ASWAGANDHARISHTAM

Vaidyaratnam P.S. Varier’s

KOTTAKKAL ARYA VAIDYA SALA,
Kottakkal – 676 503,
Malappuram Dt., Kerala State, India.
Tel. : +91 483 2808000, 2742216-19
Fax : +91 483 2742572
Email : avsho@sancharnet.in, qa@aryavaidyasala.com
Website : www.aryavaidyasala.com
 MONOGRAPH FOR ASWAGANDHARISHTAM

Aswagandharishtam is a fermented liquid preparation made with the ingredients in the

Formulation composition given below. It contains not more than 12 per cent, and not less than 4
per cent of alcohol that is self generated in the preparation over a period of time.

Name and business address of the

holder of the certificate of registration : Kottakkal Arya Vaidya Sala, Kottakkal –676 503,
Malappuram, Dist., Kerala, India
Registration number : 45/25D/87, Under Schedule I
Proprietary name and dosage form : Aswagandharishtam, liquid
Composition : Aswagandha, Musali, Manjishta, Haritaki, Rajani, Madhuka, Rasna, Vidari,
Partha, Mustha, Trivrita, Anantha, Syama, Chandana, Vacha, Chitraka,
Madhu, Dhataki, Vyosha, Trijata, Priyangu, Nagakesra.

CLASSICAL REFERENCE DETAILS

Pharmacological classification : Time tested classical formulation (Ref. Text :
Bhaishajyaratnavali), for Epilepsy, Rejuvenative, Piles,
Rheumatism
Indications : Dullness, Loss of Memory, Sluggishness, Epilepsy, Insanity and Emaciation
Contra-indications : Not Reported
Dosage and directions for use : 15 – 25 ml, as instructed by the Physician
Side effects and special precautions: Adverse reactions not reported. (Caution : Pregnancy and
Lactation)
Presentation : 450 ml PET containers
Storage Instructions : Keep in a cool, dry place

FORMULATION COMPOSITION DETAILS

Each dose of the medicine is prepared out of :

SN Botanical Name Quantity per unit dosage

1. Withania somnifera 0.617 g
2. Curculigo orchioides 0.247 g
3. Terminalia chebula 0.123 g
4. Rubia cordifolia 0.123 g
5. Curcuma longa 0.123 g
6. Glycyrrhiza glabra 0.123 g
7. Alpinia galanga 0.123 g
8. Pueraria tuberosa 0.123 g
SN Botanical Name Quantity per unit dosage
9. Terminalia arjuna 0.123 g
10. Cyperus rotundus 0.123 g
11. Operculina turpethum 0.123 g
12. Hemidesmus indicus 0.099 g
13. Ichnocarpus frutescens 0.099 g
14. Santalum album 0.099 g
15. Acorus calamus 0.099 g
16. Plumbago indica 0.099 g
17. Honey 3.704 g
18. Woodfordia fruticosa 0.198 g
19. Zingiber officinale 0.008 g
20. Piper nigrum 0.008 g
21. Piper longum 0.008 g
22. Elettaria cardamomum 0.016 g
23. Cinnamomum verum 0.016 g
24. Cinnamomum tamala 0.016 g
25. Callicarpa macrophylla 0.049 g
26. Mesua ferrea 0.025 g
 PICTURES OF RAW MATERIAL HERBS OF ASWAGANDHARISHTAM

Figure 1 Acorus calamus Figure 2 Alpinia galanga

Figure 3 Cinnamomum tamala Figure 4 Cinnamomum verum

Figure 5 Curculigo orchioides Figure 6 Curcuma longa

Figure 7 Cyperus rotundus Figure 8 Elettaria cardamomum

Figure 9 Glycyrrhiza glabra Figure 10 Hemidesmus indicus

Figure 11 Ichnocarpus frutescens Figure 12 Mesua ferrea

Figure 13 Operculina turpethum Figure 14 Piper longum

Figure 15 Piper nigrum Figure 16 Plumbago indica

Figure 17 Pueraria tuberosa Figure 18 Rubia cordifolia

Figure 19 Santalum album Figure 20 Terminalia arjuna

Figure 21 Terminalia chebula Figure 22 Withania somnifera

Figure 23 Woodfordia fruticosa Figure 24 Zingiber officinale

 PUBLISHED RESEARCH DATA OF SOME INGREDIENT RAW MATERIALS

Alpinia galanga

Modulation of antioxidant defense by Alpinia galanga and Curcuma aromatica extracts

correlates with their inhibition of UVA-induced melanogenesis
Cell Biology and Toxicology DOI 10.1007/s10565-009-9121-2
Uraiwan Panich & Kamolratana Kongtaphan & Tassanee Onkoksoong & Kannika Jaemsak &
Rattana Phadungrakwittaya & Athiwat Thaworn & Pravit Akarasereenont & Adisak
Wongkajornsilp

This is the first report representing promising findings on AG and CA extract-derived

antityrosinase properties correlated with their antioxidant potential. Inhibiting cellular oxidative
stress and improving antioxidant defenses might be the mechanisms by which the extracts yield
the protective effects on UVA-dependent melanogenesis.

Berberis aristata

Antihyperglycemic and antioxidant effect of Berberis aristata root extract and its role in
regulating carbohydrate metabolism in diabetic rats
Jyotsna Singh and Poonam Kakkar
Journal of Ethnopharmacology, Volume 123, Issue 1, 4 May 2009, Pages 22-26

BA root extract (250 mg/kg) was administered to diabetic rats and standard drug glybenclamide
(0.6 mg/kg) to group serving as positive control. Effect of extract on antioxidant and carbohydrate
metabolism regulating enzymes of liver was studied in diabetic rats along with its safety
parameters.

