CN-118465106-B - Method for establishing monascus fingerprint by high performance liquid chromatography technology and application of monascus fingerprint in Shi Guogong medicinal liquor quality detection

Abstract

The invention discloses a method for establishing a monascus fingerprint by utilizing a high performance liquid chromatography technology and application of the method in Shi Guogong medicinal liquor quality detection, wherein each chromatographic peak of the monascus fingerprint obtained by utilizing the method is better in separation, stable in base line, good in peak type, and good in precision, repeatability, specificity and stability, 7 fingerprint peaks are confirmed in total by the established monascus fingerprint, and characteristic peaks of ring-opening lovastatin and lovastatin which are main chemical components in monascus are identified by comparison of reference substances and combination of ultraviolet absorption spectrum characteristics. The method can be used for quality detection of the monascus and provides scientific basis for improving the production process and quality control of the monascus. The method can also be applied to quality detection of the monascus component in Shi Guogong medicinal liquor, and has important significance for guaranteeing the quality of the monascus in Shi Guogong medicinal liquor.

Inventors

• CHEN CAN
• TAN RUNZE
• HE JINFANG
• ZHAO WEI
• BAI LU
• TAN XIPING
• LING YONGGEN

Assignees

• 株洲千金药业股份有限公司

Dates

Publication Date

20260508

Application Date

20240430

Claims (1)

1. 1. The method for detecting the quality of the male medicated wine of the Shinewguo is characterized by comprising the following steps of: Mixing lovastatin and acetonitrile to prepare a lovastatin-acetonitrile solution with the lovastatin concentration of 0.3-0.7 mg/ml, and adding a sodium hydroxide solution with the lovastatin-acetonitrile concentration of 0.15-0.25 mol/L to the lovastatin-acetonitrile solution to obtain a mixed solution, wherein the volume ratio of the lovastatin-acetonitrile solution to the sodium hydroxide solution is 1:10-20; After ultrasonic conversion is carried out on the mixed solution for 30-60 minutes, adding a phosphoric acid aqueous solution to adjust the pH value to 6.5-7.5 to obtain a solution, and diluting the solution by 1-3 times by using acetonitrile to obtain a reference solution; The sample solution is a methanol solution extracting solution of Shi Guogong medicinal liquor; HPLC chromatographic conditions are that a Agilent ZORBAX SB-Aq chromatographic column is adopted, a phosphoric acid aqueous solution with the volume percentage of 0.08-0.12% is taken as a mobile phase A, acetonitrile is taken as a mobile phase B, and the volume percentage of the mobile phase B in a mobile phase system is changed into: 0-26 min, and increasing the mobile phase B from 1% to 15%; 26-40 min, and increasing the mobile phase B from 15% to 50%; 40-58 min, and increasing the mobile phase B from 50% to 60%; 58-60 min, and the mobile phase B is reduced from 60% to 1%; the detection wavelength is a variable wavelength detection: 0-36 min, wherein the detection wavelength is 257 nm; 36-60 min, wherein the detection wavelength is 238 nm; performing HPLC detection on the reference object solution and the sample solution according to the HPLC chromatographic conditions to obtain fingerprint patterns of Shi Guogong medicinal liquor; The Shi Guogong medicinal liquor fingerprint contains 5 fingerprint peaks, the corresponding peak of the reference solution is taken as an S peak, the relative retention time of the 5 fingerprint peaks is respectively 0.528, 0.853, 1.000, 1.066 and 1.108,5 fingerprint peaks, and the relative retention time is within +/-10% of the specified value, so that the quality of the monascus component in the Shi Guogong medicinal liquor to be detected is qualified.

