AU2019101031A4 - Compound American ginseng royal jelly oral liquid for treating deficiency of blood and preparation method thereof - Google Patents

Abstract

Abstract The present application is directed to an American ginseng royal jelly oral liquid and method of its preparation. The preparation is used for enhancing immunity.

Description

Compound American ginseng royal jelly oral liquid for treating deficiency of blood and preparation method thereof

Technical field

The invention belongs to the field of oral traditional Chinese medicine preparations, and particularly relates to a compound American ginseng royal jelly oral liquid for treating qi and blood deficiency and a preparation method thereof.

Background Art

Insufficient qi and blood is the qi deficiency and blood deficiency in Chinese medicine. The result of insufficient blood can lead to a decline in the function of the viscera, causing premature aging. Qi deficiency, that is, visceral function decline, poor resistance to disease, manifested as chills, cold limbs, spontaneous sweating, dizziness, tinnitus, apathy, fatigue, palpitations, shortness of breath, and stunting. Blood deficiency can be seen without sallow sallow, dry skin, withered hair, cracked nails, dimly lit eyes, numbness of hands and feet, insomnia, dreams, forgetfulness, and mental paralysis.

Insufficient blood is the same as blood and blood. Deficiency of qi and blood will be dystrophic, with Shenpi fatigue, shortness of breath, lazy words, pale or sallow, dizziness, pale lips, palpitations, insomnia, weak tongue and weak pulse are common syndromes.

The existing Chinese medicine preparations for treating qi and blood deficiency are complicated, and the preparation method thereof is also cumbersome.

Summary of the Invention

In view of the deficiencies in the prior art, the present invention provides a compound American ginseng royal jelly oral liquid for treating qi deficiency, and a preparation method thereof, and the compound American ginseng royal jelly oral liquid provided by the invention can prevent and/or treat low immunity caused by insufficient blood and blood. , physical weakness and so on.

The technical solution adopted by the invention is:

The invention provides a preparation method of a compound American ginseng royal jelly oral liquid.

The preparation method of the compound American ginseng royal jelly oral liquid provided by the invention comprises the following steps:

(1) 1.5 parts to 2.5 parts of American ginseng and 0.5 parts to 1.5 parts of schisandra in parts by mass, immersed in 65% to 85% ethanol for 24 to 45 hours, and then oozing for 7 to 10 times of raw medicine. In the sputum, the osmotic solution is decompressed to recover ethanol to obtain a concentrated liquid, which is an extract of Schisandra chinensis;

(2) pulverizing 0.62 parts to 1.86 parts of royal jelly, diluting with water, and filtering to obtain a component of royal jelly;

2019101031 09 Sep 2019 (3) mixing the extract of American ginseng Schisandra and the component of the royal jelly, and letting it stand for 24 to 72 hours to obtain a compound American ginseng royal jelly oral liquid.

In the step (1), the mass ratio of the ethanol to the mass of the American ginseng and the schisandra is 10 to 12:1.

In the step (1), the ethanol recovered under reduced pressure is carried out at 60 ° C to 75 ° C.

In the step (2), the water is sterile water, and the mass ratio of the royal jelly to water is 1:56 to 68.5.

In the step (2), the filtration was carried out by filtration using a 10 pm sterile filter.

In the step (3), after the mixing, the stirring and filtering steps are further included, stirred for 15 min to 30 min, and filtered through a 120 mesh sieve.

The step (1) further comprises the following steps: after obtaining the extract of the American ginseng Schisandra, the extract of the American ginseng Schisandra is mixed with citric acid, potassium sorbate, honey, boiled for 30 min, cooled, and set aside.

The compound American ginseng royal jelly oral liquid prepared by the preparation method is also within the protection scope of the present invention.

The use of the compound American ginseng royal jelly oral liquid in the preparation of a product for preventing and/or treating qi and blood deficiency is also within the scope of the present invention.

The use of the compound American ginseng royal jelly oral liquid in the preparation of an immunity-enhancing product is also within the scope of the present invention. With the preparation method of the invention, the active ingredient saponin of American ginseng and the schisandrin active ingredient of Schisandra chinensis can be better dissolved in 65%~85% ethanol, especially 75% ethanol, and the active ingredient is extracted by cold infiltration method. The extraction rate is high, the impurities are less, the loss of active ingredients is less, and the energy is saved. According to the applicant's experiment, the ethanol concentration is too high or too low, which will affect the extraction effect of the active ingredients and the interference of impurities.

