Quality control method of jujube kernel tranquilizing liquid

Abstract

The invention discloses a quality control method of a jujube kernel tranquilizing liquid. The quality control method specifically comprises the following steps: S1, preparation of a test solution: preparing a test solution 1, a test solution 2 and a test solution 3; s2, taking the test sample solutions, and detecting the three test sample solutions through an HPLC-DAD (High Performance Liquid Chromatography-Diode Array Detection) gradient elution method; respectively carrying out gradient elution on the test solution 1 and the test solution 2 by adopting acetonitrile (A) and a 0.1% glacial acetic acid solution (B) with different concentration gradients; the specificity of the sample is determined through a negative sample lacking spina date seed, lacking salvia miltiorrhiza, lacking schisandra chinensis and lacking potassium sorbate; and carrying out gradient elution on the test solution 3 by adopting a mobile phase of acetonitrile (A)-water (B), and detecting by adopting evaporative light scattering. The detection time is greatly shortened, and the detection is more accurate.

Description

Technical Field

[0001] The invention relates to the technical field of jujube seed processing, and in particular to a quality control method for jujube seed tranquilizing liquid. Background Art

[0002] Insomnia is a common disease. For the treatment of insomnia, the use of antipsychotic sedative drugs is effective, but its side effects are also obvious, mainly reflected in affecting the mental state of the next day after taking the medicine, which further affects normal work and life. Zaoren Anshen Liquid is now included in the 18th volume of Chinese medicine formula preparations of the "Ministry of Health Drug Standards", with the standard number WS3-B-3429-98. It has the effects of nourishing the heart and liver, calming the mind and improving intelligence. It is clinically used for insomnia, forgetfulness, dizziness, headache and other symptoms caused by heart and liver blood deficiency and neurasthenia. The prescription reuses sour jujube seeds to nourish the heart yin, benefit the liver blood, and calm the mind and calm the mind as the main medicine; Danshen clears the heart and cools the blood, nourishes the blood and calms the mind; Schisandra chinensis nourishes the kidney and nourishes the yin, calms the mind and calms the mind, and is a ministerial medicine. The three medicines are used together to nourish the heart and liver, calm the mind and improve intelligence.

[0003] The Zaoren Anshen Recipe currently on the market, in addition to the oral liquid dosage form, also has capsules and granules. Among them, Zaoren Anshen Capsules are made of alcohol extraction into a thick paste, and starch is added to dry it into granules and loaded into capsules. It is an improved new drug, and the effective ingredients obtained from the traditional Chinese medicine decoction are different, so the quality control method is also different. Zaoren Anshen Granules and Zaoren Anshen Liquid are both water-extracted, but the granules are directly dried to make granules from the water extract, while Zaoren Anshen Liquid adds honey as an auxiliary material. Honey itself is also a traditional Chinese medicine with certain tonic effects. In this product, it plays the role of nourishing the stomach and spleen and harmonizing yin and yang. However, the addition of honey also interferes with the quantitative detection of the effective ingredients of the three medicinal materials.

[0004] The current standard quality control item of Zaoren Anshen Liquid is the TLC method to identify Schisandra chinensis and Salvia miltiorrhiza (with protocatechuic aldehyde as the indicator ingredient) in the prescription. There is only qualitative testing, which has the problem of inadequate detection and inaccuracy. The characteristics of the overall effect of compound Chinese patent medicines determine that the research on the quality standard of Chinese medicine should actively carry out qualitative and quantitative research on multiple components of Chinese patent medicines, which is also the current development direction of the "Chinese Pharmacopoeia" standard. Summary of the invention

[0005] The purpose of the present invention is to provide a quality control method for jujube seed tranquilizing liquid, so as to solve the problem mentioned in the above background technology that the current jujube seed tranquilizing preparation adopts qualitative detection, but the detection is not in place and inaccurate problems often occur.

