A method for detecting the quality of Blumea balsamifera medicinal material

Abstract

The invention discloses a quality detection method for Blumea Balsamifera medicinal material, comprising the use of thin layer chromatography to identify Blumea Balsamifera medicinal material, mainly comprising: taking a test solution and a reference solution of Blumea Balsamifera medicinal material, spotting them on the same silica gel thin layer plate, using petroleum ether-ethyl acetate-chloroform with a volume ratio of 11:1:3 as a developing agent, spraying with a phosphomolybdic acid test solution with a mass content of 5%, and heating until the spots are clearly colored. The invention has the characteristics of accurately identifying the authenticity of Blumea Balsamifera medicinal material.

Description

Technical Field

[0001] The invention relates to a method for detecting the quality of traditional Chinese medicine, in particular to a method for detecting the quality of Blumea balsamifera medicinal material. Background Art

[0002] Blumeabalsamifera (L.) DC. is a perennial herb or subshrub of the genus Blumeabalsamifera in the Asteraceae family. The stem is stout, erect, 1-3 meters high; the lower leaves are broadly elliptical or oblong-lanceolate, with petioles, and there are 3-5 pairs of narrow linear appendages on both sides of the petioles; the upper leaves are oblong-lanceolate or ovate-lanceolate, sessile or with short petioles, and there are often 1-3 pairs of narrow linear appendages on both sides of the petioles; there are many capitula, bell-shaped involucres, honeycomb-shaped receptacles, 2-3 mm in diameter, glabrous, yellow flowers, many female flowers, thin tubular corollas, fewer bisexual flowers, tubular corollas; achenes are cylindrical, densely pubescent, and the pappus is reddish brown and rough.

[0003] The whole plant of Blumea Balsamifera is used as medicine, which has the effects of dispelling wind and dampness. It has a long history of use in ethnic minority areas such as the Li, Miao, and Zhuang. It is an important folk medicine. Its extract injected into animals can cause a drop in blood pressure, vasodilation, and inhibition of the sympathetic nervous system. It can be used for patients with excitement, insomnia, or hypertension. Blumea Balsamifera is also an aromatic plant. The wormwood oil produced by Blumea Balsamifera has a unique aromatic smell and is widely used in the spices and cosmetics industries.

[0004] The quality standards of raw medicinal materials related to "Ai Na Xiang" are included in the "Quality Standards of Traditional Chinese Medicines and Ethnic Medicinal Materials in Guizhou Province" (2003 edition: Ai Na Xiang), "Guangxi Traditional Chinese Medicine Standards·Volume 2" (Yunnan Gui Ai Na Xiang), "Hunan Traditional Chinese Medicine Standards" (2009 edition: Yunnan Gui Ai Na Xiang), "Guangdong Traditional Chinese Medicine Standards" (2011 edition: Ai Na Xiang). However, the current standards only have provisions for [Properties], [Identification] (powder, root cross section, control medicinal material thin layer, physical and chemical identification), [Inspection] (moisture, total ash content), etc., and there is no accurate identification of its quality. In the traditional Chinese medicine market, the plant traits of Senecio odoratus and Callicarpa ovata are extremely similar to those of Ai Na Xiang plants, but the effects are completely different and easy to confuse. Therefore, the development and establishment of a quality detection method for Ai Na Xiang is of great significance to controlling the overall quality of Ai Na Xiang medicinal materials and ensuring the safety and effectiveness of the clinical use of Ai Na Xiang medicinal materials. Summary of the invention

[0005] The object of the present invention is to provide a method for detecting the quality of Blumea Balsamifera medicinal material, which has the characteristics of accurately identifying the authenticity of Blumea Balsamifera medicinal material.

[0006] The technical scheme of the present invention is a method for detecting the quality of Blumea Balsamifera medicinal materials, which comprises the use of thin layer chromatography to identify Blumea Balsamifera medicinal materials, mainly comprising: taking a test solution and a reference solution of the Blumea Balsamifera medicinal materials, spotting them on the same silica gel thin layer plate, using petroleum ether-ethyl acetate-chloroform with a volume ratio of 11:1:3 as a developing solvent, spraying with a phosphomolybdic acid test solution with a mass content of 5%, and heating until the spots are clearly colored.

