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Chemical and Phytochemical Composition of Ashwagandha (Withania somnifera L.) Roots

January 2017
Asian Journal of Chemistry 29(8):1683-1686

DOI:10.14233/ajchem.2017.20536

Authors:

Susheel Gulati

Hindustan Petroleum Corporation Limited

V K Madan

Sushila Singh

Chaudhary Charan Singh Haryana Agricultural University

Isha Singh

George Brown College

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The present study was undertaken for estimation of chemical and phytochemical composition of ashwagandha (Withania somnifera L.) roots of promising genotypes viz. HWS-08-14, HWS-08-18, HWS-1228, HWS-1229 and selection-2B and varieties JA-20 and RVA-100. Chemical parameters viz. crude fibre, minerals (Fe, Cu, Zn, Mn), starch, total sugars, reducing sugars, non-reducing sugars were analyzed in ashwagandha roots. Crude fibre content ranged from 17.4 to 37.3 %, Fe content ranged from 595.83 to 983.33 ppm, Cu content ranged from 8.40 to 13.72 ppm, Zn content ranged from 16.33 to 41.00 ppm and Mn content ranged from 16.80 to 33.46 ppm. Starch, total sugars, reducing sugars and non-reducing sugars contents ranged from 7.61 to 8.22 mg/g, 5.33 to 6.89 mg/g, 0.40 to 0.64 mg/g and 4.87 to 6.33 mg/g, respectively. Phytochemical parameters viz. total alkaloids and tannins were also analyzed. Total alkaloids and tannins content ranged from 0.26 to 0.31 % and 0.66 to 0.84 mg/g, respectively.

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ASIAN JOURNAL OF CHEMISTRY

https://doi.org/10.14233/ajchem.2017.20536

INTRODUCTION

Since time immemorial, medicinal plants are of great

importance to health of individuals and communities. The

medicinal plant products, which are derived from plant parts

such as stem, bark, leaves, fruits and seeds have been part of

phytomedicine that produce a definite physiological action on

human body. The most important of these natural bioactive

constituents of plants are alkaloids, tannins, flavonoids and

phenolic compounds [1].

Among various medicinal plants, Withania somnifera is a

popular Indian medicinal plant belonging to family Solanaceae

and is also known as Ashwagandha, Indian ginseng and Winter

cherry. It is one of the most valuable plants in the traditional

Indian systems of medicine [2]. The roots are main part of the

plant that are widely used as therapeutic agents [3]. The roots

are reported to contain alkaloids, amino acids, steroids, volatile

oil, starch, reducing sugars, glycosides [4]. Ashwagandha roots

contain crude fibre 21.0 to 25.0 %, starch 6.09 to 9.46 mg/g,

tannins 0.39 to 0.82 mg/g, minerals K, Mn, Na, Fe, Zn, Cu,

Al, Ca, Cd & Ni, total sugars 2.52 to 9.52 mg/g, reducing

sugars 0.15 to 2.10 mg/g and non-reducing sugars 2.37 to 7.62

mg/g [5].

Survey of literature reveals that various field studies are

being carried out on promising genotypes of ashwagandha

but very scare information is available on chemical and phyto-

Chemical and Phytochemical Composition of Ashwagandha (Withania somnifera L.) Roots

SUSHEEL GULATI1, V. K. MADAN1,*, SUSHILA SINGH2, ISHA SINGH1 and DUSYANT1

1Medicinal, Aromatic and Potential Crops Section, Old IATTE Building, CCS Haryana Agricultural University, Hisar-125 004, India

2Department of Chemistry & Biochemistry, CCS Haryana Agricultural University, Hisar-125 004, India

*Corresponding author: E-mail: vikku60@gmail.com

Received: 23 January 2017; Accepted: 19 May 2017; Published online: 12 June 2017; AJC-18420

The present study was undertaken for estimation of chemical and phytochemical composition of ashwagandha (Withania somnifera L.)

roots of promising genotypes viz. HWS-08-14, HWS-08-18, HWS-1228, HWS-1229 and selection-2B & varieties JA-20 and RVA-100.

Chemical parameters viz. crude fibre, minerals (Fe, Cu, Zn, Mn), starch, total sugars, reducing sugars, non-reducing sugars were analyzed

in ashwagandha roots. Crude fibre content ranged from 17.4 to 37.3 %, Fe content ranged from 595.83 to 983.33 ppm, Cu content ranged

from 8.40 to 13.72 ppm, Zn content ranged from 16.33 to 41.00 ppm and Mn content ranged from 16.80 to 33.46 ppm. Starch, total

sugars, reducing sugars and non-reducing sugars contents ranged from 7.61 to 8.22 mg/g, 5.33 to 6.89 mg/g, 0.40 to 0.64 mg/g and 4.87

to 6.33 mg/g, respectively. Phytochemical parameters viz. total alkaloids and tannins were also analyzed. Total alkaloids and tannins

content ranged from 0.26 to 0.31 % and 0.66 to 0.84 mg/g, respectively.

Keywords: Crude fibre, Minerals, Starch, Total sugars, Reducing sugars, Total alkaloids, Tannins.

Asian Journal of Chemistry; Vol. 29, No. 8 (2017), 1683-1686

chemical composition of ashwagandha roots. Therefore, the

objective of present study was to estimate chemical parameters

viz. crude fibre, minerals (Fe, Cu, Zn, Mn), starch, total sugars,

reducing sugars, non-reducing sugars and phytochemical

parameters viz. total alkaloids, tannins in the promising

genotypes of ashwagandha roots.

EXPERIMENTAL

Ashwagandha (Withania somnifera L.) roots samples of

promising genotypes (HWS-08-14, HWS-08-18, HWS-1228,

HWS-1229 and Selection-2B) and two varieties (JA-20 and

RVA-100) were procured from the experimental area of Medi-

cinal, Aromatic and Potential Crops Section, Department of

Genetics and Plant Breeding, Chaudhary Charan Singh Haryana

Agricultural University, Hisar. Roots were shade dried. After

drying, roots were cut into small pieces of 2-3 inches and were

ground. Chemical parameters viz. crude fibre, minerals, starch,

total sugars, reducing sugars, non-reducing sugars and

phytochemical parameters viz. total alkaloids and tannins were

estimated from ashwagandha roots powder.

