Taxol, Vincristine, Vinblastine.pptx

Phytoconstituents

1. INDUSTRIAL PRODUCTION, ESTIMATION AND UTILIZATION OF PHYTOCONSTITUENTS Taxol, Vincristine & Vinblastine

2. T AXOL Biological Sourc e : Taxol is a diterpenoid obtained from the bark of Pacific Yew Tree, Taxus brevifolia , belonging to fam ily T axaceae . 2

3. T A X OL Production : Powdered bark extracted with methanol, filtered & evaporated 1 to dryness. 2 Partition ed with the mixture of carbon tetrachloride & water, filter & evaporated. 3 Dried CCl 4 fraction again extracted with CCl 4 : methanol, evaporate to obtain crude taxol . 3

4. HPLC Method Method: Isocratic Detector: Photodiode array detector Stationery Phase: C18 Column Mobile Phase: 0.02 M Potassium dihydrogen phosphate (buffer solution) in water (pH 4.5 adjusted with potassium hydroxide) and acetonitrile in the ratio of 60:40 v/v. Flow rate: 2ml/min. Run time: 8.0 min Detection: 230 nm. Estimation:

5. TAXOL Estimation: HPTLC method Mob phase- chloroform:methanol (7: 3 v/v ) Visualizing agent- vanillin sulphuric acid. Utilization: T r e atm e n t o f ova r i a n, l u n g , b l a d d e r , esophageal & other types of cancers. Antiproliferative agent. 5

6. VINCRISTINE & VINBLASTINE Biological Source: These are Indole alkaloid s obtained from the dried whole plant of Catharanthus roseus or Vinca rosea , belonging to family- Apocynaceae. 6

7. Production : Plant tissue culture technique. A two stage fermentation procedure was used. 500 ml Erlenmeyer flasks containing 100 ml medium (0.3%) malt extract, (1.0%) glucose, (0.3%) yeast extract and (0.5%) peptone) were inoculated with 7 days old culture and incubated at 28°C on a rotary shaker (240 rpm) for 4–5 days, which was used as seed culture. 10 ml seed culture was transferred to 500 ml Erlenmeyer flask containing 100 ml production medium called as vinca medium-1 (Glucose: 3%, Succinic acid: 1%, Sodium benzoate: 100 mg, Peptone: 1%, Magnesium sulphate: 3.6 mg, Biotin: 1 mg, Thiamine: 1 mg, Pyridoxal : 1 mg, Calcium pentothenate : 1 mg, Phosphate buffer: 1 ml (pH 6.8), L-Tryptophan: 0.1%, Geranium oil: 0.05%.) which were incubated at 28°C for 20 days as shake culture (II stage), after which it was harvested and used for further study. 8

8. Culture filtrates and mycelia were separated with the help of muslin cloth and then lyophilized. Lyophilized culture filtrate was extracted using ethyl acetate as a solvent system. The organic layer was separated from the aqueous layer using separating funnel. The extraction was repeated thrice and the solvent was dried using anhydrous sodium sulphate and concentrated under vacuum using rotavapour at 40°C in order to get crude extract. Estimation: HPLC : Mob phase- acetonitrile : 0.1 M phosphate buffer. Wavelength- 254nm.

9. Estimation: TLC: Sample: A small amount of crude extract was dissolved in ethyl acetate. Stationery Phase: silica gel-G (0.5 mm thickness) Mobile Phase: Chloroform∶methanol (8∶2) as a solvent system. Detection: Sprayed with ceric ammonium sulphate reagent. Vinca alkaloids spots produced brilliant violet color as well as purple color . Utilization: In chemotherapy regimens Childhood leukemia Immunosuppressant In Hodgkin’s disease