CN114397296A - Method for identifying ginseng age - Google Patents
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- CN114397296A CN114397296A CN202111642084.5A CN202111642084A CN114397296A CN 114397296 A CN114397296 A CN 114397296A CN 202111642084 A CN202111642084 A CN 202111642084A CN 114397296 A CN114397296 A CN 114397296A
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- 241000208340 Araliaceae Species 0.000 title claims abstract description 63
- 235000005035 Panax pseudoginseng ssp. pseudoginseng Nutrition 0.000 title claims abstract description 63
- 235000003140 Panax quinquefolius Nutrition 0.000 title claims abstract description 63
- 235000008434 ginseng Nutrition 0.000 title claims abstract description 63
- 238000000034 method Methods 0.000 title claims abstract description 16
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims abstract description 24
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 23
- 239000011521 glass Substances 0.000 claims abstract description 14
- XOSXWYQMOYSSKB-LDKJGXKFSA-L water blue Chemical compound CC1=CC(/C(\C(C=C2)=CC=C2NC(C=C2)=CC=C2S([O-])(=O)=O)=C(\C=C2)/C=C/C\2=N\C(C=C2)=CC=C2S([O-])(=O)=O)=CC(S(O)(=O)=O)=C1N.[Na+].[Na+] XOSXWYQMOYSSKB-LDKJGXKFSA-L 0.000 claims abstract description 9
- JPYHHZQJCSQRJY-UHFFFAOYSA-N Phloroglucinol Natural products CCC=CCC=CCC=CCC=CCCCCC(=O)C1=C(O)C=C(O)C=C1O JPYHHZQJCSQRJY-UHFFFAOYSA-N 0.000 claims abstract description 7
- QCDYQQDYXPDABM-UHFFFAOYSA-N phloroglucinol Chemical compound OC1=CC(O)=CC(O)=C1 QCDYQQDYXPDABM-UHFFFAOYSA-N 0.000 claims abstract description 7
- 229960001553 phloroglucinol Drugs 0.000 claims abstract description 7
- 238000004043 dyeing Methods 0.000 claims abstract description 5
- 229940126680 traditional chinese medicines Drugs 0.000 abstract description 2
- 238000010186 staining Methods 0.000 description 11
- 235000014676 Phragmites communis Nutrition 0.000 description 10
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 9
- 239000003153 chemical reaction reagent Substances 0.000 description 9
- 240000002853 Nelumbo nucifera Species 0.000 description 7
- 235000006508 Nelumbo nucifera Nutrition 0.000 description 7
- 235000006510 Nelumbo pentapetala Nutrition 0.000 description 7
- 239000002250 absorbent Substances 0.000 description 6
- 230000002745 absorbent Effects 0.000 description 6
- 239000003814 drug Substances 0.000 description 5
- 239000000463 material Substances 0.000 description 5
- 238000002474 experimental method Methods 0.000 description 4
- 238000010586 diagram Methods 0.000 description 3
- 238000002791 soaking Methods 0.000 description 3
- 238000005406 washing Methods 0.000 description 3
- 241000196324 Embryophyta Species 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- DFOATKRXQLALOL-UHFFFAOYSA-N 2-(2-hydroxyethyl)benzene-1,3,5-triol Chemical compound OCCC1=C(O)C=C(O)C=C1O DFOATKRXQLALOL-UHFFFAOYSA-N 0.000 description 1
- 230000001788 irregular Effects 0.000 description 1
Images
Classifications
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/84—Systems specially adapted for particular applications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/84—Systems specially adapted for particular applications
- G01N2021/8466—Investigation of vegetal material, e.g. leaves, plants, fruits
Abstract
The invention relates to the field of traditional Chinese medicines, in particular to a method for identifying the age of ginseng, which comprises the steps of selecting a main root part of ginseng to slice, wherein the thickness of the slice is 0.2-2 mm, placing the slice on a glass slide, dripping 200-500ul of concentrated hydrochloric acid with the weight of 37% on the surface of the slice, completely covering the surface of the whole slice by the concentrated hydrochloric acid, dripping 200-500ul of phloroglucinol alcohol solution with the weight of 25% on the surface of the slice for dyeing after 1-3 minutes, dripping 200-500ul of aniline blue with the weight of 1% on the surface of the slice for counterdyeing after 1-3 minutes, observing the dyed slice by using a body-scope, determining a central point and a cambium layer on the slice, and calculating the total number of annual rings between the central point and the cambium layer to obtain the age of the ginseng.
Description
Technical Field
The invention relates to the field of traditional Chinese medicines, in particular to a method for identifying ginseng age.
