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EluChrom Isolation
Cholesterol Acetate, Cholesterol, Cortisone,
Dextrose, L-Phe, and I-Arg. in liver and fungal culture


Graph

Extraction
25mL EluChrom Cartridge: 1 gram of sample homogenized in 75% MeOH (20mL) and charged onto cartridge.

Effluent
Cartridge eluted with MeOH (12mL), then 15mL of 2-butanone. Each eluent is collected and chromatographed.

Chromatographic conditions
LKB system running a four-step Gradient - Solvents: A: heptane, B: Me-t-Bu-ether, C: 0.1% HCOOH in MeCN, D: 0.1% HCOOH in water. Flow rate: 0.9mL/min, Column: Diol 100, Detector: SEDEX 55 Evaporative Light Scattering.

Compound retention times
Cholesterol acetate 6.3 min., Cholesterol 14.8min., Cortisone 30.2 min., Dextrose 44.5-44.8 min., L-Phe 50.1 min., I-Arg 57.9 min.

This is a normal phase separation. The most polar compounds such as the amino acids, and sugars isolate first followed by the medium polarity compounds, some fats. Lastly, the non-polar compounds, cholesterol products and fats appear.

We acknowledge Laszlo Treiber, Ali Shafiee, Tom Holt, Pilar Hernandez and Juan Bautista Garcia, of Merck & Company for the contribution of this data presented at SamPREP.



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