Method for identifying panax japonicus and rhizoma panacis majoris
Based on the peak area ratio of saponin components, the bamboo ginseng and bead ginseng were quickly identified, which solved the problem of indistinguishability between bamboo ginseng and bead ginseng in the prior art, and achieved a rapid and accurate identification effect. Reduces testing costs and is suitable for clinical applications.
Patent Information
- Application Number
- CN202411531479.1
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-10-30
- Publication Date
- 2024-12-31
AI Technical Summary
Bamboo ginseng and bead ginseng have obvious differences in plant morphology and appearance characteristics, but their chemical composition is similar, which makes it difficult to distinguish after mashing or powdering, which is easy to confuse. The existing identification methods are time-consuming and costly, and are not suitable for promotion and application.
High performance liquid chromatography was used to detect the peak area ratios of ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1, ginsenoside Ro, ginsenoside saponin IV and ginsenoside saponin IVa through high performance liquid chromatography at 200~210nm The detection time was only 75 minutes for bamboo ginseng and bead ginseng.
It achieves rapid and accurate distinction between bamboo ginseng and bead ginseng, reduces detection costs, is suitable for clinical applications, avoids misuse, is characterized by strong characteristics, and is suitable for promotion.
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Figure CN119224163A_ABST
Abstract
Description
Technical Field
[0001] The invention belongs to the technical fields of pharmacognosy and analytical chemistry, and particularly relates to a method capable of quickly identifying Panax notoginseng and Panax notoginseng. Background Art
[0002] Bamboo ginseng is the dried rhizome of Panax japonicus CAMey. It is harvested in autumn, the main root and outer bark are removed, and then dried. It is sweet, slightly bitter, and warm in nature. It has the effects of nourishing and strengthening, dispersing blood stasis and relieving pain, stopping bleeding and removing phlegm. It is used to treat cough, hemoptysis, cough with sputum, and traumatic injuries. Bamboo ginseng grows in high mountain shrubs at an altitude of 1200~1400m or under the forest on the edge of the stone in the shady valley. The most concentrated areas in my country are Shaanxi, Yunnan, Sichuan, Guizhou, Hubei Enshi, etc., and it is also distributed in Vietnam, Nepal, Myanmar, and Japan. Bamboo ginseng mainly contains triterpenoid saponins, polysaccharides, steroids, polyacetylenes and other chemical components.
[0003] Panax japonicus is the dried rhizome of Panax japonicus CA Mey. var. major (Burk.) CY Wu et KM Feng or Panax japonicus CA Mey. var. bipinnatifidus (Seem) CY Wu et KM Feng, a plant of the Araliaceae family. It is bitter, sweet and slightly cold in nature, and has the effects of nourishing the lungs, nourishing yin, activating collaterals and stopping bleeding. It is mainly used to treat symptoms such as deficiency of qi and yin, irritability, thirst, and cough due to consumptive disease. In the early 1970s, the Kunming Institute of Botany identified Panax japonicus and Panax japonicus as variants of Panax japonicus. These two plants are characterized by their thin internodes of rhizomes and nodes that swell into spherical beads. Their rhizome characteristics and chemical compositions are similar, and their medical uses are consistent. They are uniformly recorded as "Parasite" in the Chinese Pharmacopoeia. Panax notoginseng is mainly produced in Yunnan. It is one of the more precious and commonly used Chinese medicines and is a traditional medicine used by ethnic minorities such as the Bai, Naxi, Lisu and Tibetan.
[0004] Both Panax notoginseng and Panax quinquefolius are traditional precious Chinese medicinal materials in my country. Botanically, they belong to the genus Panax notoginseng of the Araliaceae family. Although the two have obvious differences in plant morphology and appearance, their chemical compositions are very similar. They are often crushed or powdered for clinical use. After crushing or powdering, Panax notoginseng and Panax quinquefolius are difficult to distinguish and are easily confused, which may even lead to the misuse of the two.
