Method for evaluating quality of different parts of sarcandra glabra based on sarcandra glabra characteristic spectrum combined with quantitative analysis of multi-components by single marker

Abstract

The invention discloses a method for evaluating the quality of different parts of sarcandra glabra based on a sarcandra glabra characteristic spectrum combined with a quantitative analysis of multi-components by single marker, which belongs to the technical field of detection methods and comprises the following steps: preparing a reference solution, preparing a test solution, establishing the characteristic spectrum and establishing the quantitative analysis of multi-components by single marker. The UPLC characteristic chromatogram established by the method and the quantitative analysis of multi-components by single marker can simultaneously evaluate different parts of the sarcandra glabra, and the method has important reference value for component analysis and quality control of the sarcandra glabra.

Description

Technical Field

[0001] The present invention relates to the technical field of detection methods, and in particular to a method for evaluating the quality of different parts of Jiujie Tea based on a Jiujie Tea characteristic spectrum combined with a one-test-multiple-evaluation method. Background Art

[0002] Jiujie Tea is the dried aerial part of Sarcandra glabra (Thunb.) Nakai, a plant of the Chrysopogonaceae family. It has the effects of clearing away heat and detoxifying, cooling blood, removing spots, removing wind and dampness, and dredge meridians to relieve pain. Jiujie Tea has a long history of use as a medicine. It was first recorded in the Ming Dynasty's "Ru'nan Garden History". The first record of the medicinal use of coral under the name of "Jiujie Tea" was seen in the Qing Dynasty's "Essentials of the Nature of Herbs". Jiujie Tea is widely distributed. The first volume of the "Guangdong Provincial Standards for Traditional Chinese Medicines" stipulates that the active ingredient is isofraquinone. Studies have shown that the main active ingredients in Jiujie Tea are sesquiterpenes, phenolic acids, flavonoids and coumarins. Among them, chlorogenic acid, isofraquinone and rosmarinic acid have strong anti-inflammatory activity; isofraquinone has strong anti-tumor activity.

[0003] Studies have shown that the chemical composition of different medicinal parts is significantly different. For example, the contents of caffeic acid, luteolin, isochlorogenic acid B, 4,5-O-dicaffeoylquinic acid, and wedelilacin in the stems of Eclipta prostrata are lower than those in leaves and flowers, while the contents of total flavonoids, baicalin, baicalin, luteolin, and apigenin in the leaves of Scutellaria barbata are higher than those in the stems, fruits, and whole herbs. With the depletion of Chinese medicinal resources, it is of great significance to study the differences in the components of different parts of Jiujiecha in order to promote the sustainable use of Chinese medicinal resources. Summary of the invention

[0004] In order to solve the above problems, the present invention provides a method for evaluating the quality of different parts of Jiujie Tea based on the characteristic spectrum of Jiujie Tea combined with a one-test-multiple-evaluation method. The method of the present invention can provide a theoretical basis for the quality control of ramie root medicinal materials. The content of the present invention is as follows: A method for evaluating the quality of different parts of Jiujie Tea based on a Jiujie Tea characteristic spectrum combined with a one-test-multiple-evaluation method, the technical point of which is that it includes the following steps: Step 1, preparation of reference solution: taking neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, caffeic acid, rosmarinic acid, and isoflurane reference substances and adding methanol to mix and dissolve to form a reference solution; Step 2, preparation of the test solution: take the above-ground part of Camellia sinensis, place it in a stoppered conical flask, add methanol, heat it under reflux, cool it, and weigh it again, make up the lost weight with methanol, shake it well, filter it, and take the filtrate to obtain the test solution; Step 3, establishment of characteristic spectrum: take the test sample solutions of the aerial part, leaves and stems of Jiujie Tea, record the characteristic spectrum of Jiujie Tea of ​​each sample under the condition of liquid chromatography, import the data into the "Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System", set the reference spectrum of the characteristic spectrum, automatically match it and then calibrate it at multiple points, perform full peak matching to generate a control overlay map, and then perform principal component and orthogonal partial least squares discrimination; Step 4, establishment of a one-test-multiple-evaluation method: Based on the results of principal component analysis and orthogonal partial least squares discriminant analysis, a one-test-multiple-evaluation quantitative characterization method was established with neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, caffeic acid, isoflurane, and rosmarinic acid as detection components.

