Gas-phase fingerprint spectrum of transverse meridian mat as well as construction method and application of gas-phase fingerprint spectrum

Abstract

The invention discloses a gas phase fingerprint spectrum of a transverse meridian mat. The gas phase fingerprint spectrum comprises a peak 1-5, and the retention time of the peak 1-5 is 16.89 + / -0.1 min, 17.36 + / -0.1 min, 17.89 + / -0.1 min, 19.37 + / -0.1 min and 19.6 + / -0.1 min in sequence; the construction method of the gas-phase fingerprint of the membranaceous beautyberry root comprises the following steps: crushing membranaceous beautyberry root to obtain membranaceous beautyberry root powder, adding water into the membranaceous beautyberry root powder, adding ethyl acetate, slightly boiling for 4 hours, naturally cooling to room temperature, standing for layering, and collecting an ethyl acetate layer as a test solution; carrying out liquid chromatography detection on the test solution; the method comprises the following steps: collecting 10 or more membranaceous beautyleaf root medicinal materials, carrying out detection operation according to the method, comparing obtained detection results, and selecting common characteristic peaks to obtain the gas-phase fingerprint spectrum of membranaceous beautyleaf root. The gas-phase fingerprint spectrum constructed by the method can be used for rapidly and accurately determining and identifying the membranaceous beautyberry root traditional Chinese medicinal material.

Description

Technical Field

[0001] The invention belongs to the technical field of detection of Hengjingxi Chinese medicinal materials, and particularly relates to a gas phase fingerprint spectrum of Hengjingxi and a construction method and application thereof. Background Art

[0002] Calophyllum membranaceum Gardn. et Champ, scientific name Calophyllum membranaceum Gardn. et Champ, is a plant belonging to the genus Calophyllum of the family Gardnaceae, commonly known as Calophyllum membranaceum. This plant mainly grows in Guangdong, Guangxi and other regions, and is a common Chinese herbal medicine. Both the root and leaves of Calophyllum membranaceum can be used as medicine, which has the functions of removing rheumatism, strengthening tendons and bones, promoting blood circulation and relieving pain.

[0003] Hengjingxi has certain similarities in appearance with other plant species of the same genus, such as Xiumaohonghouke and Honghouke. These plant species have similarities in leaf shape, surface characteristics, and powder microscopic characteristics, which makes it difficult to accurately identify Hengjingxi. In order to effectively identify Hengjingxi, researchers usually use thin layer chromatography. However, the stationary phase length of the thin layer chromatography plate is limited, which limits its separation ability. At the same time, since thin layer chromatography is an open system, it is easily affected by external factors (such as humidity and temperature), which in turn affects the accuracy of the determination. Therefore, it is still necessary to develop a simple, fast, reproducible and accurate detection method suitable for Hengjingxi. Summary of the invention

[0004] In view of the above-mentioned shortcomings, the present invention discloses a gas phase fingerprint spectrum of Hengjingxi and a construction method thereof. The gas phase fingerprint spectrum can be used to quickly and accurately determine and identify the traditional Chinese medicine of Hengjingxi.

[0005] The present invention is achieved by adopting the following technical solutions: A gas phase fingerprint spectrum of a horizontal meridian mat comprises peaks 1 to 5, wherein the retention times of the peaks 1 to 5 are 16.89±0.1 min, 17.36±0.1 min, 17.89±0.1 min, 19.37±0.1 min, and 19.6±0.1 min, respectively.

