Quality evaluation method of Curcuma aromatica based on HPLC metabolomics analysis

Abstract

The present application relates to the technical field of analyzing material properties using optical means, and specifically to a method for evaluating the quality of turmeric based on HPLC metabolomics analysis, the method comprising: collecting multiple rhizome samples of the same turmeric at different growth stages, and obtaining HPLC chromatograms and common pattern diagrams of all growth stages; performing differential analysis based on the HPLC chromatograms and common pattern diagrams of the turmeric rhizome samples at different growth stages, and constructing a growth quality evaluation model for introduced turmeric; utilizing the contribution of key metabolites at different growth stages to the quality evaluation of turmeric, a growth quality evaluation model for introduced turmeric is constructed, so as to conduct a comparative quality evaluation of the growth process of introduced turmeric. The present application aims to optimize the traditional comprehensive growth quality evaluation by combining the key metabolites of turmeric medicinal materials during the growth process, and improve the reliability of the evaluation results.

Description

Technical Field

[0001] The present application relates to the technical field of analyzing material properties using optical means, and specifically to a method for evaluating the quality of Curcuma aromatica based on HPLC metabolomics analysis. Background Art

[0002] Curcuma zedoaria is a commonly used Chinese medicinal herb derived from the dried tubers of the ginger family turmeric, Curcuma longa, Curcuma zedoaria, or Curcuma zedoariae. It has been shown to promote qi circulation, dissipate blood stasis, and relieve depression. Curcuma zedoaria's efficacy is the result of the combined effects of multiple secondary metabolites. Quality control of only one or a few of these metabolites makes it difficult to comprehensively assess the safety, efficacy, stability, and controllability of traditional Chinese medicines and their preparations.

[0003] In recent years, studies have used HPLC metabolomics analysis to evaluate the quality of Curcuma aromatica. High-performance liquid chromatography (HPLC) offers advantages such as rapid analysis, high separation efficiency, high sensitivity, and wide sample applicability, making it the most mature and widely used fingerprint technology. Constructing a fingerprint of Curcuma aromatica using HPLC can comprehensively and quantifiably reflect the chemical information contained in the material. Compared with current methods of identifying single indicator components or active ingredients, the information contained in this fingerprint is more scientific and reasonable.

[0004] When evaluating the quality of Curcuma zedoaria during its growth process based on HPLC metabolomics analysis, existing methods focus on evaluation through the similarity of shared characteristic peaks. However, the metabolic processes and component accumulation of Curcuma zedoaria at different growth stages are dynamic. Due to the dynamic changes in the types and contents of metabolites, it is difficult to accurately grasp the quality characteristics of Curcuma zedoaria at different growth stages. Summary of the Invention

[0005] In order to solve the above technical problems, the present application provides a turmeric quality evaluation method based on HPLC metabolomics analysis to solve the existing problems.

[0006] The HPLC metabolomics analysis-based turmeric quality evaluation method of this application adopts the following technical solutions:

[0007] One embodiment of the present application provides a method for evaluating the quality of Curcuma aromatica based on HPLC metabolomics analysis, the method comprising the following steps:

[0008] Multiple rhizome samples of the same Curcuma zedoaria at different growth stages were collected, and HPLC chromatograms of the rhizome samples at all growth stages were obtained using high-performance liquid chromatography. The retention time and peak area of ​​the characteristic peaks in the HPLC chromatograms were used to construct a common pattern diagram of the rhizome samples of the same Curcuma zedoaria at all growth stages.

[0009] a. Compare the HPLC chromatograms of all turmeric rhizome samples at the same growth stage with the common pattern to obtain differential patterns to screen for differentially metabolized substances at different growth stages;

[0010] b. Analyze the differences in peak changes of the same metabolite in the differential profile of any turmeric rhizome sample at adjacent growth stages to screen out key metabolites at different growth stages;

[0011] c. Determine the intensity fluctuation index of any key metabolite at the same growth stage based on the average peak change of any key metabolite in all turmeric rhizome samples at the same growth stage and the difference between the average peak change of any key metabolite in all turmeric rhizome samples and the peak change of each turmeric rhizome sample;

[0012] d. Determine the contribution of any key metabolite to the quality evaluation of turmeric at the same growth stage based on the difference between the intensity fluctuation index of several key metabolites with similar peak intensities to the peak corresponding to any key metabolite and the intensity fluctuation index of any key metabolite;

[0013] By using the contribution of key metabolites at different growth stages to the quality evaluation of Curcuma australis, a growth quality evaluation model for introduced Curcuma australis was constructed to conduct a comparative quality evaluation of the growth process of introduced Curcuma australis.

