.

2.2.2 Rectal temperature

Rectal temperature was tested on the 14th day of administration. When measuring, the mouse was first grasped to expose the anus, and the mercury tip of the thermometer was dipped in a small amount of lubricant and gently and slowly inserted into the anus of the mouse to a depth of about 1 cm. After hearing the "beep" prompt, the temperature reading was read and recorded.

2.2.3 Hot and cold plate experiment

The laboratory temperature was maintained at (23±2)℃, and the bottom plate temperature of the automatic temperature control system was set to: 20℃ for the low temperature plate and 39℃ for the high temperature plate. On the 14th day of administration, the mice in each group were placed in each channel on the hot and cold plates in a quiet environment, and their hot and cold temperature zone tendency activities of the hot and cold plates were remotely monitored by cameras, and the proportion of mice staying in the hot zone was finally obtained.

2.2.4 Experimental determination of serum cAMP, cGMP, T3, T4, Na ⁺ -ATP, and Ca ²⁺ -ATPase levels

The eyeballs of all mice were removed and blood was collected. After standing at room temperature for 2 hours, the blood was centrifuged at 4°C and 3500 r/min for 10 minutes to separate the upper serum. The levels of cAMP, cGMP, T3, and T4 in mouse serum were measured according to the instructions of the kit.

2.3 Determination of tranquilizing index (sleep experiment)

Sodium pentobarbital (55 mg kg ^-1 ) was injected intraperitoneally into mice to induce sleep. The sleep latency of mice was observed (the time from the injection of sodium pentobarbital to the appearance of righting reflex of mice was greater than 1 min). The righting reflex time of mice was greater than 1 min as the standard of falling asleep. The total sleep time of mice was also recorded (the total time from the mice falling asleep to regaining consciousness).

2.4 Determination of cough-relative efficacy indicators (ammonia-induced cough test)

Place a 500mL beaker upside down on a table with a 10cm×10cm qualitative filter paper, use a 1mL syringe to draw 0.1mL of concentrated ammonia water and inject it on the filter paper. After evaporation for 30 seconds, quickly remove the filter paper and put the mouse to be tested in. Observe the abdominal muscle contraction, chest shrinkage, open mouth and coughing of the mouse as the basis for judging cough, and record the number of coughs of the mouse within 2 minutes.

2.5 Determination of relevant indicators of expectorant efficacy (tracheal phenol red secretion test)

Inject 0.01 mg·g ^-1 of 5% phenol red saline solution into the mouse intraperitoneally. Take the same length of tracheal tissue from the mouse thyroid cartilage to the tracheal branch, place it in a centrifuge tube containing 2 mL of saline, add 0.1 mL of 0.5% sodium bicarbonate solution, ultrasonicate for 15 minutes and soak for 24 hours, centrifuge at 3000 r/min for 15 minutes, and take the supernatant. Measure the absorbance value at 546 nm, substitute it into the phenol red standard curve Y=0.1018X+0.0455, R ² =0.9992, and calculate the mouse tracheal phenol red secretion (the final result is expressed as concentration).

2.6 Determination of hemostasis index (bleeding time test by tail-cutting method)

Place the mouse in a fixator and use surgical scissors to cut the tail 0.5 cm away from the tip of the tail. Start timing when the blood overflows by itself. Use filter paper to absorb the blood drops every 30 seconds until the bleeding stops naturally (no blood is found when the filter paper is absorbed). This is the bleeding time.

3. Results

3.1 Experiment on enhancing the Yin-nourishing and heat-clearing efficacy of raw Rehmannia root slices prepared with lily bulbs

The abnormal increase of mouse water intake, serum cAMP, cAMP/cGMP, T3, T4 enzyme levels and the decrease of cGMP level are generally indicators of mouse yin deficiency syndrome, while the decrease of water intake, serum cAMP, cAMP/cGMP, T3, T4 levels and the increase of cGMP level characterize the yin nourishing effect of the drug. The present invention confirms that the raw Rehmannia slices processed by lily can reduce the levels of mouse serum cAMP, cAMP/cGMP, T3, T4 and increase the cGMP level compared with raw Rehmannia, indicating that the yin nourishing effect of the raw Rehmannia slices processed by lily is enhanced. The results are shown in Tables 1 and 2.