[0005] There are few reports on the quality difference between bamboo ginseng and pearl ginseng. Relevant studies have used thin layer chromatography to qualitatively identify bamboo ginseng and pearl ginseng (Wang Wei, Guo Lin, Feng Gaili, et al. Comparative study on the quality of bamboo ginseng and pearl ginseng [J]. Journal of Northwest University (Natural Science Edition), 2010, 40(05): 833-836.), but due to the fuzzy identification spectrum provided in the literature, it is difficult to distinguish the two, and the authors have not given a definite identification method and standard. Previous studies have found that the chemical composition of bamboo ginseng and pearl ginseng is very similar, and conventional thin layer identification methods cannot distinguish bamboo ginseng and pearl ginseng. The chemical component fingerprints of two medicinal materials were established by combining HPLC and UPLC-QTQF-MS / MS, and principal component analysis and cluster analysis were performed to classify and identify the two (Ren Bing. Study on the chemical component fingerprints of Panax notoginseng and Panax notoginseng [D]. Wuhan University of Technology, 2016.). However, this method is time-consuming and has high detection costs, which is not convenient for practical promotion and application. RAPD technology was used to obtain DNA fingerprints, and according to the specific bands in the fingerprints, a manual variety identification map (MCID) was drawn to identify the interspecific identification system of Panax notoginseng and its closely related Panax notoginseng plants (Xu Cheng, Zhang Changcheng, Li Jing, et al. Variety identification of Panax notoginseng and its closely related plants based on RAPD markers [J]. Lishizhen Traditional Chinese Medicine, 2016, 27(01):101-104.). However, the experiments in the literature used a single source of each medicinal material, so this method is not representative, and the DNA sequencing identification operation is cumbersome and the detection cost is extremely high, which is not conducive to promotion and application.
[0006] Therefore, it is necessary to establish a more rigorous, quick, simple and suitable identification method for promotion and application to identify and distinguish between Rhizoma Panacis japonicae and Rhizoma Panacis sibiricum to prevent confusion and misuse during clinical application. Summary of the invention
[0007] The present invention provides a method for identifying Panax notoginseng and Panax notoginseng, which can quickly identify Panax notoginseng and Panax notoginseng. The method is based on high performance liquid chromatography and only takes 75 minutes to complete the detection of one sample.
[0008] The technical scheme of the present invention is to provide a method for identifying Panax japonicus and Panax ginseng, using ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1, ginsenoside Ro, Panax japonicus saponin IV and Panax japonicus saponin IVa as raw materials to prepare a mixed reference solution, and the test solution and the mixed reference solution are respectively detected by high performance liquid chromatography; At any wavelength between 200 and 210 nm, if the absorption peaks of ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1, ginsenoside Ro, ginsenoside IV, and ginsenoside IVa appear, and the peak area ratio of ginsenoside IV to ginsenoside Ro is 0.55 to 0.75, and the peak area ratio of ginsenoside IVa to ginsenoside Ro is 0.20 to 0.40, then the test sample is ginseng; At any wavelength between 200 and 210 nm, if the absorption peaks of ginsenoside Rg1 and ginsenoside Re do not appear, and the absorption peaks of ginsenoside Rb1, ginsenoside Ro, bamboo ginsenoside IV, and bamboo ginsenoside IVa appear, and the peak area ratio of bamboo ginsenoside IV to ginsenoside Ro is 0.10-0.50, and the peak area ratio of bamboo ginsenoside IVa to ginsenoside Ro is 0.60-0.90, then the test sample is Panax notoginseng; At any wavelength between 200 and 210 nm, if the peak emission conditions and peak area ratio do not meet the identification criteria of either Rhizoma Panacis japonica or Rhizoma Panacis sibiricum, then the test sample is a mixed product.