[0014] Preferably, the growth stages include seedling stage, growth stage, flowering stage and fruiting stage.

[0015] Preferably, the method for constructing the shared pattern graph is:

[0016] Perform baseline correction on HPLC chromatograms; identify and label each characteristic peak in the HPLC chromatograms of all rhizome samples of the same Curcuma zedoaria at different growth stages, and determine the corresponding metabolites;

[0017] The retention time and area peak data of the common characteristic peaks in the HPLC chromatograms of all rhizome samples of the same type of Curcuma at all growth stages were sorted out to construct a common pattern diagram of the rhizome samples of the same type of Curcuma at all growth stages.

[0018] Preferably, the screening process for metabolite differences in different growth stages includes:

[0019] Compare the HPLC chromatograms of any Curcuma rhizome sample at different growth stages with the common pattern, and identify the characteristic peaks that exist in the HPLC chromatogram but are missing from the common pattern;

[0020] The HPLC chromatogram is matched with the common pattern using the retention time and relative area of ​​the identified characteristic peaks; and the HPLC chromatogram and the common pattern are normalized;

[0021] The difference spectrum was calculated using the point difference method: difference spectrum = HPLC chromatogram - common pattern; the peak point in the difference spectrum was the difference peak;

[0022] The common differential peaks present in the differential spectra of all turmeric rhizome samples at different growth stages were regarded as the differential metabolites at different growth stages.

[0023] Preferably, the method for determining the possibility that the same differential metabolite belongs to a key metabolite is:

[0024] Calculate the peak changes of any metabolite in the differential spectrum of any Curcuma rhizome sample at any growth stage;

[0025] Calculate the difference in peak changes of the same metabolite in any Curcuma rhizome sample at adjacent growth stages;

[0026] Calculate the standard deviation of the peak changes of the same metabolite in the difference maps of any Curcuma rhizome sample at all growth stages;

[0027] Determining the possibility that any metabolite difference of the Curcuma rhizome sample in any growth stage is a key metabolite based on the difference in the peak change and the standard deviation;

[0028] Metabolic differences whose probability of belonging to key metabolites was greater than or equal to the preset fluctuation threshold were recorded as key metabolites in the same growth stage.

[0029] Preferably, the peak change is determined by the difference between the peak intensity of any metabolite corresponding peak in the difference map of any Curcuma rhizome sample at any growth stage and the average peak intensity of all metabolite corresponding peaks.

[0030] Preferably, the method for determining the possibility of belonging to a key metabolite is: the cumulative sum of the ratio results of the differences in the peak changes of all turmeric rhizome samples for the same metabolite difference in adjacent growth stages and the standard deviation is used as the possibility that any metabolite difference of the turmeric rhizome sample in any growth stage belongs to a key metabolite.

[0031] Preferably, the method for determining the intensity fluctuation index of any key metabolite in the same growth stage is:

[0032] The nth growth stage The intensity fluctuation index of the key metabolites is ;

[0033]

[0034] in, represents the average peak change of the jth key metabolite in all Curcuma rhizome samples at the nth growth stage; exp represents the exponential function with the natural constant as the base; Indicates the number of turmeric rhizome samples collected; Indicates the nth growth stage The peak changes of the key metabolites in the i-th Curcuma rhizome sample.

[0035] Preferably, the method for determining the contribution of any key metabolite in the same growth stage to the quality evaluation of turmeric includes:

[0036] The nth growth stage The contribution of the key metabolites to the quality evaluation of Curcuma is recorded as ;

[0037]

[0038] Wherein, K represents the preset number of similar key metabolites of the key metabolite; Indicates the nth growth stage Intensity fluctuation index of key metabolites; Indicates the nth growth stage The intensity fluctuation index of the kth similar key metabolite of a key metabolite.

