The moisture content in dried C. nutans was gravimetrically
determined by treatment in an oven at 105 C for 24 h. Prior to the
extraction, the dried samples were grounded and sieved into
particles sizes ranging from 100 to 500mm. The prepared samples
were stored in air tight bags, swept with nitrogen to remove air and
kept in a refrigerator (8 C) until used in the extraction. Ethanol
(96%) was diluted by acidified water (pH 1 by sulfuric acid) into
several proportions, i.e., 44, 50, 65 and 86 %vol. Distilled water type
2 was boiled prior to the dilution to remove dissolved gasses.