The main constituents of root were identified as berberine, berbamine and palmatine through
HPTLC. The extract besides being safe, lowered the blood glucose significantly without any
hypoglycemic effect on their control counterparts. It increased CAT, SOD, GPx, GR activity
significantly and reduced lipid peroxidation (41.6%) and protein carbonylation (30.15%). It also
increased the glucokinase and glucose-6-phosphate dehydrogenase activities and decreased
glucose-6-phosphatase activity in diabetic rats, which play a critical role in glucose homeostasis.

The extract of Berberis sristata (root) has strong potential to regulate glucose homeostasis
through decreased gluconeogenesis and oxidative stress.

Acorus calamus

Neuroprotective effect of Acorus calamus against middle cerebral artery occlusion–induced

ischaemia in rat
Pradeep K Shukla, Industrial Toxicology Research Centre, PO Box 80, M.G. Marg, Lucknow
226001, India.

The neuroprotective potential of ethanol:water (1:1) extract of rhizomes of Acorus calamus (AC–
002) has been investigated in middle cerebral artery occlusion (MCAO)–induced ischaemia in
rats. A significant behavioural impairment in Rota–Rod performance and grid walking was
observed in rats, 72 hours after MCAO as compared to sham–operated animals. These rats also
exhibited an increase in lipid peroxidation (cortex / 157%, corpus striatum – 58%) and a decrease
in glutathione levels (cortex – 59%, corpus striatum – 34%) and superoxide dismutase (SOD)
activity (cortex – 64%, corpus striatum – 32%) as compared to sham–operated animals. Ischaemic
rats treated with AC–002 (25 mg/kg, p.o.) exhibited a significant improvement in
neurobehavioural performance viz. Rota–Rod performance and grid walking as compared to the
MCAO group. Interestingly, treatment with AC–002 in MCAO rats significantly decreased
malonaldialdehyde levels in cortex as compared to ischaemic rats. A significant increase in
reduced glutathione levels and SOD activity was observed both in cortex and corpus striatum in
MCAO rats treated with AC–002 in comparison to MCAO rats. Treatment with AC–002 in
MCAO rats also reduced the contralateral cortical infarct area (19%) as compared to MCAO rats
(33%). Neurological function score was improved in the AC–002–treated rats as compared to the
MCAO group. The results of the present study indicate the neuroprotective efficacy of A. calamus
in the rat model of ischaemia

Curcuma longa

Neuroprotective efficacy and therapeutic window of curcuma oil: in rat embolic stroke
model.
BMC Complementary and Alternative Medicine 2008
Preeti Dohare, Puja Garg, Uma sharma, NR Jagannathan and Madhur Ray

Among the naturally occurring compounds, turmeric from the dried rhizome of the plant Curcuma
longa has long been used extensively as a condiment and a household remedy all over Southeast
Asia. Turmeric contains essential oil, yellow pigments (curcuminoids), starch and oleoresin. The
present study was designed for investigating the neuroprotective efficacy and the time window for
effective therapeutic use of Curcuma oil (C.

Curcuma longa extract supplementation reduces oxidative stress and attenuates aortic fatty
streak development in rabbits,
Arterioscler Thromb Vasc Biol. 2002 Jul 1;22(7):1225-31

This study evaluates the effect of a Curcuma longa extract on the development of experimental
atherosclerosis (fatty streak) in rabbits and its interaction with other plasmatic antioxidants. Two
experimental groups of male New Zealand White rabbits, a control group and a curcuma-extract
(CU) group, were fed an atherogenic diet. Additionally, the CU group received an oral curcuma
hydroalcoholic extract. Six animals from each experimental group were killed after 10, 20, and 30
days. Compared with the CU group, the control group showed significantly higher plasma lipid
peroxide at all experimental times (10, 20, and 30 days) and significantly lower alpha-tocopherol
and coenzyme Q levels at 20 and 30 days. Histological results for the fatty streak lesions revealed
damage in the thoracic and abdominal aorta that was significantly lower in the CU group than in
the control group at 30 days. Supplementation with Curcuma longa reduces oxidative stress and
attenuates the development of fatty streaks in rabbits fed a high cholesterol diet.

Antidepressant activity of aqueous extracts of Curcuma longa in mice

Journal of Ethnopharmacology, Volume 83, Issues 1-2, November 2002, Pages 161-165
Curcuma longa (turmeric) is a well-known indigenous herbal medicine. The aqueous extracts,
when administered orally to the mice from 140 to 560 mg/kg for 14 days, were able to elicit dose-
dependent relation of immobility reduction in the tail suspension test and the forced swimming
test in mice. The effects of the extracts at the dose of 560 mg/kg were more potent than that of
reference antidepressant fluoxetine. The extracts, at the dose of 140 mg/kg or above for 14 days,
significantly inhibited the monoamine oxidize A (MAO) activity in mouse whole brain at a dose-
dependent manner, however, oral administration of the extract only at a dose of 560 mg/kg
produced observable MAO B inhibitory activity in animal brain. Fluoxetine showed only a
tendency to inhibit MAO A and B activity in animal brain in the study. Neither the extracts of C.
longa nor fluoxetine, at the doses tested, produced significant effects on locomotor activity. These
results demonstrated that C. longa had specifically antidepressant effects in vivo. The activity of
C. longa in antidepression may mediated in part through MAO A inhibition in mouse brain.

Neuroprotective role of curcumin from Curcuma longa on ethanol-induced brain damage

Phytotherapy Research, Volume 13 Issue 7, Pages 571 - 574
V. Rajakrishnan, P. Viswanathan , K. N. Rajasekharan , Dr. Venugopal P. Menon

In the present study, curcumin from Curcuma longa was screened for neuroprotective activity
using ethanol as a model of brain injury. Oral administration of curcumin to rats caused a
significant reversal in lipid peroxidation, brain lipids and produced enhancement of glutathione, a
non-enzymic antioxidant in ethanol intoxicated rats, revealing that the antioxidative and
hypolipidaemic action of curcumin isresponsible for its protective role against ethanol induced
brain injury. Copyright © 1999 John Wiley & Sons, Ltd.