[0039] Preferably, the method for constructing the growth quality evaluation model of introduced turmeric is:

[0040] The quality evaluation score of the Curcuma rhizome sample at the nth growth stage is recorded as ;

[0041]

[0042] Among them, norm represents the normalization function; represents the amount of key metabolites in the nth growth stage; represents the principal component score of the characteristic peak corresponding to the jth key metabolite in the nth growth stage in the principal component direction of the common pattern graph; represents the contribution of the jth key metabolite to the quality evaluation of Curcuma zedoaria in the nth growth stage; NJ represents the number of common characteristic peaks in the common pattern diagram; It represents the principal component score of the Jth common characteristic peak in the common pattern graph in the principal component direction;

[0043] The quality evaluation scores of turmeric rhizome samples at all growth stages were used as indicators of the growth quality evaluation model for introduced turmeric.

[0044] This application has at least the following beneficial effects:

[0045] The present application effectively identifies significantly different metabolites in different growth stages of turmeric, introduces the contribution of key metabolites to the quality evaluation of turmeric during principal component analysis, and enhances the performance characteristics of key metabolites during quality evaluation. This allows for a more accurate evaluation of the quality of introduced turmeric at different growth stages by combining the fluctuation state of key metabolites when evaluating the quality of the introduced turmeric at its growth stages. BRIEF DESCRIPTION OF THE DRAWINGS

[0046] In order to more clearly illustrate the technical solutions and advantages of the embodiments of the present application or the prior art, the following is a brief introduction to the drawings required for use in the embodiments or the description of the prior art. Obviously, the drawings described below are only some embodiments of the present application. For ordinary technicians in this field, other drawings can be obtained based on these drawings without any creative work.

[0047] Figure 1 Flowchart of the turmeric quality evaluation method based on HPLC metabolomics analysis provided in this application;

[0048] Figure 2 Schematic diagram of the process of constructing a growth quality evaluation model for introduced turmeric provided in this application. DETAILED DESCRIPTION

[0049] To further illustrate the technical means and efficacy employed by this application to achieve the intended invention objectives, the following, in conjunction with the accompanying drawings and preferred embodiments, describes in detail the HPLC metabolomics analysis-based turmeric quality evaluation method proposed in this application, including its specific implementation, structure, features, and efficacy. In the following description, references to different "one embodiment" or "another embodiment" do not necessarily refer to the same embodiment. Furthermore, specific features, structures, or characteristics of one or more embodiments may be combined in any suitable manner.

[0050] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs.

[0051] The specific scheme of the turmeric quality evaluation method based on HPLC metabolomics analysis provided in this application is described in detail below with reference to the accompanying drawings.

[0052] One embodiment of the present application provides a method for evaluating the quality of Curcuma aromatica based on HPLC metabolomics analysis.

[0053] Specifically, the following HPLC metabolomics analysis-based turmeric quality evaluation method is provided, please refer to Figure 1 , the method comprises the following steps:

[0054] Step 1: Collect multiple rhizome samples of the same type of turmeric at different growth stages, and use high performance liquid chromatography to obtain HPLC chromatograms of the rhizome samples at all growth stages; use the retention time and peak area of ​​the characteristic peaks in the HPLC chromatograms to construct a common pattern diagram of the same type of turmeric rhizome samples at all growth stages.

[0055] Root and rhizome samples of authentic turmeric were collected at different growth stages (seedling, growing, flowering, and fruiting). To minimize the impact of individual sample differences on turmeric quality evaluation and ensure statistical significance of the analyzed data, sufficient sample data was collected during the data acquisition phase. For example, 20 rhizome samples of the same turmeric variety were collected at each growth stage within the same planting area.

[0056] High performance liquid chromatography (HPLC) was used to perform qualitative and quantitative analysis on the collected Curcuma rhizome samples. HPLC chromatograms of Curcuma rhizome samples at all growth stages were obtained, and baseline correction was performed. After removing baseline drift and noise, each characteristic peak in the HPLC chromatogram was identified and marked, and the metabolites corresponding to each characteristic peak in the HPLC chromatogram were determined.

