Quality analysis method for extracting effective components of gastrodia elata by using HDES

Abstract

The invention discloses a quality analysis method for extracting effective components of gastrodia elata by using HDES. The quality analysis method comprises the following steps: (1) synthesizing HDES by using L-menthol and DL-lactic acid; (2) adding the HDES in the step (1) into gastrodia elata medicinal material powder, stirring to extract effective components, centrifuging and diluting the obtained mixture after extraction to obtain a test solution, and performing chromatographic analysis to determine the content of the effective components; (3) detecting the quality index of the test solution in the step (2); and (4) analyzing the quality of the gastrodia elata by adopting a chemometrics analysis method. The extraction method disclosed by the invention is green, simple, rapid and high in extraction efficiency, and is beneficial to extraction of effective components in medicinal and edible medicinal materials.

Description

Technical Field

[0001] The invention belongs to the technical field of chemical extraction and analysis, and particularly relates to a quality analysis method for extracting effective components of Gastrodia elata by utilizing HDES. Background Art

[0002] Gastrodia elata Bl. is the dried tuber of the orchid plant Gastrodia elata. Its rhizome is used as medicine. It is a functional food with high nutritional and medicinal value. It is one of the "Top Ten Famous Medicinal Materials" in Yunnan Province and is regarded as a "top grade" in traditional Chinese medicine. It was included in the catalog of medicine and food homology in 2019. As a traditional Chinese medicine and dietary source, Gastrodia elata mainly contains active substances such as aromatic compounds, phenols, organic acid esters, sugars and their glycosides, and sterols. Studies have shown that Gastrodia elata has a variety of medicinal effects. It is often used clinically to treat cardiovascular and cerebrovascular diseases such as ischemic dementia, ischemic myocardial injury, hypertension, atherosclerosis, cerebrovascular vertigo, and has a certain effect on neurological diseases such as Parkinson's disease, aging and migraine. Among the many active substances in Gastrodia elata, phenolic compounds have attracted more and more attention from researchers and the industry because of their potential effects such as promoting blood circulation and soothing nerves.

[0003] Before being used as medicine or food, the active ingredients of medicinal materials that are both medicinal and edible will be affected by their origin, processing methods, extraction methods, etc., which in turn affects their efficacy. The effective ingredients of most medicinal materials that are both medicinal and edible are still extracted using traditional organic solvents such as methanol, ethanol, ether, ethyl acetate, chloroform and petroleum ether. However, these traditional solvents generally have the disadvantages of long extraction time, large amount of solvent used, low extraction efficiency, easy volatility, high toxicity and non-biodegradability. In addition, the long-term use of traditional solvents will threaten human health and affect the ecological environment. Therefore, it is very important to develop a new green solvent with high extraction efficiency, low volatility and stable properties to replace traditional organic solvents.

[0004] In addition, the efficacy of edible and medicinal herbs is often the result of the combined effects of multiple components, while the content of a single component usually changes due to storage conditions and processes. Therefore, it is relatively stable and comprehensive to reflect the quality of medicinal materials by monitoring multiple components rather than a single component. Chemometric analysis plays an important role in environmental analysis, food science, drug development, and quality control with its significant advantages such as efficient data processing, accurate information extraction, and diverse and flexible methods. Therefore, a new method for the efficient extraction of three main phenolic compounds in Gastrodia elata was established. Combined with chemometric analysis, the evaluation results are more comprehensive and reasonable, which can provide a more scientific theoretical and data basis for the quality control and quality improvement of medicinal materials. Summary of the invention

[0005] The object of the present invention is to provide a mass analysis method for extracting effective components of Gastrodia elata by using HDES, which has the advantages of being green, simple, rapid and having high extraction efficiency.

