Protein estimation was done according to Lowry, Rosebrough,
Farr, and Randall (1951). A modified anthrone method of total
carbohydrate determination was followed for simple sugar estimation
(Sadasivam & Manickam, 2007). In the modified method,
0.1 mL of the coconut water was taken and the volume was made
up to 1 mL with distilled water and anthrone reagent (200 mg
anthrone in 100 mL of 95 g/L sulphuric acid) was added. Absorbance
for sugar and protein was observed at 630 nm and 660 nm
respectively in a spectrophotometer (Cecil Aquarius 7400,
England).