[0006] The method for constructing the gas phase fingerprint spectrum of the horizontal warp mat comprises the following steps: (1) Grind the horizontal warp mat and pass it through a 50-mesh sieve to obtain horizontal warp mat powder. Then, add water and ethyl acetate to the horizontal warp mat powder, and then boil it slightly for 4 hours. Then, cool it naturally to room temperature and stand it for stratification. Collect the ethyl acetate layer as the test solution; (2) The test solution obtained in step (1) was subjected to liquid chromatography detection. The conditions of the liquid chromatography detection were as follows: a ZB-264 chromatographic column was selected, the carrier gas flow rate was 1 mL / min, the split ratio was 25:1, the injection volume was 1 μL, the injection port temperature was 250°C, and the FID detector temperature was 250°C; the chromatographic column temperature was raised to an initial temperature of 80°C, maintained for 2 min, then raised to 250°C at a rate of 10°C / min, and then maintained for 6 min; (3) Collect more than 10 kinds of Hengjingxi medicinal materials and perform detection operations according to the methods described in steps (1) to (2), compare the obtained detection results, select the common characteristic peaks to obtain the gas phase fingerprint spectrum of Hengjingxi.

[0007] Furthermore, in the step (1), the mass ratio of the Hengjingxi powder to water is 20g:800mL, and the volume ratio of the Hengjingxi powder to ethyl acetate is 20g:2mL.

[0008] Furthermore, the temperature during the micro-boiling process in step (1) is controlled at 80-100°C.

[0009] Furthermore, the size of the ZB-264 chromatographic column in step (2) is 30 m×0.25 mm×1.40 μm. The ZB-264 chromatographic column is a chromatographic column produced by Phenomenex Corporation in the United States.

[0010] The gas phase fingerprint spectrum of the Hengjing Xi is used for the identification of the traditional Chinese medicine of the Hengjing Xi. Specifically, the medicinal material to be identified is tested according to the method in steps (1) to (2), and then the obtained gas chromatography results are compared with the gas phase fingerprint spectrum of the Hengjing Xi. If the peaks 1 to 5 are present, it can be preliminarily determined that the medicinal material to be tested is the Hengjing Xi.

[0011] Compared with the prior art, this technical solution has the following beneficial effects: The present invention adopts a gas chromatography method to construct a gas phase fingerprint spectrum of the Hengjingxi Chinese medicinal materials, and then detects the medicinal material samples according to the method of the present invention, and compares and analyzes the obtained results with the constructed gas phase fingerprint spectrum, which can quickly and accurately identify the authenticity and quality of the Hengjingxi medicinal materials, is conducive to screening out medicinal raw materials that do not meet the quality requirements, realizes the quality control of the Hengjingxi medicinal materials, and improves the safety and stability of the use of the Hengjingxi medicinal materials; and the method of the present invention is simple and convenient to operate, and the detection and analysis time is short, which is conducive to reducing work intensity and improving work efficiency. BRIEF DESCRIPTION OF THE DRAWINGS

[0012] Figure 1 It is a gas chromatogram of the No. 1 Hengjingxi medicinal material sample detected in Example 1.

[0013] Figure 2 It is a gas chromatogram of the No. 2 Hengjingxi medicinal material sample detected in Example 1.

[0014] Figure 3 It is a gas chromatogram of the No. 3 Hengjingxi medicinal material sample detected in Example 1.

[0015] Figure 4 It is a gas chromatogram of the No. 4 Hengjingxi medicinal material sample detected in Example 1.

[0016] Figure 5 It is a gas chromatogram of the No. 5 Hengjingxi medicinal material sample detected in Example 1.

[0017] Figure 6 It is a gas chromatogram of the No. 6 Hengjingxi medicinal material sample detected in Example 1.

[0018] Figure 7 It is a gas chromatogram of the No. 7 Hengjingxi medicinal material sample detected in Example 1.

[0019] Figure 8 It is a gas chromatogram of the No. 8 Hengjingxi medicinal material sample detected in Example 1.

[0020] Fig. 9 It is a gas chromatogram of the No. 9 Hengjingxi medicinal material sample detected in Example 1.

[0021] Fig.10 It is a gas chromatogram of the No. 10 Hengjingxi medicinal material sample detected in Example 1.

[0022] Fig.11 It is a gas chromatogram of the No. 11 Hengjingxi medicinal material sample detected in Example 1.

[0023] Fig.12 This is the gas chromatogram obtained when anhydrous ethanol solvent was used for detection in Experimental Example 1.