The commercially available chemicals from Merck, SRL

(SISCO Research Labortories), Qualigens and Sigma-Aldrich,

were used for various experimental procedures.

Estimation of moisture content: Moisture content was

estimated by the standard procedure of AOAC [6].

Estimation of crude fibre: Crude fibre was estimated by

the modified method of Maynard [7]. Two gram of moisture

and fat free powdered sample of ashwagandha roots was weighed

and transferred to the spoutless 1 L beaker and added 200 mL

of 1.25 % (w/v) sulphuric acid. The beaker was then placed

on hot plate and allowed to reflux for 30 min timed from onset

of boiling and the contents were shaked after every 5 min.

After boiling for 30 min beaker was removed from hot plate

and filtered through a muslin cloth using suction. The residue

was washed with hot water till it became free from acid, then

the material was transferred to the same beaker and added

200 mL of 1.25 % NaOH solution and the contents were again

refluxed for 30 min. It was filtered again through muslin cloth

with the help of vacuum or suction pump and the residue was

washed with hot water till it became free from alkali. The

residue was then transferred to a crucible and placed in hot

air oven, allowed to dry to constant weight at 80-110 °C and

recorded its weight. The residue was ignited in muffle furnace

at 550-660 °C for 2-3 h, then cooled and weighed again. The

loss of weight due to ignition is weight of crude fibre.

Weight of crude fibre

Crude fibre content (%) 100

Original weight of sample

=×

Estimation of starch: Starch content was estimated by

the method of McCready et al. [8]. 0.2 g of finally ground

sample of ashwagandha roots was taken in a 50 mL centrifuge

tube for extraction of sugar and starch and added 20 mL of

hot 80 % alcohol. The tubes were then shaked for 5-10 min,

centrifuged at 3000 rpm for 10 min. and supernatant was

decanted. The supernatant was free of sugars as judged by

negative test with anthrone reagent. The residue was cooled

in ice water and added 6.5 mL of 52 % perchloric acid while

stirring the contents with a glass rod. It was allowed to stand

for 15 min. with occassional stirring, centrifuged at 4 °C and

supernatant fractions were collected final volume was made

up to 100 mL with water. The above extract was diluted so

that it contains 5-20 µg of glucose per mL. Then 5 mL aliquot

of this diluted extract was taken in tubes and the tubes were

placed in cool water bath and added 10 mL freshly prepared

anthrone reagent, mixed properly and the tubes were trans-

ferred to boiling water bath for 7.5 min. After cooling the tubes

under running tap water the absorbance of these solutions was

noted at 630 nm. A standard curve was prepared using 0 to

100 µg glucose as per the procedure described above and

amount of glucose was calculated in the sample aliquot. Results

were expressed as mg/g on dry weight basis.

Estimation of minerals: Minerals content were estimated

by the method of Jackson [9] and Ruig [10]. 1 g powdered

sample of ashwagandha roots was digested with 15 mL of

diacid mixture (4HNO3:HClO4) in a conical flask by heating

on hot plate in open space till clear white precipitates settled

down at bottom of conical flask. The precipitates were dissol-

ved in 1 % HCl prepared in double glass distilled water, filtered

and final volume of filterate was made up to 50 mL with double

glass distilled water. Results were expressed as ppm on dry

weight basis.

Estimation of total sugars: Total sugars were estimated

by the modified method of Dubois et al. [11]. 1 mL of aqueous

extract was diluted with distilled water to adjust the absorbance

with in calibration limits. Then, 2 mL of phenol solution (2

%, w/v) was added followed by 5 mL concentrated sulphuric

acid. Acid was added in such a way that it directly pours on

the solution. The test tubes were allowed to cool for 30 min

and absorbance of the solution was measured at 490 nm using

UV-visible double beam spectrophotometer Model 2203

(Systronics Co.) against a blank prepared similarly but con-

taining respective solvent instead of extracts. The amount of

total sugars present in the extracts were calculated from the

standard curve and the results are expressed as milligrams per

gram on dry weight basis.

Estimation of reducing sugars: Reducing sugars were

estimated by the method of Nelson [12] as modified by Somogyi

[13]. 1 mL of aqueous extract was diluted with distilled water

to adjust the absorbance with in calibration limits. Then,

1 mL distilled water was added, followed by addition of 1 mL

alkaline copper reagent, solution was mixed, covered with

aluminum foil and heated in boiling water bath for 20 min.

The tubes were cooled to room temperature and 1 mL of

arsenomolybdate reagent was added. The contents were mixed

thoroughly and volume was made up to 10 mL with distilled

water. The absorbance of the solution was measured at 520

nm using UV-visible double beam spectrophotometer Model

2203 (Systronics Co.) against a blank prepared similarly but

containing respective solvents instead of extracts. The amount

of reducing sugars present in the extracts were calculated from

the standard curve and the results are expressed as milligrams

per gram on dry weight basis.

Estimation of non-reducing sugars: The content of non-

reducing sugars was calculated from the difference between

the content of total sugars and that of reducing sugars.

Non-reducing sugars = Total sugars – Reducing sugars

Estimation of total alkaloids: The total alkaloids content

was estimated by the method of Mishra [14]. 1 g of ashwa-

gandha roots powder was weighed and put in stoppered

test tube, then added 10 mL chloroform and three drops of

ammonia, mixed well and kept overnight. Next day after

shaking, the contents were filtered through cotton wool in a

small beaker and the residue was washed with chloroform

thrice (10 mL each), which ensured complete removal of alka-

loids. The extract was dried on water bath and added 10 mL

ethyl alcohol and mixed the contents with clean glass rod.

