The Qualitative Analysis Of The Major Constituents Of Uncaria Rhynchophylla And Its Metabolism Study In Rats By LC-MS

Uncaria rhynchophylla,is a member of Rubiaceae family,with the dry hook-like stem and branch of Uncaria rhynchophylla(Miq.)Miq.Ex Havil.,Uncaria macrophylla Wall.,Uncaria hirsute Havil.,Uncaria sinensis(Oliv.)Havil.,or Uncaria sessilifructus Roxb.as its medicinal parts.It is widely used for relieving dizziness,arreting convulsion,clearing heat and calming the liver in clinic.Our research group found that Uncaria rhynchophylla and its related compound prescription have shown good activities on the animal pharmacological model of neural degenerative diseases such as Parkinson's disease,but the efficacy of materials are not yet clear.In this paper,the analysis and identification of the major chemical constituents,the best extraction method and the metabolic process in rats of Uncaria rhynchophylla extract were preliminarily studied by LC-Q/TOF-MS/MS and have achieved the following main results:Firstly,a rapid and effective LC-Q/TOF-MS/MS method used for chemical composition analysis of Uncaria rhynchophylla extract has been successfully established.The major peaks of total ion current chromatogram of have been detected and identified by retention time,accurate molecular weight,typical fragment characteristics,coupled with the reference compounds and the previous literatures.A total of 30 compounds were temporarily identified,including 21 indole alkaloids,six flavonoids and three organic acid compounds.Secondly,an effective quantitative method was established by LC-Q/TOF-MS/MS,of which Rhynchophylline and Isorhynchophylline were used as the indicators.And the extraction factors such as extraction solvent,extraction time,times and liquid ratio were investigated.It showed that the higher levels of extraction efficiency of Rhynchophylline and Isorhynchophylline would be achieved by the following conditions:70%ethanol as solvent,the liquid ratio of 10:1,extracting 1 h for 3 times and further extracting by ether.The last,the drug-containing serum and urine of rats after dosing Uncaria rhynchophylla extracts were analyzed by LC-Q/TOF-MS/MS method.A total of eight prototypes such as 18,19-dehydrocorynoxinic acid B(PI),Isorhynchophyllic acid(P2),18,19-dehydrocorynoxinic acid(P3),Rhynchophyllic acid(P4),Isocorynoxeine(P5),Isorhynchophylline(P6),Corynoxeine(P7)and Rhynchophylline(P8)as well as four metabolistes,Glucuronidation of 18,19-dehydrocorynoxinic acid B/18,19-dehydrocorynoxinic acid(Ml),Glucuronidation of Isorhynchophyllic acid/Rhynchophyllic acid(M2),Hydroxylation of Isocorynoxeine/Corynoxeine(M3)and Glucuronidation of Rhynchophylline/Isorhynchophylline(M4)were identified in the urine,while P5-P8 and M4 were were detected in the plasma.