Taraxacum mongolicum quality control method

Abstract

The invention discloses a dandelion quality control method, and relates to the technical field of traditional Chinese medicine detection. The detection method comprises the following steps: (1) preparing a test solution: dissolving dandelion in a solvent, performing ultrasonic treatment, and taking filtrate to obtain the test solution; and (2) determining the content by using a liquid chromatography. The method disclosed by the invention is small in interference, good in durability, high in linear correlation degree, good in repeatability and stability and high in precision.

Description

Technical Field

[0001] The invention relates to the technical field of traditional Chinese medicine detection, and in particular to a quality control method for dandelions. Background Art

[0002] Dandelion is also known as Dandelion, Huanghua Diding, Huanghua Panax notoginseng, etc. Dandelion contains flavonoids, phenolic acids, terpenes, pigments, phytosterols and other compounds, which have anti-inflammatory, antibacterial, liver-protecting, anti-tumor, antioxidant, hypoglycemic effects, and are often used to treat carbuncle, breast carbuncle, red eyes, lung carbuncle, intestinal carbuncle, damp-heat jaundice and hot stranguria.

[0003] Caffeoyltartaric acid is an important phenolic acid compound in dandelion, also known as caffeoyltartaric acid and caffeoyltartaric acid. Its structural formula is as follows:

[0004]

[0005] Chinese patent application CN202111315005.X discloses a method for simultaneously detecting multiple components in multiple Chinese medicine extracts using UPLCPDA, comprising the following steps: preparing a test solution: taking a test sample, adding a solvent, ultrasonically extracting, centrifuging, filtering to obtain a supernatant, which is a test solution, for standby use; preparing a reference stock solution: taking each component to be detected as a reference, adding a solvent to prepare each reference stock solution, for standby use; UPLC-PDA detection: taking the test solution obtained in step 1 and each reference stock solution obtained in step 2, respectively, injecting them into an ultra-high performance liquid chromatograph, and performing gradient elution with a PDA detector to obtain chromatograms and spectra of the test sample and the reference respectively; by comparing and analyzing the spectra and chromatograms of the test sample and the reference, qualitative identification and quantitative determination of each component. The invention can detect chlorogenic acid, cryptochlorogenic acid, neochlorogenic acid, 4,5-di-O-caffeoylquinic acid, chicoric acid and monocaffeoyltartaric acid in Echinacea extract, dandelion extract and honeysuckle extract.

[0006] Chinese patent application CN202211063030.8 discloses a quality evaluation method for Pudilan Anti-inflammatory Tablets based on one-test-multiple-evaluation, including preparing a single reference solution, preparing a mixed reference solution, preparing a test solution, introducing the mixed reference solution into a high-performance liquid chromatograph, injecting and measuring, obtaining a liquid chromatogram, recording the peak area of ​​each component, using a multi-point correction method, taking baicalin as an internal reference, calculating the relative correction factor of the other 6 components to be tested; analyzing and determining the basis for the positioning of the chromatographic peak of the component to be tested. This invention creates a fast, accurate, and low-cost quality control and evaluation method for Pudilan Anti-inflammatory Tablets, which simultaneously determines the content of 7 index components (monocaffeoyltartaric acid, chicoric acid, baicalin, melaleucalycin A-7-0-β-D-glucuronide, wogonin, baicalein, and wogonin) through one-test-multiple-evaluation.

[0007] The current quality control method for dandelion has a complex detection process, large interference, poor repeatability and stability, and low precision. Therefore, there is currently a lack of effective quality control methods for dandelion as a raw material for traditional Chinese medicine preparations. Summary of the invention

[0008] The invention aims to provide a quality control method for dandelion, which realizes quality control by determining the content of the active ingredient monocaffeoyltartaric acid, has small interference, good durability, high linear correlation, good repeatability and stability, and high precision.