[0006] To achieve the above object, the present invention provides the following technical solution: a quality control method of jujube seed tranquilizing liquid, specifically comprising the following steps:

[0007] Step S1, preparation of test solution:

[0008] Take the Zao Ren An Shen liquid and mix it evenly. Measure and put the Zao Ren An Shen liquid into a volumetric flask after shaking evenly. Add an appropriate amount of methanol-water mixed solution. Shake it thoroughly to dissolve and dilute to the scale. Filter and take the filtrate as the test solution 1.

[0009] Take the test solution 1 and dilute it 10 times with a methanol-water mixture to make the test solution 2;

[0010] Take Zaoren Anshen liquid, shake and extract with water-saturated n-butanol for 2-4 times, 30 ml each time, combine the extracts, wash with ammonia test solution twice, 20 ml each time, combine the n-butanol solution, recover the solvent to dryness, dissolve the residue with methanol, transfer to a 5 ml volumetric flask, add methanol to the scale, shake well, filter, and take the filtrate as the test solution 3;

[0011] Step S2: taking the above test solution and testing the three test solutions by HPLC-DAD gradient elution method.

[0012] Preferably, in step S2, the specific method of taking the above-mentioned test solution and detecting the three test solutions by HPLC-DAD gradient elution method is: using acetonitrile (A)-0.1% glacial acetic acid solution (B) with different concentration gradients, gradient eluting the test solution 1 and the test solution 2 respectively, and it is found that spinosin appears at 12-14 minutes, salvianolic acid B appears at 34 minutes, schisandrae alcohol A appears at 56-58 minutes, and potassium sorbate appears at 22-24 minutes. The simultaneous analysis and determination of the three active ingredients and potassium sorbate is completed within 60 minutes, which greatly shortens the analysis time, and then the specificity is determined by the negative samples lacking spinosae seed, salvia miltiorrhiza, schisandrae chinensis, and potassium sorbate.

[0013] Preferably, the mobile phase: acetonitrile (A)-water (B) is used to perform gradient elution on the test solution 3, and evaporative light scattering detection is used, wherein jujuboside A appears at 20-21 minutes, and its negative specificity is determined. Under the above chromatographic conditions, other components in the sample have no interference with the determination of jujuboside A.

[0014] Preferably, in step S2, acetonitrile (A)-0.1% glacial acetic acid solution (B) with different concentration gradients are used to perform gradient elution on test solution 1 and test solution 2, respectively. The specific manner is: 0-16 min, 16% A; 16-30 min, 16% A→25% A; 30-40 min, 25% A; 40-41 min, 25% A→48% A; 41-60 min, 48% A→35% A; flow rate 1.0 mL·min-1; detection wavelength: 250 nm.

[0015] Preferably, in step S2, the specific manner of gradient elution of the test solution 3 using acetonitrile (A)-water (B) is: 0-15 min, 20% A→40% A; 15-28 min, 40% A; 28-30 min, 40% A→70% A; 30-32 min, 70% A→100% A; flow rate 1.0 mL·min-1.

[0016] Compared with the prior art, the present invention has the following beneficial effects: the present invention adopts HPLC-DAD gradient elution method to realize quantitative determination of four active ingredients (spinoside, jujuboside a, salvianolic acid B, schisandrae alcohol A) of three Chinese medicinal materials in the prescription of Zaoren Anshen Liquid, and realizes limited detection of antibacterial agent potassium sorbate at the same time; the method is not interfered by honey in the liquid medicine, and the separation degree can reach above 1.5, thereby ultimately ensuring higher detection precision and more accurate detection. DETAILED DESCRIPTION

[0017] The technical solutions in the embodiments of the present invention will be described clearly and completely below in combination with the embodiments of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0018] Embodiment 1:

[0019] An embodiment provided by the present invention: a quality control method of jujube seed tranquilizing liquid, specifically comprising the following steps:

[0020] Step S1, preparation of test solution:

[0021] Take the Zao Ren An Shen liquid and mix it evenly. Measure and put the Zao Ren An Shen liquid into a volumetric flask after shaking evenly. Add an appropriate amount of methanol-water mixed solution. Shake it thoroughly to dissolve and dilute to the scale. Filter and take the filtrate as the test solution 1.