Then the liquid portion was evaporated which confirms removal

of ammonia and added 10 mL of standard acid solution (0.01

N H2SO4) in the beaker and warmed slightly to dissolve the

alkaloids in acid solution. It was cooled and the unused acid

was titrated with standard NaOH (0.01 N) and note down the

volume of alkali used in titration. A blank was run to find out

the exact volume of acid neutralized by the alkaloids.

Total alkaloid (%) = 0.415 × volume of acid consumed

by alkaloids.

Estimation of tannins: Tannin contents was estimated as

catechin equivalent by vanillin-hydrochloric acid method [15].

200 mg dried plant material was taken in a 50 mL test tube

and 25 mL of methanol was added to it. The tubes were closed

with pith corks. The contents of the tubes were shaken occasio-

1684 Gulati et al. Asian J. Chem.

nally and allowed to stand overnight at 25 to 32 °C. 1 mL of

clear supernatant was then pipetted in a test tube and 5 mL of

vanillin-HCl reagent was added to it. The absorbance of

brownish red colour so produced was measured at 525 nm

after 25 min on a spectronic 20 colorimeter. A blank containing

methanol was also run simultaneously. A standard curve of

catechin was prepared simultaneously in order to calculate

amount of tannin. Results were expressed as mg/g on dry weight

basis.

RESULTS AND DISCUSSION

Moisture content: Moisture content in ashwagandha roots

of various genotypes/varieties on fresh weight basis (f.w.b.)

ranged from 6.25 to 7.42 % (Table-1) and it was maximum in

genotype HWS-1228 (7.42 %) followed by HWS-08-18

(7.18 %), Selection-2B (6.83 %), HWS-08-14 (6.34 %) and

HWS-1229 (6.25 %) in comparison to control varieties RVA-

100 (7.27 %) and JA-20 (7.10 %). Our finding is in agreement

with previous investigation which reported 5.54 % moisture

content in ashwagandha root powder [16].

TABLE-1

MOISTURE CONTENTS (%), CRUDE FIBRE (%) AND

STARCH (mg/g) IN ASHWAGANDHA ROOTS OF

VARIOUS GENOTYPES/VARIETIES

Genotypes/varieties

Moisture

content (%)

on f.w.b.

Crude fibre

(%) on d.w.b.

Starch (mg/g)

on d.w.b.

HWS-08-14 6.34 ± 0.19 37.3 ± 0.53 8.00 ± 0.06

HWS-08-18 7.18 ± 0.23 21.5 ± 0.17 8.17 ± 0.03

HWS-1228 7.42 ± 0.26 18.7 ± 0.15 7.61 ± 0.07

HWS-1229 6.25 ± 0.06 23.6 ± 0.09 8.07 ± 0.04

Selection-2B 6.83 ± 0.13 23.0 ± 0.21 7.67 ± 0.12

JA-20 (C) 7.10 ± 0.03 17.4 ± 0.29 7.78 ± 0.09

RVA-100 (C) 7.27 ± 0.09 21.7 ± 0.38 8.22 ± 0.07

Crude fibre: Crude fibre contents in ashwagandha roots

of various genotypes/varieties on dry weight basis (d.w.b.)

ranged from 17.4 to 37.3 % (Table-1) and it was maximum in

genotype HWS-08-14 (37.3 %) followed by HWS-1229

(23.6 %), Selection-2B (23.0 %), HWS-08-18 (21.5 %) and

HWS-1228 (18.7 %) in comparison to control varieties RVA-

100 (21.7 %) and JA-20 (17.4 %). Other research workers

have also reported similar findings. Crude fibre contents ranged

from 22 to 34 % at 150 day after planting and from 32.0 to

38.7 % at 210 day after planting in dry roots of five divergent

accessions of Withania somnifera collected from various

locations in India [5].

Starch: Starch content in ashwagandha roots of various

genotypes/varieties on dry weight basis (d.w.b.) ranged from

7.61 to 8.22 mg/g (Table-1) and it was maximum in genotype

HWS-08-18 (8.17 mg/g) followed by HWS-08-14 (8.00 mg/

g), HWS-1229 (8.07 mg/g), Selection-2B (7.67 mg/g) and

HWS-1228 (7.61 mg/g) in comparison to control varieties

RVA-100 (8.22 mg/g) and JA-20 (7.78 mg/g). Our finding is

in agreement with previous investigation which reported that

starch content varied between 6.09 to 9.46 mg/g in roots of

Withania somnifera (L.) Dunal [5].

