Isolation And Determination Of Anti-tumor Active Components From Scutellaria Barbata For Quality Control

Scutellaria barbata D.Don, belonging to the plant family Labiatae, has been traditionally used as a folk medicine in China for the treatment of digestive system cancer, hepatoma, lung cancer, breast cancer, chorioepithelioma, hepatitis, liver cirrhosis and other diseases. At present, summary about anti-tumor active constituences in S. barbata which have reported are still not overall. The quality control of S. barbata was so confined to scutellarin or total flavonoids, which hardly reflect the global quality of the herb, that may hold back the development exploitation of S. barbata. Therefore, in this paper, compositions and anti-tumor active components of S. barbata were extracted. A reasonable quality control method and HPLC fingerprint analysis method were established for this herb. What we did are following:1. Preliminary extraction of polarity sites and antitumor active fractions screeming of S.barbata. Five kilograms dried and minced herb of S.barbata was extracted with 75% ethanol. The extract was suspended in double distilled water, and then was extracted with petroleum, dichlormethane and ethyl acetate, respectively. By MTT assay in vitro, IC50 values show dichlormethane fraction get the best antitumor effect, and petroleum and ethyl acetate positions follow.2. GC-MS and LC-MS were used to analysis these three fractions, respectively. And seventeen components were identified from petroleum position, fifty one were identified from dichlormethane position and fifteen were identified from ethyl acetate position. Dissociation regularity of those chemicals was summaried as well. 3. Dichlormethane position and ethyl acetate position with good antitumor activities effect were chose to separate. Nine components were isolated and purified from this herbal medicine. Chemical structures of eight components were identified out by UV, IR, MS, NMR, but the other one were still not known. Seven chemicals which have been identified already, including p-coumaric acid, scutellarin, apigenin-5-O-β- glucopyranoside, luteolin, apigenin, baicalein, 4'-hydroxywogonin, were tested by MTT method with eight kinds of human tumor cells for the first time, and results show that all of these flavonoids own good antitumor effect in vitro.4. Stand up a quality control method of S. barbata. Six of these seven components own higher peak height in HPLC spectrums, which means these are main anti-tumor active compositions in S. barbata. Therefore, these seven anti-cancer active components were simultaneously determined as indexes for quality assessment of S. barbata by HPLC-UV method, which was used in the quality control successfully.5. A HPLC fingerprint analysis method was developed for the integrative quality evaluation of S. barbata. Twenty-six peaks in the HPLC spectrum were selected for characteristic peaks in HPLC spectrum of every sample. The similarity (>0.8) of ten samples was calculated by Fingerprint Similarity Calculation software. In short, the developed fingerprint analysis meets the requirements for global and systematical quality control of S. barbata.