[0022] Take the test solution 1 and dilute it 10 times with a methanol-water mixture to make the test solution 2;

[0023] Take Zaoren Anshen liquid, shake and extract with water-saturated n-butanol for 2-4 times, 30 ml each time, combine the extracts, wash with ammonia test solution twice, 20 ml each time, combine the n-butanol solution, recover the solvent to dryness, dissolve the residue with methanol, transfer to a 5 ml volumetric flask, add methanol to the scale, shake well, filter, and take the filtrate as the test solution 3;

[0024] Step S2: taking the above test solution and testing the three test solutions by HPLC-DAD gradient elution method.

[0025] Preferably, in step S2, the specific method of taking the above-mentioned test solution and detecting the three test solutions by HPLC-DAD gradient elution method is: using acetonitrile (A)-0.1% glacial acetic acid solution (B) with different concentration gradients, gradient eluting the test solution 1 and the test solution 2 respectively, and it is found that spinosin appears at 12-14 minutes, salvianolic acid B appears at 34 minutes, schisandrae alcohol A appears at 56-58 minutes, and potassium sorbate appears at 22-24 minutes. The simultaneous analysis and determination of the three active ingredients and potassium sorbate is completed within 60 minutes, which greatly shortens the analysis time, and then the specificity is determined by the negative samples lacking spinosae seed, salvia miltiorrhiza, schisandrae chinensis, and potassium sorbate.

[0026] Preferably, the mobile phase: acetonitrile (A)-water (B) is used to perform gradient elution on the test solution 3, and evaporative light scattering detection is used, wherein jujuboside A appears at 20-21 minutes, and its negative specificity is determined. Under the above chromatographic conditions, other components in the sample have no interference with the determination of jujuboside A.

[0027] Preferably, in step S2, acetonitrile (A)-0.1% glacial acetic acid solution (B) with different concentration gradients are used to perform gradient elution on test solution 1 and test solution 2, respectively. The specific manner is: 0-16 min, 16% A; 16-30 min, 16% A→25% A; 30-40 min, 25% A; 40-41 min, 25% A→48% A; 41-60 min, 48% A→35% A; flow rate 1.0 mL·min-1; detection wavelength: 250 nm.

[0028] Preferably, in step S2, the specific manner of gradient elution of the test solution 3 using acetonitrile (A)-water (B) is: 0-15 min, 20% A→40% A; 15-28 min, 40% A; 28-30 min, 40% A→70% A; 30-32 min, 70% A→100% A; flow rate 1.0 mL·min-1;

[0029] The four active ingredients of the jujube seed tranquilizing liquid of the present invention can represent the effective ingredients of the jujube seed tranquilizing liquid, wherein the main active ingredients of the Chinese jujube seed in the prescription are: Chinese jujube seed saponin, spinosin, 6'" feruloyl spinosin; the main active ingredients of salvia miltiorrhiza are: salvianolic acid B, tanshinone IIA, tanshinone I, sodium tanshinone, rosmarinic acid, and protocatechuic aldehyde; the main active ingredients of schisandra chinensis are: schisandrin A, schisandrin B, schisandrin alcohol A and other lignan components. After consulting relevant literature, it is found that spinosin and Chinese jujube seed saponin a in Chinese jujube seed can prolong sleep time, improve memory, and have anticonvulsant, anti-anxiety and anti-depression effects; according to the process of water decoction, it can be concluded that the main active ingredient of salvia miltiorrhiza in the medicine is salvianolic acid B, which has pharmacological effects such as protection of the heart and the brain; and schisandrin alcohol A, which is an important characteristic ingredient for distinguishing schisandra chinensis from southern schisandra chinensis, has the pharmacological effects of enhancing learning and memory, calming the nerves, and regulating sleep.