Minerals (Fe, Cu, Zn, Mn) content: Fe, Cu, Zn and Mn

content in ashwagandha roots of various genotypes/varieties

on dry weight basis (d.w.b.) is shown in Table-2. Iron contents

ranged from 529.16 to 983.33 ppm and it was maximum in

genotype HWS-08-18 (983.33 ppm) followed by HWS-1228

(958.33 ppm), HWS-1229 (627.08 ppm), Selection-2B (604.16

ppm) and HWS-08-14 (529.16 ppm) in comparison to control

varieties RVA-100 (795.83 ppm) and JA-20 (595.83 ppm). Cu

content ranged from 8.40 to 13.72 ppm and it was maximum

in genotype HWS-08-18 (13.72 ppm) followed by HWS-08-

14 (12.47 ppm), HWS-1229 (10.88 ppm), HWS-1228 (9.60

ppm) and Selection-2B (9.35 ppm) in comparison to control

varieties RVA-100 (8.77 ppm) and JA-20 (8.40 ppm). Zn

content ranged from 16.33 to 41.00 ppm and it was maximum

in genotype HWS-08-18 (41.00 ppm) followed by HWS-08-

14 (27.33 ppm), HWS-1228 (27.17 ppm), Selection-2B (23.50

ppm) and HWS-1229 (22.67 ppm) in comparison to control

varieties RVA-100 (25.33 ppm) and JA-20 (16.33 ppm). Mn

content ranged from 12.20 to 33.46 ppm and it was maximum

in genotype HWS-08-18 (33.46 ppm) followed by HWS-1228

(25.45 ppm), HWS-1229 (18.98 ppm), Selection-2B (17.52

ppm) and HWS-08-14 (12.20 ppm) in comparison to control

varieties RVA-100 (22.80 ppm) and JA-20 (16.80 ppm). Results

of other research workers showed 35.91 ppm Zn and 12.96

ppm Cu content in dried roots of ashwagandha [17]. Fe, Mn

and Cu content ranged from 280 to 945 ppm, from 15 to 49

ppm and from 14 to 21 ppm, respectively in roots of Withania

somnifera of Sondikola regions and from 349.5 to 602.0 ppm,

19 to 59 ppm and 18 to 42 ppm, respectively in roots of Withania

somnifera of Karthikere regions [18].

TABLE-2

MINERALS (Fe, Cu, Zn, Mn) CONTENT (ppm)

IN ASHWAGANDHA ROOTS OF VARIOUS

GENOTYPES/VARIETIES ON DRY WEIGHT BASIS

Minerals content (ppm) on dry weight basis Genotypes/

varieties Fe Cu Zn Mn

HWS-08-14 529.16 ±

15.02

12.47 ±

0.69

27.33 ±

0.93

12.20 ±

0.61

HWS-08-18 983.33 ±

45.12

13.72 ±

0.68

41.00 ±

0.50

33.46 ±

0.95

HWS-1228 958.33 ±

37.90

9.60 ±

0.19

27.17 ±

0.83

25.45 ±

0.28

HWS-1229 627.08 ±

27.56

10.88 ±

1.20

22.67 ±

1.01

18.98 ±

0.55

Selection-2B 604.16 ±

11.02

9.35 ±

0.38

23.50 ±

0.50

17.52 ±

0.52

JA-20 (C) 595.83 ±

19.87

8.40 ±

0.38

16.33 ±

0.60

16.80 ±

1.01

RVA-100 (C) 795.83 ±

13.66

8.77 ±

0.16

25.33 ±

1.01

22.80 ±

0.28

Total sugars: Total sugars content in ashwagandha roots

of various genotypes/varieties on dry weight basis (d.w.b.)

ranged from 5.33 to 6.89 mg/g (Table-3) and it was maximum

in genotype HWS-08-18 (6.89 mg/g) followed by HWS-08-

14 (6.82 mg/g), HWS-1228 (6.15 mg/g), HWS-1229 (5.51

mg/g) and Selection-2B (5.33 mg/g) in comparison to control

varieties RVA-100 (5.91 mg/g) and JA-20 (5.83 mg/g). Our

finding is in agreement with previous investigation which

reported that total sugars content varied between 2.52 to 9.72

mg/g in roots of Withania somnifera (L.) [5].

Vol. 29, No. 8 (2017) Chemical and Phytochemical Composition of Ashwagandha (Withania somnifera L.) Roots 1685

TABLE-3

TOTAL SUGARS (mg/g), REDUCING SUGARS (mg/g) AND NON-

REDUCING SUGARS (mg/g) IN ASHWAGANDHA ROOTS OF

VARIOUS GENOTYPES/VARIETIES ON DRY WEIGHT BASIS

Genotypes/

varieties

Total sugars

(mg/g)

Reducing

sugars (mg/g)

Non-reducing

sugars (mg/g)

HWS-08-14 6.82 ± 0.05 0.52 ± 0.04 6.30 ± 0.01

HWS-08-18 6.89 ± 0.04 0.56 ± 0.04 6.33 ± 0.01

HWS-1228 6.15 ± 0.04 0.64 ± 0.03 5.51 ± 0.02

HWS-1229 5.51 ± 0.06 0.54 ± 0.03 4.97 ± 0.05

Selection-2B 5.33 ± 0.03 0.46 ± 0.02 4.87 ± 0.01

JA-20 (C) 5.83 ± 0.06 0.46 ± 0.01 5.37 ± 0.05

RVA-100 (C) 5.91 ± 0.04 0.40 ± 0.01 5.51 ± 0.03

Reducing sugars: Reducing sugars content in ashwa-

gandha roots of various genotypes/varieties on dry weight basis

(d.w.b.) is shown in Table-3. Reducing sugars content ranged

from 0.40 to 0.64 mg/g and it was maximum in genotype HWS-

1228 (0.64 mg/g) followed by HWS-08-18 (0.56 mg/g), HWS-

1229 (0.54 mg/g), HWS-08-14 (0.52 mg/g) and Selection-2B

(0.46 mg/g) in comparison to control varieties JA-20 (0.46

mg/g) and RVA-100 (0.40 mg/g). Other research workers have

also reported similar findings. Reducing sugars content have

been reported to varied between 0.21 to 2.10 mg/g in roots of

Withania somnifera (L.) Dunal [5].

Non-reducing sugars: Non-reducing sugars content in

ashwagandha roots of various genotypes/varieties on dry weight

basis (d.w.b.) is shown in Table-3. It was observed that non-

reducing sugars content ranged from 4.87 to 6.33 mg/g and it

was maximum in genotype HWS-08-18 (6.33 mg/g) followed

by HWS-08-14 (6.30 mg/g), HWS-1228 (5.51 mg/g), HWS-

1229 (4.97 mg/g) and Selection-2B (4.87 mg/g) in comparison

to control varieties RVA-100 (5.51 mg/g) and JA-20 (5.37 mg/g).

Results of other research workers showed that non-reducing

sugars content content varied between 2.33 to 7.62 mg/g in

roots of Withania somnifera (L.) Dunal [5].