[0057] Chromatographic data processing software was used to process the HPLC chromatograms of turmeric rhizome samples collected at different growth stages. The characteristic peak retention time and area peak data of the HPLC chromatograms of all rhizome samples of the same turmeric at all growth stages were sorted out, and a common pattern diagram of the same turmeric rhizome samples at all growth stages was constructed.

[0058] Step 2: Combine the HPLC chromatograms and common pattern diagrams of Curcuma australis rhizome samples at different growth stages to conduct difference analysis and construct a growth quality evaluation model for introduced Curcuma australis.

[0059] Currently, when synthesizing HPLC chromatograms of several Curcuma root rhizome samples from the same growth stage to construct a shared pattern diagram reflecting the overall chemical characteristics of that growth stage, the primary consideration is shared characteristic peaks that consistently appear in Curcuma root rhizome samples at different growth stages and that reflect the quality characteristics of Curcuma. However, during the overall growth process of Curcuma, the metabolic processes and component accumulation of Curcuma at different growth stages change dynamically, and the content of metabolites may vary significantly. This dynamic change is crucial for the comprehensive evaluation of Curcuma quality. Evaluation methods that focus solely on the similarity of shared characteristic peaks tend to overlook the metabolite content of Curcuma at different growth stages.

[0060] Therefore, in this step, by comparing the composition differences of the HPLC chromatogram and the common pattern diagram of Curcuma at different growth stages, the key metabolites of the Curcuma rhizome samples at different growth stages were screened out, and the correction parameters were further constructed based on the development of key metabolites to correct the quality evaluation results of Curcuma at different growth stages.

[0061] Based on this, the present application processes the HPLC chromatograms of the collected turmeric rhizome samples, modifies the quality evaluation process of different growth stages to construct a growth quality evaluation model for introduced turmeric, and the process diagram of constructing the growth quality evaluation model for introduced turmeric is shown in the attached figure. Figure 2 As shown, specifically:

[0062] a. Compare the HPLC chromatograms of all turmeric rhizome samples at the same growth stage with the common pattern to obtain differential patterns to screen out differential metabolites at different growth stages.

[0063] Samples at different growth stages will show different metabolite peaks and expression patterns. Since some metabolites are higher in certain growth stages and extremely low or disappear in other stages, when constructing the common pattern map, these metabolites with obvious differences between different stages will be deleted. However, these metabolites can often better reflect the characteristic state of the current growth stage. Therefore, in this step, based on the differences in HPLC chromatograms, characteristic peaks with different expression between the HPLC chromatograms and the common pattern map of Curcuma rhizome samples at different growth stages are selected as metabolite differences, which can preliminarily characterize the metabolic differences in each growth stage.

[0064] Take the rhizome sample of Curcuma zedoaria in the fruiting stage as an example for analysis:

[0065] The HPLC chromatograms of all Curcuma rhizome samples at the current growth stage were compared with the common pattern map, and characteristic peaks that appeared in the HPLC chromatograms but were missing in the common pattern map were identified. These characteristic peaks were more likely to belong to metabolically differential products.

[0066] The retention time and relative area of ​​these identified characteristic peaks were used to match the HPLC chromatograms with the common pattern map to ensure that the corresponding positions of the same metabolite in the two maps were consistent; at the same time, all HPLC chromatograms were normalized with the common pattern map to make the different maps comparable.

[0067] The difference spectrum is then calculated using the point difference method: difference spectrum = HPLC chromatogram - common pattern map; in the difference spectrum, each peak point is a difference peak, indicating the difference in peak intensity at a specific retention time, reflecting the difference between the sample spectrum and the common pattern map for each metabolite.

[0068] Furthermore, the difference maps of all Curcuma rhizome samples in the fruiting stage were compared, and the common difference peaks existing in all the difference maps were identified as the metabolic differences in the current fruiting stage.

[0069] b. Analyze the differences in the peak changes of the same metabolite in the differential profile of any turmeric rhizome sample at adjacent growth stages to screen out key metabolites in different growth stages.

