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CN-121994946-A - Fingerprint detection method for monascus

CN121994946ACN 121994946 ACN121994946 ACN 121994946ACN-121994946-A

Abstract

The application discloses a fingerprint detection method of monascus. The fingerprint detection method is developed aiming at the monacolin compound in the monascus, and whether the monascus meeting the quality requirement is contained in the sample to be detected can be directly judged by comparing the similarity of the sample to be detected and the reference fingerprint, so that the detection efficiency is improved, and the detection cost is reduced.

Inventors

  • ZHOU HENG
  • MAO XIUHONG
  • CAO YIMIN
  • CHEN SUYUN
  • HU QING
  • FENG RUI
  • PAN HUIQIN
  • CHEN RUONAN
  • JI SHEN
  • WANG SHAOMIN
  • HUANG XIAOJING
  • LAN LAN

Assignees

  • 上海市食品药品检验研究院

Dates

Publication Date
20260508
Application Date
20241108

Claims (10)

  1. 1. The fingerprint detection method of the monascus is characterized by comprising the following steps of: Obtaining a fingerprint of a sample to be detected, comparing the fingerprint with a control fingerprint, and judging whether the sample to be detected contains red yeast rice or not according to the similarity between the fingerprint of the sample to be detected and the control fingerprint, wherein the control fingerprint is selected from figures 24, 25 and/or 26; the method for obtaining the fingerprint spectrum of the sample to be detected comprises the following steps: Extracting a sample to be detected by using an extracting agent to obtain a sample solution, wherein the extracting agent is a mixed solvent of acetonitrile and water in a volume ratio of 70:30, and Detecting the sample solution by using a liquid chromatography tandem triple quadrupole mass spectrometer to obtain a fingerprint of a sample to be detected, wherein the chromatographic conditions of the liquid chromatography tandem triple quadrupole mass spectrometer are as follows: aglient Poroshell EC-C18 chromatographic column; The mobile phase is acetonitrile as the organic phase and 0.1% formic acid solution as the mobile phase as the aqueous phase; The elution mode is gradient elution. The sample injection amount is 1-5 mu L; the flow rate was 0.4ml/min, and Column temperature is 20 ℃; wherein the gradient elution is performed as follows: Time (minutes) Organic phase (%) Aqueous phase (%) 0~10 5→35 95→65 10~18 35 65 18~20 35→40 65→60 20~35 40→45 60→55 35~50 45→55 55→45 50~60 55→75 45→25 60~70 75→95 25→5 70~70.1 95→5 5→95 The mass spectrum conditions of the liquid chromatography tandem triple quadrupole mass spectrum comprise selecting parent ions with charge-to-mass ratio of 283.0+ -1 and selecting child ions with charge-to-mass ratio of 223.0+ -1, and/or Selecting parent ion with charge-to-mass ratio of 285.0+ -1, and selecting child ion with charge-to-mass ratio of 225.0+ -1, and/or Selecting parent ions with charge-to-mass ratio of 287.0 +/-1, and selecting child ions with charge-to-mass ratio of 227.0+/-1; preferably, the similarity between the fingerprint of the sample to be measured and the reference fingerprint is determined by determining whether the fingerprint of the sample to be measured contains peaks as shown in fig. 24, 25 and/or 26, wherein in fig. 24, the retention time and peak area of each peak are as shown in the following table a: Table A Numbering device Retention time Peak area 1 5.426 1202 2 6.35 1747.185 3 7.019 8459.111 4 7.781 2165.741 5 7.943 7753.704 6 8.204 4982.037 7 9.197 2676.63 8 9.634 4060.63 9 9.807 1796.926 10 10.389 10850.04 11 10.934 25221.26 12 11.116 5039.111 13 11.482 2778.148 14 11.678 7681.371 15 11.843 2488.333 16 12.115 29261.74 17 12.45 8886.223 18 12.587 3076.778 19 12.875 3533.667 20 15.95 2184.259 21 17.46 6069.074 22 22.615 33118.262 23 35.961 9643.481 ; In the graph 25, the retention time and peak area of each peak are shown in the following table B: Table B ; In the graph 26, the retention time and peak area of each peak are shown in the following table C: Table C Numbering device Retention time Peak area 1 14.389 3614.178 2 15.002 114533.602 3 21.055 140844.797 4 25.458 3896.643 5 34.336 15633.64 6 41.785 10831.07 7 42.51 5761.75 8 46.203 49913.109 9 50.926 58177.609 10 59.11 342989 11 66.453 46497.18 。
  2. 2. The method according to claim 1, wherein the similarity between the fingerprint of the sample to be tested and the reference fingerprint is calculated by means of a traditional Chinese medicine chromatograph and fingerprint similarity evaluation software.