Total alkaloids: Total alkaloids content in ashwagandha

roots of various genotypes/varieties on dry weight basis (d.w.b.)

is shown in Table-4. Total alkaloids content ranged from 0.26

to 0.31 % and it was maximum in genotype HWS-08-18 (0.31

%) followed by HWS-08-14 (0.30 %), HWS-1229 (0.29 %),

HWS-1228 (0.28 %) and Selection-2B (0.26 %) in comparison

to control varieties RVA-100 (0.26 %) and JA-20 (0.26 %).

Other research workers have also reported similar findings.

Total alkaloids content in Withania somnifera (L.) roots have

been reported to varied between 0.13 to 0.31 % [19].

Tannins: Tannins content in ashwagandha roots of various

genotypes/varieties on dry weight basis (d.w.b.) is also shown

TABLE-4

TOTAL ALKALOIDS (%) AND TANNINS (mg/g)

IN ASHWAGANDHA ROOTS OF VARIOUS

GENOTYPES/VARIETIES ON DRY WEIGHT BASIS

Genotypes/varieties Total alkaloids (%) Tannins (mg/g)

HWS-08-14 0.30 ± 0.01 0.73 ± 0.04

HWS-08-18 0.31 ± 0.00 0.77 ± 0.06

HWS-1228 0.28 ± 0.01 0.84 ± 0.04

HWS-1229 0.29 ± 0.00 0.66 ± 0.06

Selection-2B 0.26 ± 0.01 0.73 ± 0.04

JA-20 (C) 0.26 ± 0.01 0.81 ± 0.04

RVA-100 (C) 0.26 ± 0.01 0.66 ± 0.06

in Table-4. It was observed that tannins content ranged from

0.66 to 0.84 mg/g and it was maximum in genotype HWS-

1228 (0.84 mg/g) followed by HWS-08-18 (0.77 mg/g), HWS-

08-14 (0.73 mg/g), Selection-2B (0.73 mg/g) and HWS-1229

(0.66 mg/g) in comparison to control varieties JA-20 (0.81

mg/g) and RVA-100 (0.66 mg/g). Our finding is in agreement

with previous investigation which reported 0.82 mg/g tannins

content in roots of Poshita variety of Withania somnifera (L.)

under in vivo conditions and 0.39 mg/g under in vitro condi-

tions, respectively [20].

Conclusion

The present study showed that among various genotypes

of Ashwagandha roots, genotypes HWS-08-18 and HWS-1228

were found better in terms of crude fibre, minerals (Fe, Cu,

Zn, Mn), starch, total sugars, reducing sugars, non-reducing

sugars, total alkaloids and tannins.

ACKNOWLEDGEMENTS

This work is financially supported by the Department of

Science and Technology, New Delhi, India.

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Effect of Supplementation of Ashwagandha (Withania somnifera) Root Powder on Body Weight Gain and Egg Quality in Layers

Article

Full-text available

Jul 2024

Background: As the demand for natural and sustainable solutions in poultry production continues to rise, ashwagandha root powder presents itself as a promising avenue for improving egg quality while aligning with consumer preferences for wholesome and nutritious products. Therefore, research trial was conducted to study the effect of supplementation of ashwagandha root powder on body weight gain and egg quality in layers during a period of 10 weeks. Methodology: One hundred and twenty White Leghorn layers were randomly divided into four treatments and reared on standard managemental conditions. The Ashwagandha root powder was supplemented with the basal diet at 0.5% level (T1), 1% (T2), 1.5 % (T3) while T0 was kept as control i.e. basal diet without ashwagandha root powder. The feed and water were offered ad libitum to experimental birds. Design used: The design of the experiment was a completely randomized design. Duncans multiple range test was used to determine significant difference among means for different treatments. Results: The result noted that the body weight change was not affected by ashwagandha root powder supplementation. Egg quality parameters viz., egg shape index, shell weight, yolk weight, yolk height, albumen weight and albumen index were not influenced by the addition of ashwagandha root powder. However, a significant (p<0.05) effect was observed on yolk index. No significant (p>0.05) difference was seen in the sensory attributes of the egg. It may be suggested that inclusion of ashwagandha root powder in the diets of layer can be improve yolk index without affecting sensory attributes of eggs in layer.

Withania somnifera a journey from traditional rejuvenator to anti-SARS-CoV-2 remedy

Article

Full-text available

Jun 2023

Ashwagandha (Withania somnifera L. Dunal) roots have been used in Ayurveda for fever, asthma, arthritis,rheumatism, inflammation, tuberculosis, mental diseases, and male sexual disorders. Phytochemical studiesrevealed that the plant mainly contains alkaloids (withanine, sominiferine, tropine, isopelletierine andanaferine), and steroidal lactones (withanolides A-Y, withaferine A, withasominiferols A-C, withanone and sitoindosides) as bioactive metabolites. These constituents and extracts have several pharmacologicalactivities, including antioxidant, antitumour, antimicrobial, antivenom, and anti-parkinsonian activities.The present review presents updated information on ashwagandha based on its phytochemistry andpharmacology. The plant and many of its secondary metabolites have been found to be effective in thetreatment of many human ailments. However, further investigations are still needed to understand the exactmechanism of their actions.

Sleep-promoting activity of amylase-treated Ashwagandha (Withania somnifera L. Dunal) root extract via GABA receptors

Article

Full-text available

Jun 2023

Ashwagandha (Withania somnifera L. Dunal), an Indian medicinal plant that has been used for centuries to treat insomnia, exhibits a variety of biological activities, such as improving cognitive function, immunity and anxiety. In this study, the effect of enzyme-treated Ashwagandha root extract (EA) and on sleep was evaluated using rodent models. Starch contained in the Ashwagandha root extract was removed by amylase treatment to prepare EA. To evaluate the sleep-promoting activity of EA, a pentobarbital-induced sleep test and electroencephalogram analysis were performed. In addition, the sleep-promoting mechanism of EA was elucidated by analyzing the expression of sleep-related receptors. In the pentobarbital-induced sleep test, EA dose-dependently increased sleep duration. Additionally, electroencephalogram analysis revealed that EA significantly increased δ-wave and non-rapid eye movement sleep times, which are involved in deep sleep, thereby improving sleep quality and quantity. EA also effectively relieved caffeine-induced insomnia symptoms. Furthermore, the γ-aminobutyric acid (GABA) content in the brain and mRNA and protein expression of GABAA, GABAB1, and serotonin receptors were significantly increased by EA compared to the normal group. In particular, EA showed sleep-promoting activity by binding to various GABAA receptor sites. Collectively, EA exhibited sleep-promoting activity through the GABAergic system and may be used as a functional material to improve sleep deprivation.

