Method for in-situ nondestructive identification of geo-geography of traditional Chinese medicinal materials

Abstract

The invention provides a method for in-situ nondestructive identification of geo-geography of traditional Chinese medicinal materials. By performing in-situ MRS / MRSI measurement on the traditional Chinese medicinal materials, comprehensive and accurate evaluation can be performed on the geo-geography of the traditional Chinese medicinal materials in combination with histomorphological characteristics and effective active component and / or metabolite characteristics of the traditional Chinese medicinal materials.

Description

Technical Field

[0001] The invention relates to the technical field of Chinese medicinal materials, and in particular to a method for screening authentic Chinese medicinal materials. Background Art

[0002] Authentic Chinese medicinal materials are synonymous with traditional high-quality Chinese medicinal materials in my country. The so-called authentic Chinese medicinal materials refer to those that have been selected through long-term clinical application of traditional Chinese medicine, are produced in a specific region, and have better quality and efficacy than the same Chinese medicinal materials produced in other regions, and have stable quality and high reputation.

[0003] Modern Chinese medicinal materials tend to be planted on a large scale and intensively. Authenticity, as a synonym for evaluating the origin and quality of medicinal materials, is increasingly valued by consumers. However, the traditional method of identifying the authenticity of Chinese medicinal materials is mainly through naked eye observation, which is highly subjective and difficult to quantify. Modern analytical technology mainly analyzes the quality of Chinese medicinal materials through the determination of the content of effective (index) components and fingerprint technology. However, the determination of the content of limited components cannot fully and accurately reflect the quality of medicinal materials, and fingerprint technology can only perform similarity evaluation after extracting common peaks, and cannot associate similarity with quality.

[0004] Therefore, there is still a lack of effective evaluation methods and systems for the authenticity of Chinese medicinal materials. Summary of the invention

[0005] In view of the above-mentioned problems in the prior art, the present invention aims to provide an in-situ, non-destructive method for screening authentic Chinese medicinal materials, which can comprehensively and accurately evaluate the authenticity of Chinese medicinal materials by combining the tissue morphological characteristics and the characteristics of effective active ingredients and / or metabolites of Chinese medicinal materials.

[0006] The technical solution of the present invention is as follows.

[0007] The present invention provides a method for in-situ and non-destructive identification of the authenticity of Chinese medicinal materials, comprising the following steps:

[0008] Step 1: In-situ measurement of magnetic resonance imaging (MRI) images of authentic Chinese medicinal materials: covering at least one part or the whole of the authentic Chinese medicinal materials with an effective imaging area of ​​a magnetic resonance apparatus; performing magnetic resonance imaging on the part or the whole of the authentic Chinese medicinal materials using a radio frequency coil and gradient encoding to obtain an MRI image composed of black, white and grayscale, wherein the selected part or the whole is divided into virtual grids to determine the tissue morphological characteristics of each grid;

[0009] Meanwhile, specific grids are selected for magnetic resonance spectroscopy and spectroscopic imaging (MRS / MRSI) measurements to obtain NMR chemical shift / signal intensity information of the effective active ingredients and / or metabolites of traditional Chinese medicine materials within the corresponding grids, thereby establishing the corresponding relationship between the NMR chemical shift / signal intensity information and the morphological characteristics of traditional Chinese medicine tissues;

[0010] Step 2. For the traditional Chinese medicine material to be tested, repeat the in-situ MRI and MRS / MRSI measurements in Step 1;

[0011] Step 3. According to the comparison of the MRI images obtained in Step 1 and Step 2, grids with the same or similar morphological characteristics of the genuine traditional Chinese medicine materials and the traditional Chinese medicine materials to be tested are obtained, and the NMR chemical shift / signal intensity information of the grids is compared, thereby identifying the genuineness of the traditional Chinese medicine materials.