[0009] Furthermore, when high performance liquid chromatography is used for detection, the chromatographic conditions are: Stationary phase: octadecylsilane bonded silica gel; Mobile phase: acetonitrile as phase A, 0.05-0.15% by volume phosphoric acid aqueous solution as phase B, preferably 0.1% phosphoric acid aqueous solution, gradient elution according to the following procedure: At 0 min, phase A accounts for 10% of the mobile phase by volume, and the rest is phase B. At 5 minutes, phase A accounts for 10% of the mobile phase by volume, and the rest is phase B. At 15 minutes, phase A accounts for 20% of the mobile phase by volume, and the rest is phase B. At 30 minutes, phase A accounts for 20% of the mobile phase by volume, and the rest is phase B. At 35 minutes, the volume percentage of phase A in the mobile phase was 26.5%, and the rest was phase B. At 40 minutes, phase A accounts for 32% of the volume of the mobile phase, and the rest is phase B. At 50 minutes, phase A accounts for 33% of the volume of the mobile phase, and the rest is phase B. At 55 minutes, phase A accounts for 48% of the mobile phase by volume, and the rest is phase B. At 65 minutes, phase A accounts for 65% of the mobile phase by volume, and the rest is phase B. At 66 minutes, phase A accounts for 10% of the mobile phase by volume, and the rest is phase B. At 75 minutes, phase A accounted for 10% of the mobile phase by volume, and the rest was phase B.
[0010] Flow rate: 1.0~1.4ml / min; preferably 1.2ml / min.
[0011] Column temperature: 28~32℃; preferably 30℃.
[0012] Detection wavelength: any wavelength between 200 and 210 nm.
[0013] Furthermore, in the mixed reference solution, ginsenoside Rg1 was 0.10~0.30 mg / mL, ginsenoside Re was 0.10~0.30 mg / mL, ginsenoside Rb1 was 0.10~0.30 mg / mL, ginsenoside Ro was 0.20~0.50 mg / mL, ginsenoside IV was 0.10~0.30 mg / mL, and ginsenoside IVa was 0.20~0.40 mg / mL.
[0014] Furthermore, the solvent of the mixed reference solution is a 60% by volume methanol aqueous solution.
[0015] Furthermore, when preparing the test solution, the sample to be tested is accurately weighed, extracted with 60% methanol aqueous solution, and then the weight loss during the extraction process is supplemented, and the filtrate is filtered to obtain the test solution.
[0016] Furthermore, ultrasonic extraction was performed for 40 min, and the sample was weighed after cooling, and the lost weight was supplemented with 60% methanol.
[0017] Furthermore, the sample to be tested is processed into a powder passing through a 50-mesh sieve.
[0018] Furthermore, the filter membrane during filtration was 0.45 µm.
[0019] The invention also relates to the application of the method in identifying Panax notoginseng and Panax notoginseng.
[0020] The present invention has the following beneficial effects: After in-depth research and a large number of experiments, the inventors found that the saponin components of Panax notoginseng and Panax ginseng, which have similar origins, have certain differences: both Panax notoginseng and Panax ginseng contain four components: ginsenoside Rb1, ginsenoside Ro, Panax notoginseng saponin IV, and Panax notoginseng saponin IVa. The two components that Panax notoginseng contains but Panax ginseng does not contain are ginsenoside Rg1 and ginsenoside Re; the main saponin components of Panax notoginseng and Panax ginseng. Further analysis and research found that the peak area ratio of ginsenoside IV to ginsenoside Ro in Rhizoma Panacis japonica was in the range of 0.55-0.75, and the peak area ratio of ginsenoside IVa to ginsenoside Ro was in the range of 0.20-0.40; the peak area ratio of ginsenoside IV to ginsenoside Ro in Rhizoma Panacis japonica was in the range of 0.10-0.50, and the peak area ratio of ginsenoside IVa to ginsenoside Ro was in the range of 0.60-0.90. Therefore, based on the differences in saponin components and their existence, with ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1, ginsenoside Ro, ginsenoside IV, and ginsenoside IVa as reference substances, a method for rapid identification of Rhizoma Panacis japonica and Rhizoma Panacis japonica was creatively established, which has strong characteristics. According to the main saponin characteristic components and peak area ratios appearing in the HPLC spectrum, Rhizoma Panacis japonica and Rhizoma Panacis japonica can be accurately and quickly identified. BRIEF DESCRIPTION OF THE DRAWINGS
[0021] Figure 1 It is the HPLC chromatogram of the mixed reference substance obtained in Example 1; in the figure, Peak 1 is the absorption peak of ginsenoside Rg1, Peak 2 is the absorption peak of ginsenoside Re, Peak 3 is the absorption peak of ginsenoside Rb1, Peak 4 is the absorption peak of ginsenoside Ro, Peak 5 is the absorption peak of ginsenoside IV, and Peak 6 is the absorption peak of ginsenoside IVa.
