Method for judging harvesting time of cinnamon medicinal material
By calculating cinnamon medicinal samples and chromatographic analysis of the extract, the harvest time is determined, and the problem of difficulty in accurately judging the harvest time of cinnamon medicinal materials in the prior art is solved, and rapid and accurate identification results are achieved.
Patent Information
- Application Number
- CN202510000775.7
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-01-02
- Publication Date
- 2025-04-11
AI Technical Summary
It is difficult for the prior art to accurately judge the harvest time of cinnamon medicinal materials, resulting in the quality of medicinal materials harvested in spring on the market being inferior to that of medicinal materials harvested in autumn, affecting the therapeutic effect.
By peeling, drying, and pulverizing the cinnamon medicinal sample, and analyzing its extract using high performance liquid chromatography (HPLC or UPLC) to determine whether it contains cinnamon acetate to determine its harvest time.
It realizes the rapid and accurate judgment of the harvest time of cinnamon medicinal materials, with reliable results, simple and easy to promote, and good reproducibility.
Smart Images
Figure CN119804708A_ABST
Abstract
Description
Technical Field
[0001] The invention relates to the technical field of chemical analysis, and in particular to a method for judging the harvesting time of cinnamon medicinal materials. Background Art
[0002] Chinese herbal medicines are used to treat diseases. Good herbal medicines can cure diseases and solve patients' pain and troubles in time. Therefore, the quality of Chinese herbal medicines is closely related to people's health and happy life.
[0003] The quality of Chinese medicinal materials is closely related to the variety of medicinal materials, the place of origin and ecological environment, cultivation technology, harvesting time, processing and storage methods, etc. The quality of medicinal materials is affected by many factors, and it is necessary to strictly control multiple links such as planting and collection, processing and storage, preparation and packaging, market circulation and quality control to ensure the quality and efficacy of medicinal materials.
[0004] Studies have shown that the quality of Chinese medicinal materials harvested at different times varies significantly, and the therapeutic effects when used to treat diseases vary greatly. Therefore, the "Chinese Pharmacopoeia" stipulates corresponding harvesting times for each medicinal material to regulate the Chinese medicinal materials market and ensure the effectiveness of Chinese medicinal materials in treating diseases.
[0005] The Chinese Pharmacopoeia stipulates that the harvesting time of cinnamon is "mostly in autumn", but in the medicinal material market, farmers mostly harvest cinnamon in spring. They feedback that cinnamon is easy to peel in spring and the trees are easy to survive after peeling.
[0006] Studies have shown that the quality of cinnamon harvested in autumn is better than that of cinnamon harvested in spring, and the therapeutic effect of cinnamon harvested in autumn is also better than that of cinnamon harvested in spring. There are both autumn-harvested and spring-harvested cinnamon on the market, and it is difficult to distinguish the two from their appearance. Therefore, there is an urgent need for a technical means to determine the harvest time of cinnamon. Summary of the invention
[0007] The invention aims to provide a method for judging the harvesting time of cinnamon medicinal materials, which has the characteristics of being simple, fast, intuitive, reproducible and easy to promote.
[0008] To achieve the above object, the present invention provides the following technical solutions:
[0009] A method for determining the harvesting time of cinnamon medicinal materials comprises the following steps:
[0010] Step 1: Sample processing
[0011] The cinnamon medicinal material sample to be identified is peeled, dried and crushed;
[0012] Step 2: Preparation of test solution
[0013] Accurately weigh appropriate amounts of cinnamon powder and place them in a conical flask, add solvent to extract, and obtain cinnamon extract; accurately draw an appropriate amount of extract, filter, and obtain the test solution;
[0014] Step 3: Preparation of reference solution
[0015] Accurately weigh an appropriate amount of reference substance and place it in a 10 mL volumetric flask, add solvent to dissolve it, and obtain a reference substance solution;
[0016] Step 4: Data Analysis
[0017] Analyze the test solution and reference solution obtained in steps 2 and 3, respectively, to obtain data for each sample solution;
[0018] Step 5: Data processing
[0019] Analyze the data obtained in step 4 to obtain a chromatogram of the cinnamon medicinal material to be identified and a chromatogram of the reference substance;
[0020] Step 6: Determine the harvesting time of cinnamon herbs
[0021] Analyze the data obtained in step 5 to determine the harvesting time of cinnamon medicinal materials.