Glycyrrhiza glabra

Neuroprotective effects of roasted licorice, not raw form, on neuronal injury in gerbil
hippocampus after transient forebrain ischemia
Acta Pharmacologica Sinica (2006) 27, 959–965; doi:10.1111/j.1745-7254.2006. In-koo Hwang,
Soon-sung Lim, Kyu-hyun Choi, Ki-yeon Yoo, Hyun-kyung Shin, Eun-ji Kim, Jung-han Yoon-
Park Tae-cheon Kang, Young-sup Kim, Dae-young Kwon, Dong-woo Kim, Won-kook Moon and
Moo-ho Won.

To observe neuroprotective effects of raw and roasted licorice against hypoxia and ischemic
damage. When elucidating the protective effects of raw and roasted licorice, we analyzed the
lactate dehydrogenase (LDH) release using PC12 cells after hypoxia in an in vitro study and after
transient forebrain ischemia in an in vivo study on Mongolian gerbils.

Raw and roasted licorice significantly reduced LDH release from PC12 cells exposed to an
hypoxic chamber for 1 h. In the roasted licorice-treated group, the decrease of LDH release was
more pronounced compared to that of the raw licorice-treated group. In roasted licorice-treated
animals, approximately 66%-71% of CA1 pyramidal cells in the ischemic hippocampus were
stained with cresyl violet compared to the control group. However, in the raw licorice-treated
animals, no significant neuroprotection against ischemic damage was shown. In addition,
ischemic animals in roasted licorice-treated group maintained the Cu, Zn-superoxide dismutase
(SOD1) activity and protein levels compared to the control group, while in raw licorice-treated
group SOD1 activity and protein levels were reduced significantly. High pressure liquid
chromatography analysis showed that non-polar compounds containing glycyrrhizin-degraded
 products, such as glycyrrhetinic acid (GA) and glycyrrhetinic acid monoglucuronide (GM), were
increased in roasted licorice.

Roasted licorice had neuroprotective effects against ischemic damage by maintaining the SOD1
levels. In addition, the difference in protective ability between raw and roasted licorice may be
associated with non-polar compounds, such as GA and GM

Protection of mitochondrial functions against oxidative stresses by isoflavans from

Glycyrrhiza glabra
Journal of pharmacy and pharmacology 2000, vol. 52, no2, pp. 219-223 (29 ref.)
Haraguchi H. ; Yoshida N.; Ishikawa H.; Tamura Y. ; Mizutani K.; Kinoshita T

Isoflavan derivatives, glabridin (1), hispaglabridin A (2), hispaglabridin B (3), 4'-O-

methylglabridin (4) and 3'-hydroxy-4'-O-methylglabridin (5), isolated from Glycyrrhiza glabra,
were investigated for their ability to protect liver mitochondria against oxidative stresses.
Mitochondrial lipid peroxidation linked to respiratory electron transport and that induced non-
enzymatically were inhibited by these isoflavans. Hispaglabridin A (2) strongly inhibited both
peroxidations and 3'-hydroxy-4'-O-methylglabridin (5) was the most effective at preventing
NADH-dependent peroxidation. 3'-Hydroxy-4'-O-methylglabridin (5) protected mitochondrial
respiratory enzyme activities against NADPH-dependent peroxidation injury.
Dihydroxyfumarate-induced mitochondrial peroxidation was also prevented by this isoflavan.
Isoflavans from G. glabra were shown to be effective in protecting mitochondrial function against
oxidative stresses.

Memory enhancing activity of Glycyrrhiza glabra in mice

Journal of Ethnopharmacology , Volume 91, Issues 2-3, April 2004, Pages 361-365
Dinesh Dhingra, , Milind Parle, , and S. K. Kulkarni

In the traditional system of medicine, the roots and rhizomes of Glycyrrhiza glabra (family:
Leguminosae) have been employed clinically for centuries for their anti-inflammatory, antiulcer,
expectorant, antimicrobial and anxiolytic activities. The present study was undertaken to
investigate the effects of Glycyrrhiza glabra (popularly known as liquorice) on learning and
memory in mice. Elevated plus-maze and passive avoidance paradigm were employed to test
learning and memory. Three doses (75, 150 and 300 mg/kg p.o.) of aqueous extract of
Glycyrrhiza glabra were administered for 7 successive days in separate groups of animals. The
dose of 150 mg/kg of the aqueous extract of liquorice significantly improved learning and
memory of mice. Furthermore, this dose significantly reversed the amnesia induced by diazepam
(1 mg/kg i.p.) and scopolamine (0.4 mg/kg i.p.). Anti-inflammatory and antioxidant properties of
liquorice may be contributing favorably to the memory enhancement effect. Since scopolamine-
induced amnesia was reversed by liquorice, it is possible that the beneficial effect on learning and
memory was due to facilitation of cholinergic-transmission in mouse brain. However, further
studies are necessitated to identify the exact mechanism of action. In the present investigation,
Glycyrrhiza glabra has shown promise as a memory enhancing agent in all the laboratory models
employed.