[0006] In order to achieve the above-mentioned invention object, the present invention adopts the following technical solutions:

[0007] The present invention provides a quality analysis method for extracting effective components of Gastrodia elata by using HDES, comprising the following steps:

[0008] (1) L-menthol and DL-lactic acid were mixed in a molar ratio of 1:1–7, and then the mixture was stirred in an oil bath at 60 °C for 30 min to obtain a homogeneous transparent liquid, which is HDES;

[0009] (2) adding the HDES prepared in step (1) to the powder of Gastrodia elata medicinal material, stirring to extract the active ingredient, and after the extraction is completed, centrifuging and diluting the obtained mixture to obtain a test solution, and performing chromatographic analysis to determine the content of the active ingredient;

[0010] (3) Detecting the quality index of the test solution in step (2);

[0011] (4) Under the optimal conditions explored, the same sample was injected six times to verify the precision of the method; the stability of the sample solution was evaluated by measuring the sample solution after 0, 2, 4, 8, 16 and 24 h at room temperature; the repeatability of the method was evaluated by measuring six sample solutions; in addition, standard solutions were added to samples with known contents of the three phenolic compounds in Gastrodia elata to determine the sample recovery rate; and the corresponding spectral data obtained were analyzed;

[0012] (5) Combining the quality indicators of step (3), chemometric analysis is used to comprehensively evaluate the quality of Gastrodia elata.

[0013] Preferably, in step (1), the molar ratio of L-menthol to DL-lactic acid is 1:4.

[0014] Furthermore, in the step (2), the solid-liquid ratio (g / mL) of Gastrodia elata medicinal material powder and HDES is 1:6-1:36.

[0015] Preferably, in step (2), the solid-liquid ratio (g / mL) of Gastrodia elata powder to HDES is 1:24.

[0016] Furthermore, the stirring extraction conditions in step (2) are: the stirring extraction speed is 1000 rpm / min, and the stirring extraction time is 10-60 min.

[0017] Preferably, the stirring extraction time in step (2) is 20 min.

[0018] Furthermore, in step (2), the chromatography is HPLC, and the conditions are: the HPLC conditions are: the chromatographic column is ZORBAXExtend-C 18 The mobile phase was acetonitrile C and 0.1% phosphoric acid solution D. The gradient elution order was: 0−20 min, 5% C, 95% D; 20−30 min, 5%−15% C, 95%−85% D; 30−40 min, 15%−20% C, 85%−80% D; 40−55 min, 20% C, 80% D; 55−60 min, 20%−5% C, 80%−95% D. The flow rate was 1.0 mL / min. The detection wavelength was 280 nm. The column temperature was 25°C and the injection volume was 10 µL.

[0019] Furthermore, in step (2), the mixture is diluted with methanol.

[0020] Furthermore, the active ingredients are gastrodin, p-hydroxybenzyl alcohol and balisonoside A.

[0021] Furthermore, the quality indicators in step (3) include moisture, total ash, alcohol-soluble extract and metal element content.

[0022] Furthermore, the chemometric analysis in step (5) includes Pearson correlation analysis, cluster analysis, principal component analysis and factor analysis.

[0023] Furthermore, when the chemometric analysis is performed in step (5), there is a significant correlation between the Cu element and the effective ingredient.

[0024] Furthermore, in the step (4), the RSD of the precision of the contents of the three phenolic compounds in the Gastrodia elata sample was 0.47–0.96%, the RSD value of the repeatability was 0.98–1.52%, the RSD value of the solution stability was 0.61–1.33%, and the sample recovery rate was 100.35–112.10%.

[0025] Compared with the prior art, the present invention has the following advantages and beneficial effects:

[0026] 1. The present invention adopts the hydrophobic deep eutectic solvent (HDES) for the first time to efficiently extract gastrodia phenolic compounds: gastrodin, p-hydroxybenzyl alcohol and balisonoside A.

[0027] 2. The present invention establishes a new method for extracting effective ingredients from edible and medicinal materials and for analyzing their quality based on chemometric analysis, which is green, simple and rapid.