[0012] In some embodiments, a spin echo pulse sequence is used for the radiofrequency coil; three gradient coils, namely X, Y, and Z, are used to achieve spatial Gx, Gy, and Gz gradient encodings respectively or simultaneously in the corresponding three dimensions. Gx, Gy, and Gz each have an MRS / MRSI spatial selection gradient and a three-dimensional MRSI imaging gradient encoding composed of N1, N2, or N3 pulse cycles at the same time sequence, where N1, N2, and N3 are integers from 1 to 128.

[0013] Through the spatial encoding sequences Gx, Gy, and Gz, a three-dimensional image of the whole or part of the traditional Chinese medicine material composed of N1×N2×N3 virtual grids can be obtained. When N3 = 1, a two-dimensional image of N1×N2 can be obtained.

[0014] In some embodiments, a structural imaging sequence is used for MRI measurements; an in-situ spectroscopy measurement sequence is used for MRS / MRSI measurements.

[0015] In some embodiments, the genuineness of the traditional Chinese medicine materials is identified based on the comparison of the NMR chemical shift / signal intensity information of the corresponding regions or parts of the genuine traditional Chinese medicine materials and the traditional Chinese medicine materials to be tested, where the corresponding regions or parts include multiple corresponding grids.

[0016] In some embodiments, the genuineness of the traditional Chinese medicine materials is identified based on the comparison of the NMR chemical shift / signal intensity information of two or more parts.

[0017] In some embodiments, the parts include roots, stems, leaves, flowers, fruits, and seeds.

[0018] In some embodiments, in Step 2, according to the differences in shape and / or size between the traditional Chinese medicine material to be tested and the genuine traditional Chinese medicine materials, the virtual grid division is adjusted in terms of the size and orientation of the grids.

[0019] In some embodiments, the grid size is selected in the range of 0.1 mm × 0.1 mm to 50 mm × 50 mm.

[0020] In some embodiments, in-situ MRI and MRS / MRSI measurements are performed on Chinese medicinal herbs at different growth stages to obtain the corresponding relationship between the NMR chemical shift / signal intensity information and the tissue morphological characteristics of Chinese medicinal herbs at different growth stages.

[0021] In some embodiments, the Chinese medicinal herbs include Dendrobium officinale, Polygonatum sibiricum, or Gastrodia elata.

[0022] In some embodiments, the Chinese medicinal herb is Dendrobium officinale, and the genuine Chinese medicinal herb is Dendrobium huoshanense; alternatively, the Chinese medicinal herb is Polygonatum sibiricum, and the genuine Chinese medicinal herb is Polygonatum cyrtonema; or the Chinese medicinal herb is Gastrodia elata, and the genuine Chinese medicinal herb is Gastrodia elata Bl. f. glauca S. Chow.

[0023] The method for identifying the genuineness of Chinese medicinal herbs of the present invention has at least one of the following advantages:

[0024] The method of the present invention is an in-situ and non-destructive method, and does not require invasive sampling treatment of Chinese herbal medicines;

[0025] The present invention can comprehensively evaluate the genuineness of Chinese medicinal herbs by combining MRI and MRS / MRSI data;

[0026] The present invention can simultaneously compare multiple parts of Chinese medicinal herbs, so as to comprehensively and accurately judge the genuineness of Chinese medicinal herbs;

[0027] The present invention can measure the MRI and MRS data of Chinese medicinal herbs at different growth stages to obtain dynamic information. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] Figure 1 is a pulse sequence diagram according to an embodiment of the present invention;

[0029] Figure 2 is a comparison diagram of MRI images and MRS of roots, stems, and leaves of different species of Dendrobium measured by the method of the present invention.

[0030] Figures 3 - 5 is a comparison diagram of MRS of roots, stems, and leaves of different species of Polygonatum sibiricum measured by the method of the present invention.

[0031] Figure 6 is a comparison diagram of MRS of tubers of different species of Gastrodia elata measured by the method of the present invention. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0032] Hereinafter, taking Dendrobium officinale, Polygonatum sibiricum, and Gastrodia elata as representative Chinese herbal medicines and referring to the accompanying drawings, the present invention will be further described in detail.