Table 1 Yin deficiency indicators (water intake, T3, T4 levels) of mice before and after intervention with raw Rehmannia slices prepared with lily bulbs

Note: Compared with the blank control group, ^* P<0.05, ^** P<0.01; compared with the raw Rehmannia root slices prepared with lily bulbs and the raw Rehmannia root group, ^# P<0.05, ^## P<0.01.

Table 2 Yin deficiency indexes (serum cAMP, cGMP, cAMP/cGMP) of mice before and after intervention with raw Rehmannia slices prepared with lily bulbs

Note: Compared with the blank control group, ^* P<0.05, ^** P<0.01; compared with the raw Rehmannia root slices prepared with lily bulbs and the raw Rehmannia root group, ^# P<0.05, ^## P<0.01.

Abnormal increase in mouse rectal temperature and abnormal decrease in the proportion of hot zones in the hot and cold plate test generally characterize internal heat syndrome in mice, and the reduction of the rectal temperature level and the increase of the proportion of hot zones by the drug can reflect the heat-clearing effect of the drug. The present invention confirms that the raw Rehmannia slices prepared by lily can reduce the rectal temperature of mice and increase the proportion of hot zones more than raw Rehmannia, indicating that the heat-clearing effect of the raw Rehmannia slices prepared by lily is enhanced. The results are shown in Table 3.

Table 3 Changes in internal heat indexes (rectal temperature, heat zone ratio) of mice before and after intervention with raw Rehmannia slices prepared with lily bulbs

Note: Compared with the blank control group, ^* P<0.05, ^** P<0.01; compared with the raw Rehmannia root slices prepared with lily bulbs and the raw Rehmannia root group, ^# P<0.05, ^## P<0.01.

3.2 Experiment on increasing the tranquilizing effect of raw Rehmannia root slices prepared with lily bulbs

The increase in the latency of sleep time and the decrease in sleep time of mice can reflect the state of insomnia. If the sleep latency of mice can be reduced or the sleep time of mice can be increased after administration, it can reflect the tranquilizing effect of the drug. The present invention confirms that the raw Rehmannia slices prepared by lily can reduce the sleep latency of mice or increase the sleep time of mice compared with raw Rehmannia. And the increase rate of the efficacy of the raw Rehmannia slices prepared by lily (Examples 1 to 6) compared with raw Rehmannia is 34.0%, 46.5%, 20.6%, 13.5%, 26.8%, and 21.1% respectively in sleep latency; the increase rate of the efficacy of the raw Rehmannia slices prepared by lily (Examples 1 to 6) compared with raw Rehmannia is 34.3%, 36.0%, 28.7%, 34.5%, 32.0%, and 30.8% respectively in sleep time. The above shows that the raw Rehmannia slices prepared by lily have increased the tranquilizing effect. The results are shown in Table 4.

Table 4 Sleep latency and sleep duration of mice before and after intervention with raw Rehmannia slices prepared with lily

Note: Compared with the raw Rehmannia root group, the raw Rehmannia root slices prepared with lily bulbs showed ^# P<0.05, ^## P<0.01.

3.3 Experiment on increasing the cough-relieving efficacy of raw Rehmannia root slices prepared with lily bulbs

The ammonia-induced cough experiment is a common experiment for testing the cough-relieving effect of drugs. If the number of ammonia-induced coughs in mice can be reduced after administration, the cough-relieving effect of the drug is reflected. The present invention confirms that the raw Rehmannia slices prepared by lily can reduce the number of coughs in mice more than raw Rehmannia. The efficacy increase rates of the raw Rehmannia slices prepared by lily (Examples 1 to 6) compared with raw Rehmannia are 42.3%, 52.9%, 30.5%, 36.7%, 38.8%, and 42.7% respectively. The above shows that the raw Rehmannia slices prepared by lily have increased cough-relieving effect. The results are shown in Table 5.