[0070] The above steps were used to obtain differential metabolites of turmeric rhizome samples during the fruiting period. Since the components gradually accumulate during the growth of turmeric, these differential metabolites exhibit different significant changes in different growth stages. There is a state in which a certain differential metabolite is absent in the seedling stage and the growth stage, but its content increases rapidly during the flowering stage or the fruiting stage.

[0071] Therefore, this step combines the dynamic accumulation process of components in turmeric at different growth stages to determine the key metabolites in the metabolic differences that affect the quality of turmeric at the current growth stage, and constructs correction parameters based on the accumulation process of key metabolites in different growth stages to more accurately determine the unique metabolic characteristics of each growth stage, which is conducive to evaluating the quality characteristics of turmeric at different growth stages.

[0072] Take the nth growth stage as an example for analysis:

[0073] Calculate the peak change of the peak corresponding to the jth metabolite difference in the difference map of the i-th turmeric rhizome sample at the n-th growth stage : ;in, represents the peak intensity of the peak corresponding to the jth metabolite difference in the difference map of the i-th Curcuma rhizome sample at the n-th growth stage; This represents the average peak intensity of all metabolite differences in the differential spectrum of the i-th turmeric rhizome sample at the n-th growth stage. By calculating the peak changes of several metabolites in each turmeric rhizome sample at each growth stage, the degree of distinctness of each metabolite is reflected. Similarly, the peak changes of the j-th metabolite in each turmeric rhizome sample at each growth stage are calculated.

[0074] Since the metabolites in the growth process of turmeric are in a dynamic change process, the possibility that the jth metabolite difference of the turmeric rhizome sample in the nth growth stage belongs to the key metabolite is calculated according to the difference in the peak change of the samples at different growth stages. : ,in, It represents the difference in the peak change of the jth metabolite in the difference map of the i-th turmeric rhizome sample at the n-th growth stage and the previous stage, It represents the standard deviation of the peak change of the jth metabolite in the difference map of all growth stages of the i-th turmeric rhizome sample. The more obvious the chaotic distribution of the difference between the jth metabolite and the adjacent stages of the current growth stage relative to the peak change of the overall metabolite, the more likely it is that the metabolite is a key metabolite. When n=1, It represents the difference in the peak change of the jth metabolite in the difference map of the i-th Curcuma rhizome sample between the n-th growth stage and the next stage.

[0075] When the changes of metabolites in the previous and next stages are more significantly different from those in the current growth stage, the metabolites are more closely related to the biological processes of the stage. The calculated value of is larger, so the difference between the metabolic differences of a single growth stage and other stages is distinguished, and the metabolic differences that are significantly different from those of other growth stages are recorded as key metabolites. This application presets a fluctuation threshold T, which is set to 0.75. Metabolite differences greater than or equal to the fluctuation threshold were recorded as key metabolites in the nth growth stage.

[0076] c. Determine the intensity fluctuation index of any key metabolite in the same growth stage based on the average peak change of any key metabolite in all turmeric rhizome samples at the same growth stage and the difference between the average peak change of any key metabolite in all turmeric rhizome samples and the peak change of each turmeric rhizome sample.

[0077] Based on the key metabolites retained in each growth stage of the turmeric rhizome sample, the similarity of the changing trends of the key metabolites and other common characteristic peaks of different turmeric rhizome samples at the same growth stage was analyzed and calculated in the original HPLC chromatogram. The key metabolites with obvious data fluctuations were analyzed. When the intensity fluctuation of the metabolite difference in the same stage is more uniform, it means that the expression state of the difference peak in the turmeric rhizome samples at the current growth stage is more concentrated, and the expression of the metabolite in this stage is more consistent. Therefore, the intensity fluctuation index of any key metabolite in the same growth stage was calculated:

[0078]

[0079] in, Indicates the nth growth stage Intensity fluctuation index of key metabolites; represents the average peak change of the jth key metabolite in all Curcuma rhizome samples at the nth growth stage; exp represents the exponential function with the natural constant as the base; Indicates the number of turmeric rhizome samples collected; Indicates the nth growth stage The peak changes of the key metabolites in the i-th Curcuma rhizome sample.