[0024] Fig.13 This is the gas chromatogram obtained when cyclohexane solvent was used for detection in Experimental Example 1.

[0025] Fig.14 This is the gas chromatogram obtained when petroleum ether solvent was used for detection in Experimental Example 1.

[0026] Fig.15 This is the gas chromatogram obtained when ethyl acetate solvent was used for detection in Experimental Example 1.

[0027] Fig.16 This is the gas chromatogram obtained when the HP-1 chromatographic column was used for detection in Experimental Example 2.

[0028] Fig.17This is the gas chromatogram obtained when using the ZB-624 chromatographic column for detection in Experimental Example 2.

[0029] Fig.18 This is the gas chromatogram obtained when the ZB-WAXplus column was used for detection in Experimental Example 2.

[0030] Fig.19 This is the gas chromatogram obtained during the blank control test in Experimental Example 3.

[0031] Fig. 20 This is the gas chromatogram obtained when parallel sample 1 was detected in Experimental Example 3.

[0032] Fig.21 This is the gas chromatogram obtained when parallel sample 2 was detected in Experimental Example 3.

[0033] Fig. 22 This is the gas chromatogram obtained when parallel sample 3 was detected in Experimental Example 3.

[0034] Fig.23 This is the gas chromatogram obtained when parallel sample 4 was detected in Experimental Example 3.

[0035] Fig.24 This is the gas chromatogram obtained when parallel sample 5 was detected in Experimental Example 3.

[0036] Fig.25 This is the gas chromatogram obtained when parallel sample 6 was detected in Experimental Example 3.

[0037] Fig.26 This is the gas chromatogram obtained when parallel sample 7 was detected in Experimental Example 3.

[0038] Fig. 27 This is the gas chromatogram obtained by experimenter 1 during the test in Experimental Example 4.

[0039] Fig.28 This is the gas chromatogram obtained by experimenter 2 during the test in Experimental Example 4.

[0040] Fig.29 This is the gas chromatogram obtained when the same Hengjingxi medicinal material sample was tested on the first day in Experimental Example 5.

[0041] Fig.30 This is the gas chromatogram obtained when the same Hengjingxi medicinal material sample was tested on the second day in Experimental Example 5.

[0042] Fig.31 This is the gas chromatogram obtained when the same Hengjingxi medicinal material sample was tested on the third day in Experimental Example 5.

[0043] Fig.32 This is the gas chromatogram obtained when the same Hengjingxi medicinal material sample was tested on the fourth day in Experimental Example 5.

[0044] Fig.33 This is the gas chromatogram obtained when the same Hengjingxi medicinal material sample was tested on the fifth day in Experimental Example 5. DETAILED DESCRIPTION

[0045] The present invention is further described by the following examples, but is not intended to be limiting of the present invention. Specific experimental conditions and methods not specified in the following examples are conventional methods commonly known to those skilled in the art.

[0046] Example 1: A vapor phase fingerprint of a horizontal meridian mat, comprising peaks 1 to 5, wherein the retention times of peaks 1 to 5 are 16.89±0.1 min, 17.36±0.1 min, 17.89±0.1 min, 19.37±0.1 min, and 19.6±0.1 min, respectively.