[0009] To achieve the above-mentioned purpose of the invention, the technical solution of the present invention is as follows:

[0010] A dandelion quality control method is provided, wherein the quality control is achieved by measuring the content of monocaffeoyltartaric acid, and the method comprises the following steps:

[0011] (1) preparing a reference solution: diluting monocaffeoyltartaric acid to obtain a reference solution;

[0012] (2) Preparation of test solution: dissolve dandelion in a solvent, perform ultrasonic treatment, and collect the filtrate to obtain the test solution;

[0013] (3) Liquid chromatography was used for content determination. The chromatographic conditions were as follows:

[0014] Chromatographic column: chromatographic column with octadecylsilane bonded silica gel as filler;

[0015] Column temperature: 30-40°C;

[0016] Flow rate: 0.6-1.0mL / min;

[0017] Elution procedure: acetonitrile as mobile phase A, 0.15-0.25% phosphoric acid as mobile phase B, gradient elution:

[0018] Table 1.

[0019] time Mobile phase A Mobile phase B 0min 6-10％ 90-94％ 30min 6-10％ 90-94％ 35min 88-92％ 8-12％ 40min 6-10％ 90-94％ 45min 6-10％ 90-94％

[0020] Detection wavelength: 310-350nm;

[0021] Injection volume: 5-15μL.

[0022] Preferably, in step (1), the monocaffeoyltartaric acid is diluted with a methanol solution, and the concentration of the reference solution is 5-15 μg / mL.

[0023] Further preferably, in step (1), the monocaffeoyltartaric acid is diluted with a 40-60% methanol solution, and the concentration of the reference solution is 8-12 μg / mL.

[0024] More preferably, in step (1), the monocaffeoyltartaric acid is diluted with a 50% methanol solution, and the concentration of the reference solution is 10 μg / mL.

[0025] Preferably, in step (2), the dandelion is dandelion powder.

[0026] Preferably, in step (2), the solvent is 20-50% by volume methanol aqueous solution or water; or 10-40% by volume methanol aqueous solution or water, more preferably 20% methanol or water, most preferably 20% methanol.

[0027] Preferably, in step (2), the conditions for the ultrasonic treatment are: power 400-600 W, frequency 40-50 KHZ, time 45-60 min; most preferably: power 500 W, frequency 45 KHZ, time 45 min.

[0028] Preferably, in step (2), the mass volume ratio of dandelion to solvent is 1:5-200, more preferably 1:100.

[0029] Preferably, in step (2), the weight of the dandelion sample is 0.2-0.5 g, most preferably 0.5 g.

[0030] As a specific embodiment of the present invention, the steps of preparing the test solution are as follows: accurately weigh dandelion powder, place it in a conical flask, accurately add 50 mL of 20% methanol, ultrasonically treat, cool, make up the lost weight with 20% methanol, shake well, filter, and take the filtrate to obtain the test solution.

[0031] Preferably, in step (3), the chromatographic column is any one of SMHADZU-Shim-pack GIST C18 column, SMHADZU Shim-pack GIST C18, Agilent ZORBAX SB-C18, and Amethyst C18-H.

[0032] Preferably, in step (3), the column temperature is 34-36°C, most preferably 35°C.

[0033] Preferably, in step (3), the flow rate is 0.7-0.9 mL / min, most preferably 0.8 mL / min.

[0034] Preferably, in step (3), the mobile phase B is 0.2% phosphoric acid.

[0035] Preferably, in step (3), the gradient elution procedure is:

[0036] Table 2.

[0037]

[0038]

[0039] Preferably, in step (3), the detection wavelength is 330 nm.

[0040] Preferably, in step (3), the injection volume is 10 μL.

[0041] As a specific embodiment of the present invention, the chromatographic conditions are as follows:

[0042] Chromatographic column: SMHADZU-Shim-pack GIST C18 column (4.6 mm × 250 mm, 5 μm);

[0043] Column temperature: 35 °C;

[0044] Flow rate: 0.8 mL / min;

[0045] Elution procedure: acetonitrile as mobile phase A, 0.2% phosphoric acid as mobile phase B, gradient elution:

[0046] Table 3.