Table 5 The number of ammonia-induced coughs in mice before and after intervention with raw Rehmannia slices prepared with lily bulbs

Note: Compared with the raw Rehmannia root group, the raw Rehmannia root slices prepared with lily bulbs showed ^# P<0.05, ^## P<0.01.

3.4 Experiment on increasing the expectorant efficacy of raw Rehmannia root slices prepared with lily bulbs

The tracheal phenol red excretion experiment is a common experiment to test the expectorant effect of drugs. If the amount of phenol red excretion in the mouse trachea can be promoted after administration, it reflects the expectorant effect of the drug. The present invention confirms that the raw Rehmannia slices prepared by lily can promote the phenol red excretion in the mouse trachea more than raw Rehmannia. The efficacy increase rates of the raw Rehmannia slices prepared by lily (Examples 1 to 6) compared with raw Rehmannia are 104.5%, 93.0%, 110.0%, 96.2%, 185.3%, and 219.3%, respectively. The above shows that the raw Rehmannia slices prepared by lily have increased the expectorant effect. The results are shown in Table 6.

Table 6 Phenol red secretion in mouse trachea before and after intervention with new Radix Rehmanniae decoction pieces

Note: Compared with the raw Rehmannia root group, the raw Rehmannia root slices prepared with lily bulbs showed ^# P<0.05, ^## P<0.01.

3.5 Experiment on enhancing the hemostatic efficacy of raw Rehmannia root slices prepared with lily bulbs

The tail-cutting method for measuring bleeding time is a common experiment for testing the hemostatic efficacy of drugs. If the bleeding time of the mouse tail can be shortened after administration, it reflects the hemostatic efficacy of the drug. The present invention confirms that the raw Rehmannia slices prepared by lily can shorten the bleeding time of the mouse tail compared with raw Rehmannia. The efficacy increase rates of the raw Rehmannia slices prepared by lily (Examples 1 to 6) compared with raw Rehmannia are 23.1%, 38.2%, 43.2%, 32.5%, 26.7%, and 49.6% respectively. The above shows that the raw Rehmannia slices prepared by lily enhance the hemostatic effect. The results are shown in Table 7.

Table 7 The bleeding time of the tail tip of mice before and after intervention with the raw Rehmannia root slices prepared by lily

Note: Compared with the blank control group, ^* P<0.05, ^** P<0.01; compared with the raw Rehmannia root slices prepared with lily bulbs and the raw Rehmannia root group, ^# P<0.05, ^## P<0.01.

3.6 Experimental study on the determination of catalpol, rehmannia glycoside D, lilyside A and lilyside B in the raw Rehmannia glutinosa slices prepared from lily of the present invention

3.6.1 Chromatographic conditions

Chromatographic conditions of Wangbaiheglycoside A and Wangbaiheglycoside B: using a Stablebond Analytical column (4.6×250mm, 5μm), the mobile phase is acetonitrile-0.1% phosphoric acid (18:82), the detection wavelength is 312nm, the flow rate is 1.0mL/min, the column temperature is 30°C, and the injection volume is 10μl.

Refer to the chromatographic conditions of catalpol in the 2020 edition of the "Chinese Pharmacopoeia": methanol-0.1% phosphoric acid (1:99) as the mobile phase, the detection wavelength is 210nm, the flow rate is 1.0mL/min, the column temperature is 30°C, and the injection volume is 10μl.

Refer to the chromatographic conditions of Rehmannia glycoside D in the 2020 edition of the "Chinese Pharmacopoeia": methanol-0.1% phosphoric acid (1:99) as the mobile phase, the detection wavelength is 203nm, the flow rate is 1.0mL/min, the column temperature is 30°C, and the injection volume is 10μl.