In vitro and in vivo neuroprotective effect and mechanisms of glabridin, a major active
isoflavan from Glycyrrhiza glabra (licorice)
Life Sci. 2008 Jan 2;82(1-2):68-78. Epub 2007 Nov 7
Yu XQ, Xue CC, Zhou ZW, Li CG, Du YM, Liang J, Zhou SF.
Stroke is a life-threatening disease characterized by rapidly developing clinical signs of focal or
global disturbance of cerebral function due to cerebral ischemia. A number of flavonoids have
been shown to attenuate the cerebral injuries in stroked animal models. Glabridin, a major
flavonoid of Glycyrrhiza glabra (licorice), possesses multiple pharmacological activities. This
study aimed to investigate whether glabridin modulated the cerebral injuries induced by middle
cerebral artery occlusion (MCAO) in rats and staurosporine-induced damage in cultured rat
cortical neurons and the possible mechanisms involved. Our study showed that glabridin at
25mg/kg by intraperitoneal injection, but not at 5mg/kg, significantly decreased the focal infarct
volume, cerebral histological damage and apoptosis in MCAO rats compared to sham-operated
rats. Glabridin significantly attenuated the level of brain malonyldialdehyde (MDA) in MCAO
rats, while it elevated the level of two endogenous antioxidants in the brain, i.e. superoxide
dismutase (SOD) and reduced glutathione (GSH). Co-treatment with glabridin significantly
inhibited the staurosporine-induced cytotoxicity and apoptosis of cultured rat cortical neurons in a
concentration-dependent manner. Consistently, glabridin significantly reduced the DNA laddering
caused by staurosporine in a concentration-dependent manner. Glabridin also suppressed the
elevated Bax protein and caspase-3 proenzyme and decreased bcl-2 induced by staurosporine in
cultured rat cortical neurons, facilitating cell survival. Glabridin also inhibited superoxide
production in cultured cortical neurons exposed to staurosporine. These findings indicated that
glabridin had a neuroprotective effect via modulation of multiple pathways associated with
apoptosis. Further studies are warranted to further investigate the biochemical mechanisms for the
protective effect of glabridin on neurons and the evidence for clinical use of licorice in the
management of cerebral ischemia.

Curculigo orchioides

Antioxidant activity of curculigo orchioides in carbon tetrachlorideinduced hepatopathy in

rats
M.R. Venukumar And M.S. Latha, Indian Journal Of Clinical Biochemistry, 2002, 17 (2) 80-87
Pharmacognosy Research Laboratory, School of Biosciences, Mahatma Gandhi University, P.D.
Hills P.O., Kottayam - 686560, Kerala.

In this study antioxidant activity of methanol extract of rhizomes of Curculigo orchioides (MEC)
was investigated using carbon tetrachloride (CCl4)- intoxicated rat liver as the experimental
model. The hepatotoxic rats were administered MEC for 90 days (daily, orally at the dose of 70
mg per kg body weight). Lipid peroxidation (LPO) in CCl4 - intoxicated rats was evidenced by a
marked increment in the levels of thiobarbituric acid reactive substances (TBARS) and diene
conjugates (CD), and also a distinct diminution in glutathione (GSH) content in the liver. In CCl4
+ MEC – treated rats these biochemical parameters attained an almost normal level.

The decreased activity of antioxidant enzymes, such as superoxide dismutase (SOD), catalase
(CAT), glutathione peroxidase (GPX) and glutathione reductase (GRD) in CCl4 –intoxicated rats,
and its retrieval towards near normalcy in CCl4 + MEC- administered rats revealed the efficacy of
MEC in combating oxidative stress due to hepatic damage. Elevated level of glutathione
transferase(GTS) observed in hepatotoxic rats too showed signs of returning towards normalcy in
MEC co-administered animals, thus corroborating the antioxidant efficacy of MEC. The findings
provide a rationale for further studies on isolation of active principles and its pharmacological
evaluation.
Elettaria cardamomum

Antioxidant and antimicrobial activities of essential oil and various oleoresins of Elettaria
cardamomum (seeds and pods)
Singh, Gurdip; Kiran, Shashi; Marimuthu, Palanisamy; Isidorov, Valery; Vinogorova, Vera.
Journal of the Science of Food and Agriculture, Volume 88, Number 2, 30 January 2008, pp. 280-
289(10)

This paper describes the chemical analysis of the essential oil and various oleoresins of Elettaria
cardamomum (seeds and pods) by gas chromatography (GC) and gas chromatography/mass
spectrometry (GC/MS) techniques. It also compares the effects of the different extraction solvents
used (chloroform, methanol, ethanol and diethyl ether) on the antioxidant and antimicrobial
activities of the essential oil and oleoresins.

Hemidesmus indicus

Antioxidant effect of Hemidesmus indicus on ethanol-induced hepatotoxicity in rats

Nadana Saravanan, Namasivayam Nalini,
Journal of medicinal food. 01/01/2008; 10(4):675-82

The antioxidant effect of the ethanolic root extract of Hemidesmus indicus, an indigenous
Ayurvedic medicinal plant used in soft drinks in India, was studied in rats with ethanol-induced
hepatotoxicity. Administering 20% ethanol (5 g/kg of body weight/day) for 60 days to male
Wistar rats resulted in significantly decreased body weight and increased liver/body weight
ratio. The liver marker enzymes, aspartate transaminase (AST), alanine transaminase (ALT),
alkaline phosphatae (ALP), gamma-glutamyl transpeptidase (GGT), and lactate dehydrogenase
(LDH), were elevated. In addition, the levels of plasma, erythrocyte, and hepatic thiobarbituric
acid-reactive substances (TBARS), hydroperoxides (LOOH), and conjugated dienes (CD) were
also elevated in ethanol-fed rats as compared to those of the experimental control rats.
Decreased activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase
(GPx), reduced glutathione (GSH), vitamin C, and alpha-tocopherol (vitamin E) were also
observed in ethanol-administered as compared to control rats. Ethanolic root extract of H.
indicus was administered at a dose of 500 mg/kg of body weight/day for the last 30 days of the
experiment to rats with ethanol-induced liver injury, which significantly increased body weight,
significantly decreased the liver/body weight ratio, AST, ALT, ALP, GGT, and LDH activities,
and also the levels of TBARS, LOOH, and CD, significantly elevated the activities of SOD,
CAT, GPx, and GSH in plasma, erythrocytes, and liver, and also increased levels of plasma and
liver vitamin C and vitamin E at the end of the experimental period as compared to those of
untreated ethanol-administered rats. Thus, our data indicate that treatment with H. indicus
extract offers protection against free radical-mediated oxidative stress in plasma, erythrocytes,
and liver of animals with ethanol-induced liver injury.