[0005] The method for evaluating the quality of different parts of Jiujie Tea based on the characteristic spectrum of Jiujie Tea combined with the one-test-multiple-evaluation method of the present invention establishes a one-test-multiple-evaluation analysis method for establishing the UPLC characteristic spectrum of Jiujie Tea and new chlorogenic acid, chlorogenic acid, cryptochlorogenic acid, caffeic acid, isofraquinone, and rosmarinic acid, and performs similarity evaluation, orthogonal partial least squares discriminant analysis (OPLS-DA) and principal component analysis. As a result, 11 common peaks were calibrated in different parts of 42 batches of Jiujie Tea, and the similarity of different parts was greater than 0.840. The aboveground parts, leaves, and stems of Jiujie Tea can be roughly divided into three categories. The chemical composition of the aboveground parts is close to that of the stems, and the difference is large with the leaves. Two principal components were obtained by principal component analysis, and the cumulative contribution rate of variance reached 80.117%. There was no significant difference in the results of the one-test-multiple-evaluation method and the external standard method. The content of chlorogenic acid and isofraquinone in the stems of Jiujie Tea was higher than that in the leaves, and the content of new chlorogenic acid, cryptochlorogenic acid, caffeic acid, and rosmarinic acid in the leaves was higher than that in the stems, and the mean value of each content in the aboveground parts was close to that in the stems. The established UPLC characteristic spectrum and one-test-multiple-evaluation method can simultaneously evaluate different parts of Jiujie Tea, which has important reference value for the component analysis and quality control of Jiujie Tea. BRIEF DESCRIPTION OF THE DRAWINGS

[0006] The drawings described herein are used to provide a further understanding of the present invention and constitute a part of the present invention. The exemplary embodiments of the present invention and their descriptions are used to explain the present invention and do not constitute an improper limitation of the present invention. In the drawings: Figure 1 It is the common peak of the characteristic spectrum of the aboveground part of Jiujie tea; Figure 2 It is the common peak of the characteristic spectrum of nine sections of tea leaves; Figure 3 It is the common peak of the characteristic spectrum of the nine-section tea stem; Figure 4 It is a comparison chart of the aerial parts, leaves, stems and mixed reference substances of Jiujiecha; Figure 5 The scree plot was analyzed for principal component analysis; Figure 6 is the OPLS-DA score graph; Figure 7It is the VIP diagram of OPLS-DA model; Figure 8 This is the cross-validation result of the OPLS-DA model. DETAILED DESCRIPTION

[0007] In order to make the purpose, technical solution and advantages of the present invention clearer, the technical solution of the present invention will be clearly and completely described below in conjunction with the specific embodiments of the present invention and the corresponding drawings. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative work are within the scope of protection of the present invention.

[0008] The technical solutions provided by various embodiments of the present invention are described in detail below in conjunction with the accompanying drawings.

[0009] This study conducted comparative analysis on different parts of the Jiujiecha medicinal material by establishing UPLC characteristic spectra and one-test-multiple-evaluation, providing a reference for the rational development and utilization, quality control and evaluation of Jiujiecha medicinal material resources.

[0010] 1. Instruments and reagents 1.1 Instrument Waters H-Class high performance liquid chromatograph (Waters Corporation), Waters BEHC18 (100 mm × 2.1 mm, 1.7 μm) chromatographic column, SQP type one-tenth balance (Sartorius Scientific Instruments Co., Ltd.); SB25-12DTS type CNC ultrasonic cleaner (Ningbo Xinzhi Biotechnology Co., Ltd.); XPR36DR / A type one-millionth balance (Mettler-Toledo Instrument Co., Ltd.).

[0011] 1.2 Drugs and reagents Methanol (Xilong Scientific Co., Ltd.) was of analytical grade, phosphoric acid (Tianjin Komiou Chemical Reagent Co., Ltd.) and acetonitrile (Merck Co., Ltd.) were of chromatographic grade; water was ultrapure water (self-made in the laboratory).