[0047] time Mobile phase A Mobile phase B 0min 8％ 92％ 30min 8％ 92％ 35min 90％ 10％ 40min 8％ 92％ 45min 8％ 92％

[0048] Detection wavelength: 330nm;

[0049] Injection volume: 10 μL.

[0050] The beneficial effects of the present invention are:

[0051] The invention provides a dandelion quality control method, which realizes quality control by measuring the content of the active ingredient monocaffeoyltartaric acid, has small interference, good durability, high linear correlation, good repeatability and stability, and high precision. BRIEF DESCRIPTION OF THE DRAWINGS

[0052] Figure 1 The chromatograms are for different extraction solvents;

[0053] Figure 2 The chromatograms are obtained at different ultrasonic extraction times;

[0054] Figure 3 The chromatograms are for different dandelion sample weights;

[0055] Figure 4 It is the chromatogram of different solvent volumes;

[0056] Figure 5 It is the standard curve diagram of Example 1;

[0057] Figure 6 is the linear overlapping chromatogram of Example 1;

[0058] Figure 7 Overlay chromatograms for precision;

[0059] Figure 8 Overlay chromatograms for stability;

[0060] Fig. 9 is a repetitive overlapping chromatogram;

[0061] Fig.10 Overlay chromatograms for recovery;

[0062] Fig.11 Example 1 Detection chromatogram;

[0063] Fig.12 Detection chromatogram for Comparative Example 1;

[0064] Fig.13 Detection chromatogram for Comparative Example 2;

[0065] Fig.14 The chromatogram was detected for Comparative Example 3. DETAILED DESCRIPTION

[0066] The following non-limiting examples can enable those skilled in the art to more fully understand the present invention, but do not limit the present invention in any way. The following content is only an exemplary description of the scope of the present invention, and those skilled in the art can make various changes and modifications to the invention of the present invention based on the disclosed content, and they should also fall within the scope of the present application.

[0067] The present invention is further described below by way of specific examples. The various chemical reagents used in the examples of the present invention are obtained through conventional commercial channels unless otherwise specified. Unless otherwise specified, the contents described below are all weight contents. Unless otherwise specified, it is understood that the process is carried out at room temperature.

[0068] Some experimental materials of the present invention are as follows:

[0069] Agilent ZORBAX SB-C18 (250×4.6mm, 5μm) chromatographic column; SHIMADZU shim-pack GISTC18 (4.6mm×250mm, 5μm); Amethyst C18-H (250×4.6mm, 5μm) chromatographic column; AG285 electronic analytical balance; KQ-500VDE ultrasonic cleaner.

[0070] Reference substance: monocaffeoyltartaric acid (batch number: 112087-202101), China Food and Drug Inspection Institute.

[0071] Chromatographic methanol from Yunnan Xinlanjing; chromatographic acetonitrile from Yunnan Xinlanjing; chromatographic phosphoric acid from Tianjin Damao Chemical Reagent Factory; Wahaha drinking purified water from Baishan Wahaha Beverage Co., Ltd.; methanol from Tianjin Hedong District Hongyan Reagent Factory; dandelion slices from Shaanxi Xingshengde Pharmaceutical Co., Ltd.

[0072] The embodiment of the present invention provides a dandelion quality control method, which realizes quality control by measuring the content of monocaffeoyltartaric acid, and comprises the following steps:

[0073] (1) preparing a reference solution: diluting monocaffeoyltartaric acid to obtain a reference solution;

[0074] (2) Preparation of test solution: dissolve dandelion in a solvent, perform ultrasonic treatment, and collect the filtrate to obtain the test solution;

[0075] (3) Liquid chromatography was used for content determination. The chromatographic conditions were as follows:

[0076] Chromatographic column: chromatographic column with octadecylsilane bonded silica gel as filler;

[0077] Column temperature: 30-40°C;

[0078] Flow rate: 0.6-1.0mL / min;

[0079] Elution procedure: acetonitrile as mobile phase A, 0.15-0.25% phosphoric acid as mobile phase B, gradient elution:

[0080] Table 4.