[0033] Embodiment 1

[0034] Dendrobium is a common Chinese herbal medicine, which has the effects of promoting the production of body fluid to benefit the stomach, nourishing yin and clearing heat, and regulating yin and yang. Different Dendrobiums vary in quality and efficacy due to different growth environments. At present, Huoshan Dendrobium is recognized as a genuine medicinal material. A large number of studies have also been carried out on the effective active ingredients of Huoshan Dendrobium.

[0035] For example, studies have shown that the rhizomes of Huoshan Dendrobium have the following chemical components:

[0036]

[0037] The relationship between different chemical components and pharmacological effects in Huoshan Dendrobium is shown in the following table.

[0038]

[0039] Although these research works have revealed some effective components and the structure-activity relationships of Dendrobium, however, due to the large number of active ingredients and the complex structure-activity relationships, it is difficult to directly use them to identify the genuineness of Dendrobium medicinal materials.

[0040] The present invention combines magnetic resonance imaging (MRI) and magnetic resonance spectroscopy and spectroscopic imaging (MRS / MRSI) measurement techniques to comprehensively analyze the tissue morphological characteristics and effective active ingredient and / or metabolite characteristics of Dendrobium, so as to give an in-situ, non-destructive and easy-to-implement identification method for judging the genuineness of Dendrobium medicinal materials by comparing the MRI images of specific parts (regions) with the NMR chemical shift / signal intensity information.

[0041] For this purpose, the present invention selects three kinds of Dendrobium medicinal materials: Huoshan Dendrobium, Tongpi Dendrobium and Tiepi Dendrobium, and conducts MRSI / MRS measurements respectively, and judges the genuineness of the medicinal materials according to the comparison results of the NMR chemical shift / signal intensity information of the corresponding parts (regions) of different Dendrobiums.

[0042] First, take different Dendrobium medicinal materials, and put the whole or part of the regions into the coil of the magnetic resonance instrument for MRSI / MRS measurement.

[0043] Figure 1 It is a pulse sequence diagram according to an embodiment of the present invention. Among them, for the radiofrequency coil, a common spin echo pulse sequence (RF sequence) is adopted; for the gradient coil, spatial encoding sequences Gx, Gy and Gz are adopted in the three dimensions of X, Y and Z. Among them, each spatial encoding sequence includes an MRS / MRSI spatial selection gradient and an MRSI imaging encoding gradient. As Figure 1As shown, the MRSI imaging encoding gradients in different dimensions have a cycle consisting of N pulses (N - order cyclic pulses) at the same time sequence. Among them, the MRSI imaging encoding gradient X has an N1 - order cyclic pulse, the MRSI imaging encoding gradient Y has an N2 - order cyclic pulse, and the MRSI imaging encoding gradient Z has an N3 - order cyclic pulse. Although Figure 1 shows that N1 = N2 = N3 = 6, however, N1, N2, and N3 can be different and can each be any integer from 1 to 128. By selecting N1, N2, and N3, a three - dimensional image composed of N1×N2×N3 virtual grids of the whole or part of the medicinal material can be obtained.

[0044] Figure 2 Figure for comparing the MRI images (left) of different species of Dendrobium and the MRS (right) of the root, stem, and leaf parts measured by the method according to the present invention. Among them, for the leaf and stem, N1 = N2 = 3 and N3 = 1 are selected, and for the root, N1 = N2 = 2 and N3 = 1 are selected. And for the root, stem, and leaf, the most representative grids are respectively selected, and the NMR information of the selected grids is compared (see the NMR spectra on the right). From Figure 2 it can be seen that for the root part, Huoshan Dendrobium has multiple significant characteristic peaks at chemical shifts near 1.25 ppm, 2 ppm, and 3.4 - 3.8 ppm, while the peaks of Cupid Dendrobium and Tiepi Dendrobium at the corresponding chemical shifts are much smaller. For the stem part, the characteristic peaks of Huoshan Dendrobium at chemical shifts of 3.4 - 3.8 ppm are still significant, while the characteristic peaks of Cupid Dendrobium and Tiepi Dendrobium at chemical shifts of 3.4 - 3.8 ppm are significantly smaller than those of Huoshan Dendrobium, but the differences in the NMR peaks of the three at chemical shifts near 1.25 ppm and 2 ppm are not obvious. For the leaf part, the characteristic peak of Tiepi Dendrobium near 1.25 ppm is significantly smaller than those of Huoshan Dendrobium and Cupid Dendrobium, and the characteristic peaks at other chemical shifts are not much different.