Phytoconstituent estimation and LC-MS studies of field grown Withania somnifera (L.) Dunal root extract in Jharkhand & Bihar

Preprint

Full-text available

Jun 2022

Background: Plants in either raw form or their root extract constituents utilized as complementary and alternative medicine in various disorders. The present study was undertaken for phytoconstituents estimation and to evaluate chemical compositions of methanolic extract of Withania somnifera (L.) Dunal (Ashwagandha) roots collected from BAU, Ranchi, Jharkhand and Sabour, Bhagalpur, Bihar. Methods: Phytochemicals viz. crude methanolic extract analyzed in Ashwagandha roots. Alcoholic extract were prepared using Soxhlet extraction apparatus. On phytochemical evaluation, total phenolic, flavonoid, total tannin and alkaloids were estimated using their respective methods. Presences of bioactive components in extract were determined by Liquid Chromatography-Mass Spectrophometry (ESI positive ion) technique. Results: Qualitative screening of extract ensured the presence of alkaloids, phenols, tannins and flavonoids. Quantitatively these phytochemicals showed TPC (66.06 mg/gm), TFC (650.0 mg/gm), TAC (102 mg/gm) and TTC (5.89 mg/gm). However, the phytochemicals and yield did not accumulate in various fractions on polarity. This result was supported by LC-MS analysis which showed 17 definite peaks in their chromatogram but only 8 peaks offer a suitable approximation. Extract revealed forty five bioactive compounds where most of them were withanolides, phenolic compounds, aryl propionic acid and cinnamide derivatives. Conclusion: Quantitative analysis of present phytochemical showed maximum quantity of TFC followed by TAC, TPC and TTC. In other hand, presence of various bioactive compounds in root extract showed plant contains very high medicinal value.

Phytoconstituent estimation and LC-MS studies of field grown Withania somnifera (L.) Dunal root extract in Jharkhand and Bihar

Article

Full-text available

Nov 2022

Plants in either raw form or their root extracts are utilized as complementary and alternative medicine in various disorders. The present study was undertaken for phytoconstituents estimation and to evaluate chemical compositions of methanolic extract of Withania somnifera (L.) Dunal (Ashwagandha) roots collected from BAU, Ranchi, Jharkhand and Sabour, Bhagalpur, Bihar. Phytochemicals through crude methanolic extract were analyzed in Ashwagandha roots. Alcoholic extract was prepared using Soxhlet extraction apparatus. On phytochemical evaluation, total phenolic, flavonoid, total tannin and alkaloids were estimated. Presence of bioactive components in extract was determined by Liquid Chromatography-Mass Spectrophotometry (ESI positive ion) technique. Qualitative screening of extract ensured the presence of alkaloids, phenols, tannins and flavonoids. Quantitatively these phytochemicals showed TPC (66.06 mg/gm), TFC (650.0 mg/gm), TAC (102 mg/gm) and TTC (5.89 mg/gm). However, the phytochemicals and yield did not accumulate in various fractions on polarity. This result was supported by LC-MS analysis which showed 17 definite peaks in their chromatogram but only 8 peaks offer a suitable approximation. Extract revealed forty five bioactive compounds where most of them were phenolic compounds, aryl propionic acid and cinnamide derivatives. Quantitative analysis of present phytochemical showed maximum quantity of TFC followed by TAC, TPC and TTC. On the other hand, presence of various bioactive compounds in root extract showed that plant is having very high medicinal value.

Antiviral perspectives of economically important Indian medicinal plants and spices

Article

Full-text available

Aug 2022

Human respiratory diseases caused by viral infections leads to morbidity. Among infectious diseases, viral infections associated with the respiratory tract remain the primary reason for global deaths due to their transmissibility. Since immemorial, traditional Indian medicinal plants, their extracts, and several phytochemicals can treat various diseases. Sources for this review paper are data derived from a peer-reviewed journal that emphasizes the economic importance of medicinal plants. Several plant-based medicines have been reported to be effective against multiple viral infections, including the Human Adenovirus, Enterovirus, Influenza virus, Hepatitis virus, etc. This review emphasizes use of the Indian medicinal plants like as Withania somnifera (Ashwagandha, Winter Cherry), Moringa oleifera (Drumstick), Ocimum tenuiflorum (Tulsi), Azadirachta indica (Neem), Curcuma longa (Turmeric), Terminalia chebula (Chebulic Myrobalan), Punica granatum (Pomegranate) and the Indian household spices (ginger, garlic and black pepper). It further describes their secondary phytoconstituents extraction procedure, mode of action and the potential application to improve clinical outcomes of neutraceuticals against various viral infections.

Chemical Composition, Biological Activity, and Health-Promoting Effects of Withania somnifera for Pharma- Food Industry Applications

Article

Full-text available

Dec 2021
J FOOD QUALITY

The Withania genus comes from the Solanaceae family and includes around 23 species, spread over some areas of the Mediterranean, Asia, and East Africa. Widely used in traditional medicine for thousands of years, these plants are rich in secondary metabolites, with special emphasis on steroidal lactones, named withanolides which are used as ingredients in numerous formulations for a plethora of diseases, such as asthma, diabetes, arthritis, impotence, amnesia, hypertension, anxiety, stress, cancer, neurodegenerative, and cardiovascular diseases, and many others. Among them, Withania somnifera (L.) Dunal is the most widely addressed species from a pharmacological and agroindustrial point of view. In this sense, this review provides an overview of the folk uses, phytochemical composition, and biological activity, such as antioxidant, antimicrobial, anti-inflammatory, and cytotoxic activity of W. somnifera, although more recently other species have also been increasingly investigated. In addition, their health-promoting effects, i.e., antistress, anxiolytic, adaptogenic, antirheumatoid arthritis, chemoprotective, and cardiorespiratory-enhancing abilities, along with safety and adverse effects are also discussed.