[0007] In the aforementioned method for detecting the quality of Blumea balsamifera medicinal materials, the method for preparing the test solution is: taking Blumea balsamifera medicinal materials, ultrasonically extracting them with ethyl acetate solution at a solid-liquid ratio of 1:20-40 ml for 15-60 min, centrifuging, and taking the supernatant as the test solution; the method for preparing the reference solution is: taking borneol reference material and adding methanol to prepare a solution containing 0.3-0.5 mg of borneol per 1 ml, as the reference solution.

[0008] In the aforementioned method for detecting the quality of Blumea balsamifera medicinal materials, the method for preparing the test solution is: taking 1 g of Blumea balsamifera medicinal materials, adding 20 ml of ethyl acetate solution, ultrasonically extracting for 30 minutes, centrifuging, and taking the supernatant as the test solution; the method for preparing the reference solution is: taking the borneol reference material and adding methanol to prepare a solution containing 0.3 mg of borneol per 1 ml, as the reference solution.

[0009] In the aforementioned method for detecting the quality of Blumea balsamifera medicinal material, 8 μl of each of the test solution and the reference solution are taken; the thin layer chromatogram of the test solution shows spots of the same color as the thin layer chromatogram of the reference solution at the corresponding positions of the thin layer chromatogram of the test solution.

[0010] In the aforementioned quality detection method for Blumea Balsamifera medicinal material, the quality detection method further comprises determining the content of eriodictyol in Blumea Balsamifera medicinal material by high performance liquid chromatography, comprising the following steps:

[0011] 1) preparing a reference solution: weighing the eriodictyol reference substance to prepare a solution containing 1 to 1.1 mg of eriodictyol per 1 ml, which is the reference solution;

[0012] 2) Prepare the test solution: weigh the Blumea balsamifera medicinal material, add 70% ethanol at a solid-liquid ratio of 1:30-70, weigh, ultrasonically treat for 30-60 minutes, cool to room temperature, and then make up the lost weight with 70% ethanol, filter, and take the filtrate to obtain the test solution;

[0013] 3) Take the reference solution and the test solution respectively, inject them into the liquid chromatograph, and determine the content of eriodictyol.

[0014] In the aforementioned method for detecting the quality of Blumea Balsamifera medicinal materials, the method for determining the content of eriodictyol in Blumea Balsamifera medicinal materials comprises the following steps:

[0015] 1) Preparation of reference solution: Weigh the eriodictyol reference substance to prepare a solution containing 1.003 mg of eriodictyol per 1 ml, which is the reference solution;

[0016] 2) Prepare the test solution: weigh 0.5 g of Blumea balsamifera, add 25 ml of 70% ethanol, weigh, ultrasonicate for 30 min, cool to room temperature, add 70% ethanol to make up the lost weight, filter, and take the filtrate to obtain the test solution;

[0017] 3) Take 10 μl of the reference solution and the test solution respectively, inject them into the liquid chromatograph and measure.

[0018] In the aforementioned method for detecting the quality of Blumea balsamifera medicinal materials, the chromatographic conditions of the liquid chromatograph are as follows: the chromatographic column is a C18 column, the detection wavelength is 287 nm, the flow rate is 1 ml / min; the column temperature is 35° C.; the mobile phase is acetonitrile and acetic acid water with a mass content of 0.1% for isocratic elution.

[0019] In the aforementioned method for detecting the quality of Blumea balsamifera medicinal materials, the volume ratio of acetonitrile to 0.1% acetic acid water in the mobile phase is 22:78.

[0020] In the aforementioned method for detecting the quality of Blumea balsamifera medicinal material, in the ultrasonic treatment in step 2), the power is 300 W and the frequency is 40 KHz.

[0021] In the aforementioned quality testing method for Blumea Balsamifera medicinal material, the Blumea Balsamifera medicinal material, calculated on the basis of dry product, shall contain no less than 0.10% eriodictyol.