[0081] time Mobile phase A Mobile phase B 0min 6-10％ 90-94％ 30min 6-10％ 90-94％ 35min 88-92％ 8-12％ 40min 6-10％ 90-94％ 45min 6-10％ 90-94％

[0082] Detection wavelength: 310-350nm; for example, the detection wavelength is 310nm, 315nm, 320nm, 325nm, 328nm, 332nm, 335nm, 340nm or 350nm, wherein the optimal embodiment is 330nm

[0083] Injection volume: 5-15 μL, for example: 5 μL, 6 μL, 7 μL, 8 μL, 9 μL, 10 μL, 11 μL, 12 μL, 13 μL, 14 μL or 15 μL, wherein the optimal embodiment is 10 μL.

[0084] In some embodiments, in step (1), the monocaffeoyltartaric acid is diluted with a methanol solution, and the concentration of the reference solution is 5-15 μg / mL.

[0085] In some embodiments, in step (1), the monocaffeoyltartaric acid is diluted with a 40-60% methanol solution, and the concentration of the reference solution is 8-12 μg / mL.

[0086] In some preferred embodiments, in step (1), the monocaffeoyltartaric acid is diluted with a 50% methanol solution, and the concentration of the reference solution is 10 μg / mL.

[0087] In some embodiments, in step (2), the dandelion is dandelion powder.

[0088] In some embodiments, in step (2), the solvent is a methanol aqueous solution or water with a volume fraction of 20-50%, more preferably 20% methanol or water, and most preferably 20% methanol.

[0089] In some embodiments, in step (2), the conditions of the ultrasonic treatment are: power 400-600 W, frequency 40-50 KHZ, time 45-60 min; most preferably: power 500 W, frequency 45 KHZ, time 45 min.

[0090] In some embodiments, in step (2), the mass volume ratio of dandelion to solvent is 1:5-200, and more preferably 1:100.

[0091] In some embodiments, in step (2), the weight of the dandelion sample is 0.2-0.5 g, most preferably 0.5 g.

[0092] As a specific embodiment of the present invention, the steps of preparing the test solution are as follows: accurately weigh dandelion powder, place it in a conical flask, accurately add 50 mL of 20% methanol, ultrasonically treat, cool, make up the lost weight with 20% methanol, shake well, filter, and take the filtrate to obtain the test solution.

[0093] In some embodiments, in step (3), the chromatographic column is any one of SMHADZU-Shim-pack GIST C18 column, SMHADZU Shim-pack GIST C18, Agilent ZORBAX SB-C18, and Amethyst C18-H.

[0094] In some embodiments, in step (3), the column temperature is 34-36°C, most preferably 35°C.

[0095] In some embodiments, in step (3), the flow rate is 0.7-0.9 mL / min, most preferably 0.8 mL / min.

[0096] In some embodiments, in step (3), the mobile phase B is 0.2% phosphoric acid.

[0097] In some embodiments, in step (3), the gradient elution procedure is:

[0098] Table 5.

[0099] time Mobile phase A Mobile phase B 0min 8％ 92％ 30min 8％ 92％ 35min 90％ 10％ 40min 8％ 92％ 45min 8％ 92％ .

[0100] In order to better understand the technical concept and beneficial effects of the present invention, the present invention will be described in detail through the following specific embodiments and comparative examples.

[0101] Example 1

[0102] A dandelion quality control method comprises the following steps:

[0103] (1) Preparation of reference solution: Take an appropriate amount of monocaffeoyltartaric acid reference substance, accurately weigh it, and add 50% methanol to make a solution containing 10 μg per 1 mL;

[0104] (2) Preparation of test solution: Take 0.5 g of dandelion powder (passed through a No. 4 sieve), accurately weigh it, place it in a stoppered conical flask, accurately add 50 mL of 20% methanol, weigh it, and treat it ultrasonically (power 500 W, frequency 45 kHz) for 45 minutes. Let it cool, weigh it again, make up the lost weight with 20% methanol, shake it well, filter it through 0.45 μm, and take the filtrate to obtain the test solution.