3.6.2 Preparation of reference solution

Accurately weigh an appropriate amount of Wangbaiheglycoside A and Wangbaiheglycoside B reference substances, place them in a 10 mL brown volumetric flask, add 80% methanol solution to prepare a reference substance solution with a concentration of 0.1 mg·mL ^-1 .

Accurately weigh an appropriate amount of catalpol reference substance, place it in a 10 mL brown volumetric flask, and add the mobile phase to prepare a 2.0 mg·mL ^-1 reference substance solution.

Accurately weigh an appropriate amount of rehmannia glycoside D reference substance, place it in a 10 mL brown volumetric flask, and add 25% methanol solution to prepare a 0.2 mg·mL ^-1 reference substance solution.

3.6.3 Preparation of test solution

Referring to the 2020 edition of the "Chinese Pharmacopoeia", accurately weigh 2.0g of raw Rehmannia slices prepared with lily, place in a round-bottom flask, add 50mL of methanol, weigh the weight, heat and reflux to extract for 1.5h, cool, weigh again, make up the lost weight with methanol, shake well, filter, accurately measure 10mL of the filtrate, concentrate to near dryness, dissolve the residue with mobile phase, transfer to a 10mL volumetric flask, and dilute to the scale with mobile phase, shake well, and pass through a 0.22μm microporous filter membrane for the determination of catalpol.

Referring to the 2020 edition of the "Chinese Pharmacopoeia", accurately weigh 1.0 g of raw Rehmannia slices prepared with lily, place in a stoppered conical flask, accurately add 25 mL of 25% methanol, weigh the weight, ultrasonically treat (power 400 W, frequency 50 kHz) for 1 hour, cool, weigh again, make up the lost weight with 25% methanol, shake well, filter, and pass through a 0.22 μm microporous filter membrane for the determination of rehmannoside D.

Accurately weigh about 2.0 g of the coarse powder of Rehmannia glutinosa slices prepared with lily, place it in a round-bottom flask, add 50 mL of 80% methanol, weigh it, heat under reflux to extract for 1.5 h, cool, weigh it again, make up the lost weight with methanol, shake well, filter, and take the filtrate to pass through a 0.22 μm microporous filter membrane for the determination of lilyside A and lilyside B.

3.6.4 Linear relationship investigation

Accurately pipette an appropriate amount of catalpol reference solution into a 10mL brown volumetric flask, add 80% methanol to dissolve and dilute to the mark, prepare reference solutions with mass concentrations of 0.05, 0.25, 0.5, 1, 2, and 3 mg·mL ^-1 , respectively, inject samples according to the chromatographic conditions in 3.6.1, plot a standard curve with the catalpol peak area as the ordinate (Y) and the catalpol mass concentration as the abscissa (X), and obtain the linear regression equation: Y = 3610.7X + 74.203, R ² = 0.9996, indicating that catalpol has a good linear relationship in the range of 0.05 to 3 mg·mL ^-1 .

Accurately pipette an appropriate amount of rehmannia glycoside D reference solution into a 10 mL brown volumetric flask, add 25% methanol to dissolve and dilute to the mark, prepare reference solutions with mass concentrations of 0.0625, 0.125, 0.25, 0.5, and 1 mg·mL ^-1, respectively, inject samples according to the chromatographic conditions under 3.6.1, plot a standard curve with the peak area of rehmannia glycoside D as the ordinate (Y) and the mass concentration of rehmannia glycoside D as the abscissa (X), and obtain the linear regression equation: Y = 3012.1X-10.349, R ² = 0.9999, indicating that catalpol has a good linear relationship in the range of 0.0625 to 1 mg·mL ^-1 .