[0022] Compared with the prior art, the present invention has the following beneficial effects:

[0023] The invention can accurately identify the authenticity of the Blumea Balsamifera medicinal material through a specific thin-layer chromatography identification method, has high sensitivity, strong specificity, good separation effect, simplicity and feasibility, strong reproducibility, and can effectively distinguish the Blumea Balsamifera medicinal material from its counterfeits and confused products such as Senecio radiata and Callicarpa macrophylla, thereby avoiding misuse and improving the safety of medicinal material use.

[0024] The present invention can also accurately and quickly determine the content of eriodictyol in the Blumea Balsamifera medicinal material through a specific high performance liquid chromatography method, thereby providing an effective qualitative and quantitative analysis method for the Blumea Balsamifera medicinal material, thereby facilitating the quality control of the Blumea Balsamifera medicinal material, effectively controlling the overall quality of the Blumea Balsamifera medicinal material, and ensuring the safety and effectiveness of the clinical use of the Blumea Balsamifera medicinal material.

[0025] The quality detection method of the Blumea Balsamifera medicinal material provided by the present invention is helpful to make up for the deficiencies of the quality detection methods in the "Quality Standards of Traditional Chinese Medicines and National Medicinal Materials of Guizhou Province" (2003 edition: Blumea Balsamifera), "Guangxi Traditional Chinese Medicine Standards Volume 2" (Diangui Blumea Balsamifera), "Hunan Traditional Chinese Medicine Standards" (2009 edition: Diangui Blumea Balsamifera), and "Guangdong Traditional Chinese Medicine Standards" (2011 edition: Blumea Balsamifera), and has strong practicality. The method of the present invention is of great significance for further quality control of Blumea Balsamifera medicinal material, and can provide a scientific basis for the formulation of the quality table of Blumea Balsamifera medicinal material. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 It is a thin layer chromatogram of different test solution preparation methods of the present invention;

[0027] Figure 2 It is a TLC survey diagram of different test sample preparation solvents;

[0028] Figure 3 It is the thin layer chromatography survey diagram of different ultrasonic extraction time;

[0029] Figure 4 It is a thin layer chromatographic survey diagram of the sample solution;

[0030] Figure 5 It is the thin layer chromatogram of borneol reference substance and 11 different medicinal materials;

[0031] Figure 6 This is the UV absorption graph of eriodictyol;

[0032] Figure 7 is the mobile phase liquid chromatogram;

[0033] Figure 8 This is the liquid chromatogram of eriodictyol standard;

[0034] Fig. 9 is the sample liquid chromatogram;

[0035] Fig.10 It is the linear relationship graph of eriodictyol. DETAILED DESCRIPTION

[0036] The present invention will be further described below in conjunction with the embodiments, but they are not intended to limit the present invention.

[0037] Example:

[0038] The instruments used are:

[0039] Shimadzu LC-20ADXR high performance liquid chromatograph, SPD-20A ultraviolet detector, Shimadzu LC-Solution workstation; MilliporeSimplicity ultrapure water purifier (Merck Millipore); AUW220D electronic analytical balance (Shanghai Tianping Instrument Factory); AB135-S one-hundred-thousandth balance (Mettler Toledo); chromatographic column: Cosmosil C18 (5μm, 4.6mm×250mm); KQ250DB CNC ultrasonic cleaner (Gongyi Yuhua Instrument Co., Ltd.); KQ5200 water bath (Kunshan Ultrasonic Instrument Co., Ltd.); DHG-9140A electric constant temperature blast drying oven (Gongyi Yuhua Instrument Co., Ltd.); 8-10 box-type resistance furnace (manufactured by Shenyang Energy-saving Electric Furnace Factory, China); Sony717 digital camera (Sony Corporation, Japan).

[0040] Reagents and test drugs

[0041] 1. Reference

[0042] Erythoracol (content greater than 98%, batch number S-076-150122, purchased from Chengdu Ruifensi Biotechnology Co., Ltd.); borneol (China Food and Drug Inspection Institute, batch number 110881-201508).