[0012] Chlorogenic acid (batch number: 110753-202119, content: 96.3%), caffeic acid (batch number: 110885-201703, content: 99.7%), rosmarinic acid (batch number: 111871-202007, content: 98.1%), and isoflurane (batch number: 110837-202009, content: 99.8%) were purchased from China Food and Drug Identification Institute; neochlorogenic acid (batch number: 906-33-2, content: 98.0%) was purchased from Wuhan Tianzhi Biotechnology Co., Ltd.; cryptochlorogenic acid (batch number: 905-99-7, content: 98.8%) was purchased from Chengdu Gelipu Biotechnology Co., Ltd.

[0013] The 14 batches of Jiujie Tea samples were identified by the Quality Department of Hunan Yifang Tianjiang Pharmaceutical Co., Ltd. as the dried aerial parts of Sarcandra glabra (Thunb.) Nakai, a plant of the Chrysanthemiaceae family. Some of the medicinal materials were selected to obtain 42 batches of aerial parts, leaves, and stems, which were numbered S1-S14, Y1-Y14, and J1-J14, respectively. The origin information is shown in Table 1.

[0014] Table 1 Sample information

[0015] 2 Methods and Results 2.1 Chromatographic conditions A Waters BEHC18 (100 mm × 2.1 mm, 1.7 μm) chromatographic column was used; the mobile phase was acetonitrile-0.1% phosphoric acid solution, and the gradient elution was: 0-2.5 min, 10% acetonitrile; 2.5-7 min, 10-16% acetonitrile; 7-14 min, 16-29% acetonitrile; 14-17 min, 29-30% acetonitrile; the detection wavelength was 330 nm; the column temperature was 25 °C; and the injection volume was 1 μL.

[0016] 2.2 Preparation of reference solution Take appropriate amount of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, caffeic acid, rosmarinic acid, and isoflurane reference substances, weigh accurately, and add methanol to prepare mixed reference substance solutions containing 5, 10, 10, 40, 20, and 10 μg per mL, respectively.

[0017] 2.3 Preparation of test solution Take about 1.0 g of the above-ground powder (S1, passed through a No. 3 sieve), accurately weigh it, place it in a stoppered conical flask, accurately add 50 mL of 60% methanol, weigh it, heat and reflux for 0.5 hour, cool, weigh it again, make up the lost weight with methanol, shake well, filter, and take the filtrate to obtain the product.

[0018] 2.4 Establishment of the characteristic map of Jiujie Tea 2.4.1 Feature map methodology review 2.4.1.1 Precision experiment The test solution of Jiujie Tea (S1) was taken and sampled and measured 6 times continuously according to the chromatographic conditions under item "2.1". The chromatographic peak of isoflurane was used as the reference peak (S). The RSD of the relative retention time of each common peak was calculated to be 0.07-0.83%, and the relative peak area was 0.31-1.82%. The results showed that the instrument had good precision.

[0019] 2.4.1.2 Stability test Take the test solution of Jiujie Tea (S1) and inject and measure at 0, 2, 4, 8, 12, 18 and 24 h according to the chromatographic conditions under "2.1". Take the chromatographic peak of isoflurane as the reference peak (S). The RSD of the relative retention time of each common peak is 0.29-0.76%, and the relative peak area is 0.18-1.27%. The results show that the test solution has good stability within 24 h.

[0020] 2.4.1.3 Repeatability test Take a Jiujie Tea sample (S1), prepare the test solution according to the method under "2.3", and repeat 6 times. Inject and measure according to the chromatographic conditions under "2.1". Take the chromatographic peak of isoflurane as the reference peak (S). The RSD of the relative retention time of each common peak is 0.24-1.63%, and the relative peak area is 0.71-1.86%. The results show that this method has good repeatability.