[0022] Preferably, in step 2, the extraction solvent used in preparing the test solution is a methanol aqueous solution with a concentration of 65-75%, the extraction method is hot reflux extraction, the extraction temperature is 50-100° C., and the extraction time is 20-60 minutes.
[0023] Preferably, in step 3, the reference substance is cinnamyl acetate and the solvent is methanol.
[0024] Preferably, in step 4, a high performance liquid chromatograph or an ultra high performance liquid chromatograph is used for detection and analysis, a silane bonded silica gel column is used for separation, acetonitrile and a buffer solution are used for gradient elution, and a UV detector or a diode array detector is used for detection.
[0025] Preferably, the judgment basis of step 6 is: cinnamon medicinal materials harvested in autumn contain cinnamyl acetate, while cinnamon medicinal materials harvested in spring do not contain cinnamyl acetate.
[0026] The beneficial effects of the present invention are as follows:
[0027] The identification method of the present invention has accurate identification results, high reliability, and is fast and efficient. The entire process only takes about 40 minutes, has good reproducibility, and is easy to promote. BRIEF DESCRIPTION OF THE DRAWINGS
[0028] Figure 1 This is the chromatogram of cinnamon bark under the HPLC conditions of Example 1;
[0029] Figure 2 This is the chromatogram of cinnamyl acetate under the HPLC conditions of Example 1;
[0030] Figure 3 This is the chromatogram of cinnamon medicinal material under UPLC conditions in Example 2;
[0031] Figure 4 This is the chromatogram of cinnamyl acetate under the UPLC conditions of Example 2;
[0032] Figure 5 This is the chromatogram of cinnamon bark under the HPLC conditions of Example 3;
[0033] Figure 6 The chromatogram of cinnamyl acetate under the HPLC conditions of Example 3;
[0034] Figure 7 This is the chromatogram of cinnamon medicinal material under UPLC conditions in Example 4;
[0035] Figure 8 This is the chromatogram of cinnamyl acetate under the UPLC conditions of Example 4. DETAILED DESCRIPTION
[0036] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.
[0037] Example 1
[0038] 1. Instruments and reagents
[0039] 1.1 Instruments
[0040] Agilent 1200 series (G1311A quaternary gradient pump, G1329B autosampler, G1316A column oven, G4212B diode array detector), rotary evaporator: N-1001, Shanghai Ailant Instrument Co., Ltd.; parallel concentrator: MPE, Ruike Group Co., Ltd.; electronic balance: AB135-S one hundred thousandth balance, Mettler Toledo.
[0041] 1.2 Samples and Reagents
[0042] 1.2.1 Standards
[0043] Cinnamyl acetate (Batch No.: 020144): purchased from Chengdu Pusi Biotechnology Co., Ltd.
[0044] 1.2.2 Experimental reagents
[0045] Formic acid and phosphoric acid were of chromatographic grade and purchased from Merck & Co., Ltd., USA; acetonitrile was purchased from Furton Scientific & Technology Co., Ltd.; methanol and ethanol were of analytical grade and purchased from Sinopharm Chemical Reagent Co., Ltd.
[0046] 1.2.3 Experimental water
[0047] Grade 1 water as specified in GB / T 6682.
[0048] 1.2.4 Experimental samples
[0049] The cinnamon medicinal material was collected by the inventor on September 15, 2023 in the Yangchun medicinal material production area in Guangdong.