Analytical Strategies to Investigate Molecular Signaling, Proteomics, Extraction and Quantification of Withanolides - A Comprehensive Review

Article

Feb 2024
CRIT REV ANAL CHEM

Withanolides are the class of steroidal molecules getting greater emphasis in recent years. Quality control throughout the manufacturing and storage period is often one of the key problems that have restricted their broad use in India's indigenous and Ayurvedic medical systems for thousands of years. Because of their diverse clinical potential, withanolides have received a great deal of scientific attention. Analytical techniques are being devised for the automated isolation, identification, and estimation of every single protein within the cell as well as in herbal extracts of withanolides, due to which now researchers are interested in determining the effects of metabolism as well as various stimuli on protein expression, which made the study easier. This study discusses the potential use of hyphenated analytical methods that are reliable in understanding the molecular signaling features, proteome evaluation and characterization of withanolides, in addition to examining existing methodological limitations. The choice of analytical techniques for the withanolides analysis, however, relies on the nature of the sample matrix, the aim of the analysis, and the sensitivity of the technique.

Acute and Sub-Acute Oral Toxicity Assessment of Mixed Extract of Trigonella Foenum-Graecum Seeds and Withania Somnifera Root in Rats

Article

Full-text available

Dec 2022

Mohammed Sadeq Al-Awar

This study aimed at assessing the safety of a mixed extract of Trigonella Foenum-graecum seeds and Withania Somnifera root (TFWS), which effectively relieves male menopausal symptoms. To this end, male and female Sprague-Dawley rats were divided into the following groups and repeatedly administered TFWS orally for 90 days: control, low-dose (500 mg/kg/day), intermediate-dose (1,000 mg/kg/day), and high-dose (2,000 mg/kg/day) groups. The animals were monitored for general symptoms; their body weights and electrolyte levels were measured; urinalysis, blood chemistry, biochemistry tests, and histopathological tests were performed to assess the toxicity of TFWS. The no-observed-adverse-effect level of TFWS was 2,000 mg/kg/day for all male and female rats. While in the TFWS-administered and control groups, most parameters were within the normal range; some rats in the high-dose group showed changes not induced by the test substance but which may be specific to an individual animal or may occur naturally. Thus, based on our findings, we consider that TFWS may be a safe, non-toxic substance for alleviating male menopausal symptoms.

Ashwagandha

Article

Full-text available

Sep 2020

Gitanjali Devi

Proximate Nutritive Values and Mineral Components of Withania Somnifera (Linn.) Dunal

Article

Full-text available

Jul 2010

Withania somnifera (Linn.) Dunal is a subtropical shrub with important medicinal properties. The nutritive value and the elemental composition of different parts of plants, Withania somnifera which are grown in two distinct geographical regions (Sondekola and Karthikere) of Karnataka have been determined. The investigation revealed that the variation of macro, micro and proximate components varied not only in the plants of different regions but also in the different parts of the same plant. Among the macro elements, Karthikere samples recorded maximum values of nitrogen, phosphorous and magnesium and Sondekola samples recorded maximum values of sodium, potassium and calcium. Among the components of micronutrients, the highest values of iron were recorded both in Sondekola and Karthikere samples. The average values of manganese, copper and zinc were more in the Karthikere samples and comparatively less in the Sondekola samples. Whereas, all the samples of Sondekola recorded maximum values of nutrition. It is believed that the dry climatic condition of the region may contribute the high values of nutrition. Further, the observations are discussed with reference to the geography, elemental composition and nutritional values. The strong and negative observations on herbal drugs and their validity, the study emphasizes the role of elemental composition, proximate components, nutritive value, habitat and geographical features which influence growth and development of Withania somnifera and also herbal products of Withania somnifera in particular and medicinal plants in general.

Methanolic extracts of Withania somnifera leaves, fruits and roots possess antioxidant properties and antibacterial activities

Article

Full-text available

Oct 2012
BMC Compl Alternative Med

Background Withania somnifera, also known as ashwagandha, is an important herb in ayurvedic and indigenous medical systems. The present study was designed to evaluate the antioxidant and antibacterial activities of an 80% aqueous methanolic extract of W. somnifera roots (WSREt), fruits (WSFEt) and leaves (WSLEt). Methods Several assays were performed to determine the antioxidant properties of this herb including 1,1-diphenyl-2-picrylhydrazyl radical (DPPH) scavenging activity, ferric reducing antioxidant power (FRAP), ferrous chelation and inhibition of β-carotene bleaching. Results The values for DPPH, FRAP, ferrous chelation and inhibition of β carotene bleaching for the three types of extracts ranged from 101.73-801.93 μg/ml, 2.26-3.29 mM Fe/kg, 0.22-0.65 mg/ml and 69.87-79.67%, respectively, indicating that W. somnifera, particularly the leaves, possesses significant antioxidant properties. The mean ascorbic acid content was 20.60-62.60 mg/100 g, and the mean anthocyanin content was 2.86-12.50 mg/100 g. Antibacterial activities were measured using the agar well diffusion method and five pathogenic Gram-negative bacteria: Escherichia coli, Salmonella typhi, Citrobacter freundii, Pseudomonas aeruginosa and Klebsiella pneumoniae. The leaf extracts displayed the highest activity against S. typhi (32.00 ± 0.75 mm zone of inhibition), whereas the lowest activity was against K. pneumoniae (19.00 ± 1.48 mm zone of inhibition). The lowest minimum inhibitory concentration value was 6.25 mg/ml, which was against S. typhi, followed by 12.5 mg/ml against E. coli. Conclusion In addition to its antioxidant properties, W. somnifera exhibited significant antibacterial activities against Gram-negative bacteria, particularly S. typhi.

