Methodology And Stability Of Hypericin Liquid Phase Analysis

Hypericin is a naturally occurring bioactive substance,which has attracted much attention due to its extensive pharmacological effects and has a broad application prospect in the prevention and treatment of cancer.At present,the main source of hypericin is extracted from Hypericum perforatum.Hypericin content in the body is low,and the content of Hypericin in Hypericum perforatum is less than 0.3%.The high cost of preparation of hypericin with high purity,and the harsh conditions of preservation and use have hindered the industrial production and drug research of hypericin.At present,a large number of studies have been carried out on the extraction and detection methods,pharmacology and clinic of hypericin,and many effective detection methods have also been reported.In this study,based on the establishment of hypericin HPLC detection method,further methodological verification was carried out.After screening conditions of hypericin HPLC detection method,a complete detection method was obtained.Methodological verification was carried out according to the relevant provisions of pharmacopoeia and quality management.Validation items include system adaptability,specificity,limits of detection,limits of quantification,linearity and range,precision(repeatability),accuracy,solution stability,and method durability.Verification results showed that the limit of detection and limit of quantification of hypericin in the current detection method were 5.2 and 10.4 mg/L,respectively,and the linear relationship between 10.4 and 124.8 mg/L was good(R~2=0.99927).The results of other verification items also proved the feasibility and adaptability of the current detection method.To provide reliable data for new drug application of hypericin.Hypericin is unstable in light,heat and oxygen.The factors affecting the stability of hypericin extraction and use were selected to conduct ultrasonic,temperature,microwave,solvent system,medium and long term preservation and degradation experiments.Ultrasound has little effect on the stability of hypericin within 120 min,which can be used as a reliable means to accelerate the dissolution of hypericin.Hypericin is sensitive to temperature,and the optimal storage temperature of hypericin solution is-20?.Hypericin solution can be stored under 60?for 48 h under short-term seal and protected from light,while hypericin is unstable above 60?.Hypericin was the most stable at 550 W power and could be used as an auxiliary extraction method.For medium and long term stability,the results of high temperature,low temperature and strong light within 10 days all showed that hypericin solution was not stable.Through the investigation of the above factors,a comprehensive evaluation result of stability factors was obtained,which provided a reference for the preparation method and follow-up preparation of hypericin.Using the optical properties of hypericin,the polymerization of hypericin under water condition was preliminarily.the degree of polymerization of hypericin was positively correlated with water content,the higher the water content,the more polymer was produced,and there was almost no relationship between the polymerization speed and the residence time in water environment.However,there is a strong correlation between the size of the polymer and the residence time,the longer the residence time,the larger the solid particles of the polymer,and the solid particles can be completely filtered through 0.2?m filter paper,which is a simple and efficient method to separate the polymer.The study of hypericin polymer provided an important reference for the optimization of extraction conditions of hypericin and the study of water-solubility and stability of downstream products.