A method for identifying jujube in Xiaoer Shengxue Gansu by thin layer chromatography

Abstract

The application provides a method for identifying jujubes in Xiaoer Shengxue syrup by adopting thin layer chromatography. The method is as follows: 10 microliters of test sample solution, 10 microliters of control medicinal material solution and 10 microliters of negative sample solution are respectively spotted on a thin layer plate, toluene-ethyl acetate-glacial acetic acid (14:4:0.5) is used as a developing agent, pre-equilibrium is carried out for 15 minutes, development is carried out, and then the thin layer plate is taken out, dried and observed under a UV light (365 nm). In the test sample chromatogram, the same color fluorescent spots appear at the corresponding positions of the control medicinal material. The method is simple in operation, fast and accurate in separation effect, good in reproducibility and strong in specificity.

Description

Technical Field

[0001] This invention belongs to the field of traditional Chinese medicine technology, specifically relating to a method for identifying jujubes in pediatric sac blood glucose syrup using thin-layer chromatography. Background Technology

[0002] The pediatric blood glucose syrup prescription consists of Rehmannia glutinosa, Dioscorea opposita, jujube, and ferrous sulfate. It has the effects of strengthening the spleen and stomach, nourishing blood and promoting body fluid production, and is used to treat nutritional anemia and iron-deficiency anemia in children. Rehmannia glutinosa can nourish blood and yin; jujube can benefit the middle jiao (spleen and stomach), nourish blood, strengthen the spleen and stomach, and calm the mind; Dioscorea opposita can nourish blood and essence, and nourish the stomach and lungs.

[0003] All of these herbs have blood-tonifying effects. The current standard for pediatric blood glucose syrup, WS3-B-2654-97, lacks thin-layer chromatography identification and content determination. A more effective thin-layer chromatography identification method is needed for more comprehensive and effective control of drug quality. Summary of the Invention

[0004] This invention provides a method for identifying jujubes in pediatric sac blood glucose syrup using thin-layer chromatography. This method is simple to operate, provides rapid and accurate separation, exhibits good stability, reproducibility, and specificity. The technical solution of this invention is as follows:

[0005] A method for identifying jujubes in pediatric sac blood glucose syrup using thin-layer chromatography includes the following steps:

[0006] (1) Preparation of the test solution:

[0007] Dilute the pediatric blood glucose syrup with water, adjust the pH to 2-3 with dilute hydrochloric acid, shake with ether, combine the ether solutions, evaporate the ether to dryness, dissolve the residue in methanol, and use it as the test solution.

[0008] (2) Preparation of control herbal solution:

[0009] Take jujube as a reference herb, decoct it in water, filter it, and prepare the reference herb solution by adding dilute hydrochloric acid to adjust the pH to 2-3, starting from step 1. (i.e., adjust the pH of the filtrate to 2-3 with dilute hydrochloric acid, shake with ether, combine the ether solutions, evaporate the ether, dissolve the residue in methanol, and prepare the reference herb solution.)

[0010] (3) Preparation of negative sample solution:

[0011] Take a negative sample that is missing jujubes and prepare a negative sample solution using the same method as in step (1);

[0012] (4) Identification by thin-layer chromatography:

[0013] Take equal volumes of the test solution, reference medicinal material solution, and negative sample solution from steps (1)-(3), and apply 5-10 μl of each to a thin-layer plate. Use toluene-ethyl acetate-glacial acetic acid (volume ratio 12-15:3.5-4.5:0.5) as the developing solvent. Pre-equilibrate for 12-20 minutes, develop, remove, air dry, and examine under ultraviolet light. In the chromatogram of the test sample, spots of the same color appear at the corresponding positions as in the chromatogram of the reference medicinal material.

[0014] Furthermore, a method for identifying jujubes in pediatric sac blood glucose syrup using thin-layer chromatography includes the following steps:

[0015] (1) Preparation of the test solution:

[0016] Take 20-30 ml of pediatric blood glucose syrup, dilute with 10 ml of water, adjust the pH to 2-3 with dilute hydrochloric acid, extract with ether 2-3 times, 30-50 ml each time, combine the ether extracts, evaporate the ether to dryness, dissolve the residue in 1-2 ml of methanol, and use it as the test solution.