[0105] (3) Accurately pipette 10 μL of the reference solution and the test solution respectively, inject into the liquid chromatograph, and measure. The chromatographic conditions are as follows:

[0106] Chromatographic column: SMHADZU-Shim-pack GIST C18 column (4.6 mm × 250 mm, 5 μm);

[0107] Column temperature: 35 °C;

[0108] Flow rate: 0.8 mL / min;

[0109] Elution procedure: Use acetonitrile as mobile phase A and 0.2% phosphoric acid as mobile phase B, and perform gradient elution according to the following table:

[0110] Table 6. Gradient elution program of Example 1

[0111] time Mobile phase A Mobile phase B 0min 8％ 92％ 30min 8％ 92％ 35min 90％ 10％ 40min 8％ 92％ 45min 8％ 92％

[0112] Detection wavelength: 330nm.

[0113] Examples 2-3, Comparative Example 1

[0114] The difference from Example 1 is that only the extraction solvent in step (2) is changed in Example 2, Example 3 and Comparative Example 1, and the remaining steps are the same. The chromatogram is shown in FIG. Figure 1 , the content determination results are as follows:

[0115] Table 7. Content determination results of Examples 1-3 and Comparative Example 1

[0116] sample solvent Content (mg / g) Example 1 20% methanol 1.063 Example 2 50% methanol 0.910 Example 3 water 1.053 Comparative Example 1 Methanol Not detected

[0117] The results showed that 20% methanol and water had higher content as solvents, and 20% methanol was better, and methanol was not detected as a solvent. After comprehensive consideration, it was determined that 20%-50% methanol was used as the extraction solvent, and 20% methanol was the best.

[0118] Embodiment 4-5

[0119] The difference from Example 1 is that only the ultrasonic extraction time in step (2) was changed in Example 4-5, and the other steps were the same. The chromatogram is shown in FIG. Figure 2 , the content determination results are as follows:

[0120] Table 8. Results of assays in Example 1 and Example 4-5

[0121] sample Ultrasonic extraction time Content (mg / g) Example 1 45 minutes 1.063 Example 4 30 minutes 1.011 Example 5 60 minutes 1.072

[0122] The results showed that the content was low after 30 minutes of ultrasound, and there was no significant difference between 45 minutes and 60 minutes of ultrasound. After comprehensive consideration, the ultrasound time was selected to be 45-60 minutes, preferably 45 minutes.

[0123] Example 6

[0124] The difference from Example 1 is that Example 7 only changes the amount of dandelion sample in step (2), and the remaining steps are the same. The chromatogram is shown in FIG. Figure 3 , the content determination results are as follows:

[0125] Table 9. Assay results of Example 1 and Example 6

[0126] sample Sample weight (g) Content (mg / g) Example 1 0.5031 1.063 Example 6 0.2012 1.079

[0127] The results showed that there was no significant difference between the sample contents of 0.2g and 0.5g. In order to ensure the representativeness of the sample and reduce the experimental error, a larger sample weight of 0.5g was selected.

[0128] Embodiment 7-8

[0129] The difference from Example 1 is that only the volume of the solvent in step (2) is changed in Examples 7-8, and the remaining steps are the same. The chromatogram is shown in FIG. Figure 4 , the content determination results are as follows:

[0130] Table 10. Content determination results of Example 1 and Example 7-8

[0131]

[0132]

[0133] The results showed that 50 mL of extraction medium had the best extraction effect.

[0134] Test example

[0135] The detection examples of the present invention tested and examined the technical solution of the present invention from aspects such as the selection of detection wavelength, linear relationship, precision, stability, repeatability, sample recovery rate, durability and detection limit.