[0080] It should be understood that The larger the value of , the smaller the peak variation of the jth key metabolite among several samples in the current growth stage, and the more obvious the difference of the key metabolites in different Curcuma rhizome samples in the same stage; The smaller the value of The larger the calculated result of , the smaller the difference between the peak changes of key metabolites in different samples and the mean values ​​of samples in the corresponding stage, indicating that the peak changes of the j-th key metabolite in the current growth stage are more consistent.

[0081] d. Determine the contribution of any key metabolite to the quality evaluation of turmeric at the same growth stage based on the difference between the intensity fluctuation indexes of several key metabolites with similar peak intensities to the corresponding peak of any key metabolite and the intensity fluctuation index of any key metabolite.

[0082] In the turmeric growth metabolic network, many metabolites may come from the same or similar biosynthetic pathways. For example, some metabolites in the current growth stage may be decomposed or synthesized by common precursors and show similar peak intensities in the HPLC chromatogram. Therefore, the K key metabolites with the closest peak intensity difference to the peak corresponding to the j-th key metabolite are obtained. The preset K value in this embodiment of the application is 3.

[0083] The synthesis of similar key substances is affected by the same physiological regulatory mechanism, and therefore produces similar changes in the same changing environment. If the intensity fluctuation index of similar metabolites in the HPLC chromatogram is highly consistent, it indicates that their production or consumption is more sensitive to environmental changes. Therefore, when performing quality analysis of turmeric growth stages, it is necessary to pay close attention to the impact of key metabolites, and the higher the contribution of the key metabolites to the quality evaluation of turmeric. The contribution of the key metabolites to the quality evaluation of Curcuma is recorded as , the expression is:

[0084]

[0085] Wherein, K represents the preset number of similar key metabolites of the key metabolite; Indicates the nth growth stage Intensity fluctuation index of key metabolites; Indicates the nth growth stage The intensity fluctuation index of the kth similar key metabolite of the key metabolite. The smaller the difference in the intensity fluctuation index of similar key metabolites of the key metabolites, and the larger the fluctuation index value of each key metabolite characteristic peak in different samples, the more susceptible the current key metabolite is to influence. The greater the contribution of a key metabolite to the quality evaluation of Curcuma.

[0086] Step 3: Using the contribution of key metabolites in different growth stages to the quality evaluation of turmeric, a growth quality evaluation model for introduced turmeric was constructed to conduct a comparative quality evaluation of the growth process of introduced turmeric.

[0087] In general, the common pattern graph contains 10-15 common characteristic peaks with the greatest similarity. Therefore, when evaluating the quality of turmeric rhizome samples, it is necessary to use the principal component analysis method to obtain the principal component direction of the common pattern graph, calculate the projection of the common characteristic peak on the principal component direction, and use it as the degree of influence of the metabolite on the quality of turmeric. This step combines the contribution of the key metabolites in different growth stages to the quality evaluation of turmeric calculated above to construct a growth quality evaluation model for introduced turmeric, evaluate the different growth stages of introduced turmeric, calculate the quality evaluation scores of turmeric rhizome samples at different growth stages, and use the quality evaluation score of turmeric rhizome samples at the nth growth stage as the final score. For example, the expression is:

[0088]

[0089] Among them, norm represents the normalization function; represents the amount of key metabolites in the nth growth stage; represents the principal component score of the characteristic peak corresponding to the jth key metabolite in the nth growth stage in the principal component direction of the common pattern graph; represents the contribution of the jth key metabolite to the quality evaluation of Curcuma zedoaria in the nth growth stage; NJ represents the number of common characteristic peaks in the common pattern diagram; It represents the principal component score of the Jth common characteristic peak in the common pattern graph in the principal component direction.

[0090] Contribution of key metabolites to the quality evaluation of Curcuma zedoaria by introducing them into principal component analysis , which enhances the performance characteristics of key metabolites during quality evaluation, so that when evaluating the quality of introduced turmeric during the growth stage, the fluctuation state of key metabolites can be combined to more accurately evaluate the quality of turmeric in the current growth stage.