Accurately pipette an appropriate amount of Wang Bailiglycoside A reference solution into a 10 mL brown volumetric flask, add 80% methanol to dissolve and dilute to the mark, prepare reference solutions with mass concentrations of 0.00625, 0.0125, 0.025, 0.05, and 0.1 mg·mL ^-1, respectively, inject according to the chromatographic conditions under 3.6.1, plot a standard curve with the peak area of Wang Bailiglycoside A as the ordinate (Y), and the mass concentration of Wang Bailiglycoside A as the abscissa (X), and obtain the linear regression equation: Y = 18147X + 2.0115, R ² = 0.9995, indicating that catalpol has a good linear relationship in the range of 0.00625 to 0.1 mg·mL ^-1 .

Accurately pipette an appropriate amount of Wang Bailiglycoside B reference solution into a 10 mL brown volumetric flask, add 80% methanol to dissolve and dilute to the mark, prepare reference solutions with mass concentrations of 0.00625, 0.0125, 0.025, 0.05, and 0.1 mg·mL ^-1, respectively, inject according to the chromatographic conditions under 3.6.1, plot a standard curve with the peak area of Wang Bailiglycoside B as the ordinate (Y) and the mass concentration of Wang Bailiglycoside B as the abscissa (X), and obtain the linear regression equation: Y = 25239X-11.929, R ² = 0.9998, indicating that catalpol has a good linear relationship in the range of 0.00625 to 0.1 mg·mL ^-1 .

3.6.5 Content determination

The results showed that the contents of catalpol and rehmannia glycoside D in the raw Rehmannia glutinosa slices prepared with lily (Examples 1 to 6) were reduced, but the components of lilyside A and lilyside B were newly added. The results are shown in Table 8.

Table 8 Contents of catalpol, rehmannia glycoside D, lilyside A and lilyside B in raw Rehmannia glutinosa slices prepared with lily

Note: Compared with the raw Rehmannia root group, ^# P<0.05, ^## P<0.01.

The above experimental data show that the raw Rehmannia slices prepared by the present invention have enhanced yin-nourishing, heat-clearing and hemostatic effects, and confirm that the raw Rehmannia slices prepared by lily have newly added tranquilizing, cough-relieving and expectorant effects, which enhances the traditional medicinal value of raw Rehmannia in nourishing yin, clearing heat and stopping bleeding, and broadens the role of raw Rehmannia in indications such as insomnia, cough, phlegm and fluid retention, changes the active ingredients of raw Rehmannia, expands the medicinal value of raw Rehmannia, makes a creative contribution to the development of new slices, and has significant economic and social benefits.

The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. For those skilled in the art, the present invention may have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims (7)