Materia Medica cloud: Western ginseng bitter and sweet, taste thick and thin, fill the lungs and reduce fire, Shengjin liquid, in addition to tiredness, virtual yin and fire are appropriate. Chinese Medicine Dictionary records: Gan Wei bitterness, benefit lung yin, clear virtual fire, Shengjin quenching thirst. Treatment of lung deficiency for a long time, blood loss, dry throat, thirst, fatigue and tired. Since the Materia Medica dependent The medical records of the past generations all gave a very high evaluation of the medicinal value of American ginseng.

Schisandra condenses the lungs, nourishes the kidneys, stimulates the body, absorbs sweat, sputum essence, soothe the nerves. Royal jelly is sweet and sour, flat; it has nourishing, strong, beneficial liver and spleen effect.

In addition to the comprehensive prescription, the compound American ginseng royal jelly oral liquid disclosed in this paper has the characteristics of nourishing and strong, nourishing blood, nourishing and nourishing, and no dryness and no heat. As the

2019101031 09 Sep 2019 medical heart of the West recorded in the cloud: Western ginseng is cool and make up, anyone who wants to use ginseng without the warmth of ginseng can be replaced by this. Therefore, the oral liquid is suitable for both young and old, has a unique health care function, and is widely used for therapeutic products.

The oral liquid has been tested to contain the following components: total saponins of American ginseng, 10-hydroxy-2-decenoic acid, schisandrin and schisandra alcohol, various trace elements and amino acids, as well as albumin and globulin, after animal testing, For mice with hemorrhagic blood deficiency, it has the effect of promoting erythropoiesis, supplementing qi and blood, and at the same time enhancing immunity.

After mixing and stirring, it is allowed to stand for 24~72h in order to make the mechanical foam generated during stirring float up, which is easy to remove before bottling.

The extraction of Schisandra and American ginseng with 75% ethanol is for the purpose of extracting the active ingredients, and on the other hand killing the bacteria, because it is well known that the concentration range of ethanol has the strongest bactericidal ability.

The invention has the following advantages:

1. The preparation steps are simple, and the Western participation in the extraction of Schisandra together simplifies the process;

2. The oral liquid prepared according to the method described herein has a significant improvement effect on the immunity caused by insufficient blood and deficiency, and has obvious improvement effect. The functional component of the oral liquid, wherein the total saponin (based on ginsenoside Re) > 30.7 mg / 100 g, 10-hydroxy-2-decenoic acid >16.1 mg / 100 g.

3. The oral liquid prepared according to the method of the present invention has high bioavailability because it is a water-soluble solution.

Detailed Description of the Preferred Embodiments

The technical solutions of the present invention are further limited in the following with reference to specific embodiments, but the scope of the claims is not limited to the description.

Reagent source:

American ginseng, Schisandra, royal jelly: purchased from Zhejiang Xinchang County Pharmaceutical Herbal Medicine Co., Ltd.;

The ethanol solution was blended with edible alcohol and purified water. Edible alcohol was purchased from Zhejiang Xinchang County Pharmaceutical Herbal Medicine Co., Ltd., and the high-quality edible alcohol content was 95%, which was in compliance with GB 10343-1989 edible alcohol standard.

Example 1. Preparation method of compound American ginseng royal jelly oral liquid 1

The preparation method of compound American ginseng royal jelly oral liquid comprises the following steps:

2019101031 09 Sep 2019 (1) 2 parts of American ginseng and 1 part of schisandra are pulverized into 60-mesh powder in mass parts, soaked in 75% of ethanol at room temperature for 24 hours, and then immersed in 7 times of the crude drug at a rate of 30-50 d/min. In the sputum, the osmotic solution is decompressed to recover ethanol, and the concentrate is obtained, which is the extract of American ginseng Schisandra; the mass ratio of ethanol to the mass ratio of the American ginseng and the schisandra is 10:1, and the vacuum is recovered at 60 ° C, vacuum. The degree is 0.06-0.1 MPa;

(2) 1.24 parts of royal jelly was ground with colloid mill for 3 min, diluted with sterile water and filtered to obtain royal jelly components; the mass ratio of royal jelly to water was 1:62.9, and the filtration pore size was 10 μιη (molecular weight cutoff % 70,000) The hollow fiber ultrafiltration machine is filtered, and the filtrate is placed in a sterile container for use;

(3) The American ginseng Schisandra extract and the royal jelly component were mixed and stirred for 15 minutes, filtered through a 120 mesh sieve, and allowed to stand for 24 hours to obtain a compound American ginseng royal jelly oral liquid.

