Page 424 - e-Book

% yield of the extract = Weight of dried crude extract in grams X 100

Weight of plant sample powder in grams

Qualitative Phytochemical Analysis:

The Zanonia indica L. fruit hexane extract (ZFHE), Zanonia fruit chloroform extract (ZFCE),

Zanonia fruit acetone extract (ZFAE), Zanonia fruit methanol extract (ZFME) and Zanonia

fruit distilled water extract (ZFWE) were subjected to various qualitative tests to determine
phytochemical constituents [Madhura et.al] [Richardson et.al].

Quantitative Phytochemical Analysis:

The total alkaloids, total phenols, total flavonoids, total terpenes were analyzed using
high performance liquid chromatography (HPLC) (Waters model no. 486; Waters Corp.,

Milford, MA, USA) on C18 column (symmetry, 4·6mm×250mm) in isocratic mode. The

mobile phase methanol and water was used in the ratio 7:3 with the RP-HPLC C-18 column at
a flow rate of 1mL/min. before using, solvents were filtered with membrane filter and then

sonicated for 15min [Caroline et.al].

HPLC analysis:

The caffein, gallic acid, rutin, tannic acid was used as standard for alkaloids, phenols,

flavonoids and terpenoids respectively with the concentration 0.1mg/mL. Sample (1mg/mL)
were dissolved in mobile phase and 20µL was injected and the elution was monitored at

230nm, 254nm, 272nm, 210nm respectively [Madhura et.al].

The amount of total alkaloids, total phenols, total flavonoids and total terpenoids present

in the sample was estimated using the formula,

Sample area x standard amount x dilution x Mean weight Standard area dilution of standard
sample amount
Anti-Oxidant Activity:

The antioxidant properties of the fruit extracts were determined using DPPH, ABTS
radical scavenging assay and FRAP assay method.

Determination of DPPH free radical-scavenging activity:

The potentiality of extracts to scavenge 1-1 -diphenyl - 2- picrylhydrazyl radicals were
determined by preparing Various concentrations of fruit extracts of the sample with methanol

414