In vitro Biosynthesis of Antioxidants from Hemidesmus indicus R. Br. Cultures.

Neeta Misra, Pratibha Misra, S. K. Datta, In Vitro Cellular and Developmental Biology - Plant
41(3):285-290. 2005

(Asclepiadaceae) were established on Murashige and Skoog medium with various hormonal
combinations. The production of antioxidants (lupeol, vanillin, and rutin) in shoot cultures, callus
cultures derived from leaf cells and root cells, was compared with root and aerial portions of the
parent plant. Shoot cultures and leaf callus cultures produced more antioxidants than root callus
cultures. In vitro culture of this species might offer an alternative method for production of these
important pharmaceuticals, which would reduce the collection pressure on this rare Shoot cultures
and callus cultures from roots and leaves of Hemidesmus indicus R. Br. Plant

Inhibition of cutaneous oxidative stress and two-stage skin carcinogenesis by Hemidesmus

indicus (L.) in Swiss albino mice
Sultana,-S; Alam,-A; Khan,-N; Sharma,-S, Indian-J-Exp-Biol. 2003 Dec; 41(12): 1416-23

Chemopreventive potential of H. indicus on 7,12-dimethyl-benz[a]anthracene (DMBA)-initiated

and 12-O-tetradecanoyl 13-phorbol acetate (TPA) promoted murine skin carcinogenesis has been
assessed. Topical application of H. indicus resulted in significant protection against cutaneous
tumorigenesis. Topical application of plant extract prior to that of TPA resulted in significant
inhibition against TPA-caused induction of epidermal ornithine decarboxylase (ODC) activity and
DNA synthesis. Application of H. indicus at a dose level of 1.5 and 3.0 mg/kg body weight in
acetone prior to that of TPA treatment resulted in significant inhibition of oxidative stress. The
level of lipid peroxidation was significantly reduced. In addition, depleted levels of glutathione
and reduced activities of antioxidant enzymes were restored respectively). The results indicate
that H. indicus is a potent chemopreventive agent in skin carcinogenesis.

Honey

Reproductive protein protects functionally sterile honey bee workers from oxidative stress
Siri-Christine Seehuus, Kari Norberg, Ulrike Gimsa, Trygve Krekling and Gro V. Amdam.

Research on aging shows that regulatory pathways of fertility and senescence are closely
interlinked. However, evolutionary theories on social species propose that lifelong care for
offspring can shape the course of senescence beyond the restricted context of reproductive
capability. These observations suggest that control circuits of aging are remodeled in social
organisms with continuing care for offspring. Here, we studied a circuit of aging in the honey bee
(Apis mellifera). The bee is characterized by the presence of a long-lived reproductive queen caste
and a shorter-lived caste of female workers that are life-long alloparental care givers. We focus on
the role of the conserved yolk precursor gene vitellogenin that, in Caenorhabditis elegans,
shortens lifespan as a downstream element of the insulin/insulin-like growth factor signaling
cascade. Vitellogenin protein is synthesized at high levels in honey bee queens and is abundant in
long-lived workers. We establish that vitellogenin gene activity protects worker bees from
oxidative stress. Our finding suggests that one mechanistic explanation for patterns of longevity in
bees is that a reproductive regulatory pathway has been remodeled to extend life. This perspective
is of considerable relevance to research on longevity regulation that builds largely on inference
from solitary model species.

Inhibition of methylnitrosourea (MNU) induced oxidative stress and carcinogenesis by

orally administered bee honey and Nigella grains in Sprague Dawely rats.
J Exp Clin Cancer Res. 2002 Sep;21(3):341-6.
Mabrouk GM, Moselhy SS, Zohny SF, Ali EM, Helal TE, Amin AA, Khalifa AA.
Oncology Diagnostic Unit, Faculty of Science, Ain Shams University, Abbassia, Cairo, Egypt.

We studied the protective effect of bee honey and Nigella grains as nutraceuticals on the oxidative
stress and carcinogenesis induced by methylnitrosourea (MNU) in Sprague Dawely rats. Four
groups of animals were used and fed ad-libitum. The first group was a control (n=8), the second
(n=8), the third (n=15) and the fourth groups (n=12) were injected MNU (single i.v. dose 50
mg/kg body weight). After one week the third and fourth groups were given orally 0.2 g ground
Nigella grains and 0.2 g Nigella with 5 g honey/rat/day, respectively. After six months all animals
were sacrificed except two from the second group (MNU-injected rats) that died one-week before
the end of the experiment. We observed that MNU injected in the second group produced a
variety of oxidative stresses ranging from severe inflammatory reaction in lung and skin to colon
adenocarcinoma in four out of six animals. There was an associated elevation of malondialdehyde
(MDA) and nitric oxide (NO) in sera obtained from animals of this group compared to the control
one. Nigella sativa grains given orally protected against MNU-induced oxidative stress and
carcinogenesis by 80% (12/15) and combated this effect by lowering MDA and NO. Whereas
honey from bees and Nigella sativa together protected 100% (12/12) against MNU-induced
oxidative stress, carcinogenesis and abolished the NO and MDA elevations shown in sera of
animals who did not receive these nutrients. These results showed that supplementation of diet
with honey and Nigella sativa has a protective effect against MNU-induced oxidative stress,
inflammatory response and carcinogenesis.

Operculina turpethum

Anbuselvam C, Vijayavel K, Balasubramanian MP

Protective effect of Operculina turpethum against 7, 12-dimethyl benz (a)anthracene induced
oxidative stress with reference to breast cancer in experimental rats.
Chem Biol Interact 2007 Apr 22

Reactive oxygen species (ROS) directly or indirectly involves in multistage process of

carcinogenesis. Antioxidant activity of methanolic extract of Operculina turpethum stems
(MEOT) on 7,12 dimethylbenz(a)anthracene (DMBA) induced breast cancer was investigated in
female Sprague-Dawley rats. Changes in the levels of lipid peroxidation and antioxidants system
was evaluated in addition to tumour development.