The total saponins of American ginseng are determined according to the determination method of total saponins in the Technical Specification for Health Food Inspection and Evaluation. The 10-hydroxy-2-decenoic acid is determined according to the general standard of GB/T9697-2002 royal jelly, and the total saponin content is determined. 30.7 mg/lOOg, 10-hydroxy-2-decenoic acid content 16.1 mg/lOOg.

Example 2 Preparation method of compound American ginseng royal jelly oral liquid 2

The preparation method of compound American ginseng royal jelly oral liquid comprises the following steps:

(1) 1.5 parts of American ginseng and 0.5 parts of schisandra were pulverized into 60-mesh powder in mass parts, soaked in 65% of ethanol at room temperature for 36 hours, and then immersed in a volume of 30-50 d/min to collect 8 times of crude drug. The percolating solution is obtained by decompressing the percolating solution to obtain ethanol, which is a extract of American ginseng Schisandra; the mass ratio of ethanol to the mass ratio of the American ginseng and the schisandra is 11:1, and the ethanol recovered under reduced pressure is 70 ° C, The vacuum is performed under the condition of 0.06 to 0.1 MPa;

(2) 0.62 parts of royal jelly was ground with colloid mill for 3 min, diluted with sterile water, and filtered to obtain royal jelly components; the mass ratio of royal jelly to water was 1:56, and the pore diameter for filtration was 10 pm (molecular weight cutoff % 70,000) The hollow fiber ultrafiltration machine is filtered, and the filtrate is placed in a sterile container for use;

(3) The American ginseng Schisandra extract and the royal jelly component were mixed and stirred for 30 minutes, filtered through a 120 mesh sieve, and allowed to stand for 36 hours to obtain a compound American ginseng royal jelly oral liquid.

The total saponins of American ginseng are determined according to the determination method of total saponins in the Technical Specification for Health Food Inspection and Evaluation. The 10-hydroxy-2-decenoic acid is determined according

2019101031 09 Sep 2019 to the general standard of GB/T9697-2002 royal jelly, and the total saponin content is determined. 30.8 mg/lOOg, 10-hydroxy-2-decenoic acid content 16.3 mg/lOOg.

Example 3 Preparation method of compound American ginseng royal jelly oral liquid 3

The preparation method of compound American ginseng royal jelly oral liquid comprises the following steps:

(1) 2.5 parts of American ginseng and 1.5 parts of schisandra were pulverized into 60-mesh powder in mass parts, soaked in 85% of ethanol at room temperature for 45 hours, and then immersed in a volume of 30-50 d/min to collect 10 times of crude drug. The percolating solution is obtained by decompressing the percolating solution to obtain ethanol, which is a extract of American ginseng Schisandra; the mass ratio of ethanol to the mass ratio of the American ginseng and the schisandra is 12:1, and the ethanol recovered under reduced pressure is 75 ° C, The vacuum is performed under the condition of 0.06 to 0.1 MPa;

(2) 1.86 parts of royal jelly was ground with colloid mill for 3 min, diluted with sterile water, and filtered to obtain the royal jelly component; the mass ratio of royal jelly to water was 1:68.5, and the filtration pore size was 10 pm (molecular weight cutoff % 70,000) The hollow fiber ultrafiltration machine is filtered, and the filtrate is placed in a sterile container for use;

(3) The American ginseng Schisandra extract and the royal jelly component were mixed and stirred for 30 minutes, filtered through a 120 mesh sieve, and allowed to stand for 72 hours to obtain a compound American ginseng royal jelly oral liquid.

The total saponins of American ginseng are determined according to the determination method of total saponins in the 2003 edition of the Technical Specifications for Health Food Inspection and Evaluation. The 10-hydroxy-2-decenoic acid is determined according to the general standard of GB/T9697-2002 royal jelly. The saponin content was 31.2 mg/100 g, and the 10-hydroxy-2-decenoic acid content was 16.8 mg/100 g.