  3. 3. The method according to claim 2, wherein the sample to be tested or the preparation raw material thereof is considered to contain monascus according to the similarity between the fingerprint of the sample to be tested and the reference fingerprint of not less than 0.8 (preferably not less than 0.85; more preferably not less than 0.86; more preferably not less than 0.87; more preferably not less than 0.88; more preferably not less than 0.89; more preferably not less than 0.90).
  4. 4. The method according to claim 3, wherein when the mass spectrometry condition of the tandem triple quadrupole mass spectrometer is that a parent ion with a charge-to-mass ratio of 283.0+ -1 is selected and an ion with a charge-to-mass ratio of 223.0+ -1 is selected, the similarity between the obtained fingerprint and FIG. 24 is calculated, and whether the sample to be measured or the preparation raw material thereof contains red rice is determined according to the fingerprint of the sample to be measured and the similarity of FIG. 24.
  5. 5. The method according to claim 3, wherein when the mass spectrometry condition of the liquid chromatography tandem triple quadrupole mass spectrometry is that a parent ion with a charge-to-mass ratio of 285.0+ -1 is selected and an ion with a charge-to-mass ratio of 225.0+ -1 is selected, the similarity between the obtained fingerprint and fig. 25 is calculated, and whether the sample to be measured or the preparation raw material thereof contains red rice is judged according to the fingerprint of the sample to be measured and the similarity of fig. 25.
  6. 6. The method according to claim 3, wherein when the mass spectrometry condition of the liquid chromatography tandem triple quadrupole mass spectrometry is that a parent ion with a charge-to-mass ratio of 287.0 + -1 is selected and an ion with a charge-to-mass ratio of 227.0+ -1 is selected, the similarity between the obtained fingerprint to be measured and fig. 26 is calculated, and whether the sample to be measured or the preparation raw material thereof contains red rice is judged according to the fingerprint of the sample to be measured and the similarity of fig. 26.
  7. 7. The method of claim 3, wherein a parent ion having a charge-to-mass ratio of 283.0+ -1 is selected and a child ion having a charge-to-mass ratio of 223.0+ -1 is selected to obtain a first fingerprint of the sample to be measured, and Selecting parent ion with charge-to-mass ratio of 285.0+ -1, selecting child ion with charge-to-mass ratio of 225.0+ -1, obtaining second fingerprint of sample to be detected, and Selecting parent ions with charge-to-mass ratio of 287.0 +/-1, and selecting child ions with charge-to-mass ratio of 227.0+/-1, so as to obtain a third fingerprint of the sample to be detected; When the similarity between the first fingerprint of the sample to be measured and the graph 24 is not lower than 0.9, the similarity between the second fingerprint of the sample to be measured is not lower than 0.9, and the similarity between the third fingerprint of the sample to be measured and the graph 26 is not lower than 0.9, the sample to be measured or the preparation raw material thereof is considered to contain red yeast.
  8. 8. The method of claim 1, wherein the mass spectrometry conditions of the liquid chromatography tandem triple quadrupole mass spectrometry comprise: the declustering voltage is 180V, the collision capacity is 24eV, parent ions with charge-to-mass ratio of 283.0+/-1 are selected, and child ions with charge-to-mass ratio of 223.0+/-1 are selected; A declustering voltage of 180V, a collision capacity of 24eV, a parent ion with a charge-to-mass ratio of 285.0+ -1, and a child ion with a charge-to-mass ratio of 225.0+ -1, and/or The declustering voltage is 200V, the collision capacity is 22eV, the parent ion with the charge-to-mass ratio of 287.0 +/-1 is selected, and the child ion with the charge-to-mass ratio of 227.0+/-1 is selected.
  9. 9. The method of claim 8, wherein the mass spectrometry conditions of the liquid chromatography tandem triple quadrupole mass spectrometry comprise: Ion source, electrospray, positive ion scanning mode ESI (+); The monitoring mode is MRM mode, and the temperature of the ion source is 200-600 ℃.
  10. 10. The method according to any one of claims 1 to 9, wherein the sample to be tested is a traditional Chinese medicine or a health care product.

