Studies On Quality Evaluation Of Uncariae Ramulus Cum Uncis

Uncaria rhynchophylla(Miq.)Miq.ex Havil.(UR),U.macrophylla(Miq.)Miq.ex Havil.(UM),U.hirsuta Havil.(UH),U.sinensis(Oliv.)Havil.(USi)and U.sessilifructus Roxb.(USe)are five commonly used medicinal species in China.Uncariae Ramulus Cum Uncis(URCU),the dried hook-bearing branches of the five plants,is a well known herbal medicine mentioned in Chinese Pharmacopoeias,and has been used to treat hypertension,fever,headache,dizziness,stroke and bilious disorders.A syatematic studies on the chemical constituents of the five species and its qualitative and quantitative methods were carried out using modem methods for plant analysis.The main results were as follows.1.The characters and microstructure of the five species were carried by morphological identification,trait identification and microscopic identification.The origin identification and herbs mainstream varieties were examined and described.2.A total of the 19 oxindole alkaloids,16 indole alkaloids and 1 flavone were identified by co-chromatography of the sample extract with authentic standards,and comparing the retention time,characteristic molecular ions and fragment ions,or tentatively identified by MS/MS determination.The accurate mass measurement for all the protonated molecules and subsequent fragment ions offers higher quality structural information for the interpretation of the fragmentation pathways of the various groups of the alkaloids.3.An ultra performance liquid chromatography(UPLC)coupled with quadrupole time-of-flight mass spectrometry(Q-TOF/MS)method has been optimized and established for the rapid analysis of the alkaloids in 22 samples originated from the five Uncaria species.The URCU contains not less than 0.03 mg/g of rhynchophylline and hirsutine,and total alkaloids not less than 0.12 mg/g(rhynchophylline and hirsutine),calculated on the dried basis.4.The potential chemical markers were identified for the classification of the U.species samples by principal component analysis(PCA)and orthogonal partial least squared discriminant analysis(OPLS-DA).The top nine leading markers mentioned above were structurally identified as IRN,RIN,ICO,COR,CNX,iso-rhynchophylline,GME,HS and HST,respectively.The results demonstrate the similarity and differences in alkaloids among the five U.species,which is helpful for the standardization and quality control of the medical materials of the URCU.5.To determine total nitrogen of U.Rhynchophylla by Kjeldah method and study on the feasibility of evaluating the quality through total Nitrogen analysis.The best parameter of digestion condition was optimized by Uniform Distribution.Ultrasound digestion by microwave and inductively coupled plasma mass spectrometry(ICP-MS)were used to determine the contents of inorganic elements in the URCU.The Ni was related to the quantility of the alkaloids in the samples and helpfut to enrich the amount.TLC and HPLC methods are established for qualitative and quantitative analysis of alkaloids in URCU,respectively.The study is quick,simple and repeatable,helpful to evaluate the quantity of URCU.