[0043] 2. Reagents

[0044] Acetonitrile (chromatographically pure, Yucheng Chemical Plant of Shandong Yuwang Industrial Co., Ltd.); methanol (analytical pure, Shanghai Zhenxing Chemical Plant No. 1); formic acid (analytical pure, Shanghai Zhenxing Chemical Plant No. 1); methanol (chromatographically pure, Yucheng Chemical Plant of Shandong Yuwang Industrial Co., Ltd.).

[0045] 3. Medicinal materials

[0046] The medicinal materials used in the experiment were provided by Guizhou Bailing Pharmaceutical Co., Ltd. and were identified as Blume abalsamifera (L.) DC. of the Asteraceae family by Professor Zhong Guoyue from the Center for Ethnic and Traditional Chinese Medicine Germplasm Resources of Jiangxi University of Traditional Chinese Medicine (see Table 1 for details).

[0047] Table 1 Batches of Blumea Balsamifera medicinal materials

[0048]

[0049] Blumea balsamifera leaves contain flavonoids: quercetin, (2R,3R)-dihydroquercetin-4′-methyl ether, (2R,3R)-dihydroquercetin-4′,7-dimethyl ether, (2R,3R)-7,5′-dimethoxy-3,5,2′-trihydroxyflavanone, (2R,3R)-5′-methyl-3,5,7,2′-tetrahydroxyflavanone, eriodictyol, Blumea balsamifera, hyperoside, etc.; volatile oils: l-borneol, borneol, isoborneol, camphor, cedarwood glycol, Blumea balsamifera lactones A, B, C, etc.

[0050] Since borneol is one of the main components of Blumea Balsamifera, TLC identification with borneol as reference material was added. Meanwhile, in the actual application of the standard, “reference medicinal materials” are not easy to obtain, so TLC identification of reference medicinal materials was cancelled.

[0051] A method for detecting the quality of Blumea balsamifera medicinal material comprises the steps of identifying the Blumea balsamifera medicinal material by thin layer chromatography and determining the content of eriochoride in the Blumea balsamifera medicinal material by high performance liquid chromatography.

[0052] Among them, the identification method of Blumea balsamifera medicinal materials according to thin layer chromatography (General Rule 0502) is:

[0053] 1) Take 1 g of Blumea Balsamifera powder and place it in a conical flask, add 20 ml of ethyl acetate solution and ultrasonically extract for 30 minutes, centrifuge, and take the supernatant as the test solution of Blumea Balsamifera; take 20 mg of borneol reference substance and add methanol to make a solution containing 0.3 mg of borneol per 1 ml as the reference substance solution;

[0054] 2) Take 8 μl of the test solution and reference solution of Blumea balsamifera and spot them on the same silica gel GF 254 On a thin layer plate (50×100 mm), use petroleum ether-ethyl acetate-chloroform with a volume ratio of 11:1:3 as the developing solvent, develop 8 cm upward, take out, dry, spray with 5% phosphomolybdic acid test solution for color development, heat in an oven at 80°C until the spots are clearly colored, and observe under sunlight. In the thin layer chromatogram of the test solution, a spot of the same color as the thin layer chromatogram of the reference solution appears at the corresponding position of the thin layer chromatogram of the reference solution.

[0055] 1. Investigation of the identification methods of Blumea balsamifera

[0056] 1.1 Investigation of extraction methods:

[0057] During the experiment, three extraction methods were compared: ultrasonic extraction of ethyl acetate for 30 minutes, reflux of ethyl acetate for 1.5 hours, and cold soaking of ethyl acetate overnight. Figure 1 The results show that there is no significant difference between the different extraction methods. Considering energy and time saving, ultrasonic extraction method was selected.

[0058] 1.2 Solvent investigation:

[0059] Ultrasonic extraction was used to investigate the extraction solvents. Methanol, ethyl acetate and anhydrous ethanol were selected for extraction. The extraction results are shown in Figure 2 The results showed that the best effect was obtained by ultrasonic extraction of methanol solution for 30 minutes.