[0050] 2 Test steps
[0051] 2.1 Preparation of reference solution
[0052] Accurately weigh 2.45 mg of the reference substance cinnamyl acetate into a 10 mL volumetric flask, dissolve it with methanol and dilute to the mark to obtain the standard stock solution. Accurately pipette 0.10 mL of the standard stock solution into a 10 mL volumetric flask, and dilute to the mark with 50% methanol to obtain the reference substance working solution.
[0053] 2.2 Preparation of test solution
[0054] Accurately weigh 0.5 g of cinnamon powder and place it in a conical flask, accurately add 25 mL of 70% methanol solution, reflux at 80°C for 40 minutes, let it stand to cool, filter, accurately measure 5 mL of the filtrate, rotary evaporate to dryness at 40°C, add 70% methanol solution to the residue and dissolve it in a 5 mL volumetric flask, make up to volume, shake well, and then filter with a 0.22 μm microporous filter membrane.
[0055] 2.3 Sample analysis
[0056] The samples were analyzed by high performance liquid chromatography, and the specific analysis conditions were as follows:
[0057] Chromatographic column: Waters T3 column, 5 μm, 4.6 × 250 mm;
[0058] Acetonitrile-0.1% phosphoric acid aqueous solution was used as the mobile phase for gradient elution. The elution gradient is shown in Table 1:
[0059] Table 1 Liquid chromatography detection elution gradient conditions
[0060]
[0061] Flow rate: 1.0 mL / min;
[0062] Detection wavelength: 250nm;
[0063] Column temperature: 30°C;
[0064] Injection volume: 10 μL.
[0065] 2.4 Data Analysis
[0066] The chromatograms of cinnamon medicinal material samples and reference solution were obtained by high performance liquid chromatography. Figure 1-2 ,from Figure 1-2 It can be seen from the chromatogram that under the above chromatographic conditions, the retention time of cinnamyl acetate is 34.3 min, and a chromatographic peak appears in cinnamon medicinal material at the retention time of 34.3 min.
[0067] 2.5 Result judgment
[0068] The chromatographic peak with a retention time of 34.3 min in the test solution of cinnamon medicinal material was identified by spectrum (the spectrum was collected simultaneously during sample analysis). Its spectrum was completely consistent with the spectrum of cinnamyl acetate. The chromatographic peak with a retention time of 34.3 min in the test solution of cinnamon medicinal material was determined to be cinnamyl acetate. Comprehensive judgment: this cinnamon medicinal material was harvested in autumn, which is consistent with the actual situation.
[0069] Example 2
[0070] 1. Instruments and reagents
[0071] 1.1 Instruments
[0072] Infinity II ultra-high performance liquid chromatograph, equipped with UV detector; rotary evaporator: N-1001, Shanghai Ailang Instrument Co., Ltd.; parallel concentrator: MPE, Ruike Group Co., Ltd.; electronic balance: AB135-S 1 / 10000 balance, METTLER TOLEDO.
[0073] 1.2 Samples and Reagents
[0074] 1.2.1 Standards
[0075] Cinnamyl acetate (Batch No.: 020144): purchased from Chengdu Pusi Biotechnology Co., Ltd.
[0076] 1.2.2 Experimental reagents
[0077] Formic acid and phosphoric acid were of chromatographic grade and purchased from Merck & Co., Ltd., USA; acetonitrile was purchased from Furton Scientific & Technology Co., Ltd.; methanol and ethanol were of analytical grade and purchased from Sinopharm Chemical Reagent Co., Ltd.
[0078] 1.2.3 Experimental water
[0079] Grade 1 water as specified in GB / T 6682.
[0080] 1.2.4 Experimental samples
[0081] The cinnamon medicinal material was collected by the inventor on September 20, 2023 at the medicinal material production area of Guigang, Guangxi.
