Extraction, Isolation And Acute Toxicity Of Iridoid Terpene Components From Spider - Like Macroporous Resin

Valeriana jatamansi Jones belongs to the genus Valeriana of the Valerianaceae family is a traditional Chinese medicine(TCM), which use the roots and rhizomes of the plant, distributed in the southwestern area of China. Valeriana jatamansi Jones, with facilitating pain relief and xiaoshi anti-diarrhea effect, is primarily employed in often abdominal distension painful, indigestion and diarrhea. Previous research of our group revealed that V. jatamansi can cure IBS by Brain-gut axis interactive.Objective:Based on the extraction and purification technics that have completed previously, for further analysis of the composition of iridoids in V. jatamansi and prepared to develop the extractive as a new drug, this thesis take the extractive as research objects, separate the chemical composition, content determination, stability and acute toxicity test. For developing a safe, effective, stability drug cure IBS lay material basis.Methods:1. The dry extract of Valeriana jatamansi Jones was acquired by reflux extraction in heating ethanol, and the chemical compositions were separated and purified by macroporous resin AB-8, silica gel, and sephadex LH-20. NMR spectral analysis were used to identify the constituents.2. Establish a HPLC method for determining 4 iridoids(11-Ethoxyviburtinal, Baldrinal,11-Methoxyviburtinal, Desacylbaldrinal) content from Valeriana jatamansi Jones’ macroporous resin eluate at same time. And 12 samples from different habitats were measured by the method.3. Affect factor of stability of 11-Ethoxyviburtinal, Baldrinal,11-Methoxyviburtinal, Desacylbaldrinal has been researched.4. For acute toxicity test, ICR male and female mice, was administered respectively of 11-Ethoxyviburtinal and Baldrinal according to the maximum dose.Results:1. Study on the chemical constituents of Valeriana jatamansi Jones’macroporous resin eluate.7 compounds were isolated and identified as 8-acetoxyl-patchouli alcohol,8-hydroxyl-patchouli alcohol, Jatamanin A, Desacylbaldrinal,11-Methoxyviburtinal, Baldrinal, 11-Ethoxyviburtinal.8-hydroxyl-patchouli alcohol were separated from Valeriana jatamansi Jones for the first time.2. A HPLC method for determining 4 iridoids contents from Valeriana jatamansi Jones’macroporous resin eluate was established. The results show that the method is stable and reliable and Valeriana jatamansi Jones from Yongping of Yunnan Province contains most of the amount of the 4 iridoids.3. Study on the affect factor of stability of 4 iridoids from Valeriana jatamansi Jones. The results show that the 4 iridoids is stabilized under the condition of high temperature and humidity.4. Acute toxicity test of iridoids from Valeriana jatamansi Jones. Compared with control group, the 11-Ethoxyviburtinal group and Baldrinal group did not appear poisoning and death obvious. Calculating the maximum tolerated dose multiples, the dose of 11-Ethoxyviburtinal and Baldrinal are equivalent to 456.6 and 1183.9 times clinical daily dose for adults. The results of acute toxicity test showed that 11-Ethoxyviburtinal and Baldrinal is safe and non-toxic in the range of dosage.Conclusion:Establish a method for determining 4 iridoids(11-Ethoxyviburtinal, Baldrinal, 11-Methoxyviburtinal, Desacylbaldrinal) content from Valeriana jatamansi Jones’ macroporous resin eluate by using HPLC for the first time, to provide an measuring indicatrix of quality control of Valeriana jatamansi Jones and process optimization. Using this method, measure changes in the 4 iridoids content of the extractive under the condition of high temperature and humidity. The results showed that the extractive is stable. The acute toxicity test of 11-Ethoxyviburtinal and Baldrinal was implemented for the first time, and the result showed that 11-Ethoxyviburtinal and Baldrinal is non-toxic. Iridoids from Valeriana jatamansi Jones is safe, stable and suitable for use in drug development.