Experimental Example 1. Toxicological test (The following test samples are all the oral liquids shown in Example 1, and the recommended daily intake for adults is 20 mL/60 kgbw)

First, the test method:

1. Acute toxicity test in mice (maximum tolerated dose method): 20 healthy Kunming mice of 20~22g, 10 males and 10 females, with a dose of 30.00g/kgbw (100 times recommended intake), interval After 4 hours, after three times of intragastric administration, continuous observation for 14 days, if the mice did not die, determine the maximum tolerated dose (MTD).

2, rat acute toxicity test (maximum tolerated dose method): 180g ~ 220g healthy Wistar rats (purchased from Nanjing Junke Biological Engineering Co., Ltd.) 20, 10 male and 10 each tested, with 30.00g / kgbw The dose (100 times recommended), the rat gastric perfusion volume 20mL / kgbw, 4 hours apart, after three times of intragastric administration, continuous observation for 14 days, if not caused by rat

2019101031 09 Sep 2019 death, determine the maximum tolerated dose (MTD) .

3. Sperm malformation test in mice: 25 male sexually mature male mice weighing 25-30 g were randomly divided into 5 groups. The dose of 40 mg/kg bw of cyclophosphamide (peritoneal injection) was used as a positive control, and the sample solvent was negative. For the control, the maximum gavage capacity was used once, and the experimental samples were set to 2.50, 5.00, 10.00 g/kg bw, and the test samples were freshly prepared when used. The rats were intragastrically administered once a day for 5 days. The animals were sacrificed 30 days after the last gavage. The epididymis tablets were taken and stained with eosin. Five animals were counted in each group. Each animal counted 1000 intact spermatozoa to calculate the abnormal spermatogenesis. Rate (in percentage) and statistical processing.

Second, the test results:

1. Maximum Tolerance Test in Mice: As shown in Table 1, the test samples were administered with mice of two sexes at a dose of 30.00 g/kg bw at intervals of 4 hours and three times for 14 days. After the gavage, the animals slightly bloated and returned to normal after 24 hours without death. The experimental sample had a maximum oral tolerated dose (MTD) of more than 30.00 g/kg bw for both sexes. According to the acute toxicity grading standards, the test samples are non-toxic.

Table 1 Test results of maximum tolerance of mice

Sex	route	Dose g/kgbw	number of animals	deaths	MTD g/kgbw
Male	oral	30.00	10	0	>30.00
Female	oral	30.00	10	0	>30.00

2. Rat maximum tolerated dose test: It can be seen from Table 2 that the test samples were administered with mice of two sexes at a dose of 30.00 g/kg bw at intervals of 4 hours and three times, and observed for 14 days. After the gavage, the animals slightly bloated and returned to normal after 24 hours without death. The experimental sample had a maximum oral tolerated dose (MTD) of greater than 30.00 g/kg bw for both sexes. According to the acute toxicity grading standards, the test samples are non-toxic.

Table 2 Test results of maximum tolerated rats

Sex	route	Dose g/kgbw	number of animals	deaths	MTD g/kgbw
Male	oral	30.00	10	0	>30.00
Female	oral	30.00	10	0	>30.00

2019101031 09 Sep 2019

3, mouse sperm abnormality test: As can be seen from Table 3, the incidence of sperm abnormality in the experimental sample of each dose group compared with the sample solvent control group, no significant difference (P> 0.05); dicyclophosphamide group mouse sperm There was a significant difference in the incidence of malformation compared with the sample solvent group (P<0.05). No test samples were found to have significant damage to male mouse germ cells.

Table 3 mouse sperm abnormality test

Name	Dose g/kgbw	numbe r of animal s	Obser ving the numb er of sperm	sperm type of sperm abnormality
No hook head	amor phous head	banan a head	other malfo rmati on	def or mity	defor mity rate /%	P value
Solve nt Contr ol		5	5000	42	32	32	1	107	2.14
Test sampl e	2.50	5	5000	43	30	30	0	103	2.06	0.78
5.00	5	5000	45	37	31	1	114	2.28	0.63
10.00	5	5000	44	36	33	1	114	2.28	0.63
Cyclo phosp hamid e	0.04	5	5000	120	111	154	40	425	8.5	<0.01

The above experiment proves that the oral liquid prepared by the preparation method of the compound American ginseng royal jelly oral liquid of Example 1 has no acute toxicity to mice and rats, and is a non-toxic substance, and has no reproductive toxicity to male mouse germ cells.

The oral liquid prepared by the preparation method of the compound American ginseng royal jelly oral liquid of Example 2-3 was not significantly different from the toxicological test result of the product of Example 1, that is, no acute toxicity was found to the mice and the rats, and it was non-toxic. Substance and no reproductive toxicity to male mouse germ cells.