[0091] The quality evaluation scores of turmeric rhizome samples at all growth stages were used as indicators in the growth quality evaluation model for introduced turmeric, which was used to screen high-quality samples or assess the quality consistency between different batches. Using the growth quality evaluation model for introduced turmeric, the quality evaluation scores of introduced turmeric rhizome samples at each growth stage were calculated for different batches of introduced turmeric rhizome samples. These scores were compared with the quality evaluation scores of authentic turmeric rhizome samples at the same growth stage to identify batches of introduced turmeric with significant quality differences. Based on these differences, introduced turmeric was classified into three categories: high-quality (the absolute difference in the quality evaluation scores between introduced and authentic turmeric rhizome samples at the same growth stage was greater than 0.7 or higher), medium-quality (the absolute difference in the quality evaluation scores between introduced and authentic turmeric rhizome samples at the same growth stage was within the range of 0.3-0.7), and low-quality (the absolute difference in the quality evaluation scores between introduced and authentic turmeric rhizome samples at the same growth stage was less than or equal to 0.3). This provides a scientific basis for optimizing the cultivation and improving the quality of introduced turmeric.

[0092] The above technical features constitute the best embodiment of the present application, which has strong adaptability and optimal implementation effect. Non-essential technical features can be added or removed according to actual needs to meet the requirements of different situations.

Claims

1. A method for evaluating the quality of Curcuma aromatica based on HPLC metabolomics analysis, characterized in that: The method comprises the following steps: Multiple rhizome samples of the same Curcuma zedoaria at different growth stages were collected, and HPLC chromatograms of the rhizome samples at all growth stages were obtained using high-performance liquid chromatography. The retention time and peak area of ​​the characteristic peaks in the HPLC chromatograms were used to construct a common pattern diagram of the rhizome samples of the same Curcuma zedoaria at all growth stages. a. Compare the HPLC chromatograms of all turmeric rhizome samples at the same growth stage with the common pattern to obtain differential patterns to screen for differentially metabolized substances at different growth stages; b. Analyze the differences in peak changes of the same metabolite in the differential profile of any turmeric rhizome sample at adjacent growth stages to screen out key metabolites at different growth stages; c. Determine the intensity fluctuation index of any key metabolite at the same growth stage based on the average peak change of any key metabolite in all turmeric rhizome samples at the same growth stage and the difference between the average peak change of any key metabolite in all turmeric rhizome samples and the peak change of each turmeric rhizome sample; d. Determine the contribution of any key metabolite to the quality evaluation of turmeric at the same growth stage based on the difference between the intensity fluctuation index of several key metabolites with similar peak intensities to the peak corresponding to any key metabolite and the intensity fluctuation index of any key metabolite; By using the contribution of key metabolites at different growth stages to the quality evaluation of Curcuma australis, a growth quality evaluation model for introduced Curcuma australis was constructed to conduct a comparative quality evaluation of the growth process of introduced Curcuma australis.

2. The turmeric quality evaluation method based on HPLC metabolomics analysis as claimed in claim 1, wherein The growth stages include seedling stage, growth stage, flowering stage and fruiting stage.

3. The turmeric quality evaluation method based on HPLC metabolomics analysis as claimed in claim 1, wherein The method for constructing the common pattern diagram is: Perform baseline correction on HPLC chromatograms; identify and label each characteristic peak in the HPLC chromatograms of all rhizome samples of the same Curcuma zedoaria at different growth stages, and determine the corresponding metabolites; The retention time and area peak data of the common characteristic peaks in the HPLC chromatograms of all rhizome samples of the same type of Curcuma at all growth stages were sorted out to construct a common pattern diagram of the rhizome samples of the same type of Curcuma at all growth stages.

