Screen And Study On Anti-cancer Extractive Of Momordica Seed With A LC-MS Method

Momordica seed is the dried ripe seeds of Momordica cochinchinesis(Lour.)spreng. Single and compound recipe of it has been widely used clinically and has demonstrated significant effects. It is particularly worth mentioning is that as the effect of detumescence and wound treatment, Momordica seed is used in compound recipe of anti-tumor. Although Momordica seed has been used in anti-tumor compound, and occupies a central position in the prescription, the study of material basis is still blank, so looking for the active composition of anti-tumor in Momordica seed, confirming its material basis, are helpful to further evaluate efficacy, and explore its mechanism. However, Momordica seed contains a large number of components, which need to find a variety of quick and effective screening method. High Performance Liquid Chromatography combined with mass spectrometry (HPLC-MS) is a widely used, simply operational, easy automated and highly sensitive. It has very important significance that this technology has been employed into traditional Chinese medicine active component of screening and activity determination of natural products research.This project through the establishment of a new HPLC-MS method and its optimization and validation, carried out the method to explore the anti-tumor activity of Momordica seed extract. The method is economical and effective. It lays a good foundation for the further search for the active compounds and provides a good reference for screening of active natural products. Specific works are summarized as following:1. A novel LC-MS method has been established to achieve the detection and quantitative analysis for the polar molecule, ATP, by adding 5mM ammonium acetate in mobile phase. The method is rapid, simple, sensitive and reliable, and the LLOQ was 5μg/mL. ATP can be detected for a variety of environments.2. Based on LC-MS quantitative analysis of the ATP which is a substrate of hexokinase, the activity of hexokinase has been detected. The measure results matched the theoretical value; the activity of hexokinase inhibitor,3-bromo pyruvate also has been measured; extracts of Momordica seed (MBZA,MBZB, MBZC and MBZD) have been screened for the activity of hexokinase inhibition; the inhibiting activity of the MBZA which was obtain for the screening, has been measured and calculated. IC50 was 85μg/mL.3. Cell model of human gastric cancer cell strain, MKN-45 and a nude mouse model have been established, and MBZA cytotoxicity and inhibition rate have also been investigated. The results show that MBZA obtains a strong antitumor effect in vivo.