Experimental example 2: qi and blood animal experiment

This experiment is to determine the effect of American ginseng royal jelly on the number of Hb content RBC in mice with hemorrhagic blood deficiency. 50 males of 20-24 g Kunming mice were randomly divided into 5 groups, and about 0.6 mL of blood was collected from each eyelid. At the same time, test Hb and RBC. After 24 hours of blood loss, blood was taken for Hb and RBC, and then administered to the stomach (according to the preparation method of the compound American ginseng royal jelly oral liquid preparation method of Example 1), the dosage is shown in Table

2019101031 09 Sep 2019

4, continuous administration for 10 days, Blood was taken on day 10 to measure Hb and RBC. Before blood loss and blood loss, the drug group was compared with the blank control group, and statistical analysis was performed to compare the difference. The experiment showed that each dose of oral American ginseng royal jelly had an effect on the Hb content of mice with hemorrhagic blood deficiency, and the corresponding RBC number increased by P<0.001.

Table 4 Effect of American ginseng royal jelly on the number of Hb content RBC in mice with hemorrhagic blood deficiency

Group	Dose g/kg.d	numb er of anim als	Hb(g/dl)
Before blood loss	after blood loss	after administratio n	the number of RBC increases
American	0.35X10	10	16.110.40	9.211.47’	15.011.13	4.9411.38
ginseng	0.70X10	10	15.710.45	9.411.39’	15.111.17	5.711.55Λ
royal jelly	1.00X10	10	15.510.40	8.711.19’’’	15.410.57	6.8 + 1.20ΛΛΛ
Bazhen	3.00X10	10	16.010.54	8.511.15’’’	15.111.65	6.6+ 1.54ΛΛΛ
Yimu Pills
Distilled		10	15.910.24	9.811.20’’’	14.110.91	4.311.07
water

The experiment proves that the oral liquid prepared by the preparation method of the compound American ginseng royal jelly oral liquid of the embodiment 1 has the effect of promoting erythropoiesis and tonifying blood and blood for the mice with hemorrhagic blood deficiency.

The oral liquid prepared by the preparation method of the compound American ginseng royal jelly oral liquid of Example 2-3 has no significant difference with the experimental result of the product of the blood supply of the blood of the animal of the first embodiment, that is, the blood cell of the blood-deficient blood-deficiency mouse promotes erythropoiesis and qi The role of blood.

Experimental Example 3: Enhanced immunity functional animal experiment The experiment is based on the 2003 edition of the Technical Specifications for Health Food Inspection and Evaluation.

According to the standard, the test results were determined as follows: Under the laboratory conditions, 18~22g Kunming male mice were used as experimental animals, and the oral liquid prepared by the compound American ginseng royal jelly oral liquid preparation method of Example 1 was divided into 0.60g/ Kgbw, 0.40g/kgbw, 0.20g/kgbw three experimental groups and sample solvent control group, once a day, three days after continuous measurement of various immune indicators.

Experimental results:

1. Measurement results of organ/body weight ratio: As shown in Table 5, the thymus/body weight ratio, gland/body weight ratio of each experimental group was not significantly different from that of the sample solvent control group (P>0.05).

2019101031 09 Sep 2019

Table 5 Results of mouse organ/body weight ratio measurement unit mg/g

Dose g/kgbw	number of animals	thymus / body weight ratio	Spleen / body weight ratio
X ± sd	P value	X +sd	P value
0.60	10	2.06	0.39	0.32	5.95	1.36	0.41
0.40	10	1.91	0.46	0.11	5.45	1.19	0.99
0.20	10	2.14	0.60	0.51	5.34	1.67	0.85
Solvent control	10	2.29	0.61		5.45	0.94

2. ConA-induced spleen lymphocyte transformation test results: As shown in Table 6, the initial body weight, end-of-test weight, weight gain and the sample solvent control group were not significantly different (P>0.05); 7 It can be seen that the difference in optical density of each experimental group is significantly different from that of the sample solvent control group (P<0.05).