[0022] Figure 2 It is the HPLC chromatogram of the japonicus obtained in Example 2; wherein 2A is the control chromatogram of the japonicus, and 2B is the HPLC chromatograms of japonicus from different batches of japonicus from different origins; in the figure, Peak 1 is the absorption peak of ginsenoside Rg1, Peak 2 is the absorption peak of ginsenoside Re, Peak 3 is the absorption peak of ginsenoside Rb1, Peak 4 is the absorption peak of ginsenoside Ro, Peak 5 is the absorption peak of japonicus saponin IV, and Peak 6 is the absorption peak of japonicus saponin IVa.
[0023] Figure 3 It is the HPLC chromatogram of the Panax notoginseng medicinal material obtained in Example 3; wherein 3A is the control chromatogram of Panax notoginseng, and 3B is the HPLC chromatogram of Panax notoginseng from different batches of Panax notoginseng from different origins; Peak 3 in the figure is the absorption peak of ginsenoside Rb1, Peak 4 is the absorption peak of ginsenoside Ro, Peak 5 is the absorption peak of Panax notoginseng saponin IV, and Peak 6 is the absorption peak of Panax notoginseng saponin IVa.
[0024] Figure 4~Figure 6 13 batches of samples (S1-S13) in Example 4; Figure 4 For samples S1~S5, Figure 5 For samples S6~S9, Figure 6 For samples S10~S13; Figure 4~Figure 6 Peak 1 is the absorption peak of ginsenoside Rg1, peak 2 is the absorption peak of ginsenoside Re, peak 3 is the absorption peak of ginsenoside Rb1, peak 4 is the absorption peak of ginsenoside Ro, peak 5 is the absorption peak of bamboo ginsenoside IV, and peak 6 is the absorption peak of bamboo ginsenoside IVa. The spectrum information of the adulterated products S6 (Panax notoginseng), S9 (Ginseng), S11 (Ginseng root), S13 (Ginger-shaped Panax notoginseng) is obviously different from that of bamboo ginseng and pearl ginseng. DETAILED DESCRIPTION
[0025] The present invention is described in detail below in conjunction with specific embodiments. This embodiment is implemented based on the technical solution of the present invention, and provides a detailed implementation method and a specific operation process, but the protection scope of the present invention is not limited to the following embodiments.
[0026] The experimental methods in the following examples are conventional methods unless otherwise specified. The medicinal materials, reagents, etc. used in the following examples are commercially available products unless otherwise specified.
[0027] The instruments, reagents and medicinal materials used in the following examples and their sources are: 1. Instrument U3000 HPLC, including Chameleon workstation, DAD detector, UV detector, automatic injection system, etc.; 1 / 100,000 electronic analytical balance (model: XSE205DU, Mettler-Toledo Instruments (Shanghai) Co., Ltd.); 1 / 10,000 electronic balance (model: ME204T / 02, Mettler-Toledo Instruments (Shanghai) Co., Ltd.); Ultrasonic cleaner (model: DTC-27, Dingtai Biochemical Technology Equipment Manufacturing Co., Ltd.); InfinityLab poroshell HPH (4.6 × 250 mm, 5 μm) column.
