Toxicity And Pharmacokinetic Of Diester Alkaloids For Multiple Dose In Fuzi

ObjectiveRecent years,there are many reports about toxicity or adverse reactions in clinical when patients are taking Fuzi and Fuzilizhong pills(FLP).The toxic signs are various and occur mainly in the nervous system and heart.In this paper,we aim to study the toxic effects of Fuzi and FLP in rats,and find out the effect of key substances on neurotransmitters in certral nervous system.Then the pharmacokinetic and tissue distribution for the active ingredients of three diester-type alkaloids(aconitine(AC),Hypaconitine(HC)and Mesaconitine(MC))in Fuzi and FLP are also investigated.Methods1 The toxicity of Fuzi and FLP.(1)Toxicity:the acute toxicity evaluation of ether extract of Fuzi;the evaluation of acute and long-term toxicity of FLP.(2)To determine the influence of Fuzi and FLP to rat heart rate and locomotor activity;Determination for amino acids and acetylcholine neurotransmitters in rat,s brain by HPLC-RF after oral administration of Fuzi and FLP.2 LC-MS/MS method for determination of three diester alkaloids in biological samplesTo devolop a sensitive and reliable LC-MS/MS methods for determination of three diester alkaloids in biological samples(plasma and tissue),and Evaluate the methods of specificity,sensitivity,linear range,recovery,precision,matrix effects,stability etc.3 The pharmacokinetic of three diester alkaloids in Fuzi and FLP.To study the pharmacokinetic of three diester alkaloids after a single dose Fuzi and FLP in rats.To detect the changes of the body' s diester-type alkaloids at different time,after repeated dose of the Fuzi and FLP,according to the single-dose half-life obtained.4 To study the tissue distribution of Fuzi in rats.SD rats were administered crude extract of Fuzi and prepared extract Fuzi,and the rats were killed in different time,access to the heart,liver,spleen,lung,kidney,brain,stomach,duodenum,and testis quickly,then homogenate and the supernament were measured by LC-MS/MS.Results1.FLP has no acute toxic effects,but cause some influences to liver and kidney function after the long-term administration.Treatments with Fuzi extract can significantly increase the release of glutamate,?-aminobutyric acid and acetylcholine in the brain tissue and not only reduce activity of acetyl cholinesterase,but also reduce activity of the rats' heart rate and locomotors.Moreover,it can reduce the release of aspartate,glutamate and gamma-aminobutyric acid and activity of acetylcholinesterase in brain tissue after administration of FLP at the dose of 15 g/kg.2.While a single dose of crude Fuzi and prepared Fuzi are taken,diester-type alkaloids showed a double peaks phenomenon in rats,the three alkaloid concentration were lower at different time;After equal interval and multiple dose,the drug can accumulate in the body,reach steady state after about 6-8 time internal,but the concentration maintain still low;The mean residence time of three diester-type alkaloids of prepared Fuzi is 2-fold than that of crude Fuzi.In FLP,three diester-type alkaloids were below the detection limit after administration of a single dose,but hypaconitine can be detected after equal interval and multiple doses,with the increase of the frequency of administration while elevated.3.Crude Fuzi and prepared Fuzi can quickly distribute to the organizations after administration.The three alkaloids are mainly distributed in the gastrointestinal,liver,spleen,and lung.There are slight differences between two Fuzi.The percentage of the three alkaloids in the liver,spleen,and lung was significantly higher after prepared aconite root is taken.ConclusionIn this paper,the analysis method for determination of three diester-type alkaloids by LC-MS/MS analysis method which meets the requirements of the determination of biological samples is developed.It can be applied to the study of related pharmacokinetics and tissue distribution.Diester-type alkaloids can be quickly distributed to the appropriate organizations and toxic accumulated within the organization after Fuzi is taken,resulting in the lower blood concentration in rats.In addition,the drugs are show double peaks phenomenon after Fuzi administration,which may be caused by type three kind of diester alkaloids in the high levels of stomach and duodenum released into the blood resulting in observation of the blood medicine bimodal curve,then again distributed back to various tissues and organs,and ultimately leads those chemicals to show double peaks phase in plasma and various tissues.