[0028] 3. Compared with the traditional extraction method, the extraction rates of gastrodin, p-hydroxybenzyl alcohol and balisonoside A in the present invention are respectively increased by 2.8, 2.5 and 16.1 times that of methanol extraction, so the extraction efficiency of this method is significantly improved.

[0029] 4. The present invention verifies that the gastrodin content is significantly correlated with the Cu content. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] Figure 1 It is the infrared spectrum of L-menthol, DL-lactic acid and HDES.

[0031] Figure 2 This is the single factor optimization diagram for the extraction of three phenolic compounds from Gastrodia elata using HDES, where Figure A is the optimization diagram for the extraction method; Figure B is the optimization diagram for the type of HDES; Figure C is the optimization diagram for the extraction time; Figure D is the optimization diagram for the solid-liquid ratio; and Figure E is the optimization diagram for the molar ratio of HDES.

[0032] Figure 3 This is a graph showing the determination results of moisture, total ash and alcohol-soluble extracts of 15 batches of Gastrodia elata samples.

[0033] Figure 4 This is a heat map showing the correlation between phenolic compounds, moisture, total ash, alcohol-soluble extracts and metal element contents in Gastrodia elata.

[0034] Figure 5 Figure 1 is the average linkage (between-group) pedigree diagram of 15 batches of Gastrodia elata, in which Figure A uses gastrodin, p-hydroxybenzyl alcohol, balisonoside A, moisture, total ash, alcohol-soluble extracts and 7 metal element contents as the data matrix; Figure B uses gastrodin, p-hydroxybenzyl alcohol, balisonoside A, moisture, total ash, alcohol-soluble extracts and Cu element content as the data matrix; Figure C uses gastrodin, p-hydroxybenzyl alcohol, balisonoside A, moisture, total ash and alcohol-soluble extract contents as the data matrix.

[0035] Figure 6 These are principal component analysis diagrams of 15 batches of Gastrodia elata, among which Figure A performs principal component analysis on the contents of gastrodin, p-hydroxybenzyl alcohol, balisonoside A, moisture, total ash, alcohol-soluble extracts and 7 metal elements; Figure B performs principal component analysis on the contents of gastrodin, p-hydroxybenzyl alcohol, balisonoside A, moisture, total ash, alcohol-soluble extracts and Cu element; Figure C performs principal component analysis on the contents of gastrodin, p-hydroxybenzyl alcohol, balisonoside A, moisture, total ash and alcohol-soluble extracts. DETAILED DESCRIPTION

[0036] The technical solution of the present invention is further described in detail with reference to the following specific examples.

[0037] In the following examples, unless otherwise specified, the experimental methods used are all conventional methods, and the materials and reagents used can be purchased from biological or chemical reagent companies.

[0038] Example 1

[0039] 1. Preparation and characterization of hydrophobic deep eutectic solvent (HDES)

[0040] L-menthol was used as HBA and DL-lactic acid was used as HBD. HBA and HBD were mixed in a round-bottom flask at a molar ratio of 1:4, and then the mixture was stirred in an oil bath at 60 °C for 30 min to obtain a uniform and transparent liquid HDES.

[0041] In order to confirm the formation of hydrogen bonds, the FT-IR spectra of pure L-menthol, DL-lactic acid and HDES were compared, as shown in Fig. Figure 1 The hydroxyl stretching band (O–H) of L-menthol starts at 3261 cm −1 Blue shift to 3407 cm −1 , DL-lactic acid at 1720 cm −1 The characteristic band at 1720 cm −1 Red shifted to 1717 cm −1 , proving that intermolecular hydrogen bonds were formed between L-menthol and DL-lactic acid, indicating that the HDES was successfully prepared. The characteristic peaks of L-menthol and DL-lactic acid are both shown in the HDES spectrum. In addition, the FT-IR spectrum of HDES was compared with the FT-IR spectra of its components, and no new peaks disappeared or appeared, indicating that no reaction occurred during the preparation of HDES.