[0028] 2. Reagents Acetonitrile (chromatographically pure) Specification: 4L, Hubei Futon Science and Technology Co., Ltd.; Methanol (chromatographic grade) specification: 4L, Hubei Futon Science and Technology Co., Ltd.; Phosphoric acid (analytical grade) Specification: 500mL, Tianjin Komiou Chemical Co., Ltd.; Deionized water, homemade.
[0029] 3. Reference Ginsenoside Rg1 (batch number: 110704-202331, China Food and Drug Inspection Institutes), ginsenoside Re (batch number: 110703-202235, China Food and Drug Inspection Institutes), ginsenoside Rb1 (batch number: 110704-202331, China Food and Drug Inspection Institutes), ginsenoside Ro (batch number: 111903-202106, China Food and Drug Inspection Institutes), bamboo ginsenoside IV (batch number: 200319, Beijing Century Aoke Biological), bamboo ginsenoside IVa (batch number: 111861-201102, China Food and Drug Inspection Institutes).
[0030] 4. Samples to be tested The bamboo ginseng was collected from many medicinal material planting bases such as Leshan, Sichuan, Ebian, Sichuan, Zhaotong, Yunnan, and Enshi, Hubei. It was identified by Professor He Haibo of the Three Gorges University as the dried rhizome of the bamboo ginseng Panax japonicus CAMey.
[0031] The pearl ginseng was collected from many medicinal material planting bases such as Zhaotong, Yunnan and Ebian, Sichuan. It was identified by Professor He Haibo of the Three Gorges University as the dried rhizomes of the pearl ginseng Panax japonicus CA Mey.var.major (Burk.)CYWuetK.M.Feng and the feather-leaved Panax notoginseng Panax japonicus CA Mey.var.bip.
[0032] Confusing products: Ginseng, Panax notoginseng, ginseng root and ginger-shaped Panax notoginseng were purchased from the Anguo Chinese herbal medicine market in Hebei Province. They were identified by Professor He Haibo of the Three Gorges University as the dried roots and rhizomes of Panax ginseng CA Mey. of the Araliaceae family, the dried roots and rhizomes of Panax notoginseng (Burk.) FH Chen of the Araliaceae family, the dried lateral roots and fibrous roots of the cultivated product of Panax ginseng CA Mey. (commonly known as garden ginseng) of the Araliaceae family, and the dried roots and rhizomes of Panax zingiberensis of the Araliaceae family.
[0033] Example 1 Establishment of HPLC Chromatogram of Reference Substance 1. Chromatographic conditions Column: InfinityLab poroshell HPH (4.6×250 mm, 5μm) column.
[0034] Mobile phase: acetonitrile was used as mobile phase A, and 0.10% by volume aqueous phosphoric acid solution was used as phase B for gradient elution. The absorption peaks of each reference substance had good peak shape, high separation, and stable baseline, so phase B was preferably 0.1% aqueous phosphoric acid solution. The mobile phase gradient elution program is shown in Table 1.
[0035] Table 1. Gradient elution program
[0036] Flow rate: 1.0~1.4mL / min; After testing, it was found that the separation effect was best when the flow rate was 1.2mL / min, so the preferred flow rate is 1.2mL / min.
[0037] Column temperature: 28~32℃, preferably 30℃.
[0038] Detection wavelength: any wavelength between 200 and 210 nm. Specifically in this embodiment, the detection wavelength is 203 nm.
[0039] Elution time: 75min.
[0040] 2. Preparation of mixed reference solution The mixed reference substance of the present invention can be prepared by the following method: accurately weigh ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1, ginsenoside Ro, ginsenoside IV, and ginsenoside IVa reference substances, and add 60% methanol (methanol aqueous solution with a methanol volume percentage concentration of 60%) to prepare a mixed reference substance solution containing 0.10-0.30 mg of ginsenoside Rg1, 0.10-0.30 mg of ginsenoside Re, 0.10-0.30 mg of ginsenoside Rb1, 0.20-0.50 mg of ginsenoside Ro, 0.10-0.30 mg of ginsenoside IV, and 0.20-0.40 mg of ginsenoside IVa per 1 ml to obtain the mixed reference substance solution.