[0136] 1. Selection of detection wavelength

[0137] Take an appropriate amount of monocaffeoyltartaric acid reference substance, add 50% methanol to prepare a reference substance solution of 20 μg / ml, and perform spectral scanning at 200nm-650nm on an ultraviolet spectrophotometer. The result shows that monocaffeoyltartaric acid has a maximum absorption near 330nm. Therefore, the test wavelength of the present invention is determined to be 310nm-350nm, and the optimal choice is 330nm.

[0138] 2. Linear relationship

[0139] Accurately weigh 10.30 mg of monocaffeoyltartaric acid reference substance (98.3%), place it in a 50 ml volumetric flask, add 50% methanol to dissolve and dilute to the scale, shake well, and use it as the reference solution stock solution. Accurately pipette 0.5, 1.0, 2.5, 5.0, and 7.5 ml of the reference solution stock solution into 10 ml volumetric flasks, add 50% methanol to dissolve and dilute to the scale, shake well, and use it as the reference solution. Accurately pipette 10 μl of the above reference solution into the liquid chromatograph, and draw a standard curve with the reference solution injection concentration (μg / ml) as the horizontal axis (X) and the peak area (A) as the vertical axis (Y). The results show that the monocaffeoyltartaric acid injection concentration in the range of 10.12-151.88 μg / ml has a good linear relationship with the peak area; regression equation: Y=46.218X-0.1472; correlation coefficient: R 2 =1. For specific results, see Figure 5-6 And the table below.

[0140] Table 11. Linear relationship determination results

[0141] Injection concentration (μg / mL) Peak area (A) 10.12 458.266 20.25 948.520 50.62 2350.346 101.25 4650.557 151.88 7034.036

[0142] 3. Precision test

[0143] The reference solution was accurately aspirated and injected 6 times, 10 μl each time, and the retention time and peak area were examined. The RSD% were 0.09% and 0.15% respectively, indicating that the method had good precision. Figure 7 And the table below.

[0144] Table 12. Precision test results

[0145] Injection times Retention time (minutes) Peak area (A) 1 17.081 510.761 2 17.117 512.448 3 17.098 511.860 4 17.110 512.959 5 17.087 512.427 6 17.117 512.316 RSD% 0.09 0.15

[0146] 4. Stability test

[0147] For the same sample, the peak area was measured at 0, 3, 6, 9, 12, 24, and 36 hours, and the RSD% was 1.23%, indicating that the sample solution had good stability within 36 hours. Figure 8 And the table below.

[0148] Table 13. Stability test results

[0149] Storage time (h) Peak area 0 455.327 3 453.398 6 458.417 9 462.953 12 464.177 24 465.605 36 468.772 RSD% 1.23

[0150] 5. Repeatability

[0151] For the same batch of samples, 6 test solutions were prepared in parallel and the content was determined. The RSD% was 0.69%, indicating that the method had good repeatability. Fig. 9 And the table below.

[0152] Table 14. Repeatability test results

[0153]

[0154] 6. Sample recovery test

[0155] Using the sample addition recovery method, accurately weigh 0.25g of the same batch of samples with known content, and add 1ml of monocaffeoyltartaric acid reference solution with a concentration of 0.2025mg / ml to each of 6 parallel portions, then accurately add 50ml of 20% methanol, weigh the weight, ultrasonically treat (power 500W, frequency 45KHZ) for 45 minutes, cool, weigh again, make up the lost weight with 20% methanol, shake well, filter, take the filtrate, and determine it according to the chromatographic conditions determined above. The average recovery rate was 92.02% and the RSD was 1.78%, which fully complies with the standard requirements of "Content of components to be determined in samples and recovery limit" under "9101 Guidelines for Validation of Analytical Methods" in Part IV of the "Chinese Pharmacopoeia" (2020 edition), that is, when the content of the component to be determined is 1mg / g, the recovery limit is 90%-108%. This indicates that the recovery rate is good, as shown in the results. Fig.10 and the table below.

