This study evaluated the antioxidant activity, total phenolics, flavonoids, and micronutrient content of seven varieties of fenugreek seed extracts. Various antioxidant assays were performed, including DPPH radical scavenging, ferrous ion chelating ability, and determination of total phenols and flavonoids. PEB variety showed the highest DPPH radical scavenging activity and total phenol content, while Rmt-361 exhibited the highest ferrous ion chelating ability. Micronutrient analysis found copper, iron, magnesium, and zinc present in the extracts. The study provides information on the antioxidant and nutritional properties of different fenugreek varieties.

1. EVALUATION OF ANTIOXIDANT
ACTIVITY, TOTAL PHENOLICS
FLAVONOIDS &
MICRONUTRIENT CONTENT IN
SELECTED VARIETIES OF
FENUGREEK ( Trigonella
foenum-graecum L.) SEED
EXTRACT.
Nisha* & P.B. Rao.
Department of Biological Sciences,
C.B.S.H., G.B Pant University of
Agriculture & Technology ,Pantnagar-
263145, Uttarakhand, India

2. Introduction
 Antioxidants are our first line of defence against
free radical damage, and are critical for
maintaining optimum health and wellbeing.
‘Or’
 An antioxidant is a molecule capable of slowing
or preventing the oxidation of other molecules

3. antioxidants
-Nutrient derived
-Enzymatic
-Metal chelating

4. Material andmethod
Trigonella
foenum-graecum

5. Classification…
Bentham and Hooker (1862)
Class - Dicotyledons
Subclass - Polypetalae
Series - Calyciflorae
Order - Rosales
Family - Leguminosae
Subfamily - Papilionoideae (Faboideae)
Genus- Trigonella
Species- T foenum-graecum

6. Economic importance
 It has the various medicinal properties
like anticholestrolemic, anti-
inflammatory, antitumour, cardio tonic,
carminative, demulcent, diuretic,
emollient, expectorant, febrifuge,
galactogogue, hypoglycemic,
hypotensive and laxative have been
attributed to this plant in the
traditional system of Indian medicine.

7. Varieties of fenugreek…
 AFG-1 Ajmer Fenugreek
 Am-1- Azad methi
 GM2- Gujrat Methi
 HS- Hisar Sonali
 PEB- Pusa early bunching
 PR- Pant Ragini
 RMT-361 Rajasthan methi

8. Continue……
Antioxidants were evaluated by the following
methods.
DPPH assay (Brand & Williams et al .,1995)
Ferrous ion chelating activity (Hsu et al ., 2003)
Total phenols (Wolfe, 2003)
Total flavonoids (Ordon, 2006)
Statistical analysis

9. Methodology
The crude extract was stored, and used for further experimentation
The extract was filtered, the filtrate was evaporated in a water bath
The seed powder is extracted for 5 days in aqua-methanol (20 % and 80 %)
Seeds of different varieties are procured from different places and are grinded
to make fine powder

10. Radical scavenging activity by DPPH
assay
Different concentrations of extract (0.2 to 1.0 ml) was
taken into a test tube
To this 2.0 ml (0.04%) DPPH solution was added and
placed in dark for 30 min. the absorbance was
measured at 517 nm.
Flavonoids
To the 0.5 ml extract, 0.5 ml of 2 % Aluminium chloride was added,
absorbance was measured at 420 nm, after 1 hour incubation, against
control.
Yellow color indicate the presence of flavovoids.

11. Determination of total phenolic content
According to the method the Folin -Ciocalteau
To the extract (0.5 ml), 0.2 ml of FCR was added
After 5 min. 7 % Sodium bicarbonate solution was added
Absorbance was recorded at 765 nm after 1 hr incubation
Chelating
effect
To 1 ml of different concentrations (0.2-1.0 µg/ml) of each extract, 0.1 ml
(2mM) FeCl2.4H2O and 0.2 ml (0.5) Ferrozine were added.
The final volume make upto 3 ml, vortexed,
Absorbance was measured at 562 nm after 10 min of incubation.

12. Continue…..
 Micronutrient content by atomic
absorption spectrophotometer(AAS)

13. RESULTS

14. DPPH radical scavenging activity in different varieties of fenugreek
Concentration(µg/ml)
Varieties 0.2 0.4 0.6 0.8 1.0
PEB 52.67±0.68 64.16±0.24 78.93±0.75 88.81±1.20 93.29±0.79
HS 51.53±0.46 59.33±0.28 66.82±0.78 75.04±0.54 83.19±0.30
PR 45.13±0.51 57.18±1.06 67.87±0.72 78.74±0.46 85.97±0.51
Afg-1 31.57±0.62 41.92±0.68 51.49±1.09 65.46±1.05 74.73±0.50
Am-1 53.46±0.77 68.64±.041 82.83±0.43 85.62±0.52 93.82±0.13
GM-2 54.90±0.31 72.10±0.33 81.02±0.23 89.63±0.25 93.97±0.27
Rmt-361 33.39±0.18 43.62±0.22 49.76±0.01 57.89±0.13 67.10±0.15

15. Ferrous ion (Fe2+) chelating ability
Concentration(µg/ml)
Varieties 0.2 0.4 0.6 0.8 1.0
Am-1
74.40±0.030 73.17±0.010 72.79±0.012 72.94±015 72.16±00.24
Afg-1
75.87±0.015 74.78±0.021 73.63±0.023 72.68±0.08 71.70±0.028
HS 74.29±0.35 72.50±0.23 71.18±0.13 70.29±0.25 68.91±0.21
PR 75.46±0.21 73.74±0.23 70.14±0.10 69.14±0.33 68.33±0.10
GM-2 75.17±0.21 72.90±0.38 71.43±0.38 70.35±0.27 68.62±0.26
PEB 74.46±0.22 72.33±0.29 71.98±0.21 71.70±0.35 69.66±0.13
Rmt-361 72.04±0.28 73.34±0.10 74.46±0.15 76.90±0.10 78.80±0.25

16. Total Phenols in fenugreek varieties…..
Varieties Concentration (0.5 µg/ml)
PEB 3.17±0.48
HS 1.41±0.73
PR 1.94±0.40
Rmt-361 2.79±0.49
GM-2 2.84±0.59
AM-1 1.80± 0.09
AFG-1 1.75±0.07
Phenolic content Gallic acid equivalent

17. Flavonoids Content In Different Varieties Of
Fenugreek
Varieties Concentration (0.5µg/ml)
AM-1
1.36±0.17
Afg-1 1.24±0.25
HS 1.61±0.37
PR 1.52±0.17
GM-2 1.72±0.17
PEB 1.46 ±0.21
Rmt-361 1.26±0.45
Results indicated as Quercitin equivalent

18. Micronutrient content (µg/gm)
0
5
10
15
20
25
30
Cu Fe Mg Zn

19. Acknowledgement
University Grant Commission for financial
assistance.
19