[0042] 2. Extraction of active ingredients from Gastrodia elata

[0043] 0.5000 g of Gastrodia elata powder was accurately weighed, 12 mL of the synthesized HDES was added, and the mixture was stirred at 1000 rpm / min for 20 min at room temperature. After the extraction was completed, the mixture was centrifuged at 8000 rpm for 5 min. The final sample was diluted 3 times with methanol to reduce the viscosity, and the test solution was prepared. Finally, HPLC analysis was performed to extract the active ingredients gastrodin, p-hydroxybenzyl alcohol and barisin A.

[0044] HPLC conditions: chromatographic column: ZORBAX Extend-C 18(4.6 mm ´ 250 mm,5 µm). The mobile phase was acetonitrile (C) and 0.1% phosphoric acid solution (D); the gradient elution order was: 0−20 min, 5% C, 95% D; 20−30 min, 5%−15% C, 95%−85% D; 30−40 min, 15%−20% C, 85%−80% D; 40−55 min, 20% C, 80% D; 55−60 min, 20%−5% C, 80%−95% D; the flow rate was 1.0 mL / min; the detection wavelength was 280 nm; the column temperature was 25℃, and the injection volume was 10 µL.

[0045] Accurately weigh appropriate amounts of gastrodin, p-hydroxybenzyl alcohol and balisonoside A, add methanol to dissolve, and prepare mixed reference solution and reference solution for determination.

[0046] The established method was validated in terms of linearity and range, precision, solution stability, repeatability, and spike recovery. For example, the same sample was injected six times to validate the method’s precision; the stability of the sample solution was evaluated by measuring the sample solution after 0, 2, 4, 8, 16, and 24 h at room temperature; the repeatability of the method was evaluated by measuring six sample solutions; in addition, standard solutions were added to samples with known contents of three phenolic compounds in Gastrodia elata to determine sample recovery.

[0047] The RSD of precision of the three phenolic compounds in Gastrodia elata samples was 0.47–0.96%, the RSD of repeatability was 0.98–1.52%, the RSD of solution stability was 0.61–1.33%, and the recovery rate was 100.35–112.10%.

[0048] 3. Single Factor Investigation

[0049] 1. Extraction method

[0050] Common methods for extracting active ingredients include ultrasound, stirring, microwave, cold soaking, reflux and Soxhlet extraction. Ultrasonic extraction and stirring extraction have the advantages of low energy consumption, simple operation, high extraction efficiency and wide application, so this study mainly compares these two extraction methods. 0.5000 g of Gastrodia elata medicinal material (S2) was accurately weighed in a centrifuge tube, HDES (L-menthol: DL-lactic acid) was used as the extraction solvent, and ultrasound (power 500 W) and stirring were performed at room temperature for 60 min to extract the three components (Gastrodin, p-hydroxybenzyl alcohol and Balisonoside A). After the extraction was completed, the mixture was centrifuged at 8000 rpm for 5 min. The final sample was diluted 3 times with methanol to reduce the viscosity, and finally analyzed by HPLC.

[0051] The results are as follows Figure 2 A shows that the extraction volume is larger when the stirring extraction method is used. Therefore, the subsequent experiments were carried out using the stirring extraction method.

[0052] 2. Selection of HDES types

[0053] Different HDES have different components, which will significantly affect the polarity and hydrogen bond network of HDES, thereby affecting its solubility. Therefore, different HDES can extract effective components with different polarities. Take Gastrodia elata medicinal materials, use HDES-1 (L-menthol: hexanoic acid, 1:1), HDES-2 (L-menthol: nonanoic acid, 1:1), HDES-3 (L-menthol: decanoic acid, 1:1), HDES-4 (L-menthol: octanol, 1:1) and HDES-5 (L-menthol: DL-lactic acid, 1:1) as extraction solvents to extract three phenolic compounds, obtain the test solution, and finally perform HPLC analysis to calculate the extraction amount of the three components.