[0041] Specifically in this embodiment, the mixed reference solution was prepared by the following method: Accurately weigh 2.1, 2.5, 2.2, 4.2, 2.0 and 3.2 mg of ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1, ginsenoside Ro, panax ginsenoside IV and panax ginsenoside IVa reference substances, respectively, put them into a 10 mL volumetric flask, add 60% methanol to dissolve and dilute to the scale to make a mixed reference substance solution containing 0.21 mg of ginsenoside Rg1, 0.25 mg of ginsenoside Re, 0.22 mg of ginsenoside Rb1, 0.42 mg of ginsenoside Ro, 0.20 mg of panax ginsenoside IV and 0.32 mg of panax ginsenoside IVa per 1 ml.
[0042] 3. Determination Accurately pipette 5 μL of the mixed reference solution, inject it into the liquid chromatograph for determination, and record the chromatogram at 203 nm from 0 to 75 min. Figure 1 .from Figure 1 It can be seen that the separation degree of each reference substance is good and there is basically no interference between each other, which indicates that the chromatographic conditions of the present invention are suitable for the separation of the above-mentioned mixed reference substances.
[0043] The absorption peaks of ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1, ginsenoside Ro, ginsenoside IV, and ginsenoside IVa are marked as peak 1 to peak 6. The peak conditions and retention times of each reference substance at a wavelength of 203 nm are shown in Table 2.
[0044] Table 2. Peak results of reference substances
[0045] Note: + indicates that the absorption peak was detected; - indicates that the absorption peak was not detected.
[0046] Example 2 Establishment of HPLC Chromatogram of Panax notoginseng 1. Chromatographic conditions The preferred chromatographic conditions are the same as those in Example 1, wherein the detection wavelength is 203 nm.
[0047] 2. Preparation of mixed reference solution Same as the "Preparation of Mixed Reference Solution" in Example 1.
[0048] 3. Preparation of test solution Weigh 0.5 g of dry Panax notoginseng powder, weigh accurately, place in a stoppered conical flask, add 25 mL of 60% methanol, stopper, weigh, extract by ultrasonic for 40 min, cool, weigh again, make up the lost weight with 60% methanol, shake well, filter through a 0.45 µm filter membrane, and obtain the filtrate.
[0049] 4. Determination Accurately pipette 5 μL of the mixed reference solution prepared in step 2 and the test solution prepared in step 3, respectively, and inject them into the high performance liquid chromatograph. Under the chromatographic conditions, record the liquid chromatogram from 0 to 75 min at a wavelength of 203 nm. Figure 2 The peak elution conditions and retention times of peaks 1 to 6 are shown in Table 3. The peak area ratios of ginsenoside IVa and ginsenoside Ro in 15 batches of ginseng from different origins are shown in Table 4.
[0050] Compared with the chromatogram of the mixed reference solution, judging from the retention time of the absorption peaks, the absorption peaks of ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1, ginsenoside Ro, ginsenoside IV, and ginsenoside IVa appeared in Japonica ginseng at 203nm, and the peak area ratio of ginsenoside IV to ginsenoside Ro was in the range of 0.55-0.75, and the peak area ratio of ginsenoside IVa to ginsenoside Ro was in the range of 0.20-0.40.
[0051] Table 3. Peaks of Panax japonicus
[0052] Note: + indicates that the absorption peak is detected; - indicates that the absorption peak is not detected.
[0053] The peak area ratio of ginsenoside IV to ginsenoside Ro is 0.72, within the range of 0.55-0.75; the peak area ratio of ginsenoside Ⅳa to ginsenoside Ro is 0.22, within the range of 0.20-0.40.