4. The turmeric quality evaluation method based on HPLC metabolomics analysis as claimed in claim 1, characterized in that, The screening process of the metabolites at different growth stages includes: Compare the HPLC chromatograms of any Curcuma rhizome sample at different growth stages with the common pattern, and identify the characteristic peaks that exist in the HPLC chromatogram but are missing from the common pattern; The HPLC chromatogram is matched with the common pattern using the retention time and relative area of ​​the identified characteristic peaks; and the HPLC chromatogram and the common pattern are normalized; The difference spectrum was calculated using the point difference method: difference spectrum = HPLC chromatogram - common pattern; the peak point in the difference spectrum was the difference peak; The common differential peaks present in the differential spectra of all turmeric rhizome samples at different growth stages were regarded as the differential metabolites at different growth stages.

5. The turmeric quality evaluation method based on HPLC metabolomics analysis as claimed in claim 1, characterized in that, The method for determining the possibility that the same metabolite belongs to a key metabolite is: Calculate the peak changes of any metabolite in the differential spectrum of any Curcuma rhizome sample at any growth stage; Calculate the difference in peak changes of the same metabolite in any Curcuma rhizome sample at adjacent growth stages; Calculate the standard deviation of the peak changes of the same metabolite in the difference maps of any Curcuma rhizome sample at all growth stages; Determining the possibility that any metabolite difference of the Curcuma rhizome sample in any growth stage is a key metabolite based on the difference in the peak change and the standard deviation; Metabolic differences whose probability of belonging to key metabolites was greater than or equal to the preset fluctuation threshold were recorded as key metabolites in the same growth stage.

6. The method for evaluating the quality of Curcuma aromatica based on HPLC metabolomics analysis as claimed in claim 5, wherein: The peak change is determined by the difference between the peak intensity of a peak corresponding to any metabolite difference in the difference map of any Curcuma rhizome sample at any growth stage and the average peak intensity of the peaks corresponding to all metabolites difference.

7. The method for evaluating the quality of Curcuma aromatica based on HPLC metabolomics analysis as claimed in claim 5, wherein: The method for determining the possibility of belonging to a key metabolite is: the cumulative sum of the ratio results of the differences in the peak changes of all turmeric rhizome samples for the same metabolite difference in adjacent growth stages and the standard deviation is used as the possibility that any metabolite difference of the turmeric rhizome sample in any growth stage is a key metabolite.

8. The method for evaluating the quality of Curcuma aromatica based on HPLC metabolomics analysis as claimed in claim 1, wherein The method for determining the intensity fluctuation index of any key metabolite in the same growth stage is: The nth growth stage The intensity fluctuation index of the key metabolites is ; ;in, represents the average peak change of the jth key metabolite in all Curcuma rhizome samples at the nth growth stage; exp represents the exponential function with the natural constant as the base; Indicates the number of turmeric rhizome samples collected; Indicates the nth growth stage The peak changes of the key metabolites in the i-th Curcuma rhizome sample.

9. The method for evaluating the quality of Curcuma aromatica based on HPLC metabolomics analysis as claimed in claim 1, wherein The method for determining the contribution of any key metabolite in the same growth stage to the quality evaluation of turmeric includes: The nth growth stage The contribution of the key metabolites to the quality evaluation of Curcuma is recorded as ; ; Wherein, K represents the preset number of similar key metabolites of the key metabolite; Indicates the nth growth stage Intensity fluctuation index of key metabolites; Indicates the nth growth stage The intensity fluctuation index of the kth similar key metabolite of a key metabolite.

10. The method for evaluating the quality of Curcuma aromatica based on HPLC metabolomics analysis as claimed in claim 3, wherein: The construction method of the growth quality evaluation model of introduced turmeric is: The quality evaluation score of the Curcuma rhizome sample at the nth growth stage is recorded as ; ; Among them, norm represents the normalization function; represents the amount of key metabolites in the nth growth stage; represents the principal component score of the characteristic peak corresponding to the jth key metabolite in the nth growth stage in the principal component direction of the common pattern graph; represents the contribution of the jth key metabolite to the quality evaluation of Curcuma zedoaria in the nth growth stage; NJ represents the number of common characteristic peaks in the common pattern diagram; It represents the principal component score of the Jth common characteristic peak in the common pattern graph in the principal component direction; The quality evaluation scores of turmeric rhizome samples at all growth stages were used as indicators of the growth quality evaluation model for introduced turmeric.

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