[0054] The results are as follows Figure 2 B shows that the extraction volume was the largest when HDES-5 (L-menthol: DL-lactic acid, 1:1) was used as the extraction solvent. Therefore, HDES-5 was used for extraction in subsequent content determination experiments.

[0055] 3. Influence of extraction time

[0056] Extraction time has an important influence on the amount of active ingredients extracted. Generally speaking, too short an extraction time may result in the inability of the active ingredients to be completely dissolved or transferred into the solution, resulting in low extraction efficiency; while too long an extraction time will lead to unnecessary energy waste and may introduce other ingredients. Take Gastrodia elata and perform stirring extraction for different times of 10–60 min to obtain the test solution.

[0057] The results are as follows Figure 2 C shows that after the stirring time exceeds 20 min, the extraction amount of each component tends to change slowly. Based on the principle of simple operation and less time consumption, the optimal extraction time is selected as 20 min.

[0058] 4. Influence of material-liquid ratio

[0059] Too high or too low a solid-liquid ratio may affect the extraction efficiency and the purity of the extracted substance. When the solid-liquid ratio is too high, it may lead to solvent waste, low extraction efficiency or the introduction of impurities; when the solid-liquid ratio is too low, the solvent may not be able to fully cover the raw materials, thus affecting the extraction effect. Take Gastrodia elata medicinal materials, add HDES at different solid-liquid ratios (g / mL) of 1:6-1:36, obtain the test solution and measure it.

[0060] The results are as follows Figure 2D shows that the extraction rate gradually increases when the solid-liquid ratio (g / mL) is 1:6-1:24. With the increase of liquid volume, the extraction amount of each index component does not change much. Insufficient liquid volume will lead to incomplete extraction, while excessive liquid volume will cause waste of extractant. Therefore, a solid-liquid ratio (g / mL) of 1:24 was selected for subsequent experiments.

[0061] 5. Effect of HDES molar ratio

[0062] Different molar ratios of HDES will result in different properties such as structural stability, viscosity, polarity and melting point. An appropriate amount of Gastrodia elata was taken and 7 different HDES molar ratios of 1:1-7 were selected to extract three phenolic compounds and obtain the test solution.

[0063] The results are as follows Figure 2 E shows that when the molar ratio of L-menthol to DL-lactic acid is 1:4, the extraction amount of the three index components is the largest.

[0064] Example 2

[0065] 1. Comparison of extraction efficiency between HDES and traditional methanol

[0066] In order to compare the effects of HDES and methanol in extracting three phenolic compounds from Gastrodia elata, the powder of Gastrodia elata was prepared according to the steps of Example 1 to obtain a test solution, which was determined by HPLC. Methanol extraction was carried out according to the method reported in the literature, that is, 2.0000 g of Gastrodia elata powder was accurately weighed in a 50 mL glass conical flask with a stopper, 25 mL of 60% methanol was added, the mass was weighed, and ultrasonication was performed for 60 min. After cooling, 60% methanol was used to make up the lost mass, shake well, and filter with a microporous filter membrane (0.45 μm) to obtain the test solution and determine it. The corresponding extraction rate was calculated and the extraction rates of the two extraction solvents were compared.

[0067] The results are shown in Table 1. It can be seen from the results in the table that the extraction rates of gastrodin, p-hydroxybenzyl alcohol and balisonoside A by HDES were 2.8, 2.5 and 16.1 times higher than those by methanol extraction, respectively. It can be seen that the extraction rates of the three main phenolic compounds in Gastrodia elata by HDES were greatly improved compared with the traditional methanol extraction, especially balisonoside A.

[0068] Table 1 Comparison of extraction rates between HDES and methanol

[0069]

[0070] 2. Determination of active ingredient content

[0071] 0.5000 g of Gastrodia elata powder from 15 origins (for details, see Table 2) was accurately weighed to prepare the test solution, which was injected for determination. The contents of gastrodin, p-hydroxybenzyl alcohol and barisin A were calculated using the external standard method.