[0060] 1.3 Extraction time investigation:

[0061] The ultrasonic extraction method was used to investigate the extraction time. Three portions of 1g of the medicinal material were placed in a conical flask, and 20ml of methanol was added. The extraction was ultrasonicated for 15 minutes, 30 minutes, and 1 hour respectively, and then filtered for use. The extraction results are shown in the figure. Figure 3 The results showed that the ultrasonic extraction of methanol solution for 30 minutes had the best effect.

[0062] 1.4 Sampling quantity inspection:

[0063] According to the thin layer chromatography test, take the above-mentioned reference solution borneol, and the test solution 2μL, 4μL, 8μL, and 10μL after ultrasonication for 30 minutes, and spot them on the same thin layer chromatography plate respectively for inspection. The results are as follows Figure 4 When the sample solution was 8 μL, the spots were clearest, so the final spotting volume was 8 μL.

[0064] 1.5 Thin layer chromatograms of different batches of medicinal materials:

[0065] According to the above method, different batches of Blumea Balsamifera were tested, and the results are shown in Figure 5 .

[0066] The original standard did not contain any provisions for the determination of the content of Blumea Balsamifera. Blumea Balsamifera mainly contains flavonoids, volatile oils and other ingredients. Studies have shown that the flavonoids contained in Blumea Balsamifera have significant antioxidant effects, which is one of its mechanisms of protection against acute liver damage, and the antioxidant activity of flavonoids containing free hydroxyl groups is greater than that of methylated compounds, among which eriodictyol is one of its antioxidant active ingredients. Through the study of the chemical composition of Blumea Balsamifera, it was found that the content of eriodictyol in its flavonoid components is relatively high; at the same time, Blumea Balsamifera as a medicinal material is different from "Ai Tablets", and in the [Identification] item, borneol is used as a reference for thin layer identification, so eriodictyol (C 15 H 12 O6) was used as an indicator component to conduct content determination research and establish a content determination method.

[0067]

[0068] The present invention determines the content of eriodictyol in the medicinal material Blumea balsamifera according to high performance liquid chromatography (General Rule 0512 of Part 4 of the 2015 edition of the Chinese Pharmacopoeia), and the determination method is:

[0069] 1) Preparation of reference solution: Weigh the eriodictyol reference substance to prepare a solution containing 1.003 mg of eriodictyol per 1 ml, which is the reference solution;

[0070] 2) Prepare the test solution: weigh 0.5 g of Blumea balsamifera powder, place it in a stoppered conical flask, add 25 ml of 70% ethanol, weigh it, and perform ultrasonic treatment for 30 min at an ultrasonic power of 300 W and an ultrasonic frequency of 40 KHz; cool it to room temperature, then make up the lost weight with 70% ethanol, filter it, and take the filtrate to obtain the test solution;

[0071] 3) Take 10 μl of the reference solution and the test solution respectively, inject them into the liquid chromatograph and measure.

[0072] Calculated on the basis of dry product, the content of eriodictyol in Blumea balsamifera must not be less than 0.10%.

[0073] Among them, the chromatographic column of the high performance liquid chromatography method is a Cosmosil C18 column (5 μm, 4.6×250 mm), the detection wavelength is 287 nm, the flow rate is 1 ml / min; the column temperature is 35° C.; the mobile phase is acetonitrile and 0.1% acetic acid water for isocratic elution, and the volume ratio of acetonitrile to 0.1% acetic acid water in the mobile phase is 22:78.

[0074] 2. Investigation of the determination method of eriodictyol content in Blumea Balsamifera:

[0075] 2.1 Detection wavelength investigation:

[0076] The above method is used to scan the wavelength using the wavelength scanning function of the diode array detector in the high performance liquid chromatograph to obtain the ultraviolet absorption spectrum of the eriodictyol reference substance (such as Figure 6 ). Figure 6 It can be seen that eriodictyol has maximum absorption at 287nm, so 287nm is selected as the detection wavelength.