Background
At present, the traditional method for years identification of ginseng, which flows for thousands of years, determines the age limit of the ginseng according to the reed head length of the ginseng and the number of reed bowls, so that the age limit identification is subjective, and the method for observing the reed bowls with reed heads is difficult to operate due to the fact that the types of the reed heads of the ginseng are various, namely round reed, stacked reed flowers, two sections of reed, contracted reed and the like, and the experience requirements of an identifier are extremely high and the method does not have universality. The market needs a method for identifying the age limit of the ginseng, which is relatively accurate, strong in operability, simple and rapid.
Disclosure of Invention
The invention aims to provide a method for identifying the age of ginseng.
Specifically, the first aspect of the invention provides a method for identifying the age of ginseng, which comprises the steps of selecting a main root part of ginseng to be sliced, wherein the thickness of the slice is 0.2-2 mm, placing the slice on a glass slide, dripping 200-500ul of 37% wt concentrated hydrochloric acid on the surface of the slice, completely covering the surface of the whole slice by the concentrated hydrochloric acid, dripping 200-500ul of 25% wt phloroglucinol ethanol solution on the surface of the slice for dyeing after 2-3 minutes, dripping 200-500ul of 1% wt aniline blue on the surface of the slice for counterdyeing after 2-3 minutes, observing the dyed slice by using a body mirror, determining a central point and a cambium on the slice, and calculating the total number of annual rings between the central point and the cambium to obtain the age of the ginseng.
In a preferred embodiment, the ginseng is selected from the group consisting of common garden ginseng, wild ginseng, or mountain ginseng.
In another preferred example, the magnification of the stereoscope is 2-4 times.
The details of various aspects of the invention are set forth in subsequent sections. The features, objects, and advantages of the invention will be apparent from the description and from the claims.
Drawings
FIG. 1: cambium, annual ring and center point of ginseng slice are shown schematically.
FIG. 2: the staining of cross-sectional slices of Yuansheng produced in Jiangxi is shown schematically, S is cambium, and A-E represent annual ring number of 5 years in total.
FIG. 3: the cross section of the ginseng from Jilin garden is stained schematically, S is cambium, and A-F represent annual ring number of 6 years in total.
FIG. 4: the cross section of the wild ginseng is dyed in a schematic view in 1-3 years under the forest, S is a cambium, and A-C represent the number of turns of different annual rings; wherein:
4a is a schematic cross-sectional slice dyeing diagram of the mountain ginseng under a 1-year-old forest, S is a cambium, and A represents the number of annual rings of 1 year;
4b is a schematic cross-sectional slice staining diagram of the mountain ginseng under the 2-year-old forest, S is a cambium, and A, B represents the number of turns of the 2-year annual ring;
4c is a schematic cross-sectional section staining diagram of the mountain ginseng under the 3-year-old forest, S is a cambium, and the number of rounds of the 3-year-old generation is A, B, C.
FIG. 5: schematic cross section staining of 9-year-old mountain ginseng.
FIG. 6: schematic cross section staining of 17-year-old mountain ginseng in forest.
Detailed Description
The invention will be further illustrated with reference to the following specific examples. It should be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention. The experimental procedures, in which specific conditions are not noted in the following examples, are generally carried out according to conventional conditions or according to conditions recommended by the manufacturers. All percentages, ratios, proportions, or parts are by weight unless otherwise specified.
Those skilled in the art will know how to determine the centroids and cambium of sliced ginseng according to the teaching of the textbook of medicinal plant science in the field (natural pharmacology (second edition) Roohai, Yao Ronglin; medicinal plant science (Chinese medicine speciality) Dingjing and Suo Shuo, Shanghai science and technology Press, 1987).
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. In addition, any methods and materials similar or equivalent to those described herein can be used in the methods of the present invention. The preferred embodiments and materials described herein are intended to be exemplary only.
The features mentioned above with reference to the invention, or the features mentioned with reference to the embodiments, can be combined arbitrarily. All the features disclosed in this specification may be combined in any combination and each feature disclosed in this specification may be replaced by alternative features serving the same, equivalent or similar purpose. Thus, unless expressly stated otherwise, the features disclosed are merely generic examples of equivalent or similar features.
Example one identification of growth years of general Yuanshen
Experimental materials: collecting 22 garden ginseng samples of 4 places related to Jiangxi, Heilongjiang, Liaoning and Jilin.
Laboratory instruments and reagents: nikon SM2800N stereomicroscope (Shanghai university of medicine laboratory platform center), ethanol, concentrated hydrochloric acid, phloroglucinol reagent, aniline blue were purchased from the exploration reagent website.