[0072] Table 2 Numbers of Gastrodia elata samples from different origins

[0073]

[0074] The results are shown in Table 3. The contents of gastrodin, p-hydroxybenzyl alcohol and barisin A are 3.4-18.8 mg / g, 0.8-3.4 mg / g and 49.6-167.8 mg / g, respectively, indicating that the contents of the index components of gastrodia elata from different origins are different. In terms of the extraction amount of the three phenolic compounds extracted by HDES, the extraction amount of barisin A is the largest, and the extraction amount of p-hydroxybenzyl alcohol is the least. Among them, the content of p-hydroxybenzyl alcohol compound extracted by HDES in S14 is the highest, and the content of gastrodin and barisin A compounds extracted by S9 is the highest. In general, the contents of the three index components in S6, S9 and S12 are relatively high.

[0075] Table 3 Results of determination of active ingredient contents in Gastrodia elata from different origins

[0076]

[0077] 3. Determination of moisture, total ash and alcohol-soluble extracts

[0078] The determination of moisture, total ash and alcohol-soluble extracts is of great significance for accurately evaluating the quality of edible and medicinal materials and ensuring their stable efficacy during use. An appropriate amount of Gastrodia elata powder from 15 different origins was taken and the moisture, total ash and alcohol-soluble extracts were determined according to the corresponding methods of the four general rules of the 2020 edition of the Chinese Pharmacopoeia.

[0079] The results are as follows Figure 3 As shown, the moisture content of 15 batches of Gastrodia elata samples was less than 15.0%, the total ash content was less than 4.5%, and the alcohol-soluble extract content was greater than 15.0%, indicating that the determination results of the moisture, total ash and alcohol-soluble extract content of Gastrodia elata samples from 15 different origins were all qualified.

[0080] 4. Determination of metal elements in Gastrodia elata

[0081] During the planting and processing of Chinese herbal medicines, they may be contaminated by heavy metals due to problems such as irregular planting, abuse of various pesticide preparations or immature technology, which in turn affects their quality and safety. Therefore, the analysis and determination of metal elements is also an important part of evaluating the quality of Chinese herbal medicines. In this paper, the contents of seven metal elements in Gastrodia elata from 15 different origins were determined by ICP-OES.

[0082] The results are shown in Table 4. The results of the determination of metal element contents in Gastrodia elata from different origins are different, and the contents of some metal elements vary greatly, which also verifies that the content of metal elements is closely related to the origin.

[0083] Table 4 Determination results of metal elements

[0084]

[0085] ND*: Not Detected

[0086] Example 3: Chemometric Analysis

[0087] 1. Pearson correlation analysis

[0088] Pearson correlation analysis was performed between the "quality indicators" in Gastrodia elata, such as the contents of three phenolic compounds, moisture, total ash, and alcohol-soluble extracts and the contents of seven metal elements.

[0089] Results Figure 4 As can be seen from the figure, there is a significant difference between the contents of the three phenolic compounds and the contents of the other ten variables, the content of gastrodin and the content of Cu element, and there is no significant correlation between the contents of the other two components and the contents of the other variables; there is a significant difference between the total ash content and the contents of Cd and Pb metal elements; the contents of the seven metal elements mostly have a certain correlation between each other.