[0077] 2.2 Extraction solvent investigation:

[0078] Weigh three portions of Blumea balsamifera powder, 0.5 g each, and place them in a stoppered conical flask. Add 25 ml of methanol, ethanol, and water, respectively, weigh them, treat them with ultrasound for 30 min, cool them, make up the lost weight, filter them, and take the filtrate. Inject the samples according to the above chromatic conditions and analyze them. The results are shown in Table 1.

[0079] Table 1 Investigation results of solvents for extracting Blumea balsamifera

[0080]

[0081]

[0082] The results of the investigation of the above three solvents showed that the extraction efficiency of methanol and ethanol was similar, while water could not extract eriodictyol. Therefore, we continued to investigate different concentrations of methanol and ethanol to determine the extraction solvent.

[0083] 2.3 Investigation of different concentrations of ethanol and methanol:

[0084] Weigh four portions of Blumea Balsamifera powder, 0.5 g each, and place them in a stoppered conical bottle. Add 25 ml of 90% ethanol, 70% ethanol, 50% ethanol, and 30% ethanol, respectively, weigh them, treat them with ultrasound for 30 minutes, cool them, make up the lost weight, filter them, and take the filtrate. Inject the samples according to the above chromatographic conditions and analyze them. The results are shown in Table 2.

[0085] Table 2 Results of investigation on different concentrations of ethanol

[0086]

[0087] Weigh four portions of Blumea balsamifera powder, 0.5 g each, and place them in stoppered conical bottles. Add 25 ml of 90% methanol, 70% methanol, 50% methanol, and 30% methanol, respectively, weigh them, and treat them with ultrasound for 30 minutes. Cool them, make up the lost weight, filter them, and take the filtrate. Inject the samples according to the above chromatographic conditions and analyze them. The results are shown in Table 3.

[0088] Table 3 Results of investigation on different concentrations of methanol

[0089]

[0090] The results of the investigation on the extraction effects of different concentrations of methanol and ethanol showed that the extraction efficiency of 70% ethanol and 70% methanol for eriodictyol was similar. After comprehensive consideration, 70% ethanol was selected as the extraction solvent.

[0091] 2.4 Investigation of different extraction methods:

[0092] Weigh three portions of Blumea Balsamifera powder, 0.5 g each, and place them in a stoppered conical flask. Add 25 ml of 70% ethanol and weigh them. Consider three extraction methods: reflux for 30 min, ultrasonic for 30 min, and cold soak for 1 h. Cool, make up for the lost weight, filter, and take the filtrate. Inject and measure according to the above chromatographic conditions. The results are shown in Table 4.

[0093] Table 4 Results of investigation of different extraction methods

[0094]

[0095] From the results of the three extraction methods, it can be seen that the contents of eriodictyol in reflux extraction and ultrasonic extraction are similar. Considering the simplicity and energy saving of ultrasonic extraction, ultrasonic extraction method was selected.

[0096] 2.5 Extraction time investigation:

[0097] Weigh three portions of Blumea Balsamifera powder, 0.5 g each, accurately weigh, place in round-bottom flasks, add 25 ml of 70% methanol, weigh, ultrasonicate for 15, 30, and 60 min, cool, make up for the lost weight, filter, and take the filtrate. Inject and measure according to the above chromatographic conditions. The results are shown in Table 5.

[0098] Table 5 Extraction time investigation results

[0099]

[0100] The above table shows that the extraction efficiencies of ultrasonic extraction for 30 min and 60 min are similar, so the ultrasonic extraction time is determined to be 30 min.

[0101] 2.6 Investigation of solvent dosage

[0102] Weigh three portions of Blumea Balsamifera powder, 0.5 g each, accurately weigh, place in round-bottom flasks, add 15, 25, 35 ml of 70% ethanol, weigh, ultrasonicate for 30 min, cool, make up for the lost weight, filter, and take the filtrate. Inject and measure according to the above chromatographic conditions. The results are shown in Table 6.