[0082] 2 Test steps
[0083] 2.1 Preparation of reference solution
[0084] Accurately weigh 2.45 mg of the reference substance cinnamyl acetate into a 10 mL volumetric flask, dissolve it with methanol and dilute to the mark to obtain the standard stock solution. Accurately pipette 0.10 mL of the standard stock solution into a 10 mL volumetric flask, and dilute to the mark with 50% methanol to obtain the reference substance working solution.
[0085] 2.2 Preparation of test solution
[0086] Accurately weigh 0.5 g of cinnamon powder and place it in a conical flask, accurately add 25 mL of 70% methanol solution, reflux at 80°C for 40 minutes, let it stand to cool, filter, accurately measure 5 mL of the filtrate, rotary evaporate to dryness at 40°C, add 70% methanol solution to the residue and dissolve it in a 5 mL volumetric flask, make up to volume, shake well, and then filter with a 0.22 μm microporous filter membrane.
[0087] 2.3 Sample analysis
[0088] Ultra-high performance liquid chromatography was used to analyze the samples. The specific analysis conditions were as follows:
[0089] Chromatographic column: SB-C18 column, 1.8 μm, 2.1 × 100 mm;
[0090] Acetonitrile-0.1% formic acid aqueous solution was used as the mobile phase for gradient elution. The elution gradient is shown in Table 2:
[0091] Table 2 Ultra-high performance liquid chromatography fingerprint detection elution gradient conditions
[0092]
[0093]
[0094] Flow rate: 0.300 mL / min;
[0095] Detection wavelength: 250nm;
[0096] Column temperature: 35 °C;
[0097] Injection volume: 2 μL.
[0098] 2.4 Data Analysis
[0099] The chromatograms of cinnamon medicinal material samples and reference solution were obtained by ultra-high performance liquid chromatography. Figure 3-Figure 4 It can be seen from the chromatogram that under the above chromatographic conditions, the retention time of cinnamyl acetate is 42.1 min, and a chromatographic peak appears in cinnamon medicinal material at the retention time of 42.1 min.
[0100] 2.5 Result judgment
[0101] The retention time of the chromatographic peak with a retention time of 42.1 min in the cinnamon medicinal material test solution is consistent with the retention time of cinnamyl acetate. It can be determined that the cinnamon medicinal material test solution contains cinnamyl acetate. Comprehensive judgment: this cinnamon medicinal material is harvested in autumn, which is consistent with the actual harvesting time.
[0102] Example 3
[0103] 1. Instruments and reagents
[0104] 1.1 Instruments
[0105] Agilent 1200 series (G1311A quaternary gradient pump, G1329B autosampler, G1316A column oven, G4212B diode array detector), rotary evaporator: N-1001, Shanghai Ailant Instrument Co., Ltd.; parallel concentrator: MPE, Ruike Group Co., Ltd.; electronic balance: AB135-S one hundred thousandth balance, Mettler Toledo.
[0106] 1.2 Samples and Reagents
[0107] 1.2.1 Standards
[0108] Cinnamyl acetate (Batch No.: 020144): purchased from Chengdu Pusi Biotechnology Co., Ltd.
[0109] 1.2.2 Experimental reagents
[0110] Formic acid and phosphoric acid were of chromatographic grade and purchased from Merck & Co., Ltd., USA; acetonitrile was purchased from Furton Scientific & Technology Co., Ltd.; methanol and ethanol were of analytical grade and purchased from Sinopharm Chemical Reagent Co., Ltd.
[0111] 1.2.3 Experimental water
[0112] Grade 1 water as specified in GB / T 6682.
[0113] 1.2.4 Experimental samples
[0114] The cinnamon medicinal material was collected by the inventor on April 5, 2024 in the Yangchun medicinal material production area in Guangdong.
[0115] 2 Test steps
[0116] 2.1 Preparation of reference solution
[0117] Accurately weigh 2.45 mg of the reference substance cinnamyl acetate into a 10 mL volumetric flask, dissolve it with methanol and dilute to the mark to obtain the standard stock solution. Accurately pipette 0.10 mL of the standard stock solution into a 10 mL volumetric flask, and dilute to the mark with 50% methanol to obtain the reference substance working solution.