[0045] It can be seen that different Dendrobium species have similar NMR characteristic peak positions (similar chemical shifts of the characteristic peaks) in the same / similar regions (grids) of the tissue morphological characteristics of the same part, but the intensities of the characteristic peaks are different. Among them, the intensity of the characteristic peaks of Huoshan Dendrobium is significantly higher than that of Cupid Dendrobium and Tiepi Dendrobium, especially in the root and stem parts. Generally speaking, from the three parts, the weighted characteristic peak intensity of Huoshan Dendrobium is also the highest, that is, the content of the effective active ingredients and / or metabolites corresponding to the characteristic peaks is also the highest. This is consistent with the record in the Chinese Pharmacopoeia that Huoshan Dendrobium is a genuine medicinal material.

[0046] Therefore, by comparing the same / similar regions (grids) of the tissue morphological characteristics of the same part of the three Dendrobium species, the differences between different Dendrobium varieties can be shown very intuitively, thus giving a criterion for judging the genuineness of medicinal materials.

[0047] It should be noted that for different varieties of Dendrobium, more than two regions (grids) of the same part can be selected for comparison to more accurately determine the authenticity of medicinal materials.

[0048] Embodiment 2

[0049] In the same way as in Embodiment 1, MRS measurements were performed on the roots, stems, and leaves of three different species of Polygonatum sibiricum (Polygonatum kingianum, Polygonatum cyrtonema, and Polygonatum sibiricum). Figures 3 - 5 It shows the MRS comparison diagrams of the roots, stems, and leaves of different species of Polygonatum sibiricum measured by the method of the present invention.

[0050] The regions (grids) with the same / similar tissue morphological characteristics in the same part of different Polygonatum sibiricum have similar NMR characteristic peak positions (similar chemical shifts of the characteristic peaks), but the intensities of the characteristic peaks are different. Among them, at the rhizome, the main medicinal part of Polygonatum sibiricum, the intensity of the characteristic peaks of Polygonatum cyrtonema is significantly higher than that of Polygonatum kingianum and Polygonatum sibiricum. This is consistent with the record in the Chinese Pharmacopoeia that Polygonatum cyrtonema is a genuine medicinal material.

[0051] Embodiment 3

[0052] In the same way as in Embodiment 1, MRS measurements were performed on the tubers (medicinal parts) of three different species of Gastrodia elata (Gastrodia elata Bl. f. glauca S. Chow, Gastrodia elata Bl. f. elata, and Gastrodia elata Bl. f. alba). Figure 6 It shows the MRS comparison diagrams of the tubers of different species of Gastrodia elata.

[0053] The regions (grids) with the same / similar tissue morphological characteristics in the same part of different Gastrodia elata have similar NMR characteristic peak positions (similar chemical shifts of the characteristic peaks), but the intensities of the characteristic peaks at different chemical shifts are significantly different. Among them, the characteristic peaks of Gastrodia elata Bl. f. elata near 3.8 ppm are significantly higher than those of Gastrodia elata Bl. f. glauca S. Chow and Gastrodia elata Bl. f. alba. It is judged that the characteristic peaks near 3.8 ppm represent the main active ingredients of Gastrodia elata.

[0054] The MRSI / MRS measurement method of the present invention has the characteristics of in-situ, non-destructive, and fast, can be widely applied to various Chinese medicinal materials, and has the feasibility of carrying out operations in the fields such as Chinese medicinal material planting bases and forests in the wild, providing an effective and accurate method for identifying the authenticity of medicinal materials.

