Calendula Officinalis Extract Chemical Transformation And Biotransformation Preparation Of Lutein

The hydrolyzing lutein esters in the extraction of Marigold flowers to obtain lutein was studied in the paper. The research include six aspects as following.(1) The reviewing of the research on lutein in the world. Reviewed the reports about lutin and lutein esters in the past twentyyears and summarized the method of analysis, extraction, separation, transformation and purification of lutein and lutein esters.(2) Studying method of HPLC determining lutein and lutein esters synchronously.Lutein and lutein esters in marigold flowers was quantitatively determined by high performance chromatography (HPLC) with ODS-C18 column. A mixture of CH₃CN-CH₃OH -CH₃COOCH₂CH₃(volume ratio 55:1:44) was used as mobile phase at a flow rate of 1.0mL/min and detection was carried out at 460nm , the column temperature was about 25℃.(3) Studying the chemical method in transforming lutein esters to lutein.Effect of solvent variety, KOH concentration, reaction time and reaction temperature on the saponification were studied. The optimal reaction condition was adding 10.0g lutein esters into 50mL saturated KOH solution, reacting under room temperature for 4 hours.(4) Studying the method of separation and purification of lutein Using 5℃ cold water, the filter remainder of saponified washed until the remainder was neutral. Collecting the water and adjusting it to neuter, filtering the water and drying the filter remainder we can obtain luteinproduct with the purify more than 22.0%.Dried the neutral remainder and used 20mL hexane washed it twice. Then using CH2C12 dissolved the remainder and obtained supersaturation solution. Added the hexane into the solution slowly until deposition appeared, then placed it in to icebox with 5°C. After about 12 hours, added 0.4 times of CH2C12 hexane in the solution again, in about 12 hours filtered the solution, drying the remainder we can obtain lutein product with the purity more than 90.0%.(5) Screening of bacterium for lutein esters biotransforming Studied on the biotransformation of lutein esters by Penicillium ontabulate revulsive culture medium. Porule of and Aspergillus niger Penicillium could transform lutein esters into lutein, but biotransformation of Penicillium was better than Aspergillus niger. Porule of Pleurotus ostreatus could not grow on revulsive culture medium, but Pleurotus ostreatus could grow on the culture medium and transformed lutein esters into lutein.(6) Research of Shaking flash fermentation of Penicillium for trans -forming lutein esters.When pH is 6.0 , quantity of incubation is 2 bacteria plugs and fermentation time is 24 hours ,the transformation is completed and the content of lutein is 91.6%(HPLC)O Prolonging the transformation time will improve the percent of lutein but the content of lutein decreases.