[0047] The method for constructing the gas phase fingerprint spectrum of the horizontal warp mat comprises the following steps: (1) Take the horizontal warp mat and grind it and pass it through a 50-mesh sieve to obtain the horizontal warp mat powder. Then, add water to the horizontal warp mat powder, then add ethyl acetate, and then slightly boil it for 4 hours. Then, naturally cool it to room temperature and stand it for stratification. Collect the ethyl acetate layer as the test solution; the temperature during the slight boiling process is controlled at 90±2°C; the mass ratio of the horizontal warp mat powder to the volume of water is 20g:800mL, and the mass ratio of the horizontal warp mat powder to the volume of ethyl acetate is 20g:2mL; (2) The test solution obtained in step (1) was subjected to liquid chromatography detection. The conditions of the liquid chromatography detection were as follows: a ZB-264 chromatographic column was selected, the carrier gas flow rate was 1 mL / min, the split ratio was 25:1, the injection volume was 1 μL, the injection port temperature was 250°C, and the FID detector temperature was 250°C; the chromatographic column temperature was raised to 80°C at the initial temperature, maintained for 2 min, then raised to 250°C at a rate of 10°C / min, and then maintained for 6 min; the size of the ZB-264 chromatographic column was 30 m×0.25 mm×1.40 μm; the detection instrument model was Agilent 7890A; (3) Collect 11 kinds of Hengjingxi herbs (see Table 1) and perform the detection operation according to the method described in steps (1) to (2). The obtained detection results (see Table 2 and Figures 1 to 11 ) were compared and the common characteristic peaks were selected to obtain the gas phase fingerprint spectrum of the horizontal mat.

[0048] The gas phase fingerprint spectrum of the Hengjing Xi described in this embodiment is used for the identification of the traditional Chinese medicine of the Hengjing Xi. Specifically, the medicinal material to be identified is tested according to the method in steps (1) to (2), and then the obtained gas chromatography results are compared with the gas phase fingerprint spectrum of the Hengjing Xi. If the peaks 1 to 5 are present, it can be preliminarily determined that the medicinal material to be tested is the Hengjing Xi.

[0049] Table 1 Collection of Chinese medicinal materials for different horizontal jing mats

[0050] Table 2 Retention time results of different horizontal Chinese medicinal materials (min)

[0051] Experimental Example 1: The same Hengjingxi medicinal material sample was tested according to Example 1 and the following three methods. The test results are shown in Tables 3 and Figures 12 to 15 .

[0052] Method 1: Take the Hengjingxi powder and crush it to obtain the Hengjingxi powder, add it into anhydrous ethanol, shake it for 30 minutes, ultrasonicate it for 30 minutes, filter it to obtain the filtrate as the test solution, and the mass ratio of the Hengjingxi powder to the volume of anhydrous ethanol is 1g:5mL; Method 2: Take the Hengjing mat and crush it to obtain the Hengjing mat powder, add cyclohexane, shake for 30 minutes, ultrasonicate for 30 minutes, filter and obtain the filtrate as the test solution, the mass ratio of the Hengjing mat powder to the volume ratio of cyclohexane is 1g:5mL; Method 3: Take the Hengjingxi mat and crush it to obtain the Hengjingxi mat powder, add petroleum ether, shake for 30 minutes, ultrasonicate for 30 minutes, filter and obtain the filtrate as the test solution, the mass ratio of the Hengjingxi mat powder to the volume ratio of petroleum ether is 1g:5mL; Table 3 Retention time results of different detection methods (min)

[0053] According to Table 3 and Figures 12 to 15 It can be seen from the results that only by detecting according to the method of the present invention can a good peak effect be obtained, thereby realizing the detection and identification of horizontal warp mats.

[0054] Experimental Example 2: The same sample of Hengjingxi medicinal material was tested according to the method described in step (1) and step (2) of Example 1, wherein different chromatographic columns were used for testing, and the chromatographic columns were HP-1 (30m×0.25mm×0.25μm), ZB-624 (30m×0.25mm×1.40μm), and ZB-WAXplus (30m×0.25mm×0.25μm). The test results are shown in Figures 16 to 18 .according to Figures 16 to 18The results show that the ZB-624 column selected in the present invention has better separation and no tailing of the peak.

[0055] Experimental Example 3: According to the method described in step (1) and step (2) of Example 1, the Hengjingxi medicinal material sample was tested and divided into 7 samples for parallel testing. At the same time, the test solution without the Hengjingxi medicinal material sample was used as a blank control for testing. The test results are shown in Tables 4 and Figures 19 to 26 .

[0056] Table 4 Parallel test results

[0057] According to Table 4 and Figures 19 to 26 The results show that the method of the present invention has good specificity, and the method is stable and has good reproducibility.

