ArticlePDF Available

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

Authors:

Abstract

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.
ASIAN JOURNAL OF CHEMISTRY
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.
REFERENCES
1. A.F. Hill, Economic Botony: A Textbook of Useful Plants and Plant
Products, McGraw Hill Book Company Inc; New York, edn 2, pp. 205
(1952).
2. N. Alam, M. Hossain, M.A. Mottalib, S.A. Sulaiman, S.H. Gan and M.I.
Khalil, BMC Complement. Altern. Med., 12, 1175 (2012);
https://doi.org/10.1186/1472-6882-12-175.
3. N. Alam, M. Hossain, M.I. Khalil, M. Moniruzzaman, S.A. Sulaiman and
S.H. Gan, BMC Complement. Altern. Med., 11, 65 (2011);
https://doi.org/10.1186/1472-6882-11-65.
4. Q. Uddin, L. Samiulla, V.K. Singh and S.S. Jamil, J. Appl. Pharm. Sci.,
2, 170 (2012).
5. P.K. Khanna, A. Kumar, A. Ahuja and M.K. Kaul, Asian J. Plant Sci.,
5, 1061 (2006);
https://doi.org/10.3923/ajps.2006.1061.1063.
6. AOAC, Official Methods of Analysis. Association of Official Analytical
Chemists, Washington, DC, edn 17 (2000).
7. A.J. Maynard, Method in Food Analysis, Academic Press, New York,
pp. 176 (1970).
8. R.M. McCready, J. Guggolz, V. Silviera and H.S. Owens, Anal. Chem.,
22, 1156 (1950);
https://doi.org/10.1021/ac60045a016.
9. M.L.C. Jackson, Soil Chemical Analysis, Prentice Hall of India Pvt.
Ltd., New Delhi (1973).
10. W.G.D. Ruig, J. Assoc. Off. Anal. Chem., 69, 1009 (1986).
11. M. DuBois, K.A. Gilles, J.K. Hamilton, P.A. Rebers and F. Smith, Anal.
Chem., 28, 350 (1956);
https://doi.org/10.1021/ac60111a017.
12. N. Nelson, J. Biol. Chem., 153, 375 (1944).
13. M. Somogyi, J. Biol. Chem., 195, 19 (1952).
14. S.N. Mishra, Quick Method for Estimation of Total Alkaloid in
Asgandh, In: Biennial Report of All India Coordinated Research Project
on Medicinal and Aromatic Plants, NRCM&AP (Anand), pp. 254 (1996).
15. R.E. Burns, Agron. J., 63, 511 (1971);
https://doi.org/10.2134/agronj1971.00021962006300030050x.
16. A. Rani, N.R. Baranwal and R.K. Nema, Asian J. Biochem. Pharm. Res.,
4, 195 (2012).
17. M. Garg and J. Ravikant, Pharmacogn. Phytochem., 2, 66 (2014).
18. S.R. Krishnamurthy and P. Sarala, E-J. Chem., 7, 985 (2010);
https://doi.org/10.1155/2010/616851.
19. S. Johri, U. Jamwal, S. Rasool, A. Kumar, V. Verma and G.N. Qazi,
Plant Sci., 169, 1014 (2005);
https://doi.org/10.1016/j.plantsci.2005.05.015.
20. M.O. Viji, M.M. Mathew and R. Parvatham, Vistas, 2, 143 (2013).
1686 Gulati et al. Asian J. Chem.
... Crude fiber content reported to range from 18.7 to 35.8% (150 DAS). Similar results were also reported byGulati et al. (2017) 22 andKumar et al. (2011) ...
... Crude fiber content reported to range from 18.7 to 35.8% (150 DAS). Similar results were also reported byGulati et al. (2017) 22 andKumar et al. (2011) ...
Article
Full-text available
Withania somnifera (L.) Dunal (Fam. Solanaceae) commonly known as Ashwagandha has been used in traditional Indian medicine system from ancient times. Roots of the plant are being used in the preparation of many Ayurvedic medicines for treatment of many diseases. Many compounds have been reported and isolated from the roots, which mainly consists of withanolides, (steroidal lactones). In present study, the roots of 25 diverse W. somnifera genotypes were collected and evaluated for morphological parameters, such as root length, root diameter and dry root yield. Total alkaloids, crude fibers and starch content were also estimated from the dried roots of these accessions. TLC and HPLC revealed the presence of withanolide A in the methanolic extract of roots from diverse genotypes. Quantification through reverse-phase HPLC revealed varied concentrations of withanolide A in different genotypes grown under field conditions. Accumulation of withanolide A was reported highest in the genotype UWS-59 makes this genotype superior for medicinal use.
... As stated before, Ashwagandha (Withania somnifera (L.) Dunal) has long been used in the traditional Ayurvedic medicine, although its mineral content has also been studied by Krishnamurthy and Sarala (2010) and Gulati et al. (2017). For example, samples of Withania somnifera were collected from two different locals in India (Krishnamurthy and Sarala 2010) Gulati et al. (2017) when studying the mineral composition of four different genotypes and three varieties of Ashwagandha roots, concluded that the Fe content ranged from 595.83 to 983.33 μg/g, Cu from 8.40 to 13.72 μg/g, Zn from 16.33 to 41.00 μg/g and Mn content ranged from 16.80 to 33.46 μg/g, whose average values agree with our results. ...