[0054] Table 4. Peak areas of ginsenoside IV, ginsenoside IVa and ginsenoside Ro in 15 batches of ginsenosides from different origins
[0055] Example 3 Establishment of HPLC Chromatogram of Ginseng Root 1. Chromatographic conditions The preferred chromatographic conditions are the same as those in Example 1, wherein the detection wavelength is 203 nm.
[0056] 2. Preparation of mixed reference solution Same as the "Preparation of Mixed Reference Solution" in Example 1.
[0057] 3. Preparation of test solution Weigh 0.5 g of dry Panax notoginseng powder, weigh accurately, place in a stoppered conical flask, add 25 mL of 60% methanol, stopper, weigh, extract by ultrasonic for 40 min, cool, weigh again, make up the lost weight with 60% methanol, shake well, filter through a 0.45 µm filter membrane, and obtain the filtrate.
[0058] 4. Determination Accurately pipette 5 μL of the mixed reference solution prepared in step 2 and the test solution prepared in step 3, respectively, and inject them into the high performance liquid chromatograph. Under the chromatographic conditions, record the liquid chromatogram from 0 to 75 min at a wavelength of 203 nm. Figure 3 The peak elution conditions and retention times of peaks 1 to 6 are shown in Table 5. The peak area ratios of ginsenoside IVa and ginsenoside Ro in 15 batches of Rhizoma Panacis japonicae from different origins are shown in Table 6.
[0059] Compared with the chromatogram of the mixed reference solution, judging by the retention time of the absorption peaks, the absorption peaks of ginsenoside Rg1 and ginsenoside Re did not appear in Ginseng chinensis at 203nm, but the absorption peaks of ginsenoside Rb1, ginsenoside Ro, bamboo ginsenoside IV, and bamboo ginsenoside IVa appeared, and the peak area ratio of bamboo ginsenoside IV to ginsenoside Ro was in the range of 0.10-0.50, and the peak area ratio of bamboo ginsenoside IVa to ginsenoside Ro was in the range of 0.60-0.90.
[0060] Table 5. Peaks of Ginseng chinensis
[0061] Note: + indicates that the absorption peak was detected; - indicates that the absorption peak was not detected.
[0062] The peak area ratio of ginsenoside IV to ginsenoside Ro is 0.41, ranging from 0.10 to 0.50; the peak area ratio of ginsenoside Ⅳa to ginsenoside Ro is 0.72, ranging from 0.60 to 0.90.
[0063] Table 6 Peak areas of ginsenoside IV, ginsenoside IVa and ginsenoside Ro in 15 batches of Ginseng from different origins
[0064] Example 4 Identification of Radix Panacis Sinensis, Radix Panacis Sinensis and Mixed Medicinal Materials Thirteen samples of bamboo joint ginseng, pearl ginseng and easily confused products collected from different channels were identified by properties (appearance and microscopic properties of medicinal materials) to determine their medicinal material varieties, as shown in Table 7. The above medicinal materials were crushed and handed over to the analysis experimenter, who randomly numbered them as S1 to S13 (without telling them what kind of medicinal materials they were), and the chromatograms of the mixed reference and each sample at 203 nm were obtained according to the methods and steps of Example 2, as shown in Table 7. Figure 4~Figure 6 .
[0065] The chromatogram of each sample was compared with the corresponding chromatogram of the mixed reference substance, and the sample was judged to be Panax japonicus, Panax ginseng, or a confused substance according to the peak of the reference substance. The results are shown in Table 7.
[0066] Table 7. Identification results of Panax notoginseng, Panax notoginseng and mixed products
[0067] As can be seen from Table 7, the rapid identification method of the present invention is used to identify the medicinal materials of bamboo joint Panax notoginseng and pearl Panax notoginseng, and the results are consistent with the classic property identification results. In addition, when the medicinal materials are crushed and cannot be identified by the property identification method, the method of the present invention can also quickly identify whether the medicinal material is bamboo joint Panax notoginseng, pearl Panax notoginseng, or a confused product.