Table 6 ConA-induced mouse spleen lymphocyte transformation experiment body weight measurement unit: g

Dose g/kgbw	num ber of anim als	Initial weight	Test end weight	Weight gain
X + sd	P value	X+ sd	P value	X + sd	P value
0.60	10	20.17	1.19	0.94	34.66	3.19	0.77	14.49	2.10	0.69
0.40	10	20.16	1.06	0.92	34.72	3.15	0.80	14.56	2.14	0.75
0.20	10	20.25	1.91	0.94	34.60	3.52	0.74	14.35	2.45	0.59
Solvent control	10	20.21	1.18		35.08	2.87		14.87	1.73

Table 7 Results of optical density measurement of induced mouse spleen lymphocyte test

2019101031 09 Sep 2019

Dose g/kgbw	number of animals	difference in density	optical
X d	z sd	P value
0.60	10	0.098	0.026	0.0K
0.40	10	0.067	0.024	0.03
0.20	10	0.069	0.018	0.03
Solvent	10	0.062	0.013
control

3. DNFB induced mouse DTH test results

It can be seen from Table 8 that the initial body weight, test body weight, and body weight gain of each experimental group were not significantly different from those of the sample solvent control group (P>0.05); as shown in Table 9, the left and right ear weight difference and sample solvent of each experimental group The difference was significant in the control group (P<0.05).

Table 8 DNFB-induced mouse DTH test body weight results

Dose g/kgbw	number of animal s	Initial weight	Test end weight	Weight gain
X + sd	P value	x+	sd	P value	X + sd	P value
0.60	10	20. 19	1.19	0.99	34.95	3.52	0.86	14.76	2.45	0.78
0.40	10	20. 22	1.24	0.94	34.40	2.97	0.60	14.18	1.83	0.41
0.20	10	20. 23	1.23	0.93	34.60	3.50	0.69	14.37	2.30	0.52
Solvent control	10	20. 18	1.36		34.23	3.92		15.05	2.61

Table 9 DNFB-induced DTH test in the left and right ear weight difference measurement results

Dose g/kgbw

number of

left and right ear weight difference

2019101031 09 Sep 2019

	animals	X	± sd	P value
0.60	10	0.0195		0.0033	0.03
0.40	10	0.0202		0.0063	0.01
0.20	10	0.0195		0.0064	0.04
Solvent	10	0.0171		0.0050
control

4. Antibody-producing cell detection (Jerne modified slide method)

It can be seen from Table 10 that the initial body weight, test body weight, and body weight gain of each experimental group were not significantly different from those of the sample solvent control group (P>0.05); as shown in Table 11, the hemolytic plaque number of each experimental group and the sample solvent control group were observed. The difference was significant (P < 0.05).

Table 10 Mouse antibody-producing cells to measure body weight measurement results

Dose g/kgbw	num ber of anim als	Initial weight	Test end weight	Weight gain
X + sd	P value	X+ sd	P value	X + sd	P value
0.60	10	20.19	1.28	0.93	34.56	3.35	0.68	14.37	2.18	0.49
0.40	10	20.22	1.16	0.94	34.16	3.38	0.88	14.88	2.32	0.85
0.20	10	20.28	1.22	0.97	34.66	3.02	0.63	14.44	1.89	0.45
Solvent control	10	20.24	1.22		34.94	3.46		13.70	2.27

Table 11 Results of measurement of plaque number in mouse antibody-producing cells

Dose g/kgbw	number of animals	plaque value
X	± sd	P value
0.60	10	2.07	0.54	0.01
0.40	10	1.61	0.33	0.02
0.20	10	1.74	0.56	0.04

2019101031 09 Sep 2019

Solvent	10	1.50	0.41
control

5. Mouse serum hemolysin determination results (blood coagulation method) It can be seen from Table 12 that there is no significant difference between the initial system, the end body weight and the weight gain of the experimental group in the experimental group (P>0.05). From Table 13, the anti-volume of each experimental group is compared with the sample solvent. The difference was significant (P<0.05).

Table 12 mouse serum hemolysin test body weight measurement results

Dose g/kgbw	num ber of anim als	Initial weight	Test end weight	Weight gain
X + sd	P value	X+ sd	P value	X + sd	P value
0.60	10	20.22	1.19	0.96	34.50	2.80	0.97	14.28	1.77	0.94
0.40	10	20.25	1.26	0.91	34.56	3.10	0.99	14.31	1.94	0.96
0.20	10	20.18	1.19	0.99	34.62	3.46	0.96	14.44	2.31	0.94
Solvent control	10	20.19	1.25		34.55	3.84		13.36	2.70

Table 13 Results of test results of mouse serum hemolysin test

Dose g/kgbw	number of animals	anti-volume
X	± sd	P value
0.60	10	148.20		13.76	<0.01
0.40	10	155.90		17.40	<0.01
0.20	10	124.20		21.82	0.03
Solvent	10	119.70		16.99
control

6. Mouse carbon clearance test results

It can be seen from Table 14 that the initial system, the weight at the end of the experiment, and the weight gain at the end of the experiment were not significantly different from the sample solvent control group (P>0.05). From Table 15, the phagocytic index of each experimental group and the sample solvent control group

2019101031 09 Sep 2019 were observed. The difference was significant (P < 0.05).