[0103] Table 6 Solvent dosage investigation results

[0104]

[0105]

[0106] The results show that most of the components can be extracted using 25 ml of solvent. Considering cost savings, the amount of solvent is determined to be 25 ml.

[0107] Based on the above methodological investigation results, the preparation method of the test solution was determined as follows: accurately weigh 0.5 g of Blumea balsamifera powder, place it in a stoppered conical flask, add 25 ml of 70% ethanol, weigh it, and ultrasonically treat it for 30 minutes with an ultrasonic power of 300 W and an ultrasonic frequency of 40 kHz; cool it, then make up the lost weight with 70% ethanol, filter it, and take the filtrate to obtain the test solution.

[0108] 3. Methodological Validation:

[0109] 3.1 System adaptability:

[0110] The determination was carried out by high performance liquid chromatography, wherein the liquid chromatogram of the mobile phase is shown in Figure 7 The liquid chromatogram of the standard substance of eriodictyol is shown in Figure 8 , the liquid chromatogram of the Blumea balsamifera sample is shown in Fig. 9 , indicating that acetonitrile-0.1% acetic acid aqueous solution (volume ratio of 22:78) was well separated.

[0111] 3.2 Linear relationship investigation:

[0112] Six portions of different concentrations of the eriodictyol reference solution were accurately taken, and 10 μL of each was injected for determination. The peak area (y) was used as the ordinate and the injection reference concentration (x) was used as the abscissa for linear regression. The linear regression equation of eriodictyol was: y=36.694x+44967, R2=0.9994. The results are shown in Table 7 and Fig.10 .

[0113] Table 7 Linear relationship investigation results

[0114]

[0115] The results showed that the concentration of eriodictyol in the range of 1.5875 to 250ug·ml-1 had a good linear relationship with the peak area.

[0116] 3.3 Precision test

[0117] The same portion of eriodictyol reference solution was sampled six times continuously and its RSD value was calculated. The results are shown in Table 8.

[0118] Table 8 Precision test results

[0119]

[0120] 3.4 Repeatability test

[0121] According to the method for preparing the test solution determined above, 6 portions of Blumea balsamifera medicinal material test solution were prepared in parallel for determination, and their RSD values ​​were calculated. The results are shown in Table 9.

[0122] Table 9 Repeatability test results

[0123]

[0124] The results showed that 6 samples of Blumea balsamifera medicinal material test solution were prepared in parallel for determination, and the RSD value of eriodictyol was 1.032%, indicating that the method had good repeatability.

[0125] 3.5 Solution stability test

[0126] Take the same sample solution of Blumea balsamifera and measure it after 0, 3, 6, 9 and 12 hours respectively, record the peak area and calculate the RSD value. The results are shown in Table 10.

[0127] Table 10 Solution stability test results

[0128]

[0129]

[0130] The results showed that 10 μl of the same Blumea balsamifera sample solution was taken for determination at 0, 3, 6, 9 and 12 h, respectively, and the RSD value of the eriodictyol peak area was 0.408%, indicating that the solution had good stability under this method.

[0131] 3.6 Sample recovery rate

[0132] Accurately weigh 6 samples of the same batch of Blumea balsamifera with known content, accurately add appropriate amount of eriodictyol, and measure according to the method under the test sample. The results are shown in Table 11.

[0133] Table 11 Sample recovery test results (n = 6)

[0134]

[0135] The results showed that the recovery rate of eriochoride in the same batch of Blumea Balsamifera was 100.35% (RSD=0.48%), indicating that the recovery rate of the established method was good.

[0136] 3.7 Sample content determination

[0137] Take 0.5 g of each of 10 batches of Blumea balsamifera medicinal materials, accurately weigh them, prepare them according to the preparation method of the test solution, take 10 μl of each sample for injection, record the peak area, and calculate the mass fraction of each component using the regression equation. The results are shown in Table 12.

[0138] Table 1210 batch Blumea Balsamifera content determination results (mg / g)

[0139]

[0140]

[0141] The experimental results showed that the content of Artemisia balsamifera in the medicinal material ranged from 0.10% to 0.18%, with an average of 0.13%. The average was reduced by 20%, which was 0.10%. All 10 batches of samples were qualified, so the limit of the content of Artemisia balsamifera was determined to be no less than 0.10%.