[0021] 2.4.2 Feature Spectrum Analysis and Evaluation Take 42 batches of Jiujie Tea aerial parts, leaves and stems, prepare the test solution according to the method under "2.3", and inject and measure according to the chromatographic conditions under "2.1". Import the chromatogram results into the "Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System" (2012 Edition), set S1 as the reference spectrum, and perform multi-point correction after automatic matching, and perform full spectrum peak matching. 14 batches of Jiujie Tea aerial parts and their leaves and stems are superimposed, and a total of 11 characteristic peaks are confirmed, see Figure 1-3 By comparing with the reference substance, a total of 6 chromatographic peaks were identified, peak 1 was neochlorogenic acid, peak 2 was chlorogenic acid, peak 3 was cryptochlorogenic acid, peak 5 was caffeic acid, peak 6 was isofraquinone, and peak 9 was rosmarinic acid. Figure 4 Taking peak No. 6 isoflurane with known composition, good peak shape and separation, and moderate retention time as the reference peak (S), the similarities of the aerial parts, leaves, and stems of 14 batches of Jiujie Tea were evaluated, and the similarities were 0.903-994, 0.873-997, and 0.848-0.991, respectively. The similarities were all greater than 0.840, and they were consistent with the control characteristic spectrum, indicating that the quality of the aerial parts, leaves, and stems of Jiujie Tea was stable, and the differences between the same parts were small. The results are shown in Table 2.

[0022] Table 2 Similarity evaluation results of different parts of 42 batches of Jiujie Tea

[0023] 2.4.3 Principal Component Analysis The common peak areas of the characteristic spectra of the aerial parts, leaves and stems of 42 batches of Jiujie Tea were used as variables. The principal component analysis was performed using SPSS20.0 software, and the eigenvalues ​​and variances of the correlation matrix were obtained (Table 3). The scree plot ( Figure 5), as shown in Table 3, according to the eigenvalue greater than 1, two principal components were obtained, and the cumulative contribution rate to the variance reached 80.117%, which is representative and can characterize most of the common peak information of the characteristic spectra of the aboveground parts, leaves and stems of Jiujie Tea. Component matrix (Table 4), as shown in Table 4, the first principal component mainly reflects peak 1 (neochlorogenic acid), peak 2 (chlorogenic acid), peak 3 (cryptochlorogenic acid), peak 4, peak 5 (caffeic acid), peak 7, peak 9 (rosmarinic acid), peak 11, and the second principal component mainly reflects peak 6 (isofraxinidin), peak 8, and peak 10. The principal component scores of different parts of Jiujie Tea were calculated (Table 5). Overall, the scores of principal component 2 of the aboveground parts and stems were higher, and the score of principal component 1 of the leaves was higher. There are large differences in the scores of principal components in different parts.

[0024] Table 3 Principal component eigenvalues ​​and variances

[0025] Table 4 Principal component analysis matrix

[0026] Table 5 Principal component scores

[0027] OPLS-DA analysis was performed on 42 batches of samples using the relative peak areas of 11 chromatographic peaks of the characteristic spectra of different parts of Jiujie Tea as variables. The R2X (cum), R2y (cum), and Q2 (cum) of the established model were all greater than 0.5, indicating that the model fitting results were acceptable. After 200 permutation tests, Figure 7 As shown in the figure, the intersection point of the Q2 regression line and the vertical axis is less than 0, indicating that the model does not fit and the model verification is effective. It is believed that this result can be used for the identification of different parts of Jiujie Tea. From the VIP value diagram of 11 variables ( Figure 8 ) It can be seen that with VIP>1.0 as the significance, a total of 3 differential markers were found, and their significant order was peak 8>peak 6>peak 10, indicating that these chemical components have a greater contribution to distinguishing different parts of Jiujie Tea. OPLS-DA analysis can better distinguish samples from different parts of Jiujie Tea ( Figure 6 ).

[0028] 2.5 Establishment of One Test, Multiple Evaluations Method According to the results of principal component and orthogonal partial least squares discriminant analysis, peak 1 (neochlorogenic acid), peak 2 (chlorogenic acid), peak 3 (cryptochlorogenic acid), peak 5 (caffeic acid), peak 6 (isofraxidin) and peak 9 (rosmarinic acid) were used as detection components, and a one-test-multiple-evaluation quantitative characterization method for the above six components was established.

[0029] 2.5.1 Methodological Review of the One Test, Multiple Evaluations Method 2.5.1.1 Precision investigation The test solution of Jiujie Tea (S1) was taken and sampled and measured 6 times continuously according to the chromatographic conditions under "2.1". The RSDs of the peak areas of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, caffeic acid, isofraquinone, and rosmarinic acid were 0.17%, 0.83%, 0.28%, 0.48%, 0.71%, and 0.29%, respectively, indicating that the instrument has good precision.