[0017] (2) Preparation of control herbal solution:

[0018] Take 2-3g of jujube reference material, add 50-90ml of water, decoct for 25-45 minutes, filter, and prepare the reference material solution from the step 1 ("adjusting the pH to 2-3 with dilute hydrochloric acid") using the same method as the test sample. (That is, adjust the pH of the filtrate to 2-3 with dilute hydrochloric acid, extract with ether 2-3 times, 30-50ml each time, combine the ether extracts, evaporate the ether to dryness, dissolve the residue in 1-2ml of methanol, and prepare the reference material solution.)

[0019] (3) Preparation of negative sample solution:

[0020] Take a negative sample that is missing jujubes and prepare a negative sample solution using the same method as in step (1);

[0021] (4) Identification by thin-layer chromatography:

[0022] Take equal volumes of the test solution, control medicinal material solution, and negative sample solution from steps (1)-(3), and take 10 μl of each. Spot them onto a thin-layer plate. Use toluene-ethyl acetate-glacial acetic acid with a volume ratio of (12-15:3.5-4.5:0.5) as the developing solvent. Pre-equilibrate for 12-20 minutes, develop, remove, air dry, and examine under a UV lamp (365 nm).

[0023] Furthermore, a method for identifying jujubes in pediatric sac blood glucose syrup using thin-layer chromatography includes the following steps:

[0024] (1) Preparation of the test solution:

[0025] Take 20ml of pediatric blood glucose syrup, dilute with 10ml of water, adjust the pH to 2 with dilute hydrochloric acid, extract with 30ml of ether three times by shaking, combine the ether liquids, evaporate the ether to dryness, dissolve the residue in 1ml of methanol, and use it as the test solution.

[0026] (2) Preparation of control herbal solution:

[0027] Take 2g of jujube as a reference material, add 60ml of water, decoct for 30 minutes, filter, and prepare the reference material solution by adjusting the pH to 2 with dilute hydrochloric acid, starting from step 1; (i.e., adjust the pH to 2 with dilute hydrochloric acid, extract with ether three times, 30ml each time, combine the ether extracts, evaporate the ether to dryness, dissolve the residue in 1ml of methanol, and prepare the reference material solution;)

[0028] (3) Preparation of negative sample solution:

[0029] Take a negative sample that is missing jujubes and prepare a negative sample solution using the same method as in step (1);

[0030] (4) Identification by thin-layer chromatography:

[0031] Take equal volumes of the test solution, control medicinal material solution, and negative sample solution from steps (1)-(3), and take 10 μl of each. Spot them onto a thin-layer plate. Use toluene-ethyl acetate-glacial acetic acid with a volume ratio of 14:4:0.5 as the developing solvent. Pre-equilibrate for 15 minutes, develop, remove, air dry, and examine under a UV lamp (365 nm).

[0032] Preferably, the thin-layer plate mentioned in step (4) of the method of the present invention is a silicone GF254 thin-layer plate.

[0033] Preferably, the volume ratio of the developing agent toluene-ethyl acetate-glacial acetic acid in step (4) of the method of the present invention is 14:4:0.5.

[0034] The beneficial effects of this invention are: the method is simple to operate, produces clear spots, has a fast and accurate separation effect, good stability, good reproducibility, and strong specificity, and can be used as an identification method for jujubes in pediatric blood glucose syrup. Attached Figure Description

[0035] Figure 1 A is a thin-layer chromatography identification image of Comparative Example 1, and B is a thin-layer chromatography identification image of Example 1.

[0036] Figure 2 A is a thin-layer chromatography identification image of Comparative Example 1 after spotting for 3 minutes; B is a thin-layer chromatography identification image of Example 1 after spotting for 3 minutes.

[0037] Figure 3 A is a thin-layer chromatography identification image of Comparative Example 1, and B is a thin-layer chromatography identification image of Comparative Example 1.

