Separation and purification of glucosinolates from crude plant homogenates by high-speed counter-current chromatography.
Glucosinolates are anionic, hydrophilic plant secondary metabolites which are of particular interest due to their role in the prevention of cancer and other chronic and degenerative diseases. The separation and purification of glucosinolates from a variety of plant sources (e.g. seeds of broccoli, arugula and the horseradish tree), was achieved using high-speed counter-current chromatography (HSCCC). A high-salt, highly polar system containing 1-propanol-acetonitrile-saturated aqueous ammonium sulfate-water (1:0.5:1.2:1), was run on a semi-preparative scale and then transferred directly to preparative scale. Up to 7 g of a concentrated methanolic syrup containing about 10% glucosinolates was loaded on an 850-ml HSCCC column, and good separation and recovery were demonstrated for 4-methylsulfinylbutyl, 3-methylsulfinylpropyl, 4-methylthiobutyl, 2-propenyl and 4-(rhamnopyranosyloxy)benzyl glucosinolates. Multiple injections (5 to 6 times) were performed with well-preserved liquid stationary phase under centrifugal force. Pooled sequential runs with broccoli seed extract yielded about 20 g of its predominant glucosinolate, glucoraphanin, which was produced at > 95% purity and reduced to powdered form.
Fahey
Jed W
JW
Lewis B. and Dorothy Cullman Cancer Chemoprotection Center, Department of Pharmacology and Molecular Sciences, School of Medicine, Johns Hopkins University, 406 WBSB, 725 North Wolfe Street, Baltimore, MD 21205-2185, USA. jfahey@jhmi.edu
Wade
Kristina L
KL
Stephenson
Katherine K
KK
Chou
F Edward
FE
D001937:Brassica / Q000737:chemistry; D019607:Brassicaceae / Q000737:chemistry; D003377:Countercurrent Distribution / Q000379:methods*; D005961:Glucosinolates / Q000302:isolation & purification*; D029588:Moringa oleifera / Q000737:chemistry; D010936:Plant Extracts / Q000737:chemistry*; D012639:Seeds / Q000737:chemistry
D005961:Glucosinolates; D010936:Plant Extracts