[0068] The above embodiments describe the preferred implementation of the present invention, but the present invention is not limited thereto. Within the technical concept of the present invention, the technical solution of the present invention can be subjected to a variety of simple modifications, including the combination of various technical features in any other manner, and these simple modifications and combinations should also be regarded as the contents disclosed by the present invention and belong to the protection scope of the present invention. Therefore, the protection scope of the patent of the present invention shall be subject to the attached claims.
Claims
1. A method for distinguishing Panax notoginseng from Panax notoginseng, characterized in that: A mixed reference solution was prepared with ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1, ginsenoside Ro, ginsenoside Ⅳ and ginsenoside Ⅳa as raw materials, and the test solution and the mixed reference solution were detected by high performance liquid chromatography respectively; At any wavelength between 200 and 210 nm, if the absorption peaks of ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1, ginsenoside Ro, ginsenoside IV, and ginsenoside IVa appear, and the peak area ratio of ginsenoside IV to ginsenoside Ro is 0.55 to 0.75, and the peak area ratio of ginsenoside IVa to ginsenoside Ro is 0.20 to 0.40, then the test sample is ginseng; At any wavelength between 200 and 210 nm, if the absorption peaks of ginsenoside Rg1 and ginsenoside Re do not appear, and the absorption peaks of ginsenoside Rb1, ginsenoside Ro, bamboo ginsenoside IV, and bamboo ginsenoside IVa appear, and the peak area ratio of bamboo ginsenoside IV to ginsenoside Ro is 0.10-0.50, and the peak area ratio of bamboo ginsenoside IVa to ginsenoside Ro is 0.60-0.90, then the test sample is Panax notoginseng; At any wavelength between 200 and 210 nm, if the peak emission conditions and peak area ratio do not meet the identification criteria of either Rhizoma Panacis japonica or Rhizoma Panacis sibiricum, then the test sample is a mixed product.
2. The method according to claim 1, characterized in that When high performance liquid chromatography is used for detection, the chromatographic conditions are: Stationary phase: octadecylsilane bonded silica gel; Mobile phase: acetonitrile as phase A, 0.05-0.15% volume percent phosphoric acid aqueous solution as phase B, gradient elution according to the following program: ； Flow rate: 1.0~1.4ml / min; Column temperature: 28~32℃; Detection wavelength: any wavelength between 200 and 210 nm.
3. The method according to claim 2, characterized in that: Phase B is a 0.10% volume percent phosphoric acid aqueous solution; the flow rate is 1.2 ml / min; and the column temperature is 30°C.
4. The method according to claim 1, characterized in that: In the mixed reference solution, ginsenoside Rg1 was 0.10~0.30 mg / mL, ginsenoside Re was 0.10~0.30 mg / mL, ginsenoside Rb1 was 0.10~0.30 mg / mL, ginsenoside Ro was 0.20~0.50 mg / mL, ginsenoside IV was 0.10~0.30 mg / mL, and ginsenoside IVa was 0.20~0.40 mg / mL.
5. The method according to claim 4, characterized in that: The solvent of the mixed reference solution is a 60% by volume methanol aqueous solution.
6. The method according to any one of claims 1 to 5, characterized in that: When preparing the test solution, accurately weigh the sample to be tested, extract it with 60% methanol aqueous solution, then make up the weight lost during the extraction process, and filter the filtrate to obtain the test solution.
7. The method according to claim 6, characterized in that: Ultrasonic extraction was used for 40 min, and the sample was weighed after cooling, and the lost weight was made up with 60% methanol.
8. The method according to claim 6, characterized in that: The sample to be tested was processed into a powder passing 50 mesh.
9. The method according to claim 6, characterized in that: The filter membrane during filtration is 0.45µm.
10. Application of the method according to any one of claims 1 to 9 in the identification of Panax notoginseng and Panax notoginseng.