The experimental method comprises the following steps:
1) collecting Ginseng radix slices returned to different pharmacies of a laboratory, and soaking slices of about 0.2-2 mm in 75% wt ethanol for about half an hour.
2) Taking out the soaked ginseng slices, sucking off the redundant alcohol solution by using absorbent paper, and then flatly spreading the ginseng slices on a glass slide for later use.
3) Sucking about 200-500ul of prepared concentrated hydrochloric acid (37 wt% concentration) with a glass dropper to completely cover the ginseng slices, and after 2 minutes, sucking about 200-500ul of 25 wt% phloroglucinol solution with a glass dropper to further dye the ginseng slices. Wait for 2 minutes.
4) And dripping 1 wt% aniline blue solution on the ginseng slices, waiting for 2 minutes, quickly washing off redundant dye by using 95 wt% ethanol solution after the time, wiping the slices cleanly, and placing the slices on glass slides. After staining for 2 minutes, the excess solution was blotted off with absorbent paper, a drop of glycerol was dropped, a cover slip was covered, and the slide was placed under a scope (2-4 times) for observation.
5) Firstly, confirming a cambium on a slice, observing and confirming a central point, counting the annual ring number formed by red catheter bundle cells one by one from the cambium to the central area, and obtaining the number obtained by counting, namely the growth period of the observed ginseng sample.
The experimental results are as follows: except for the fuzzy recognition, all the other samples meet the growth period, and the total meeting rate is 95.4%, which is shown in the following table.
TABLE 1
| Year of cultivation | Number of samples | Collecting the source | Color matched number | Remarks for note |
| 5-6 years old | 6 | (Jiangxi) | 6 | Example of the drawings 2 |
| 5-6 years old | 5 | Heilongjiang | 4 | 1 case of fuzzy recognition |
| 5-6 years old | 7 | Liaoning medicine | 7 | Conform to |
| 5-6 years old | 4 | (Jilin) | 4 | Example of the drawings 3 |
Example two identification of growth years of 1-3 year old wild ginseng under forest
Experimental materials: taking ginseng samples which grow for 1-3 years year one by one in the early stage of the northeast ginseng base, and carrying out section staining. The aim is to observe and verify the scientificity and feasibility of the method.
Laboratory instruments and reagents: nikon SM2800N stereomicroscope (Shanghai university of medicine laboratory platform center), ethanol, concentrated hydrochloric acid, phloroglucinol reagent, aniline blue were purchased from the exploration reagent website.
The experimental method comprises the following steps:
1. collecting Ginseng radix slices of different pharmacies in a laboratory of northeast Ginseng radix base, wherein the growth period of the slices is about 0.2-2 mm in 1 year, 2 years and 3 years, and soaking the slices in 75 wt% ethanol for about half an hour.
2. Taking out soaked Ginseng radix slice, removing excessive alcohol solution with absorbent paper, cutting into slices with thickness of about 500um-1mm with thin slices, and spreading on glass slide.
3. The prepared concentrated hydrochloric acid (37% wt concentration) was pipetted with a glass pipette at about 200-. Wait for 3 minutes.
4. And dripping 1 wt% aniline blue solution on the ginseng slices, waiting for 2 minutes, quickly washing off redundant dye by using 95 wt% ethanol solution after the time, wiping the slices cleanly, and placing the slices on glass slides.
5. After 3 minutes of staining, excess solution was blotted with absorbent paper, a drop of glycerol was dropped, a cover slip was covered, and the slide was placed under a scope (2-4 times) for observation.
6. The cambium of the slice is confirmed, the center point is observed and confirmed, the annual ring number formed by red ductal tract cells is counted one by one from the cambium to the center area, and the counted number is the growth age of the observed ginseng sample (for example, fig. 4a, fig. 4b and fig. 4 c).
The experimental results are as follows: in total, 34 samples, 32 in total, met actual year with a 94.1% match rate, except that one pattern was not standard and one staining failed to result in poor recognition. See table below:
TABLE 2
| Year of cultivation | Number of samples | Collecting the source | Color matched number | Remarks for note |
| 1 | 15 | Northeast | 15 | Conform to |
| 2 | 12 | Northeast | 11 | 1 example graph is not standard |
| 3 | 7 | Northeast | 6 | 1 example is unclear |
EXAMPLE III identification of growth years of under-forest Ginseng
Experimental materials: taking 10-20 years of wild ginseng from a Leyun Shandong mountain ginseng base, slicing, observing, and extracting the wild ginseng from the Leyun Shandong forest for different years.