[0038] Figure 4 A is a thin-layer chromatography identification image of Comparative Example 1 after 10 minutes of repeated spotting; B is a thin-layer chromatography identification image of Example 1 after 10 minutes of repeated spotting.

[0039] In the figure, 1-jujube reference material; 2-test sample (230401); 3-test sample (230701); 4-test sample (240301); 5-negative control without jujube. Detailed Implementation

[0040] Example 1

[0041] A method for identifying jujubes in pediatric sac blood glucose syrup using thin-layer chromatography includes the following steps:

[0042] (1) Preparation of the test solution:

[0043] Take 20ml of pediatric blood glucose syrup, dilute with 10ml of water, adjust the pH to 2 with dilute hydrochloric acid, extract with 30ml of ether three times by shaking, combine the ether liquids, evaporate the ether to dryness, dissolve the residue in 1ml of methanol, and use it as the test solution.

[0044] (2) Preparation of control herbal solution:

[0045] Take 2g of jujube reference material, add 60ml of water, decoct for 30 minutes, filter, add dilute hydrochloric acid to adjust the pH of the filtrate to 2, extract with ether three times, 30ml each time, combine the ether liquids, evaporate the ether, add 1ml of methanol to dissolve the residue, and prepare the reference material solution.

[0046] (3) Preparation of negative sample solution:

[0047] Take a negative sample that is missing jujubes and prepare a negative sample solution using the same method as in step (1);

[0048] (4) Identification by thin-layer chromatography:

[0049] Take equal volumes of the test solution, control drug solution, and negative sample solution from steps (1)-(3), and apply 10 μl of each to the same silica gel GF. 254 On a thin-layer chromatography plate, a toluene-ethyl acetate-glacial acetic acid solution (volume ratio 14:4:0.5) was used as the developing solvent. The plate was pre-equilibrated for 15 minutes, developed, removed, air-dried, and examined under ultraviolet light (365 nm). Figure 1-4 As can be seen from both, the test sample chromatogram shows spots of the same color at the corresponding positions as the reference medicinal material chromatogram.

[0050] Comparative Example 1

[0051] A method for identifying jujubes in pediatric sac blood glucose syrup using thin-layer chromatography includes the following steps:

[0052] (1) Preparation of the test solution:

[0053] Take 20ml of pediatric blood glucose syrup, dilute with 10ml of water, adjust the pH to 2 with dilute hydrochloric acid, extract with 30ml of ether three times by shaking, combine the ether liquids, evaporate the ether to dryness, dissolve the residue in 1ml of methanol, and use it as the test solution.

[0054] (2) Preparation of control herbal solution:

[0055] Take 2g of jujube as a reference herb, add 60ml of water, decoct for 30 minutes, filter, and prepare the reference herb solution by adding dilute hydrochloric acid to adjust the pH to 2, starting from the same treatment method as the test sample.

[0056] (3) Preparation of negative sample solution:

[0057] Take a negative sample that is missing jujubes and prepare a negative sample solution using the same method as in step (1);

[0058] (4) Identification by thin-layer chromatography:

[0059] Take equal volumes of the test solution, control medicinal material solution, and negative sample solution from steps (1)-(3), and take 10 μl of each. Spot them onto a thin-layer plate, pre-equilibrate with toluene-ethyl acetate-glacial acetic acid at a volume ratio of 14:4:0.5 for 15 minutes, develop, remove, air dry, and examine under a UV lamp (254 nm).

[0060] from Figure 1-4 It can be seen that by comparing the spectra of the jujube reference material, the test sample, and the negative sample, a relatively obvious bluish-purple fluorescent main spot appeared on the spectrum of the jujube reference material under 254nm wavelength ultraviolet light. However, this feature spot was very unstable and became difficult to observe after about 3 minutes. However, when examined under 365nm wavelength ultraviolet light, a relatively obvious and stable fluorescent feature spot was visible in both the jujube reference material and the test sample solution.