Piper longum

Protective effect of Piper longum L. on oxidative stress induced injury and cellular
abnormality in adriamycin induced cardiotoxicity in rats.
Indian J Exp Biol. 2008 Jul;46(7):528-33. Wakade AS, Shah AS, Kulkarni MP, Juvekar AR.

Effect of methanolic extract of fruits of P. longum (PLM) on the biochemical changes, tissue
peroxidative damage and abnormal antioxidant levels in adriamycin (ADR) induced
cardiotoxicity in Wistar rats was investigated. The results indicate that PLM administration offers
significant protection against ADR induced oxidative stress and reduces the cardiotoxicity by
virtue of its antioxidant activity.

Protective effect of Piper longum fruits against experimental myocardial oxidative stress
induced injury in rats
Wakade, Alok S., Mrugaya P. Kulkarni and Archana R. Juvekar, Journal of Natural Remedies 9,1,
43 – 50, Piperine modulation of carcinogen induced oxidative stress in intestinal mucosa
Molecular and Cellular Biochemistry, Volume 189, Numbers 1-2 / December, 1998
113-118, Annu Khajuria, Neelima Thusu, Ushu Zutshi and K.L. Bedi

Reactive oxygen species (ROS) and reactive metabolic intermediates generated from various
chemical carcinogens are known to play an important role in cell damage and in the initiation and
progression of carcinogenesis. Many radical scavengers, interestingly naturally occuring
antioxidants have been found to be effective in inhibiting the induction of carcinogenesis by a
wide variety of chemical carcinogens. Studies have also indicated that various spice principles
form an important group as antioxidants. In the present study our goal was to investigate whether
piperine an pungent principle of black and long peppers was able to inhibit or reduce the oxidative
changes induced by chemical carcinogens in rat intestinal model. Carcinogenesis was initiated in
intestinal lumen of male rats with 7,12, dimethyl benzanthracene, dimethyl amino-methyl
azobenzene and 3-methyl cholenthrene. Oxidative alterations were assessed by determining
thiobarbituric reactive substances, mainly malonaldehyde (as a measure of lipid peroxidation),
thiol status and expression of -GT and Na+-K+-ATPase activity in intestinal mucosa. Data
indicated that carcinogens treatment induced GSH depletion with substantial increase in
thiobarbituric reactive substances and enzyme activities. Piperine treatment with carcinogens
resulted in inhibition of thiobarbituric reactive substances. It mediated a significant increase in the
GSH levels and restoration in -GT and Na+-K+-ATPase activity. The studies thus indicate a
protective role of piperine against the oxidative alterations by carcinogens. It may be suggested
that piperine modulates the oxidative changes by inhibiting lipid peroxidation and mediating
enhanced synthesis or transport of GSH thereby replenishing thiol redox. Piperine - Piper
species - 7,12,dimethyl benzanthracene - dimethyl amnino-methyl azobenzene - 3-methyl
cholenthrene - radical scavenger - cytoprotection

Protective effects of Piper nigrum and Vinca rosea in alloxan induced diabetic rats
M. Kaleem, Sheema, H. Sarmad And B. Bano, Indian J Physiol Pharmacol 2005; 49 (1) : 65–71

In the present study aqueous extract of Piper nigrum seeds and Vinca rosea flowers were
administered orally to alloxan induced diabetic rats once a day for 4 weeks. These treatments lead
to significant lowering of blood sugar level and reduction in serum lipids. The levels of
antioxidant enzymes, catalase and glutathione peroxidase decreased in alloxan induced diabetic
rats however these levels returned to normal in insulin, P. nigrum and V. rosea treated rats. There
was no significant difference in superoxide dismutase activity in all groups compared to controls.
Lipid peroxidation levels were significantly higher in diabetic rats and it was slightly increased in
insulin, P. nigrum and V. rosea treated rats as compared to control rat. These results suggest that
oxidative stress
plays a key role in diabetes, and treatment with P. nigrum and V. rosea are useful in controlling
not only the glucose and lipid levels but these components may also be helpful in strengthening
the antioxidants potential.

Antioxidant efficacy of black pepper (Piper nigrum L.) and piperine in rats with high fat diet induced
oxidative stress
R.S. Vijayakumar; D. Surya; N. Nalini, Redox Report, Volume 9, Number 2, April 2004 , pp.
105-110(6)

The present study was aimed to explore the effect of black pepper (Piper nigrum L.) on tissue
lipid peroxidation, enzymic and non-enzymic antioxidants in rats fed a high-fat diet. Thirty male
Wistar rats (95–115 g) were divided into 5 groups. They were fed standard pellet diet, high-fat
diet (20% coconut oil, 2% cholesterol and 0.125% bile salts), high-fat diet plus black pepper (0.25
g or 0.5 g/kg body weight), high-fat diet plus piperine (0.02 g/kg body weight) for a period of 10
weeks. Significantly elevated levels of thiobarbituric acid reactive substances (TBARS),
conjugated dienes (CD) and significantly lowered activities of superoxide dismutase (SOD),
catalase (CAT), glutathione peroxidase (GPx), glutathione-S-transferase (GST) and reduced
glutathione (GSH) in the liver, heart, kidney, intestine and aorta were observed in rats fed the high
fat diet as compared to the control rats. Simultaneous supplementation with black pepper or
piperine lowered TBARS and CD levels and maintained SOD, CAT, GPx, GST, and GSH levels
to near those of control rats. The data indicate that supplementation with black pepper or the
active principle of black pepper, piperine, can reduce high-fat diet induced oxidative stress to the
cells.