[0030] 2.5.1.2 Repeatability study Take Jiujie Tea sample (S1), prepare the test solution according to the method under "2.3", and make 6 parallel samples. The samples were injected and determined according to the chromatographic conditions under "2.1". The contents of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, caffeic acid, isofraquinone and rosmarinic acid were calculated. The RSDs of the contents of the six components were 1.21%, 1.31%, 0.89%, 1.79%, 0.78% and 1.38%, respectively. The RSDs were all less than 3%, indicating that the sample preparation process had good repeatability.

[0031] 2.5.1.3 Stability study Take the test solution of Jiujie Tea (S1) and inject the sample at 0, 2, 4, 8, 12, 18 and 24 hours according to the chromatographic conditions under "2.1". The peak areas RSD of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, caffeic acid, rosmarinic acid and isoflurane are all less than 3%, indicating that the test solution is stable within 24 hours.

[0032] 2.5.1.4 Investigation of linear relationship Take an appropriate amount of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, caffeic acid, isofraquinone, and rosmarinic acid reference substances, accurately weigh them, place them in a volumetric flask, and add methanol to prepare mixed reference substance solutions with concentrations of 24.70, 101.90, 170.90, 30.20, 184.30, and 209.30 μg / mL for neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, caffeic acid, isofraquinone, and rosmarinic acid, respectively.

[0033] Accurately pipette 1mL of the above mixed stock solution into 5, 10, 20, 50, and 100mL volumetric bottles, add methanol to the scale, and inject and measure the above reference stock solution in sequence according to the chromatographic conditions under "2.1", and record the peak area of ​​the chromatographic peak. Use the peak area as the ordinate (y) and the reference concentration as the abscissa (x), and draw a standard curve (see Table 6). The results show that the six components have a good linear relationship within the linear range.

[0034] Table 6 Regression equation and correlation coefficient

[0035] 2.5.1.5 Investigation of sample recovery rate Take about 0.5g of the powder of the aboveground part of the determined content of Jiujie Tea (S1), accurately weigh it, and add chlorogenic acid, chlorogenic acid, cryptochlorogenic acid, caffeic acid, isofraquinone, and rosmarinic acid reference substances in a ratio of 1:1 in 6 parallel groups; prepare the test solution according to the method under "2.3", inject and determine according to the chromatographic conditions under "2.1", calculate the recovery rate of the sample, and the average recovery rate of the six components is in the range of 95.63-101.84%, and the RSD% is less than 3%, which is in line with the provisions of General Principles 9101 of Part IV of the 2020 edition of the Chinese Pharmacopoeia. This shows that the determination method has good accuracy.

[0036] 2.5.2 Establishment of One Test, Multiple Evaluations Method 2.5.2.1 Calculation of relative correction factor Because isofraquinone has a medium retention time, a high content and easy access to reference substances, the calculation formula for the relative correction factor of isofraquinone as the internal reference (s) and other components to be tested (i) is:

[0037] Among them, f represents the relative correction factor, s represents the internal reference, i represents other components to be measured, A represents the peak area, and C represents the concentration.

[0038] 2.5.2.2 Durability test The relative retention time method was used, and isofraquinone was used as the reference peak to calculate the relative retention time of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, caffeic acid, and rosmarinic acid (ris=tRi / tRs (where tRs is the retention time of the reference peak and tRi is the retention time of the component to be measured). The chromatographic peaks were located, and the sample solution under "2.5.1.1" was taken to investigate the effects of different column temperatures (20, 25, 30°C), flow rates (0.25, 0.30, 0.35 mL / min), chromatographic columns (Waters HSST3, Waters BEHC18), and chromatographs (Waters H-Class, Aglient 1290) on the relative retention time (Table 6). The RSD was less than 5%, indicating that the method was robust and the relative retention time value could be used to locate the chromatographic peaks.

[0039] Table 7 Durability test results (relative retention time)

[0040] 2.5.2.3 Reproducibility study of relative correction factor The samples under item "2.5.1.1" were taken, and different column temperatures (20, 25, 30℃), flow rates (0.25, 0.30, 0.35mL / min), chromatographic columns (WatersHSST3, WatersBEHC18), and chromatographs (WatersH-Class, Aglient1290) were investigated respectively. The correction factors of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, caffeic acid, and rosmarinic acid were calculated (Table 8). The RSD was less than 2%, indicating that the correction factors had good applicability.