Table 14 Mouse carbon clearance test body weight measurement results

Dose g/kgbw	numb er of anim als	Initial weight	Test end weight	Weight gain
X + sd	P value	x+	sd	P value	X + sd	P value
0.60	10	20.32	1.14	0.88	34.44	2.98	0.86	14.12	1.93	0.85
0.40	10	20.19	1.29	0.93	34.19	3.36	0.99	14.00	2.12	0.96
0.20	10	20.26	1.23	0.97	34.62	2.87	0.75	14.37	1.73	0.63
Solvent control	10	20.24	1.15		34.97	2.87		13.96	1.81

Table 15 Results of phagocytic index measurement in mouse carbon clearance test

Dose g/kgbw	number of animals	phagocytic index a
X	± sd	P value
0.60	10	4.07		0.85	0.01
0.40	10	4.28		0.77	0.01
0.20	10	4.71		0.77	0.02
Solvent	10	4.51		0.54
control

7. Mouse peritoneal macrophage phagocytosis chicken red blood cell test results (half-in vivo method)

It can be seen from Table 16 that the initial system, the weight at the end of the experiment, and the body weight gain at the end of the experiment were not significantly different from the sample solvent control group (P>0.05). Table 17 shows the percentage of phagocytosis, phagocytic index and sample in each experimental group. The difference was significant in the solvent control group (P<0.05).

Table 16 mouse serum hemolysin test body weight measurement results

Dose	num	Initial weight	Test end weight	Weight gain
g/kgbw	ber	X + sd	P	Χ + sd	P	X + sd	P value
	of		value		value

2019101031 09 Sep 2019

	anim als
0.60	10	20.24	1.20	0.87	34.05	3.36	0.78	13.81	2.20	0.61
0.40	10	20.19	1.15	0.94	34.46	2.78	0.99	1427	1.68	0.96
0.20	10	20.21	1.20	0.91	34.57	3.54	0.95	14.36	2.51	0.97
Solvent	10	20.15	1.26		34.47	3.66		13.32	2.44
control

Table 17 Percentage of phagocytosis of mouse peritoneal macrophages phagocytic chicken red blood cells, phagocytic index measurement results

Dose	num	percentage of	inverse sine function	phagocytic index
g/kgbw	ber	phagocytosis	conversion value
	of	X ±	sd	P	X	t sd	P	X ±	sd	P value
	anim			value			value
	als
0.60	10	43.50	4.58	<0.01	41.25	2.64	<0.01	0.50	0.08	<0.01
0.40	10	41.50	7.96	<0.01	40.05	4.66	<0.01	0.50	0.13	<0.01
0.20	10	31.30	8.30	0.36	33.85	5.17	0.37	0.33	0.09	0.30
Solvent	10	28.50	5.17		32.19	3.27		0.31	0.06
control

8. Results of mouse NK cell activity assay

It can be seen from Table 18 that the initial system, the weight of the experimental end, the weight gain of the experimental group and the sample solvent control group were not significantly different (P>0.05); as shown in Table 19, the NK cell activity of each experimental group was compared with the sample solvent. The difference was significant (P<0.05).

Table 18 Mouse NK cell activity test body weight measurement results

Dose g/kgbw	num ber of anim als	Initial weight	Test end weight	Weight gain
X + sd	P value	X+ sd	P value	X + sd	P value

2019101031 09 Sep 2019

0.60	10	20.27	1.19	0.94	34.22	2.44	0.90	13.95	1.33	0.81
0.40	10	20.29	1.14	0.91	34.33	2.57	0.97	14.04	1.56	0.98
0.20	10	20.27	1.22	0.94	34.82	3.49	0.68	13.55	2.31	0.49
Solvent	10	20.23	1.24		34.39	3.68		14.16	2.47
control