[0156]

[0157] Table 15. Results of sample recovery test

[0158]

[0159]

[0160] 7. Durability

[0161] 255 The column temperature, different chromatographic columns, and different types of liquid chromatographs were investigated, and the results showed that

[0162] This method has good durability and can adapt to different column temperatures, different chromatographic columns and different models of liquid chromatographs.

[0163] The specific investigation results are as follows:

[0164] 7.1 Effect of column temperature

[0165] The same sample solution and the same HPLC were used. The column temperature was adjusted to 30℃, 35℃, 260℃ and 40℃ respectively. Other chromatographic conditions remained unchanged and the results were determined according to the above experimental method. The results showed that the change of column temperature had little effect on the determination results, with an RSD of 1.53%. The ideal separation was achieved and only the retention time was affected. The results are as follows.

[0166] Table 16. Effect of column temperature

[0167] Column temperature Retention time (minutes) Content (mg / g) 30℃ 27.22 1.127 35℃ 23.35 1.145 40℃ 19.33 1.162 RSD% / 1.53

[0168] 7.2 Investigation of different chromatographic columns and liquid phases

[0169] 265 chromatographic column selected different chromatographic columns and liquid phases, and the results were all able to achieve satisfactory determination results, with the content RSD being 1.13%, which had little impact on the results. The results are as follows.

[0170] Table 17. Effects of different chromatographic columns and liquid phases

[0171] serial number Liquid Brand Chromatographic column brand Content (mg / g) 1 Agilent1260 SHMADZU Shim-pack GIST C18 1.108 2 Agilent1260 Agilent ZORBAX SB-C18 1.145 3 ThermoFisher U3000 Amethyst C18-H 1.139 RSD% / / 1.13

[0172] 8. Determination of the content of dandelion medicinal materials

[0173] According to the above-determined chromatographic conditions and test sample preparation method, 5 batches of sample content were determined, and the results are as follows.

[0174] Table 18. Results of assays of different batches

[0175] Sample batch number Sample content (mg / g) Sample 1 1.353 Sample 2 0.879 Sample 3 0.751 Sample 4 0.918 Sample 5 1.220 Average content 1.024

[0176] 9. Limit determination

[0177] After testing the 5 batches of dandelion samples collected, the average content of monocaffeoyltartaric acid in the 5 batches of dandelions was 1.024 mg / g, or 0.102%. Considering that the quality of Chinese medicinal materials is greatly affected by factors such as origin, harvest time, and processing methods, the limit of monocaffeoyltartaric acid content in dandelions is determined to be "this product, calculated on a dry basis, contains monocaffeoyltartaric acid (C13 H 12 O9) shall not be less than 0.05%.

[0178] Comparative Example 2

[0179] Different from Example 1, the test solution preparation method is as follows:

[0180] Take about 0.5g of dandelion powder (passed through No. 4 sieve), weigh it accurately, put it in a stoppered conical flask, add 50mL of water accurately, weigh it, heat and reflux for 60min, cool, weigh it again, make up the lost weight with water, shake well, filter, and take the filtrate. The rest are the same.

[0181] The results are as follows Fig.12 , the peak shape is the same as that in Example 1 ( Fig.11 ) is relatively poor.

[0182] Comparative Example 3

[0183] The difference from Example 1 is that acetonitrile is used as mobile phase A, 0.2% phosphoric acid solution is used as mobile phase B (10:90), and isocratic elution is used. The rest is the same.

[0184] The results are as follows Fig.13 , the peak shape is the same as that in Example 1 ( Fig.11 ) is relatively poor.

[0185] Comparative Example 4

[0186] The difference from Example 1 is that the gradient elution procedure is as follows, and the rest is the same:

[0187] Table 19. Comparative Example 4 Gradient elution program

[0188] Time (min) A(％) B(％) 0 5 95 30 20 80 40 90 10 45 90 10 46 5 95 50 5 95

[0189] The results are as follows Fig.14 The separation degree of the monocaffeoyltartaric acid peak in the sample is 2.4, but after amplifying the chromatographic peak, there is a peak-enveloping phenomenon.