Description

Fingerprint detection method for monascus Technical Field The invention relates to the field of analysis and detection, in particular to a fingerprint detection method of monascus. Background Red rice originates from China, is called Danqu in ancient times, is also called red rice, and is a dry rice grain obtained by inoculating and fermenting rice with red aspergillus (Monascus spp.). Since the 20 th century 70 th Japanese scientist Yuanteng chapter separated from Monascus purpureus went for the first time, the hypolipidemic component monacolin K (lovastatin) was obtained, after lovastatin was developed as a hypolipidemic drug, monascus purpureus went as a research hotspot of pharmacy, and has higher economic value. However, it takes time and effort to identify the red rice or the ingredients derived from the red rice added in the traditional Chinese medicine or the health care product, which makes the quality control of the medicine or the health care product containing the red rice very difficult. The red rice contains abundant secondary metabolites, the development of a fingerprint detection method is very difficult, and no fingerprint detection method of red rice is developed at present. The monacolin compound is a main source of the hypolipidemic activity of the monascus, the monascus can produce a plurality of monacolin compounds, the content distribution of the compound is directly related to the hypolipidemic effect of the monascus, and if a fingerprint detection method based on monacolin in the monascus is developed, the method is not only beneficial to the identification and quality control of the monascus, but also beneficial to the comprehensive analysis and detection of the monascus Qu Zhongmo monacolin compound. Therefore, there is a need in the art to develop a fingerprint detection method for monascus. Disclosure of Invention The invention aims to provide a fingerprint spectrum detection method of monascus. In order to solve the technical problems, according to a first aspect of the present invention, there is provided a method for detecting a fingerprint of monascus, the method comprising the steps of: Obtaining a fingerprint of a sample to be detected, comparing the fingerprint with a control fingerprint, and judging whether the sample to be detected contains red yeast rice or not according to the similarity between the fingerprint of the sample to be detected and the control fingerprint, wherein the control fingerprint is selected from figures 24, 25 and/or 26; the method for obtaining the fingerprint spectrum of the sample to be detected comprises the following steps: Extracting a sample to be detected by using an extracting agent to obtain a sample solution, wherein the extracting agent is a mixed solvent of acetonitrile and water in a volume ratio of 70:30, and Detecting the sample solution by using a liquid chromatography tandem triple quadrupole mass spectrometer to obtain a fingerprint of a sample to be detected, wherein the chromatographic conditions of the liquid chromatography tandem triple quadrupole mass spectrometer are as follows: aglient Poroshell EC-C18 chromatographic column; The mobile phase is acetonitrile as the organic phase and 0.1% formic acid solution as the mobile phase as the aqueous phase; The elution mode is gradient elution. The sample injection amount is 1-5 mu L; the flow rate was 0.4ml/min, and Column temperature is 20 ℃; wherein the gradient elution is performed as follows: Time (minutes) Organic phase (%)Aqueous phase (%)0~105→3595→6510~18356518~2035→4065→6020~3540→4560→5535~5045→5555→4550~6055→7545→2560~7075→9525→570~70.195→55→95 The mass spectrum conditions of the liquid chromatography tandem triple quadrupole mass spectrum comprise selecting parent ions with charge-to-mass ratio of 283.0+ -1 and selecting child ions with charge-to-mass ratio of 223.0+ -1, and/or Selecting parent ion with charge-to-mass ratio of 285.0+ -1, and selecting child ion with charge-to-mass ratio of 225.0+ -1, and/or Parent ions with a charge-to-mass ratio of 287.0 + -1 are selected, and daughter ions with a charge-to-mass ratio of 227.0+ -1 are selected. In some preferred embodiments, the similarity between the fingerprint of the sample to be measured and the reference fingerprint is determined by determining whether the fingerprint of the sample to be measured contains peaks as shown in fig. 24, 25 and/or 26. In some preferred embodiments, the sample to be tested or its preparation is considered to contain monascus when the peak in the fingerprint of the sample to be tested matches the peak 80% (preferably 85%, more preferably 90%) before the peak area values in fig. 24, 25 and/or 26. In fig. 24, retention times and peak areas of the respective peaks are shown in the following table a: Table A Numbering deviceRetention timePeak area15.426120226.351747.185 37.0198459.111 47.7812165.741 57.9437753.704 68.2044982.037 79.1972676.63 89.6344060.63 99.8071796.926 1010.38910850.04 11