[0041] Table 8 Effect of different conditions on relative correction factor

[0042] 2.5.2.4 Sample determination Take 42 batches of samples from different parts of Jiujie Tea, prepare the test solution according to the method under "2.3", inject and measure according to the chromatographic conditions under "2.1", and record the peak area. Using isoflurane as a reference, the contents of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, caffeic acid, and rosmarinic acid were calculated by the one-measurement-multiple-evaluation method. The results are shown in Table 9. The content determinations of the two methods are close, with no significant difference, indicating that one-measurement-multiple-evaluation can be used for the quality evaluation of samples from different parts of Jiujie Tea.

[0043] The results of the comparison of the contents of samples from different parts of Jiujie Tea are shown in Table 10. The average total content of the aboveground part is 2.398 mg / g, the average total content of the leaves is 2.645 mg / g, and the average total content of the stems is 2.394 mg / g. From the average total content, the leaves>aboveground parts≈stems. From the mean content, the contents of chlorogenic acid and isoflurane in the stems are higher than those in the leaves, and the contents of neochlorogenic acid, cryptochlorogenic acid, caffeic acid, and rosmarinic acid in the leaves are higher than those in the stems. The mean contents of the aboveground parts of Jiujie Tea are close to those in the stems.

[0044] Table 9 Determination results of content in different parts of Jiujie Tea

[0045]

[0046] Table 10 Comparison of content in different parts of Jiujie Tea

[0047]

[0048] 3 Discussion and Conclusion In this study, characteristic spectrum analysis was performed on 42 batches of aboveground parts, leaves, and stems of Jiujie Tea, and a total of 11 common peaks were determined, of which 6 characteristic peaks, including neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, caffeic acid, isofraquinone, and rosmarinic acid, were identified. The similarity of the aboveground parts, leaves, and stems of 42 batches of Jiujie Tea was evaluated, and the similarity of the medicinal materials in the same part was above 0.840, indicating that the similarity of the medicinal materials in the same part of different batches was good. The principal component analysis of the common peaks of the characteristic spectrum of different parts of Jiujie Tea was performed by SPSS20.0 software, and 2 principal components were obtained. The principal component scores between different parts were calculated. The principal component 2 of the aboveground part and stem of Jiujie Tea had a higher score, and the principal component 1 of the leaf had a higher score. From the analysis of the principal component scores, the aboveground part and stem of Jiujie Tea were more similar. Through orthogonal partial least squares discriminant analysis, the OPLS-DA analysis of the characteristic spectrum of different parts of Jiujie Tea can distinguish the aboveground part, leaves, and stems, which is consistent with the principal component analysis and similarity results of the characteristic spectrum.

[0049] Through the one-test-multiple-evaluation method, a correction factor was established with isofraquinone as the internal reference and the components to be tested being neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, caffeic acid and rosmarinic acid. The durability of the correction factor and the relative retention time was investigated, and the contents of 6 components in different parts of 42 batches of Jiujie Tea were determined. There was no significant difference between the content determination results of the QAMS method and the ESM method. Therefore, the QAMS method can be used instead of the ESM method to determine the content of Jiujie Tea. The QAMS method can be used to calculate the contents of other components using one internal reference (isoraquinone), which saves the detection cost, increases the detection convenience, and can quickly analyze the content of Jiujie Tea.

[0050] Different parts of medicinal materials have different chemical compositions, contents and medicinal effects. The effective ingredients of plant medicinal materials are mostly derived from plant secondary metabolites. Age, harvest period and environmental factors will affect the effective ingredients of plants, and may also affect the accumulation of chemical components in different parts. This study shows that the contents of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, caffeic acid, isoflurane and rosmarinic acid in different medicinal parts of Jiujie Tea are quite different. The contents of chlorogenic acid and isoflurane are higher than those in leaves, and the contents of neochlorogenic acid, cryptochlorogenic acid, caffeic acid and rosemary in leaves are higher than those in stems. The average contents of each part of the aboveground part of Jiujie Tea are close to those in the stems, mainly because the weight proportion of the stems of the aboveground part of Jiujie Tea is much higher than that of the leaves. There are differences in the content trends of the aboveground parts, leaves and stems of some batches of Jiujie Tea, which may be related to the growth environment, age, harvest period and processing methods of the medicinal materials. This study established a quantitative analysis method combining the characteristic spectrum of Jiujie Tea with one test and multiple evaluations, which can be used for the process research and quality evaluation of Jiujie Tea and Jiujie Tea Chinese medicine preparations. Further qualitative and quantitative analysis of different medicinal parts of Jiujie Tea was carried out, which can provide data support for the rational use of Jiujie Tea medicinal resources.