[0190] Comparative Example 5

[0191] The difference from Example 1 is that the mobile phase A is a 0.1% phosphoric acid solution in acetonitrile, and the mobile phase B is a 0.1% phosphoric acid solution in water, and the rest are the same.

[0192] Comparative Example 6

[0193] The difference from Example 1 is that methanol is used as mobile phase A and 0.2% phosphoric acid is used as mobile phase B, and the rest are the same.

[0194] Comparative Example 7

[0195] The difference from Example 1 is that acetonitrile is used as mobile phase A and 0.1% phosphoric acid is used as mobile phase B, and the rest are the same.

[0196] The results are as follows:

[0197] Table 20. Comparative Examples 5-7 Test Results

[0198]

[0199] The results show that the change of the type and concentration of the mobile phase has a significant effect on the determination of the content of monocaffeoyltartaric acid in dandelion. After changing the mobile phase, the linear correlation coefficient value decreases, the precision and repeatability decrease, the precision RSD% value increases from 0.15% to 0.25-0.36%, and the repeatability RSD% value increases from 0.69% to 0.75-0.91%. Similarly, the performance in the durability test is also significantly worse than that of the mobile phase in Example 1, and the content detection limit increases.

[0200] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. A method for quality control of dandelions, It is characterized in that Quality control by determining the content of monocaffeoyltartaric acid includes the following steps: (1) preparing a reference solution: diluting monocaffeoyltartaric acid to obtain a reference solution; (2) Preparation of test solution: dissolve dandelion in a solvent, perform ultrasonic treatment, and collect the filtrate to obtain the test solution; (3) Liquid chromatography was used for content determination. The chromatographic conditions were as follows: Chromatographic column: chromatographic column with octadecylsilane bonded silica gel as filler; Column temperature: 30-40°C; Flow rate: 0.6-1.0mL / min; Elution procedure: acetonitrile as mobile phase A, 0.15-0.25% phosphoric acid as mobile phase B, gradient elution: Detection wavelength: 310-350nm; Injection volume: 5-15μL.

2. The quality control method according to claim 1, It is characterized in that In step (1), the monocaffeoyltartaric acid is diluted with a methanol solution, and the concentration of the reference solution is 5-15 μg / mL.

3. The quality control method according to claim 2, It is characterized in that In step (1), the monocaffeoyltartaric acid is diluted with a 40-60% methanol solution, and the concentration of the reference solution is 8-12 μg / mL.

4. The quality control method according to claim 1, It is characterized in that In step (2), the solvent is a methanol aqueous solution or water with a volume fraction of 20-50%.

5. The quality control method according to claim 1, It is characterized in that In step (2), the mass volume ratio of dandelion to solvent is 1:5-200.

6. The quality control method according to claim 1, It is characterized in that In step (2), the weight of the dandelion sample is 0.2-0.5 g.

7. The quality control method according to claim 1, It is characterized in that The steps of preparing the test solution are as follows: Accurately weigh dandelion powder, place in a conical flask, accurately add 50 mL of 20% methanol, ultrasonically treat, cool, make up the lost weight with 20% methanol, shake well, filter, and take the filtrate to obtain the test solution.

8. The quality control method according to claim 1, It is characterized in that In step (3), the chromatographic column is any one of SMHADZU-Shim-pack GIST C18 column, SMHADZU Shim-pack GIST C18, Agilent ZORBAX SB-C18, and Amethyst C18-H.

9. The quality control method according to claim 1, It is characterized in that In step (3), the detection wavelength is 330 nm.

10. The quality control method according to claim 1, It is characterized in that The chromatographic conditions are as follows Chromatographic column: SMHADZU-Shim-pack GIST C18 column (4.6 mm × 250 mm, 5 μm); Column temperature: 35°C; Flow rate: 0.8 mL / min; Elution procedure: acetonitrile as mobile phase A, 0.2% phosphoric acid as mobile phase B, gradient elution: Detection wavelength: 330nm; Injection volume: 10 μL.

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