[0055] The above are only the preferred embodiments of the present invention, and do not limit the protection scope of the invention. Any modification, replacement, or improvement within the spirit and principle of the present invention is included in the protection scope of the present invention.

Claims

1. A method for in-situ, non-destructive identification of the authenticity of Chinese medicinal materials, comprising the following steps: Step 1: In-situ measurement of magnetic resonance imaging (MRI) images of authentic Chinese medicinal materials: using an effective imaging area of ​​a magnetic resonance imager to cover at least one part or the whole of the authentic Chinese medicinal materials; using a radio frequency coil and gradient encoding to perform magnetic resonance imaging on the part or the whole of the authentic Chinese medicinal materials to obtain an MRI image composed of black, white and grayscale, wherein the selected part or the whole is divided into virtual grids to determine the tissue morphological characteristics of each grid; At the same time, a specific grid is selected to perform magnetic resonance spectroscopy and spectral imaging (MRS / MRSI) measurements to obtain the nuclear magnetic resonance (NMR) chemical shift and corresponding signal intensity information of the effective active ingredients and / or metabolites of the Chinese herbal medicines in the corresponding grid, thereby establishing the corresponding relationship between the NMR chemical shift / signal intensity information and the histomorphological characteristics of the Chinese herbal medicines; Step 2: Repeat the in situ MRI and MRS / MRSI measurements in step 1 for the Chinese medicinal materials to be tested; Step 3: Based on the comparison of the MRI images obtained in step 1 and step 2, a grid with the same or similar tissue morphological characteristics of the authentic Chinese medicinal material and the Chinese medicinal material to be tested is obtained, and the NMR chemical shift / signal intensity information of the grid is compared to identify the authenticity of the Chinese medicinal material.

2. The method according to claim 1, wherein: The radio frequency coil adopts a spin echo pulse sequence and three gradient coils X, Y and Z to realize spatial Gx, Gy and Gz gradient encoding in the corresponding three dimensions respectively or simultaneously, Gx, Gy and Gz each have MRS / MRSI spatial selection gradient and three-dimensional MRSI imaging gradient encoding composed of N1, N2 or N3 pulse cycles at the same time sequence, N1, N2, N3 are integers from 1 to 128, and Structural imaging sequence was used for MRI measurement, and in situ spectroscopy sequence was used for MRS / MRSI measurement.

3. The method according to claim 1, wherein: The authenticity of the Chinese medicinal materials is identified based on the comparison of NMR chemical shift / signal intensity information of the corresponding regions or parts of the authentic Chinese medicinal materials and the Chinese medicinal materials to be tested, wherein the corresponding regions or parts include a plurality of corresponding grids.

4. The method according to claim 1, wherein: The authenticity of Chinese medicinal materials can be identified by comparing the NMR chemical shift / signal intensity information of two or more parts.

5. The method according to claim 1, wherein: The parts include roots, stems, leaves, flowers, fruits and seeds.

6. The method according to claim 1, wherein: In step 2, the virtual grid division is adjusted in terms of the size and orientation of the grid according to the difference between the Chinese medicinal materials to be tested and the authentic Chinese medicinal materials in terms of shape and / or size.

7. The method according to claim 6, wherein the grid size is selected in the range of 0.1 mm x 0.1 mm to 50 mm x 50 mm.

8. The method according to claim 1, wherein: In situ MRI and MRS measurements were performed on Chinese medicinal materials at different growth stages to obtain the correspondence between the NMR chemical shift / signal intensity information and the tissue morphological characteristics of Chinese medicinal materials at different growth stages.

9. The method according to claim 1, wherein: The Chinese medicinal materials include dendrobium, polygonatum, or gastrodia.

10. The method according to claim 9, wherein: The Chinese medicinal material is Dendrobium officinale, and the authentic Chinese medicinal material is Dendrobium officinale from Huoshan; or The Chinese herbal medicine is polygonatum, and the authentic Chinese herbal medicine is polygonatum multiflorum; or The Chinese medicinal material is Gastrodia elata, and the authentic Chinese medicinal material is Black Gastrodia elata.

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