[0118] 2.2 Preparation of test solution
[0119] Accurately weigh 0.5 g of cinnamon powder and place it in a conical flask, accurately add 25 mL of 70% methanol solution, reflux at 80°C for 40 minutes, let it stand to cool, filter, accurately measure 5 mL of the filtrate, rotary evaporate to dryness at 40°C, add 70% methanol solution to the residue and dissolve it in a 5 mL volumetric flask, make up to volume, shake well, and then filter with a 0.22 μm microporous filter membrane.
[0120] 2.3 Sample analysis
[0121] The samples were analyzed by high performance liquid chromatography, and the specific analysis conditions were as follows:
[0122] Chromatographic column: Waters T3 column, 5 μm, 4.6 × 250 mm;
[0123] Acetonitrile-0.1% phosphoric acid aqueous solution was used as the mobile phase for gradient elution. The elution gradient is shown in Table 3:
[0124] Table 3 Liquid chromatography detection elution gradient conditions
[0125]
[0126] Flow rate: 1.0 mL / min;
[0127] Detection wavelength: 250nm;
[0128] Column temperature: 30°C;
[0129] Injection volume: 10 μL.
[0130] 2.4 Data Analysis
[0131] The chromatograms of cinnamon medicinal material samples and reference solution were obtained by high performance liquid chromatography. Figure 5-6 It can be seen from the chromatogram that under the above chromatographic conditions, the retention time of cinnamyl acetate is 34.3 min, and no chromatographic peak appears in cinnamon medicinal material at the retention time of 34.3 min.
[0132] 2.5 Result judgment
[0133] No chromatographic peak appeared at a retention time of 34.3 min in the test solution of cinnamon medicinal materials. Under the above conditions, the substance with a retention time of 34.3 min was cinnamyl acetate. It was determined that the test solution of cinnamon medicinal materials did not contain cinnamyl acetate. Comprehensively judging: this cinnamon medicinal material was harvested in spring, which was consistent with the actual situation.
[0134] Example 4
[0135] 1. Instruments and reagents
[0136] 1.1 Instruments
[0137] Infinity II ultra-high performance liquid chromatograph, equipped with UV detector; rotary evaporator: N-1001, Shanghai Ailang Instrument Co., Ltd.; parallel concentrator: MPE, Ruike Group Co., Ltd.; electronic balance: AB135-S 1 / 10000 balance, METTLER TOLEDO.
[0138] 1.2 Samples and Reagents
[0139] 1.2.1 Standards
[0140] Cinnamyl acetate (Batch No.: 020144): purchased from Chengdu Pusi Biotechnology Co., Ltd.
[0141] 1.2.2 Experimental reagents
[0142] Formic acid and phosphoric acid were of chromatographic grade and purchased from Merck & Co., Ltd., USA; acetonitrile was purchased from Furton Scientific & Technology Co., Ltd.; methanol and ethanol were of analytical grade and purchased from Sinopharm Chemical Reagent Co., Ltd.
[0143] 1.2.3 Experimental water
[0144] Grade 1 water as specified in GB / T 6682.
[0145] 1.2.4 Experimental samples
[0146] The cinnamon medicinal material was collected by the inventor on April 10, 2024 at the medicinal material production area of Guigang, Guangxi.
[0147] 2 Test steps
[0148] 2.1 Preparation of reference solution
[0149] Accurately weigh 2.45 mg of the reference substance cinnamyl acetate into a 10 mL volumetric flask, dissolve it with methanol and dilute to the mark to obtain the standard stock solution. Accurately pipette 0.10 mL of the standard stock solution into a 10 mL volumetric flask, and dilute to the mark with 50% methanol to obtain the reference substance working solution.