Description

Method for establishing monascus fingerprint by high performance liquid chromatography technology and application of monascus fingerprint in Shi Guogong medicinal liquor quality detection Technical Field The invention relates to the technical field of traditional Chinese medicine identification, in particular to a method for establishing a monascus fingerprint by utilizing a high performance liquid chromatography technology and application of the method in Shi Guogong medicinal liquor quality detection. Background Shi Guogong medicated liquor comprises rhizoma Polygonati Odorati, carapax Trionycis, rhizoma Atractylodis Macrocephalae, radix Achyranthis bidentatae, herba Taxilli, faeces Bombycis, rhizoma Chuanxiong, radix Saposhnikoviae, fructus Chaenomelis, radix Angelicae sinensis, flos Carthami, radix Glycyrrhizae, rhizoma Et radix Notopterygii, radix Angelicae Pubescentis, radix Dipsaci, colla cornus cervi, and red yeast rice. Has effects of dispelling pathogenic wind, removing dampness, promoting blood circulation, and dredging collaterals, and can be used for treating arthralgia due to wind-cold-dampness, bone joint pain, and numbness of limbs. The monascus contained in Shi Guogong medicinal liquor is a food and medicine double-effect fermentation product prepared by inoculating Aspergillus purpureus strain on rice (peeled kernel) and artificially culturing, has good lipid-lowering activity, is a raw material of natural statin lipid-lowering drugs such as blood lipid health and zhibituo, and is clinically used for treating hyperlipidemia, atherosclerosis and other diseases. Research shows that the main functional component of the red yeast rice lipid-lowering effect is lovastatin which is a secondary metabolite. Although the prior art discloses methods for measuring the content of lovastatin components in monascus by HPLC (Gao Wenming, lin Yuesong, dai Guoliang, zhao Lingang. HPLC method simultaneously measures the content of 3 components such as adenosine in monascus of different sources [ J ]. Pharmaceutical and clinical research, 2022,30 (2): 140-142), lovastatin has both acid form (closed loop) and lactone form (open loop), both of which exist in monascus at the same time. Therefore, the quality and efficacy of monascus cannot be comprehensively and accurately evaluated in the prior art. The efficacy and safety of Shi Guogong medicated wine depend on the quality and proportion of each component, and the quality and content of monascus as a key component directly influence the curative effect of Shi Guogong medicated wine. However, at present, there is no method for establishing a monascus fingerprint by adopting high performance liquid chromatography, mainly because compared with ultra-high performance liquid chromatography, the high performance liquid chromatography has challenges for detecting trace components in the monascus, and the complexity and uncertainty for establishing the fingerprint are increased. But the high performance liquid chromatography has the advantages of high separation efficiency and high analysis speed, and the instrument and the operation method of the high performance liquid chromatography are relatively simple and popular, so that the method for establishing the fingerprint is simple, convenient and efficient for the traditional Chinese medicine industry, and is beneficial to improving the quality control level of the traditional Chinese medicine and promoting the healthy development of the traditional Chinese medicine industry. Because the prior art cannot fully reflect the characteristics of various components in the monascus, such as the acid form and the lactone form of lovastatin and other potential active ingredients, the quality control and comparison between drug batches cannot be further carried out, and the consistency and stability of the monascus quality in the Shi Guogong medicinal liquor are further evaluated. Therefore, in order to further detect the quality of monascus in Shi Guogong medicinal liquor, control the overall medicinal effect of Shi Guogong medicinal liquor, reveal the interaction relation between each component, promote the quality control of Shi Guogong medicinal liquor, the field needs to establish a fingerprint of monascus in Shi Guogong medicinal liquor and be applied to the high performance liquid chromatography for Shi Guogong medicinal liquor detection. Disclosure of Invention The invention aims to overcome the defects of the prior art and provides a method for establishing a monascus fingerprint by utilizing a high performance liquid chromatography technology and application of the method in Shi Guogong medicinal liquor quality detection. The first object of the invention is to provide a method for establishing a monascus fingerprint by utilizing high performance liquid chromatography technology. The second object of the invention is to provide a method for establishing a red yeast rice contrast fingerprint. The third object of the in