[0058] Experimental Example 4: Two experimenters were arranged to use two instruments of the same model to test the same Hengjingxi medicinal material sample according to the method described in step (1) and step (2) in Example 1. The test results are shown in Tables 5 and Figures 27-28 .

[0059] Table 5 Retention time results detected by different experimenters (min)

[0060] According to Table 5 and Figures 27-28 It can be seen from the results that the results of the tests performed by different experimenters according to the method of the present invention have good repeatability and accuracy.

[0061] Experimental Example 5: The same sample of Hengjingxi medicinal material was tested for 5 consecutive days according to the method described in steps (1) and (2) in Example 1. The test results are shown in Tables 6 and Figures 29 to 33 .

[0062] Table 6 Test results for 5 consecutive days

[0063] According to Table 6 and Figures 29 to 33 It can be seen from the results that the present invention has good stability.

[0064] In addition, it should be understood that although the present specification is described according to implementation modes, not every implementation mode contains only one independent technical solution. This description of the specification is only for the sake of clarity. Those skilled in the art should regard the specification as a whole. The technical solutions in each embodiment may also be appropriately combined to form other implementation modes that can be understood by those skilled in the art.

Claims

1. A gas phase fingerprint spectrum of a horizontal warp mat, characterized in that: The gas phase fingerprint spectrum of the horizontal mat includes peaks 1 to 5, and the retention times of the peaks 1 to 5 are 16.89±0.1 min, 17.36±0.1 min, 17.89±0.1 min, 19.37±0.1 min, and 19.6±0.1 min, respectively.

2. The method for constructing the gas phase fingerprint of the horizontal warp mat as claimed in claim 1, characterized in that: The following steps are involved: (1) Grind the horizontal warp mat and pass it through a 50-mesh sieve to obtain horizontal warp mat powder. Then, add water and ethyl acetate to the horizontal warp mat powder, and then boil it slightly for 4 hours. Then, cool it naturally to room temperature and stand it for stratification. Collect the ethyl acetate layer as the test solution; (2) The test solution obtained in step (1) was subjected to liquid chromatography detection. The conditions of the liquid chromatography detection were as follows: a ZB-264 chromatographic column was selected, the carrier gas flow rate was 1 mL / min, the split ratio was 25:1, the injection volume was 1 μL, the injection port temperature was 250°C, and the FID detector temperature was 250°C; the chromatographic column temperature was raised to an initial temperature of 80°C, maintained for 2 min, then raised to 250°C at a rate of 10°C / min, and then maintained for 6 min; (3) Collect more than 10 kinds of Hengjingxi medicinal materials and perform detection operations according to the methods described in steps (1) to (2), compare the obtained detection results, select the common characteristic peaks to obtain the gas phase fingerprint spectrum of Hengjingxi.

3. The method for constructing the gas phase fingerprint of the horizontal warp mat according to claim 2, characterized in that: In the step (1), the mass ratio of the horizontal warp mat powder to water is 20 g:800 mL, and the mass ratio of the horizontal warp mat powder to ethyl acetate is 20 g:2 mL.

4. The method for constructing the gas phase fingerprint of the horizontal warp mat according to claim 2, characterized in that: The temperature during the micro-boiling process in step (1) is controlled at 80-100°C.

5. The method for constructing the gas phase fingerprint of the horizontal warp mat according to claim 2, characterized in that: The size of the ZB-264 chromatographic column in step (2) is 30m×0.25mm×1.40μm.

6. The application of the gas phase fingerprint spectrum of the horizontal warp mat as claimed in claim 1, characterized in that: The gas phase fingerprint spectrum of the Hengjing Xi is used to identify the Chinese medicinal materials of the Hengjing Xi, specifically, the medicinal materials to be identified are tested according to the methods in steps (1) to (2), and then the obtained gas chromatography results are compared with the gas phase fingerprint spectrum of the Hengjing Xi. If the peaks 1 to 5 are present, it can be preliminarily determined that the medicinal material to be tested is the Hengjing Xi.

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