Larvicidal Activity of Isobutylamides Identified in Piper nigrum Fruits

Agil-Kwon Park, Sang-Gil Lee, Sang-Chul Shin, Ji-Doo Park, and Young-Joon Against Three
Mosquito Species, School of Agricultural Biotechnology, Seoul National University, Suwon 441-
744, Republic of Korea, and Korea Forest Research Institute, Seoul 130-012, Republic of Korea,
J. Agric. Food Chem., 2002, 50 (7), pp 1866–1870.

The insecticidal activity of materials derived from the fruits of Piper nigrum against third instar
larvae of Culex pipiens pallens, Aedes aegypti, and A. togoi was examined and compared with
that of commercially available piperine, a known insecticidal compound from Piper species. The
biologically active constituents of P. nigrum fruits were characterized as the isobutylamide
alkaloids pellitorine, guineensine, pipercide, and retrofractamide A by spectroscopic analysis.
Retrofractamide A was isolated from P. nigrum fruits as a new insecticidal principle. On the basis
of 48-h LC50 values, the compound most toxic to C. pipiens pallens larvae was pipercide (0.004
ppm) followed by retrofractamide A (0.028 ppm), guineensine (0.17 ppm), and pellitorine (0.86
ppm). Piperine (3.21 ppm) was least toxic. Against A. aegypti larvae, larvicidal activity was more
pronounced in retrofractamide A (0.039 ppm) than in pipercide (0.1 ppm), guineensine (0.89
ppm), and pellitorine (0.92 ppm). Piperine (5.1 ppm) was relatively ineffective. Against A. togoi
larvae, retrofractamide A (0.01 ppm) was much more effective, compared with pipercide (0.26
ppm), pellitorine (0.71 ppm), and guineensine (0.75 ppm). Again, very low activity was observed
with piperine (4.6 ppm). Structure−activity relationships indicate that the N-isobutylamine moiety
might play a crucial role in the larvicidal activity, but the methylenedioxyphenyl moiety does not
appear essential for toxicity. Naturally occurring Piper fruit-derived compounds merit further
study as potential mosquito larval control agents or as lead compounds.

Stimulation of mouse melanocyte proliferation by Piper nigrum fruit extract and its main
alkaloid, piperine.
Lin Z, Hoult JR, Bennett DC, Raman A. Planta Med. 1999 Oct; 65(7):600-3.

During a herbal screening programme to find potential repigmenting agents for the treatment of
vitiligo, Piper nigrum L. fruit (black pepper) extract was found to possess growth-stimulatory
activity towards cultured melanocytes. Its aqueous extract at 0.1 mg/ml was observed to cause
nearly 300% stimulation of the growth of a cultured mouse melanocyte line, melan-a, in 8 days (p
< 0.01). Piperine (1-piperoylpiperidine), the main alkaloid from Piper nigrum fruit, also
significantly stimulated melan-a cell growth. Both Piper nigrum extract and piperine induced
morphological alterations in melan-a cells, with more and longer dendrites observed. The
augmentation of growth by piperine was effectively inhibited by RO-31-8220, a selective protein
kinase C (PKC) inhibitor, suggesting that PKC signalling is involved in its activity. This is the
first full report on such an activity of black pepper and piperine
Zingiber officinale

Effects of combining extracts (from propolis or Zingiber officinale) with clarithromycin on

Helicobacter pylori
A. Nostro, L. Cellini, S. Di Bartolomeo, M.A. Cannatelli, E. Di Campli, F. Procopio, R. Grande,
L. Marzio, V. Alonzo, Pharmaco-Biological Department, Section of Microbiology, University of
Messina, Messina, Italy, Departments of Biomedical Sciences and Medicine, University of Chieti,
Italy.

Propolis and Zingiber officinale have been shown to be specifically targeted against Helicobacter
pylori strains, to possess antiinflammatory, antioxidant and antitumoral activity and to be used in
traditional medicine for the treatment of gastrointestinal ailments. Considering that these natural
products could potentially serve as novel therapeutic tools also in combination with an antibiotic,
the aim of this work was to evaluate their effect when combined with clarithromycin on clinical
H. pylori isolates (n = 25), characterized in respect to both clarithromycin susceptibility and the
presence of the cagA gene. The results showed that the combinations of propolis extract +
clarithromycin and Z. officinale extract + clarithromycin exhibited improved inhibition of H.
pylori with synergistic or additive activity. Interestingly, the susceptibility to combinations was
significantly independent of the microbial clarithromycin susceptibility status. Only one H. pylori
strain showed antagonism towards the Z. officinale extract + clarithromycin combination. The
data demonstrate that combinations of propolis extract + clarithromycin and Z. officinale extract +
clarithromycin have the potential to help control H. pylori-associated gastroduodenal disease.
Copyright © 2006 John Wiley & Sons, Ltd.

Rubia cordifolia

In vivo evaluation of antioxidant activity of alcoholic extract of Rubia cordifolia linn. And its
influence on ethanol-induced immunosuppression
Indian journal of pharmacology 2003; 35: 232-236, A.A. Joharapurkar, S.P. Zambad, M.M.
Wanjari, S.N. Umathe

To evaluate the in vivo antioxidant activity of alcoholic extract of the roots of Rubia cordifolia
Linn. (RC) and to study its influence on ethanol-induced impairment of immune responses.

Chronic administration of ethanol decreased the humoral and cell-mediated immune response,
phagocytosis, phagocytosis index, TLC, GSH, CAT and SOD activities and increased the LPO.
These influences of ethanol were prevented by concurrent daily administration of RC and the
effect was comparable with that of the combination of vitamin E and C.

The ethanol-induced immunosuppression is due to oxidative stress and Rubia cordifolia can
prevent the same by virtue of its in vivo antioxidant property.
Ethyl alcohol impaired immunity oxidative stress

In vitro evaluation of free radical scavenging activity of Rubia cordifolia L.