... As stated before, Ashwagandha (Withania somnifera (L.) Dunal) has long been used in the traditional Ayurvedic medicine, although its mineral content has also been studied by Krishnamurthy and Sarala (2010) and Gulati et al. (2017). For example, samples of Withania somnifera were collected from two different locals in India (Krishnamurthy and Sarala 2010) Gulati et al. (2017) when studying the mineral composition of four different genotypes and three varieties of Ashwagandha roots, concluded that the Fe content ranged from 595.83 to 983.33 μg/g, Cu from 8.40 to 13.72 μg/g, Zn from 16.33 to 41.00 μg/g and Mn content ranged from 16.80 to 33.46 μg/g, whose average values agree with our results. Also Raju et al. (2006), using particle induced X-ray emission (PIXE) observed high levels of Cr (63.2 μg/g) and Ni (19.3 μg/g) in Ashwagandha roots, whose elements were not detected in our samples although their levels of Ca (0.95%) were close to our values-1.35% ...
Article
Full-text available
The aim of this study is to evaluate the elemental composition of six food supplements of plant origin, commonly sold in the Portuguese market, by energy dispersive X-ray fluorescence. The presence of arsenic in all the Maca, Ashwagandha, Camu-Camu and Hemp protein samples (except the generic form) is a reason of concern due to the long-term effects of As mainly in its inorganic form. Thus, great caution must be taken on some food supplements, particularly the cases of Moringa from Egypt and Yellow/Xpresso Maca, whose inorganic As concentrations are in line with the upper bound concentration for the 95th dietary exposure according to the European Food Safety Authority which is 0.64 μg/kg bw/day. In what regards Hemp protein, if the supplier’s daily intake recommendation (30 g) is followed, values as high as 1.75 μg/kg bw/day of inorganic As will be consumed, which are dangerously above the upper bound. In this case this specific supplement lot should be removed from the market. Also the consumption of Hemp protein leads to a daily intake of Mn above the Daily Reference Intake (DRI) and Adequate Intakes (AIs) for adults. The contamination of Goji berries by Pb is a reason for concern—organic berries contained 11.3 μg/g while berries derived from conventional agriculture 11.6 μg/g, leading to daily intake doses of 315.3 μg and 324.8 μg, respectively, if the recommended daily intake of 28 g is followed. Our findings point out to an inadequacy of the recommended intakes by the supplier vis a vis the concentrations observed, greatly increasing the risk for public health.
... Estimation of total sugars content: Total sugars content was determined by phenol sulphuric acid method [8] using glucose as standard. Detailed method is discussed in previously published paper [9]. The total sugars content present in various extracts was calculated and results were expressed as milligrams per gram (mg/g). ...
... Estimation of reducing sugars content: Reducing sugars content was determined by Nelson-Somogyi method [10] using glucose as standard. The detailed method is discussed in published article [9]. The reducing sugars content present in various extracts was calculated and results were expressed as milligrams per gram (mg/g). ...
Article
For the standardization of herbal products, selection of a suitable extraction technique is important. This study was conducted to observe the effect of various extraction techniques viz. maceration followed by mechanical shaking, refluxing, Soxhlet extraction and centrifugation and solvents viz. acetone, ethanol and water on extract yield and sugars content of clove buds. In clove buds, highest extract yield (27.64 g/100 g), total sugars (28.91 mg/g), reducing sugars (28.50 mg/g) and non-reducing sugars (0.41 mg/g) contents were found in extracts obtained by Soxhlet technique followed by refluxing, mechanical shaking/centrifugation. Amongst solvents, extract yield (34.29 g/100 g), total sugars (29.58 mg/g), reducing sugars (29.18 mg/g) and non-reducing sugars (0.40 mg/g) were highest in water extracts followed by ethanol and acetone extracts. Thus, the extract yield and sugars content of clove are dependent on the kind of extraction technique used and solvent polarity.
... Estimation of total sugars content: Total sugars content was determined by phenol sulphuric acid method [11,12] using glucose as standard. The total sugars content present in various extracts was calculated and results were expressed as milligrams per gram (mg/g). ...
... Estimation of reducing sugars content: Reducing sugars content was determined by Nelson-Somogyi method [12,13] using glucose as standard. The reducing sugars content present in various extracts was calculated and results were expressed as milligrams per gram (mg/g). ...