1. A method for preparing Rehmannia glutinosa using lily, which is characterized in that it includes the following steps: a. Take raw rehmannia glutinosa slices, remove impurities, and set aside; b. Take the lily pieces, add 12 times the weight and volume of the lily pieces in water and decoct three times for 30 minutes each time. Combine the filtrate and concentrate it to a lily decoction with a mass concentration of 0.50 to 0.83g/mL; The weight volume mentioned above refers to solids measured in grams and liquids measured in mL; c. Take the raw rehmannia glutinosa decoction pieces in step a, add the lily decoction in step b, the mass fraction ratio of raw rehmannia glutinosa to lily is 100:15~25, mix well, and moisten until the medicine is completely absorbed; d. Process and dry the moistened rehmannia glutinosa slices in step c to obtain the finished product; The processing and drying is slow fire frying and drying, or freeze drying at -80°C. 2. The method for preparing rehmannia glutinosa with lily according to claim 1, characterized in that it includes the following steps: a. Take raw rehmannia glutinosa slices, remove impurities, and set aside; b. Weigh 15.0g of lily pieces, add water 12 times the weight and volume of the lily pieces each time, decoct three times, 30 minutes each time, combine the decoctions, and concentrate to a lily water decoction with a concentration of 0.50g/mL; c. Take 100g of the raw rehmannia glutinosa pieces after removing impurities in step a, add the lily decoction in step b, mix well, and moisten until the medicine is completely absorbed; d. Place the moistened Rehmannia glutinosa slices in step c in a wok, stir-fry over low heat until the section of the Rehmannia glutinosa is brown to black or jet black with slight burn spots, then dry it in a 60°C oven and take it out. Let cool and serve. 3. The method for preparing rehmannia glutinosa with lily according to claim 1, characterized in that it includes the following steps: a. Take raw rehmannia glutinosa slices, remove impurities, and set aside; b. Weigh 20.0g of lily pieces, add water 12 times the weight and volume of the lily pieces each time, decoct three times, 30 minutes each time, combine the decoctions, and concentrate to a lily water decoction with a concentration of approximately 0.67g/mL. ; c. Take 100g of the raw rehmannia glutinosa pieces after removing impurities in step a, add the lily decoction in step b, mix well, and moisten until the medicine is completely absorbed; d. Place the moistened Rehmannia glutinosa slices in step c in a wok, stir-fry over low heat until the cross section of the Rehmannia glutinosa is brown to black or jet black, with slight burn spots, then dry it in an oven at 60°C, take it out, and let it cool. , that’s it. 4. The method for preparing rehmannia glutinosa with lily according to claim 1, characterized in that it includes the following steps: a. Take raw rehmannia glutinosa slices, remove impurities, and set aside; b. Weigh 25.0g of lily pieces, add water 12 times the weight and volume of the lily pieces each time, decoct three times, 30 minutes each time, combine the decoctions, and concentrate to a lily water decoction with a concentration of approximately 0.83g/mL. ; c. Take 100g of the raw rehmannia glutinosa pieces after removing impurities in step a, add the lily decoction in step b, mix well, and moisten until the medicine is completely absorbed; d. Place the moistened Rehmannia glutinosa slices in step c in a wok, stir-fry over low heat until the cross section of the Rehmannia glutinosa is brown to black or jet black, with slight burn spots, then dry it in an oven at 60°C, take it out, and let it cool. , that’s it. 5. The method for preparing rehmannia glutinosa with lily according to claim 1, characterized in that it includes the following steps: a. Take raw rehmannia glutinosa slices, remove impurities, and set aside; b. Weigh 15.0g of lily pieces, add water 12 times the weight and volume of the lily pieces each time, decoct three times, 30 minutes each time, combine the decoctions, and concentrate to a lily water decoction with a concentration of approximately 0.50g/mL. ; c. Take 100g of the raw rehmannia glutinosa pieces after removing impurities in step a, add the lily decoction in step b, mix well, and moisten until the medicine is completely absorbed; d. Place the moistened rehmannia glutinosa slices in step c into a freeze dryer at -80°C and freeze-dry them, and then take them out to obtain the result. 6. The method for preparing rehmannia glutinosa with lily according to claim 1, characterized in that it includes the following steps: b. Weigh 20.0g of lily pieces, add water 12 times the weight and volume of the lily pieces each time, decoct three times, 30 minutes each time, combine the decoctions, and concentrate to a lily water decoction with a concentration of approximately 0.67g/mL. ; c. Take 100g of the raw rehmannia glutinosa pieces after removing impurities in step a, add the lily decoction in step b, mix well, and moisten until the medicine is completely absorbed; d. Place the moistened rehmannia glutinosa slices in step c into a freeze dryer at -80°C and freeze-dry them, and then take them out to obtain the result. 7. The method for preparing rehmannia glutinosa with lily according to claim 1, characterized in that it includes the following steps: a. Take raw rehmannia glutinosa slices, remove impurities, and set aside; b. Weigh 25.0g of lily pieces, add water 12 times the weight and volume of the lily pieces each time, decoct three times, 30 minutes each time, combine the decoctions, and concentrate to a lily water decoction with a concentration of approximately 0.83g/mL. ; c. Take 100g of the raw rehmannia glutinosa pieces after removing impurities in step a, add the lily decoction in step b, mix well, and moisten until the medicine is completely absorbed; d. Place the moistened rehmannia glutinosa slices in step c into a freeze dryer at -80°C and freeze-dry them, and then take them out to obtain the result.