Claims

1. A method for detecting the quality of Blumea Balsamifera medicinal material, characterized in that: The method comprises the following steps: taking a test solution and a reference solution of the herb, spotting them on the same silica gel thin layer plate, using petroleum ether-ethyl acetate-chloroform with a volume ratio of 11:1:3 as a developing solvent, spraying with a phosphomolybdic acid test solution with a mass content of 5%, and heating until the spots are clearly colored; The preparation method of the test solution is as follows: take the medicinal material of Blumea balsamifera, ultrasonically extract it with ethyl acetate solution at a solid-liquid ratio of 1:20~40ml for 15~60min, centrifuge, and take the supernatant as the test solution; the preparation method of the reference solution is as follows: take the borneol reference substance and add methanol to prepare a solution containing 0.3~0.5mg of borneol per 1ml, which is used as the reference solution; The quality detection method also includes determining the content of eriodictyol in the Blumea Balsamifera medicinal material by high performance liquid chromatography, comprising the following steps: 1) Preparation of reference solution: Weigh the eriodictyol reference substance to prepare a solution containing 1-1.1 mg of eriodictyol per 1 ml, which is the reference solution; 2) Prepare the test solution: weigh the Herba Blumea Balsamifera, add 70% ethanol at a solid-liquid ratio of 1:30-70, weigh, ultrasonicate for 30-60 min, cool to room temperature, add 70% ethanol to make up the lost weight, filter, and take the filtrate to obtain the test solution; 3) Take the reference solution and the test solution respectively, inject them into the liquid chromatograph, and determine the content of eriodictyol; The chromatographic conditions of the liquid chromatograph are as follows: the chromatographic column is a C18 column, the detection wavelength is 287 nm, the flow rate is 1 ml / min; the column temperature is 35°C; the mobile phase is an isocratic elution of acetonitrile and 0.1% acetic acid water in a volume ratio of 22:

78.

2. A method for detecting the quality of Blumea Balsamifera medicinal material according to claim 1, characterized in that: The preparation method of the test solution is as follows: take 1 g of Blumea balsamifera medicinal material, add 20 ml of ethyl acetate solution, ultrasonically extract for 30 minutes, centrifuge, and take the supernatant as the test solution; the preparation method of the reference solution is as follows: take borneol reference and add methanol to make a solution containing 0.3 mg of borneol per 1 ml, which is used as the reference solution.

3. A method for detecting the quality of Blumea Balsamifera medicinal material according to claim 1, characterized in that: Take 8 μl of each of the test solution and the reference solution; the TLC of the test solution should show spots of the same color as those of the reference solution at the corresponding positions.

4. A method for detecting the quality of Blumea Balsamifera medicinal material according to claim 1, characterized in that: The method for determining the content of eriodictyol in Blumea Balsamifera medicinal material comprises the following steps: 1) Preparation of reference solution: Weigh the eriodictyol reference substance to prepare a solution containing 1.003 mg of eriodictyol per 1 ml, which is the reference solution; 2) Prepare the test solution: weigh 0.5 g of Blumea balsamifera, add 25 ml of 70% ethanol, weigh, ultrasonicate for 30 min, cool to room temperature, add 70% ethanol to make up the lost weight, filter, and take the filtrate to obtain the test solution; 3) Take 10 μl of the reference solution and the test solution respectively, inject them into the liquid chromatograph, and determine the content of eriodictyol.

5. A method for detecting the quality of Blumea Balsamifera medicinal materials according to claim 1 or 4, characterized in that: In the ultrasonic treatment in step 2), the power is 300 W and the frequency is 40 KHz.

6. A method for detecting the quality of Blumea Balsamifera medicinal materials according to claim 1 or 4, characterized in that: Calculated on the basis of dry product, the content of eriodictyol in Blumea balsamifera must not be less than 0.10%.

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