Article
Moisture content in a plant matrix is an important factor as the nature of phyto-constituents present in spices may be altered by the presence of moisture. The present study was conducted to observe the effect of moisture levels viz. 5 % moisture level (more than normal), 10 % moisture level (more than normal) and normal moisture level (control) on phyto-constituents and antioxidant activities of clove buds. Contents of total phenols, flavonoids, sugars, DPPH free radical scavenging activity and antioxidant activity decreased under the influence of moisture conditions during storage. The contents of total phenols in samples with 10 % and 5 % and normal moisture level after 64 days of storage were 70.89, 73.76 and 96.07 mg GAE/g, flavonoids (21.02, 23.37 & 38.24 mg CE/g), total sugars (12.73, 13.88 & 23.01 mg/g), reducing sugars (12.59, 13.73 and 22.77 mg/g), non-reducing sugars (0.14, 0.15 and 0.24 mg/g), IC50 values from DPPH free radical scavenging activity (253.0, 231.7 and 124.9 μg/mL) and antioxidant activity (51.05, 55.19 and 84.67 %). The contents of various phytochemicals and antioxidant activity decreased more at 10 % moisture level in comparison to 5 % moisture level.
... Various compounds, including withanolide B, rosifoliol, and phytol, were reported [58]. Gulati et al. [59] studied the chemical composition of various extracts from W. somnifera roots of different genotypes and stated several metals in its composition, along with different concentrations of total sugars, alkaloids, and tannins. Bhatia et al. [60], studying the effect of chemotype variations in the chemical composition of W. somnifera fruits using GC-MS and nuclear magnetic resonance (NMR) spectroscopy, stated clear variations in metabolites contents in different chemotypes. ...
Article
Full-text available
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.
... 663 General chemical composition of roots was determined. 664 First isolation and characterization of isopelletierine ( Figure 27). 665 Seven new withanolide glycosides were isolated from the methnolic root extract. ...
Article
Full-text available
Solanaceae is one of the most studied among all plant families. Plants of this family are very rich with medicinally active natural products, some of them, with nutritional importance. Some of these compounds are psychoactive and mind altering. Alkaloids, physalins and withanolides are the major notable active natural products present in these plants. In this review article, we will present the vast majority of published research about these plants. This work will include biological and medicinal activities, extended presentation of natural products, their activities and in some cases, their synthesis, biosynthesis and production, along with other chemical aspects. Despite all the wideness of this article, we have introduced limited (but not ignored. presentations of two important aspects of this plant family, mainly due to the large size of this article. One, ethnobotany and ethnomedicine uses of these plants, and two, the influence of various cultivation conditions on these plants, and the results of these conditions.
... The dose used in the present experimental study was calculated on the basis of in vitro antibacterial effect of WSR extract on E. coli (Kumari and Gupta, 2015). Results of other workers revealed that total alkaloids content of WSR collected from the same source as that of present study varied between 0.26 and 0.31%, and tannins content varied between 0.66 and 0.84 mg/g (Gulati et al., 2017). Experimental Design and Sampling The 1-day-old chicks were divided into 2 groups viz. ...
Article
Full-text available
The present study was undertaken to investigate the effect of aqueous Withania somnifera root (WSR) extract in broiler chicks experimentally infected with Escherichia coli O78 @ 10⁷ CFU/0.5 ml intraperitoneally. Clinical signs and mortality due to colibacillosis observed in infected chicks were mild and lasted for short duration in WSR extract supplemented group as compared with the nonsupplemented group. A significant increase in serum alanine transaminase, aspartate transaminase, lactate dehydrogenase, and creatine phosphokinase activities and a decrease in total protein and albumin concentrations were observed in the infected groups, though these changes were of lower magnitude in WSR extract supplemented group. A significantly higher activity of oxidative blood parameters such as superoxide dismutase, catalase, glutathione reductase, and glutathione-S-transferase enzymes were noticed in WSR extract supplemented group. The WSR extract supplemented group revealed significantly higher E. coli–specific antibody titer and enhanced lymphocyte proliferation response as compared with the nonsupplemented group. The gross and histopathological lesions of colibacillosis were mild in the WSR extract–supplemented infected group as compared with the nonsupplemented infected group. Withania somnifera root extract supplementation produced 31.48 and 34.38% protection in the gross and histopathological lesions in E. coli infected chicks, respectively. It is concluded that supplementation of 20% WSR extract @ 20 ml/L of water caused a reduction in the severity, mortality, and recovery period of E. coli infection and enhanced the humoral and cellular immune responses suggesting its protective effect on limiting the pathology of E. coli infection in broiler chickens.
Article
Full-text available
Article
Full-text available
Ashwagandha (Withania somnifera (L.) is a perennial medicinal plant that belongs to family Solanaceae. This is popularly known as Indian ginseng as it has rejuvenating and stimulating potential. The rapid and indiscriminate use of antimicrobial synthetic drugs, have necessitated the need to use ashwagandha as an antimicrobial source plant. The plant is a "sleep-inducer and it is highly medicinal. The leaves are useful in painful swelling, the flowers are aphrodisiac and the seeds are anti-helminthic. This has a wide spectrum of actions and applications against diseases. The roots are useful in promoting health and longevity, the plants are rich source of phytochemicals such as steroids,
Article
Full-text available
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.
Article
Full-text available
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.
Article
Full-text available
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.
Article
Full-text available
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.
Article
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.
Article
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 © . .
Article
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.
Article
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.