[0150] 2.2 Preparation of test solution
[0151] Accurately weigh 0.5 g of cinnamon powder and place it in a conical flask, accurately add 25 mL of 70% methanol solution, reflux at 80°C for 40 minutes, let it stand to cool, filter, accurately measure 5 mL of the filtrate, rotary evaporate to dryness at 40°C, add 70% methanol solution to the residue and dissolve it in a 5 mL volumetric flask, make up to volume, shake well, and then filter with a 0.22 μm microporous filter membrane.
[0152] 2.3 Sample analysis
[0153] Ultra-high performance liquid chromatography was used to analyze the samples. The specific analysis conditions were as follows:
[0154] Chromatographic column: SB-C18 column, 1.8 μm, 2.1 × 100 mm;
[0155] Acetonitrile-0.1% formic acid aqueous solution was used as the mobile phase for gradient elution. The elution gradient is shown in Table 4:
[0156] Table 4 ULTRA PERFORMANCE LIQUID CHROMATOGRAPHIC FINGERPRINT DETECTION ELUTATION GRADIENT CONDITIONS
[0157]
[0158] Flow rate: 0.300 mL / min;
[0159] Detection wavelength: 250nm;
[0160] Column temperature: 35 °C;
[0161] Injection volume: 2 μL.
[0162] 2.4 Data Analysis
[0163] The chromatograms of cinnamon medicinal material samples and reference solution were obtained by ultra-high performance liquid chromatography. Figure 7-8 It can be seen from the chromatogram that under the above chromatographic conditions, the retention time of cinnamyl acetate is 41.98 min, and no chromatographic peak appears in cinnamon medicinal material at the retention time of 41.98 min.
[0164] 2.5 Result judgment
[0165] There is no chromatographic peak at the retention time of 42.1min in the cinnamon medicinal material test solution. The substance at the retention time of 42.1min is cinnamyl acetate. It can be determined that the cinnamon medicinal material test solution does not contain cinnamyl acetate. Comprehensive judgment: This cinnamon medicinal material was harvested in spring, which is consistent with the actual harvesting time.
Claims
1. A method for determining the harvesting time of cinnamon medicinal materials, characterized in that: The following steps are involved: Step 1: Sample processing The cinnamon medicinal material sample to be identified is peeled, dried and crushed; Step 2: Preparation of test solution Accurately weigh appropriate amounts of cinnamon powder and place them in a conical flask, add solvent to extract, and obtain cinnamon extract; accurately draw an appropriate amount of extract, filter, and obtain the test solution; Step 3: Preparation of reference solution Accurately weigh an appropriate amount of reference substance and place it in a 10 mL volumetric flask, add solvent to dissolve it, and obtain a reference substance solution; Step 4: Data Analysis Analyze the test solution and reference solution obtained in steps 2 and 3, respectively, to obtain data for each sample solution; Step 5: Data processing Analyze the data obtained in step 4 to obtain a chromatogram of the cinnamon medicinal material to be identified and a chromatogram of the reference substance; Step 6: Determine the harvesting time of cinnamon herbs Analyze the data obtained in step 5 to determine the harvesting time of cinnamon medicinal materials.
2. The method according to claim 1, characterized in that In step 2, the extraction solvent used in the preparation of the test solution is a methanol aqueous solution with a concentration of 65-75%, the extraction method is hot reflux extraction, the extraction temperature is 50-100° C., and the extraction time is 20-60 minutes.
3. The method according to claim 1, characterized in that In step 3, the reference substance is cinnamyl acetate and the solvent is methanol.
4. The method according to claim 1, characterized in that: In step 4, high performance liquid chromatography or ultra high performance liquid chromatography is used for detection and analysis, a silane bonded silica gel column is used for separation, acetonitrile and buffer solution are used for gradient elution, and a UV detector or a diode array detector is used for detection.
5. The method according to claim 1, characterized in that The judgment basis of step 6 is that the cinnamon medicinal materials collected in autumn contain cinnamyl acetate, while the cinnamon medicinal materials collected in spring do not contain cinnamyl acetate.