[0090] 2. Cluster analysis

[0091] Using IBM SPSS Statistics 23 software, with "quality index" and the contents of seven metal elements as data matrix, the intergroup homogeneity method was adopted, the square Euclidean distance was selected as the scale, and the pedigree diagram was used to represent the phylogenetic analysis of Gastrodia elata samples from 15 production areas. Figure 5A, can intuitively reflect the classification of different Gastrodia samples in this study. The results of cluster analysis show that when the square Euclidean distance is 5, 15 batches of Gastrodia samples can be clearly clustered into 5 groups. The first group (3 origins): S2, S11, S15; the second group (3 origins): S1, S4, S7; the third group (5 origins): S9, S12, S6, S13, S14; the fourth group (3 origins): S5, S10, S3; the fifth group (1 origin): S8. However, from the above correlation analysis results, it can be seen that among the many variables, only the gastrodin content and the Cu element content have a significant correlation. Therefore, in order to verify the results of the correlation analysis, a systematic cluster analysis was performed with the "quality index" content and the Cu element content as the data matrix and the "quality index" content as the data matrix, and the clustering results obtained by the two were the same, see. Figure 5 B and 5C. Among them, S2, S3, S1 and S11, S5 in the first group of samples were clustered into a small category, and S4, S5, S7, S10, S13 and S8, S14, S6 in the second group of samples were clustered into a small category, and the samples showed certain intra-group differences; the clustering of the third group of samples was relatively concentrated, and the sample quality consistency was good.

[0092] In summary, compared with the contents of other metal elements, the distribution pattern of Cu element content was the same as that of three phenolic compounds, moisture, total ash and alcohol-soluble extracts that closely affected the quality of Gastrodia elata, and the results of cluster analysis were consistent with those of Pearson correlation analysis.

[0093] 3. Principal Component Analysis

[0094] The principal component analysis was performed on the "quality index" and the contents of seven metal elements in the samples of Gastrodia elata from 15 origins. According to the analysis, the results of the Kaiser-Meyer-Olkin (KMO) and Bartlett's sphericity test were KMO=0.359, P=0.000, which did not meet the KMO>0.6 condition required for the data applicability of the principal component analysis application program. Figure 6It can be seen from A that the samples are relatively scattered, and the classification results obtained are quite different from the cluster analysis results, indicating that the data corresponding to the above variables are not suitable for principal component analysis. According to the results of cluster analysis and Pearson correlation analysis, principal component analysis was performed on the "quality index" content, Cu element content data and "quality index" content data. The KMO value and Bartlett's sphericity test results showed that KMO>0.6, P<0.05, and the data correlation was significant, which was suitable for principal component analysis and factor analysis. In the analysis results of two groups of different variable data, two principal components (PC1 and PC2) with eigenvalues ​​greater than 1 were selected, and the cumulative contribution rates were 63.75% and 70.05%, respectively, indicating that the first two principal components can represent 63.75% of the information of the seven components of Gastrodia elata and 70.05% of the information of the six components of Gastrodia elata, respectively, so the two principal components can basically reflect the information of the corresponding components measured. By Figure 6 As shown in Figures B and 6C, gastrodin, p-hydroxybenzyl alcohol, balisonoside A, alcohol-soluble extracts and total ash showed a large positive correlation with PC1, while moisture and Cu element content showed a large positive correlation with PC2; in addition, there was also a strong correlation between alcohol-soluble extracts and p-hydroxybenzyl alcohol; the 15 batches of samples were divided into 3 categories, and the results were basically consistent with the results of cluster analysis, which further verified the classification results of cluster analysis.

[0095] 4. Factor Analysis

[0096] In factor analysis, the higher the comprehensive score, the better the product quality. Factor analysis was performed on the "quality index" content and Cu element content data and the "quality index" content data. The eigenvector was multiplied by the standardized data to obtain two principal component scores (F1 and F2). The ratio of the eigenvalue corresponding to each principal component to the sum of the eigenvalues ​​of the extracted principal components was used as the weight. The calculation method is as shown in formula (1) to obtain the principal component comprehensive model. The comprehensive scores (F1 and F2) of 15 batches of Gastrodia elata samples were calculated according to formula (2). j ), see Tables 5 and 6. According to the calculation results, in the two cases, the main component comprehensive scores of the samples from 15 origins were -1.043−1.352 and -1.029−1.353, respectively. The comprehensive scores in the two cases were sorted, and the results were the same, with S9, S12 and S6 ranking in the top three, indicating that the quality of Gastrodia elata from these three origins was relatively good, and the results were consistent with the results of the determination of the content of the indicator components. It can be seen that factor analysis can be used to evaluate the quality differences of Gastrodia elata samples from different origins.

