Study On Pharmacokinetics And Metabolites Of Magnolol In Goldfish

Magnolol is one of the main active ingredients of traditional Chinese medicine Magnolia officinalis.Studies have shown that the compound has anti-inflammatory,antibacterial and anti-tumor effects.It has recently been reported that it also has a good inhibitory effect on various fish pathogens such as Grass Carp Reovirus and Ichthyophthirius multifiliis.Because of the good biological activity of magnolol and its simple chemical structure,it has broad prospects for development as a new drug,and the development of new drugs can not be separated from metabolic kinetics,tissue distribution and Study of metabolites.However,at present,only mammals have reported the research on the metabolism kinetics and metabolites of magnolol.No reports have been made on aquatic animals,so the study of metabolic kinetics,tissue distribution and metabolites in aquatic animals is a necessary link for its further application in aquaculture.This study established and validated a high-efficiency liquid chromatography-mass spectrometry method for detecting magnolol in goldfish plasma and various tissues.This method was applied to the pharmacokinetics of goldfish after bathing,injection,and oral magnolol.And liver,kidney,intestinal tract,muscle,gill and other tissue distribution and metabolites,while using metabolic kinetics results to verify the inhibitory effect of goldfish oral honokiol on the infection of small melon insects,providing a basis for subsequent drug development of honoki.The results obtained are as follows:1.Chromatographic conditions and methodological verificationBy optimizing the honokiol extraction and extraction method and verifying the detection method,by verifying the extraction effect of different reagents,different ratios of reagents,different ratios of hydrochloric acid in the reagents,etc.,it was determined that 1 gram of tissue or 1 ml of serum was used in sample processing.When 5 m L of hydrochloric acid was added simultaneously with m L of methanol and ethyl acetate extractant(v/v7: 3),the recovery rate in each tissue reached more than 85%,and the precision within the day and between the days was less than 5%.The degradation of aquinol has no effect on the pharmacokinetic results.When methanol-water is used as the mobile phase gradient for HPLC detection,there is no interference before and after the peak time,indicating that the method established in this research can accurately detect magnolol..2.Pharmacokinetics of magnolol and its inhibitory effect on resistance to small melon insects(1)Utilizing the above-established liquid chromatography-mass spectrometry method for detection of magnolol in goldfish plasma and various tissues,bathing,oral,and single injections at concentrations of 1.5 mg/L,30 mg/Kg,and 30 mg/Kg,respectively.After the administration,the metabolism and tissue distribution of the goldfish were measured.The results showed that the magnolol metabolism in the three modes of administration conformed to the one-compartment open model;the magnolol absorption and metabolism in the bath administration Slow,the retention time in the blood reaches 96 hours,and the apparent volume of distribution in the blood is 44.456 L,indicating that honokiol is widely distributed in the blood of goldfish,and the concentration in each tissue is gills> kidney> liver> muscle> intestinal tract;injection Both the absorption and metabolism are rapid during administration,the content in the blood is low for 48 hours,and the apparent volume of distribution in the blood is 120.594 L,indicating that honokiol is widely distributed in the tissues of goldfish,and the concentration in each tissue The situation is liver> intestine> muscle> kidney,and the elimination in the liver is slow and easy to accumulate;the highest content in blood,the fastest elimination rate when administered orally,the retention time is 72 hours,and the apparent distribution volume in blood is 129.438 L Indicate thick Phenol goldfish widely distributed in various tissues,in each case the concentration of intestinal tissue> liver> muscle> kidney,elimination rate in various tissues in the body are not easy and rapid enrichment.(2)Validation of oral magnolol,it was found that the number of goldfish infected with small melon insects decreased after oral administration of 30 and 90 mg/Kg magnolol,and the number of small melon insects infected by each fish decreased significantly.The control group increased by 50% and 62.5%,respectively.3.Detection of metabolites in goldfishA study of magnolol metabolites revealed that eight metabolites were detected in goldfish,including three redox reaction products,three sulfate ester compounds,one glucuronic acid conjugate,and one glutamic acid conjugate.In summary,based on the established LC-MS method for the detection of magnolol in plasma and tissues of goldfish,this study determined the magnolol’s metabolic kinetics,tissue distribution,and metabolites,and identified magnolol The metabolic model conforms to the one-compartment open model,and it is easy to produce enrichment in the liver and kidneys,and a total of 8 metabolites were detected.The anti-inhibition effect of anti-small worm infection was verified.Oral magnolol could inhibit the small worm infection and increase infection of goldfish Survival rate.The above results provide an important basis for the further development of magnolol.