Claims

1. A method for identifying jujubes in pediatric sac blood glucose syrup using thin-layer chromatography, characterized in that... Includes the following steps: (1) Preparation of the test solution: Dilute the pediatric blood glucose syrup with water, adjust the pH to 2-3 with dilute hydrochloric acid, shake with ether, combine the ether solutions, evaporate the ether to dryness, dissolve the residue in methanol, and use it as the test solution. (2) Preparation of control herbal solution: Boil jujube as a reference material in water, filter, adjust the pH of the filtrate to 2-3 with dilute hydrochloric acid, shake with ether, combine the ether solutions, evaporate the ether, dissolve the residue in methanol to prepare the reference material solution. (3) Preparation of negative sample solution: Take a negative sample that is missing jujubes and prepare a negative sample solution using the same method as in step (1); (4) Identification by thin-layer chromatography: Take equal volumes of the test solution, control medicinal material solution, and negative sample solution from steps (1)-(3), and apply 5-10 μl of each to a thin-layer plate. Use toluene-ethyl acetate-glacial acetic acid (volume ratio 12-15:3.5-4.5:0.5) as the developing solvent. Pre-equilibrate for 12-20 minutes, develop, remove, air dry, and examine under ultraviolet light.

2. The method according to claim 1, characterized in that... Includes the following steps: (1) Preparation of the test solution: Take 20-30 ml of pediatric blood glucose syrup, dilute with 10 ml of water, adjust the pH to 2-3 with dilute hydrochloric acid, extract with ether 2-3 times, 30-50 ml each time, combine the ether extracts, evaporate the ether to dryness, dissolve the residue in 1-2 ml of methanol, and use it as the test solution. (2) Preparation of control herbal solution: Take 2-3g of jujube reference material, add 50-90ml of water, decoct for 25-45 minutes, filter, adjust the pH of the filtrate to 2-3 with dilute hydrochloric acid, extract with ether 2-3 times, 30-50ml each time, combine the ether extracts, evaporate the ether, add 1-2ml of methanol to dissolve the residue, and prepare the reference material solution. (3) Preparation of negative sample solution: Take a negative sample that is missing jujubes and prepare a negative sample solution using the same method as in step (1); (4) Identification by thin-layer chromatography: Take equal volumes of the test solution, control medicinal material solution, and negative sample solution from steps (1)-(3), and take 10 μl of each. Spot them onto a thin-layer plate. Use toluene-ethyl acetate-glacial acetic acid with a volume ratio of (12-15:3.5-4.5:0.5) as the developing solvent. Pre-equilibrate for 12-20 minutes, develop, remove, air dry, and examine under a UV lamp (365 nm).

3. The method according to claim 1 or 2, characterized in that... Includes the following steps: (1) Preparation of the test solution: Take 20ml of pediatric blood glucose syrup, dilute with 10ml of water, adjust the pH to 2 with dilute hydrochloric acid, extract with 30ml of ether three times by shaking, combine the ether liquids, evaporate the ether to dryness, dissolve the residue in 1ml of methanol, and use it as the test solution. (2) Preparation of control herbal solution: Take 2g of jujube reference material, add 60ml of water, decoct for 30 minutes, filter, add dilute hydrochloric acid to adjust the pH of the filtrate to 2, extract with ether three times, 30ml each time, combine the ether liquids, evaporate the ether, add 1ml of methanol to dissolve the residue, and prepare the reference material solution. (3) Preparation of negative sample solution: Take a negative sample that is missing jujubes and prepare a negative sample solution using the same method as in step (1); (4) Identification by thin-layer chromatography: Take equal volumes of the test solution, control medicinal material solution, and negative sample solution from steps (1)-(3), and take 10 μl of each. Spot them onto a thin-layer plate. Use toluene-ethyl acetate-glacial acetic acid with a volume ratio of 14:4:0.5 as the developing solvent. Pre-equilibrate for 15 minutes, develop, remove, air dry, and examine under a UV lamp (365 nm).

4. The method according to any one of claims 1-3, characterized in that, The thin-layer plate is silicone GF. 254 Thin-layer plate.

5. The method according to claim 1 or 2, characterized in that, The volume ratio of the developing solvent toluene-ethyl acetate-glacial acetic acid is 14:4:0.5.

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