Table 19 Results of mouse NK cell activity assay

Dose g/kgbw	number of animals	NK cell activity %	P value
0.60	10	9.6±1.2	0.02
0.40	10	9.411.2	0.02
0.20	10	8.911.4	0.03
Solvent control	10	7.411.2	-

The test results showed that the oral liquid prepared by the preparation method of the compound American ginseng royal jelly oral liquid of Example 1 had no significant effect on the body weight, body weight gain, chest weight/body weight ratio and spleen weight/body weight ratio of the experimental animals. Each experimental group can enhance the ability of mouse peritoneal macrophages to phagocytose chicken red blood cells; each experimental group can increase the degree of hemagglutination in mice; each experimental group can enhance the lymphocyte proliferation ability of mice, and can increase the number of antibody-producing cells. The experimental group can promote the formation of delayed-type allergic reaction (DTH) induced by dinitrofluorobenzene (DNFB) in mice. Each experimental group can enhance the carbon clearance ability of mice and can display the activity of mouse NK cells. The product prepared in Example 1 has an effect of enhancing immunity. The oral liquid prepared by the preparation method of the compound American ginseng royal jelly oral liquid of Example 2-3 has no significant difference with the results of the product of the first embodiment, that is, the body weight, body weight gain, chest weight/weight of the experimental animals of each test. The ratio, spleen weight/body weight ratio had no significant effect. Each experimental group can enhance the ability of mouse peritoneal macrophages to phagocytose chicken red blood cells; each experimental group can increase the degree of hemagglutination in mice; each experimental group can enhance the lymphocyte proliferation ability of mice, and can increase the number of antibody-producing cells. The experimental group can promote the formation of delayed-type allergic reaction (DTH) induced by dinitrofluorobenzene (DNFB) in mice. Each experimental group can enhance the carbon clearance ability of mice and can display the activity of mouse NK cells. The products prepared in Example 2-3 have an immunopotentiating effect.

Claims (10)

1. (1) 1.5 parts to 2.5 parts of American ginseng and 0.5 parts to 1.5 parts of schisandra in parts by mass, immersed in 65% to 85% ethanol for 24 to 45 hours, and then oozing for 7 to 10 times of raw medicine. In the sputum, the osmotic solution is decompressed to recover ethanol to obtain a concentrated liquid, which is an extract of Schisandra chinensis;

   1. The preparation method of compound American ginseng royal jelly oral liquid, comprising the following steps:
2. 2. The method for preparing a compound American ginseng royal jelly oral liquid according to claim 1, wherein in the step (1), the mass ratio of the ethanol to the mass of the American ginseng and the schisandra is 10 to 12:1. .

   (2) pulverizing 0.62 parts to 1.86 parts of royal jelly, diluting with water, and filtering to obtain a component of royal jelly;
3. 3. The method for preparing a compound American ginseng royal jelly oral liquid according to claim 1, wherein in the step (1), the vacuum recovery of ethanol is carried out at 60 ° C to 75 ° C.

   (3) mixing the extract of American ginseng Schisandra and the component of the royal jelly, and letting it stand for 24 to 72 hours to obtain a compound American ginseng royal jelly oral liquid.
4. 4. The method for preparing a compound American ginseng royal jelly oral liquid according to claim 1, wherein in the step (2), the water is sterile water, and the mass ratio of the royal jelly to water is 1:56. -68.5.
5. 5. The method for preparing a compound American ginseng royal jelly oral liquid according to claim 1, wherein in the step (2), the filtering is performed by using a 10 pm sterile filter.
6. 6. The method for preparing a compound American ginseng royal jelly oral liquid according to claim 1, wherein in the step (3), after the mixing, the step of stirring and filtering is further included, stirring for 15 min to 30 min, and 120 mesh sieve filtration. .
7. 7. The method for preparing a compound American ginseng royal jelly oral liquid according to claim 1, wherein the step (1) further comprises the following steps: after obtaining the extract of American ginseng Schisandra, the extract of the American ginseng Schisandra and citric acid Mix potassium sorbate and honey, boil for 30 minutes, cool, and set aside.
8. 8. The compound American ginseng royal jelly oral solution prepared by the method according to any one of claims 1-7.
9. 9. The compound American ginseng royal jelly oral solution according to claim 8 for use in the preparation of a product for preventing and/or treating qi and blood deficiency.
10. 10. The use of the compound American ginseng royal jelly oral liquid according to claim 8 for the preparation of a product for enhancing immunity.