Questioning And Arguable Research On Edible Hemerocallis Citrina Containing Colchicine

Belonging to Liliaceae family,Hemerocallis citrina Baroni（HCB）has a long history in China.It contains rich protein,carbohydrates,fat,calcium,iron,vitamin B,niacin and other nutrients.Several cases have been reported that having the effects of antidepressant,peaceful sleepinging and lactation promotion,HCB is an optimal raw material to develope functional foods.People often regard HCB containing colchicine as the main component of diarrhea and poisoning.However,in the pre-study,the presence of colchicine in HCB could not be detected.In order to accurately analyze whether edible HCB contains colchicine,in this thesis,we focus on the detection methods of colchicine in edible HCB,the determination of HCB samples from different sources and identification of metabolites in the biosynthetic pathway of colchicine.The main results are as following:1.We establish a highly efficient HPLC-DAD-Q/TOF MS method for colchicine detection in plants by applying and operating Electrospray ionization-（ESI）source in positive mode,Xaqua C18 chromatographic column with 0.1%formic acid-0.1%formic acid acetonitrile by gradient elution.In this method,colchicine in the range of 1⁵00ng/mL linear performs better with limit of detection 0.25 ng/mL,the recovery 97.10%,and the precision and stability RSD less than 5%.2.By using HPLC-DAD-Q/TOF MS to detect the contents of colchicine HCB in both HCB and autumnale samples of different origins,parts,and techniques of handling,the results show that colchicine can be obviously found in C.autumnale but not in 173 different origins HCB samples of their roots,scapes and leaves.3..By using HPLC-DAD-Q/TOF MS to test the intermediate metabolites of colchicine biosynthetic pathway（IMCBP）in HCB and C.autumnale samples,it shows that seven of which can be obviously found in C.autumnale but not be detected in HCB samples which proves no colchicine biosynthetic pathways in HCB samples.4.From liquid chromatography of HCB samples,there is an unknown peak very close to the retention time of colchicine reference material.By analyzing this peak,it can initially be recognized as the common flow part of various compounds.Among them,the major ion and mass ratio is m/z:455.1455(speculative molecular formula of C₂₃H₂₂N₂O₈)different from both the molecular formula of colchicine and its intermediate metabolites.The experiment indicates that the edible HCB includes no HCB colchicine at all and the traditional view is debatable that diarrhea caused by eating fresh HC is due to colchicine.