23 Folin–Ciocalteu reagent. They were mixed well and then allowed to stand 6 min before
1.25 ml of a 7 sodium carbonate solution was added. The mixture was diluted to 3 ml with deionized water. The color was developed for 90 minutes at room temperature
and the absorbance was measured at 760 nm using a spectrophotometer. The measurement was compared to a standard curve of prepared gallic acid solutions and
expressed as mg of gallic acid equivalents per litre extract.
6.3. Antioxidant capacity Sharma and Bhat 2009
A total of + 1 gram green grass jelly was extracted with 7 ml of methanol, homogenized, and centrifuged at 3000 rpm for 15 minutes, until the supernatants were
obtained. Supernatants were filtered. Then, 2 ml solution of DPPH 0.25 mM added, homogenized, and incubated at the dark room temperature for 30 minutes. The
absorbance then measured at 517 nm using a spectrophotometer. Ascorbic acid standard curve made with the preparation of such samples.
6.4. Dietary Fiber Asp. 1983
Sample preparation Samples were freeze dried. A total of 1.0 g of sample was added into the
erlenmeyer, 25 ml of sodium phosphate buffer pH 6.00 added and made into a suspension. Then 100 µL enzyme termamyl added, closed, and incubated at 80
o
C for 15 minutes with shaking incubator, then it was removed and cooled. Then conducted
adjusting the pH to 1.5 by addition of 4 N HCl. Samples then added with 1 ml enzyme pepsin 0.1 gml, incubated at 37°C, while agitated for 120 minutes.
pH adjustion was conducted to reach pH of 6.8 by addition of 4 N NaOH, then added 1 ml enzyme pancreatin 0.1 gml, closed, and incubated at 37°C for 120
minutes while agitated. pH was then adjusted to 4.5 by adding 4 N HCl, then filtered using Whatman filter paper No. 40 which has been dried in oven 105
o
C for 3 hours and obtained constant weight B. Filtering was conducted by using a vacuum pump and
flushing water destilata as much as 2 x 10 ml, so that residue and filtrate obtained. Determination of Insoluble Dietary Fibre
The residue obtained was washed with 2 x 10 ml of 78 ethanol and 2 x 10 ml of acetone pro analysis. The mixture solution of the residue dried at 105
o
C, until a constant weight for 3 hours C = constant weight after analysis and dried. Porcelain
bowl was heated in an oven at 105
o
C for 1 hour, cooled, and weighed D = weight of porcelain bowls. Filter paper and residue in the furnace incinerated at 500
o
C for 5 hours, cooled, and weighed E = weight after incineration.
Determination of Soluble Dietary Fibre The filtrate was added by 500 ml of 95 ethanol warm 60°C and
precipitated for 24 hours. The precipitate was filtered with filter paper which has dried in oven 105oC for 3 hours and has constant weight F = weight of filter paper, washed
24 with 2 x 10 ml of 78 ethanol and 2 x 10 ml of acetone. The precipitate was dried at a
temperature of 105°C for 3 hours, cooled in a desiccator, and weighed G = weight after analyzed and dried. Porcelain bowl was heated in an oven 105
o
C for 13 hours, cooled in a desiccator, and weighed H = weight of porcelain bowls, then filter paper
and residue incinerated at temperatures about 550°C for 5 hours, cooled in a desiccator, and weighed I = weight after incinerated.
Determination of Total Dietary Fibre Level of total dietary fiber was obtained by sum the value insoluble dietary fiber with
soluble dietary fiber. Blank was done without samples, the calculations are: 1. Insoluble dietary fiber IDF in dry base
IDF =
{CB ED} x 100 A
2. Soluble dietary fiber SDF in dry base SDF =
{GF IH} x 100 A
3. Total dietary fiber = IDF + SDF Where:
A = dry sample weight g
B, F = filter paper weight g C, G = filter paper and residue weight after dried g
D, H = porcelain bowl weight g E, I
= porcelain bowl and ash weight after incinerated g
6.5. Moisture Content AOAC 2000