[0051] The above description is only an embodiment of the present invention and is not intended to limit the present invention. For those skilled in the art, the present invention may have various modifications and variations. Any modification, equivalent substitution, improvement, etc. made within the spirit and principle of the present invention should be included in the scope of the claims of the present invention.

Claims

1. A method for evaluating the quality of different parts of Jiujie Tea based on the characteristic spectrum of Jiujie Tea combined with a one-test-multiple-evaluation method, characterized in that: The following steps are involved: Step 1, preparation of reference solution: taking neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, caffeic acid, rosmarinic acid, and isoflurane reference substances and adding methanol to mix and dissolve to form a reference solution; Step 2, preparation of the test solution: take the above-ground part of Camellia sinensis, place it in a stoppered conical flask, add methanol, heat it under reflux, cool it, and weigh it again, make up the lost weight with methanol, shake it well, filter it, and take the filtrate to obtain the test solution; Step 3, establishment of characteristic spectrum: take the test sample solutions of the aerial part, leaves and stems of Jiujie Tea, record the characteristic spectrum of each sample Jiujie Tea under liquid chromatography conditions, import the data into the "Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System", set the reference spectrum of the characteristic spectrum, automatically match it and then perform multi-point correction, perform full peak matching to generate a control overlay map, and then perform principal component analysis and orthogonal partial least squares discriminant analysis; Step 4, establishment of a one-test-multiple-evaluation method: Based on the results of principal component analysis and orthogonal partial least squares discriminant analysis, a one-test-multiple-evaluation quantitative characterization method was established with neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, caffeic acid, isoflurane, and rosmarinic acid as detection components.

2. The method for evaluating the quality of different parts of Jiujie Tea based on the Jiujie Tea characteristic spectrum combined with the one-test-multiple-evaluation method according to claim 1, characterized in that: Each 1 mL of the reference solution in step 1 contains 5 μg, 10 μg, 10 μg, 40 μg, 20 μg and 10 μg of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, caffeic acid, rosmarinic acid and isoflurane, respectively.

3. The method for evaluating the quality of different parts of Jiujie Tea based on the Jiujie Tea characteristic spectrum combined with the one-test-multiple-evaluation method according to claim 1, characterized in that: The preparation of the test solution in step 2 is as follows: take about 1.0 g of the aerial part of Camellia ternata, accurately weigh it, put it in a stoppered conical flask, accurately add 50 mL of 60% methanol, weigh it, heat and reflux for 0.5 hour, cool it, weigh it again, make up the lost weight with methanol, shake it well, filter it, and take the filtrate to obtain the solution.

4. The method for evaluating the quality of different parts of Jiujie Tea based on the Jiujie Tea characteristic spectrum combined with the one-test-multiple-evaluation method according to claim 1, characterized in that: In the liquid chromatography conditions in step 3, mobile phase A is acetonitrile and mobile phase B is 0.1% phosphoric acid solution.

5. The method for evaluating the quality of different parts of Jiujie Tea based on the characteristic spectrum of Jiujie Tea combined with the one-test-multiple-evaluation method according to claim 4, characterized in that: The gradient elution program in the liquid chromatography conditions in step 3 is: 0-2.5 min, 10% acetonitrile; 2.5-7 min, 10-16% acetonitrile; 7-14 min, 16-29% acetonitrile; 14-17 min, 29-30% acetonitrile.

6. The method for evaluating the quality of different parts of Jiujie Tea based on the Jiujie Tea characteristic spectrum combined with a one-test-multiple-evaluation method according to claim 5, characterized in that: The liquid chromatography conditions in step 3 are as follows: the detection wavelength is 330 nm; the column temperature is 25° C.; and the injection volume is 1 μL.

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