[0097] (1)

[0098] (2)

[0099] Among them, W tis the relative weight of the tth principal component, C t is the variance contribution rate of the tth principal component, C z is the cumulative variance contribution rate. F1 and F2 are the corresponding principal component scores, F j For the comprehensive score.

[0100] Table 5 Principal component scores and rankings of 15 batches of Gastrodia elata samples

[0101]

[0102] Data matrix analysis was performed on phenolic compounds, moisture, total ash, alcohol-soluble extracts and Cu contents of 15 batches of GE samples.

[0103] Table 6 Principal component scores and rankings of 15 batches of Gastrodia elata samples

[0104]

[0105] Data matrix analysis was performed on the contents of phenolic compounds, moisture, total ash and alcohol-soluble extracts of 15 batches of GE samples.

[0106] The above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit the same. Although the present invention has been described in detail with reference to the aforementioned embodiments, it is still possible for a person skilled in the art to modify the technical solutions described in the aforementioned embodiments, or to replace some of the technical features therein by equivalents. However, these modifications or replacements do not deviate the essence of the corresponding technical solutions from the spirit and scope of the technical solutions claimed to be protected by the present invention.

Claims

1. A quality analysis method for extracting effective components of Gastrodia elata using HDES, characterized in that: The steps include: (1) Synthesis of HDES using L-menthol and DL-lactic acid; (2) adding the HDES prepared in step (1) to the powder of Gastrodia elata medicinal material, stirring to extract the active ingredient, and after the extraction is completed, centrifuging and diluting the obtained mixture to obtain a test solution, and performing chromatographic analysis to determine the content of the active ingredient; (3) Detecting the quality index of the test solution in step (2); (4) In combination with the quality indicators of step (3), the quality of Gastrodia elata is analyzed by chemometric analysis.

2. The method according to claim 1, characterized in that The specific steps of synthesizing HDES in step (1) are: mixing L-menthol and DL-lactic acid in a molar ratio of 1:1-7, and then stirring the mixture in an oil bath at 60° C. to obtain a uniform transparent liquid, which is HDES.

3. The method according to claim 1, characterized in that In the step (2), the solid-liquid ratio of Gastrodia elata powder to HDES is 1:6-1:

36.

4. The method according to claim 1, characterized in that The chromatography in step (2) is HPLC, and the conditions are: the HPLC conditions are: the chromatographic column is ZORBAX Extend-C 18 ; The mobile phase was acetonitrile C and 0.1% phosphoric acid solution D; the gradient elution sequence was: 0−20 min, 5% C, 95% D; 20−30 min, 5%−15% C, 95%−85% D; 30−40 min, 15%−20% C, 85%−80% D; 40−55 min, 20% C, 80% D; 55−60 min, 20%−5% C, 80%−95% D; flow rate was 1.0 mL / min; detection wavelength was 280 nm; column temperature was 25°C, and injection volume was 10 µL.

5. The method according to claim 1, characterized in that The stirring extraction conditions in step (2) are: the stirring extraction speed is 1000 rpm / min, and the stirring extraction time is 10-60 min.

6. The method according to claim 1, characterized in that In the step (2), the mixture is diluted with methanol.

7. The method according to claim 1, characterized in that The effective ingredients are gastrodin, p-hydroxybenzyl alcohol and parisonoside A.

8. The method according to claim 1, characterized in that The quality indicators in step (3) include moisture content, total ash content, alcohol-soluble extract and metal element content.

9. The method according to claim 1, characterized in that: The chemometric analysis in step (4) includes Pearson correlation analysis, cluster analysis, principal component analysis and factor analysis.

10. The method according to claim 1, characterized in that When the chemometric analysis is performed in step (4), there is a significant correlation between the Cu element and the effective ingredients.

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