Biochemical Composition of Roots of Withania somnifera (L.) Dunal

Article

Full-text available

Jun 2006

Contents of soluble protein, total amino acids, reducing sugars, non-reducing sugars, starch were examined in fresh roots of five selective accessions viz. AGB-002, AGB-009, AGB-015, AGB-025 and AGB-030 of Withania somnifera (L.) Dunal while crude fiber was analyzed in dry roots during young and maturity stages. All the five accessions followed a uniform pattern of maximum biochemical constituent accumulation during maturity. The accession AGB-002 appears to be more efficient in accumulation and synthesis of biochemical constituents during the two stages of root growth.

High catechin concentrations detected in Withania somnifera (ashwagandha) by high performance liquid chromatography analysis

Article

Full-text available

Aug 2011
BMC Compl Alternative Med

Withania somnifera is an important medicinal plant traditionally used in the treatment of many diseases. The present study was carried out to characterize the phenolic acids, flavonoids and 1,1-diphenyl-2-picrylhydrazyl radical (DPPH) scavenging activities in methanolic extracts of W. somnifera fruits, roots and leaves (WSFEt, WSREt and WSLEt). WSFEt, WSREt and WSLEt was prepared by using 80% aqueous methanol and total polyphenols, flavonoids as well as DPPH radical scavenging activities were determined by spectrophotometric methods and phenolic acid profiles were determined by HPLC methods. High concentrations of both phenolics and flavonoids were detected in all parts of the plant with the former ranging between 17.80 ± 5.80 and 32.58 ± 3.16 mg/g (dry weight) and the latter ranging between 15.49 ± 1.02 and 31.58 ± 5.07 mg/g. All of the three different plant parts showed strong DPPH radical scavenging activities (59.16 ± 1.20 to 91.84 ± 0.38%). Eight polyphenols (gallic, syringic, benzoic, p-coumaric and vanillic acids as well as catechin, kaempferol and naringenin) have been identified by HPLC in parts of the plant as well. Among all the polyphenols, catechin was detected in the highest concentration (13.01 ± 8.93 to 30.61 ± 11.41 mg/g). The results indicating that W. somnifera is a plant with strong therapeutic properties thus further supporting its traditional claims. All major parts of W. somnifera such as the roots, fruits and leaves provide potential benefits for human health because of its high content of polyphenols and antioxidant activities with the leaves containing the highest amounts of polyphenols specially catechin with strong antioxidant properties.

Soil Chemical Analysis

Book

Jan 1958

M. L. Jackson

Purification and characterization of peroxidases from Withania somnifera (AGB 002) and their ability to oxidize IAA

Article

Dec 2005
PLANT SCI

Four types of class 111 peroxidases (EC 1.11.1.7) were detected in Withania somnifera (AGB 002) roots of 5 months old plant by zymogram. One of them showed relatively slow mobility on polyacrylamide gels under non-denaturating conditions in comparison to other three enzymes. All the four peroxidases designated as WS1, WS2, WS3 and WS4 were purified from cell free extract by FPLC using ion exchange, affinity and hydrophobic columns. The purity of peroxidases was ascertained by SDS-PAGE and spectral analysis. Purified peroxidases were shown to be monomer glycoproteins with molecular weights between 34 and 48kDa and pI between 3.6 and 4.8. All the peroxidases were optimally active at pH 5.0 and highly thermotolerant and stable in the pH range of 3–9. Three of the four peroxidases exhibited indole-3-acetic acid oxidase activity with different rates and needed 2,4-DCP and manganese as cofactors but did not require H2O2 for the activity. In addition, all the peroxidases were capable of oxidizing general phenolic substrates like guaiacol, ABTS, o-dianisidine, aminoantipyrine, euginol and tyrosine.

Method for Estimation of Tannin in Grain Sorghum1

Article

May 1971

Robert E. Burns

The acidified vanillin method of tannin analysis, widely used for forage crops, was modified to permit rapid estimation of relative tannin content of grain sorghum [Sorghum bicolor (L.) Moench]. The method involves overnight extraction of ground grain with methanol at room temperature. An aliquot of the extract is added to a solution of vanillin and hydrochloric acid and the resultant color is read on a colorimeter at 500-525 mµ The reagent is specific for astringent compounds in plants and the rcsuhs were highly correlated with digestibility. Please view the pdf by using the Full Text (PDF) link under 'View' to the left. Copyright © . .

Determination of Starch and Amylose in Vegetables

Article

Sep 1950

Sugars are extracted from peas with 80% ethyl alcohol and the starch is solubilized with dilute perchloric acid. Starch is then determined colorimetrically, without previous acid hydrolysis, by means of the sugar-anthrone-sulfuric acid reaction. Amylose is estimated colorimetrically with iodine. Both total starch and amylose can be determined in about 30 minutes in sugar-free solutions. The accuracy and precision are at least equal to those of standard methods involving acid hydrolysis. Analyses of smooth and wrinkled dried peas and fresh wrinkled peas are presented.

Calorimetric Dubois Method for Determination of Sugar and Related Substances

Article

May 2002

Simple sugars, oligosaccharides, polysaccharides, and their derivatives, including the methyl ethers with free or potentially free reducing groups, give an orange-yellow color when treated with phenol and concentrated sulfuric acid. The reaction is sensitive and the color is stable. By use of this phenol-sulfuric acid reaction, a method has been developed to determine submicro amounts of sugars and related substances. In conjunction with paper partition chromatography the method is useful for the determination of the composition of polysaccharides and their methyl derivatives.

Economic botany : a textbook of useful plants and plant products / Albert F. Hill

Article