[0030] Therefore, the present invention adopts the HPLC-DAD gradient elution method to achieve the quantitative determination of the four active ingredients (spinoside, jujuboside a, salvianolic acid B, and schisandrin A) of the three Chinese medicinal materials in the prescription of Zaoren Anshen Liquid, and at the same time achieves limited detection of the antibacterial agent potassium sorbate. This method is not interfered by the honey in the liquid medicine, and its separation degree can reach more than 1.5, ultimately ensuring higher detection accuracy and more accurate detection.

[0031] The invention adopts the quality control method to measure the content of spinosin to be 100 μg / ml, the content of jujuboside a to be 20 μg / ml, the content of salvianolic acid B to be 100 μg / ml, the content of schisandrae alcohol A to be 50 μg / ml, and the content of potassium sorbate to be 0.2%.

[0032] Although the present invention has been described in detail with reference to the aforementioned embodiments, it is still possible for those skilled in the art to modify the technical solutions described in the aforementioned embodiments, or to make equivalent substitutions for some of the technical features therein. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the protection scope of the present invention.

Claims

1. A quality control method for jujube seed tranquilizing liquid, characterized in that: The specific steps include: Step S1, preparation of test solution: Take the Zao Ren An Shen liquid and mix it evenly. Measure and put the Zao Ren An Shen liquid into a volumetric flask after shaking evenly. Add an appropriate amount of methanol-water mixed solution. Shake it thoroughly to dissolve and dilute to the scale. Filter and take the filtrate as the test solution 1. Take the test solution 1 and dilute it 10 times with a methanol-water mixture to make the test solution 2; Take Zaoren Anshen liquid, shake and extract with water-saturated n-butanol for 2-4 times, 30 ml each time, combine the extracts, wash with ammonia test solution twice, 20 ml each time, combine the n-butanol solution, recover the solvent to dryness, dissolve the residue with methanol, transfer to a 5 ml volumetric flask, add methanol to the scale, shake well, filter, and take the filtrate as the test solution 3; Step S2: taking the above test solution and testing the three test solutions by HPLC-DAD gradient elution method.

2. The quality control method of the Zao Ren An Shen Liquid according to claim 1, characterized in that: In step S2, the specific method of taking the above-mentioned test solution and detecting the three test solutions by HPLC-DAD gradient elution method is: using acetonitrile (A)-0.1% glacial acetic acid solution (B) with different concentration gradients, gradient eluting the test solution 1 and the test solution 2 respectively, and it is found that spinosin appears at 12-14 minutes, salvianolic acid B appears at 34 minutes, schisandrae alcohol A appears at 56-58 minutes, and potassium sorbate appears at 22-24 minutes. The simultaneous analysis and determination of the three active ingredients and potassium sorbate are completed within 60 minutes, which greatly shortens the analysis time, and then the specificity is determined by the negative samples lacking spinosae seed, salvia miltiorrhiza, schisandrae chinensis, and potassium sorbate.

3. The quality control method of a Zao Ren An Shen Liquid according to claim 2, characterized in that: The mobile phase: acetonitrile (A)-water (B) was used to perform gradient elution on the test solution 3, and evaporative light scattering detection was used, wherein jujuboside A appeared at 20-21 minutes, and its negative specificity was determined. Under the above chromatographic conditions, other components in the sample had no interference with the determination of jujuboside A.

4. The quality control method of a Zao Ren An Shen Liquid according to claim 2, characterized in that: In step S2, acetonitrile (A)-0.1% glacial acetic acid solution (B) with different concentration gradients are used to perform gradient elution on test solution 1 and test solution 2, respectively. The specific manner is: 0-16 min, 16% A; 16-30 min, 16% A→25% A; 30-40 min, 25% A; 40-41 min, 25% A→48% A; 41-60 min, 48% A→35% A; flow rate 1.0 mL·min-1; detection wavelength: 250 nm.

5. The quality control method of the Zaoren Anshen Liquid according to claim 3, characterized in that: In step S2, the specific method of gradient elution of the test solution 3 using acetonitrile (A)-water (B) is: 0-15 min, 20% A→40% A; 15-28 min, 40% A; 28-30 min, 40% A→70% A; 30-32 min, 70% A→100% A; flow rate 1.0 mL·min-1.