Laboratory instruments and reagents: nikon SM2800N stereomicroscope (Shanghai university of medicine laboratory platform center), ethanol, concentrated hydrochloric acid, phloroglucinol reagent, aniline blue were purchased from the exploration reagent website.
The experimental method comprises the following steps:
1) collecting sun-dried ginseng from the forest of different years, cutting with guillotine, collecting main root part of Ginseng radix with length of 2CM at the position of 1/2-1/3 between the reed head and the branch leg, and soaking in 75% wt ethanol for about 2 hr.
2) Taking out the soaked ginseng sample, manually slicing the ginseng sample to a slice thickness of 0.2-2 mm, and cutting the ginseng sample by using a thin blade to ensure that the slice is as light and thin as possible and is easy to color. Absorbing the excessive alcohol solution by using absorbent paper, and then spreading the solution on a glass slide for later use.
3) Sucking about 200-500ul of prepared concentrated hydrochloric acid (37 wt% concentration) with a glass dropper to completely cover the ginseng slices, sucking about 200-500ul of 25 wt% phloroglucinol solution with a glass dropper after 3 minutes, further dyeing, and waiting for 3 minutes.
4) And dripping 1 wt% aniline blue solution on the ginseng slices, waiting for 2 minutes, quickly washing off redundant dye by using 95 wt% ethanol solution after the time, wiping the slices cleanly, and placing the slices on glass slides.
5) After 3 minutes of staining, excess solution was blotted with absorbent paper, a drop of glycerol was dropped, a cover slip was covered, and the slide was placed under a scope (2-4 times) for observation.
6) Firstly, confirming a cambium on a slice, observing and confirming a central point, counting the annual ring number formed by red catheter bundle cells one by one from the cambium to the central area, and obtaining the number obtained by counting, namely the growth period of the observed ginseng sample.
The experimental results are as follows: except for one case of difficulty in interpretation caused by irregular annual ring characters, the counting of another 2 cases has errors, and the total coincidence rate is 90.3%, which is shown in the following table. A partial staining picture is shown in fig. 5.
TABLE 3
| Sample classes | Year of cultivation | Collecting the source | Number of samples | Year identification |
| WG-10 | 10 | Liaoning seed of lotus | 4 | 1 example 9 years old |
| WG-12 | 12 | Liaoning seed of lotus | 5 | Conform to |
| WG-13 | 13 | Liaoning seed of lotus | 6 | 1 example 12 years old |
| WG-15 | 15 | Liaoning seed of lotus | 2 | Conform to |
| WG-17 | 17 | Liaoning seed of lotus | 6 | 1 example of shape difficult to interpret |
| WG-19 | 19 | Liaoning seed of lotus | 5 | Conform to |
| WG-20 | 20 | Liaoning seed of lotus | 3 | Conform to |
The various aspects of the invention are addressed above. It should be understood, however, that equivalent changes and modifications may be made thereto by those skilled in the art without departing from the spirit of the present invention, and that such changes and modifications are intended to be covered by the appended claims.
Claims (3)
1. A method for identifying the age of ginseng comprises the steps of selecting a main root part of ginseng to be sliced, wherein the thickness of the sliced part is 0.2-2 mm, placing the sliced part on a glass slide, dropwise adding 200-500ul of 37% wt concentrated hydrochloric acid on the surface of the sliced part, wherein the concentrated hydrochloric acid completely covers the surface of the whole sliced part, dropwise adding 200-500ul of 25% wt phloroglucinol alcohol solution on the surface of the sliced part for dyeing after 1-3 minutes, dropwise adding 200-500ul of 1% wt aniline blue on the surface of the sliced part for counterdyeing after 1-3 minutes, observing the dyed sliced part by using a body-scope, determining a central point and a cambium on the sliced part, and calculating the total number of annual rings between the central point and the cambium to obtain the age of the ginseng.
2. The method of claim 1, wherein the ginseng is selected from the group consisting of common garden ginseng, wild ginseng, or mountain ginseng.
3. The method according to claim 1, wherein the magnification of the stereoscope is 2-4 times.
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Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1994003065A1 (en) * | 1992-07-31 | 1994-02-17 | The University Of Sydney | Biocontrol of fungal infection using pseudomonas strains |
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| CN110174298A (en) * | 2019-04-30 | 2019-08-27 | 长江大学 | A kind of colouring method of observation of plant internal microstructure |
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-
2021
- 2021-12-29 CN CN202111642084.5A patent/CN114397296A/en active Pending
Patent Citations (8)
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| US20090049568A1 (en) * | 2004-07-28 | 2009-02-19 | Plant Research International B.V. | Method for preventing dehiscence and altering plant lignification |
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