To evaluate the free radical scavenging activity of Rubia cordifolia L.in vitro.MethodsThe
extract/fractions viz 80% methanol extract (RME),hexane fraction (RHF),chloroform fraction
(RCF),ethyl acetate fraction (REA),n-butanol fraction (RBF) and precipitates (RPP) were isolated
from the roots of Rubia cordifolia L.and tested for the free radical scavenging activity by 2,2-
diphenyl-1-picrylhydrazyl (DPPH) method.The data were analyzed for statistical significance
using analysis of variance (one way ANOVA).Results. The RHF and REA showed maximum free
radical scavenging activity of 96.74% and 94.76% respectively at a concentration of 1 mg/ml.
From the dose response curves, present study showed very high correlation coefficient of the
concentration used and radical scavenging activity of all the extract/fractions. The phytochemical
studies show the presence of anthraquinones and their glycosides. Conclusion The tested
extract/fractions of Rubia cordifolia possess constituents, which can be effectively exploited in
food industry as chemopreventive agents.

Neuroprotective studies of Rubia cordifolia Linn. On β-amyloid Induced Cognitive

Dysfunction in Mice
Chitra V., Pavan Kumar K. Department Of Pharmacology, International Journal of PharmTech
Research, Vol.1, No.4, pp 1000-1009,

Rubia cordifolia is a medicinal plant with antioxidant properties. IntraCerebroVentricular

injection of Aβ 25-35 induced impairment of memory assessed by passive avoidance and Morris
water tests. Aβ 25-35 has the potential to induced oxidative stress in the brain cholinergic hypo
function, elevation of AChE and MAO.

Terminalia arjuna

Effect of Terminalia arjuna on antioxidant defense system in cancer

Biomedical and Life Sciences, Volume 36, Number 1 / January, 2009, 159-164, Nibha Verma and
Manjula Vinayak

Constant production of reactive oxygen species (ROS) during aerobic metabolism is balanced by
antioxidant defense system of an organism. Although low level of ROS is important for various
physiological functions, its accumulation has been implicated in the pathogenesis of age-related
diseases such as cancer and coronary heart disease and neurodegenerative disorders such as
Alzheimer’s disease. It is generally assumed that frequent consumption of phytochemicals derived
from vegetables, fruits, tea and herbs may contribute to shift the balance towards an adequate
antioxidant status. The present study is aimed to investigate the effect of aqueous extract of
medicinal plant Terminalia arjuna on antioxidant defense system in lymphoma bearing AKR
mice. Antioxidant action of T. arjuna is monitored by the activities of catalase, superoxide
dismutase and glutathione S transferase which constitute major antioxidant defense system by
scavenging ROS. These enzyme activities are low in lymphoma bearing mice indicating impaired
antioxidant defense system. Oral administration of different doses of aqueous extract of T. arjuna
causes significant elevation in the activities of catalase, superoxide dismutase and glutathione S
transferase. T. arjuna is found to down regulate anaerobic metabolism by inhibiting the activity of
lactate dehydrogenase in lymphoma bearing mice, which was elevated in untreated cancerous
mice. The results indicate the antioxidant action of aqueous extract of T. arjuna, which may play a
role in the anti carcinogenic activity by reducing the oxidative stress along with inhibition of
anaerobic metabolism

Effect of Terminalia arjuna stem bark on antioxidant status in liver and kidney of alloxan
diabetic rats
B. Raghavan And S. Krishna Kumar, Indian J Physiol Pharmacol 2006; 50 (2) : 133–142
Free radicals and associated oxidative stress induced by alloxan are implicated in eliciting
pathological changes in diabetes mellitus. Terminalia arjuna bark, an indigenous plant used in
ayurvedic medicine in India, primarily as a cardiotonic is also used in treating diabetes, anemia,
tumors and hypertension. The present study examined the effect of ethanolic extract (250 and 500
mg/kg body weight) of Terminalia arjuna stem bark in alloxan induced diabetic rats and its lipid
peroxidation, enzymatic and nonenzymatic activity was investigated in the liver and kidney
tissues. The extract produced significant (P<0.05) reduction in lipid peroxidation (LPO). The
effect of oral T. arjuna at the dose of 500 mg/kg body weight was more than the 250 mg/kg body
weight. The extract also causes a significant (P<0.05) increase in superoxide dismutase, catalase,
glutathione peroxidase, glutathione-s-transferase glutathione reductase and glucose-6-phosphate
dehydrogenase, reduced glutathione, vitamin A, vitamin C, vitamin E, total sulfhydryl groups
(TSH) and non protein sulfhydryl groups (NPSH) in liver and kidney of alloxan induced diabetic
rats, which clearly shows, the antioxidant property of T. arjuna bark. The result indicates that the
extract exhibit the antioxidant activity through correction of oxidative stress and validates the
traditional use of this plant in diabetic animals.

Terminalia chebula

Antioxidant Effect of Terminalia chebula Aqueous Extract on Age-related Oxidative Stress

in Heart
Ramalingam Mahesh and Vava Mohaideen Hazeena Begum, Received May 21, 2007; Revised
August 22, 2007; Accepted November 20, 2007, Iranian Journal of Pharmacology & Therapeutics
1735-2657/07/62-197-201

Reactive oxygen species (ROS) are generated via normal metabolic processes or as the products
of exogenous insults. They are capable of damaging essential biomolecules and accelerating
cancer, cardiovascular diseases and neurodegenerative diseases. In the present study, the
antioxidant role of Terminalia chebula aqueous extract was evaluated against age-related
oxidative stress in heart tissues of young and aged rats. Young and aged rats were treated with T.
chebula aqueous extract at a dose of 200mg/kg body weight in 1.5ml sterile water orally for 4
weeks. Control young and aged rats were received sterile water only. In aged rats, the increased
content of malondialdehyde (MDA) was observed. The antioxidants, superoxide dismutase
(SOD), catalase (CAT), glutathione peroxidase (GPx) activities, reduced glutathione (GSH),
vitamin C and E levels were decreased in heart tissues of aged control rats. Administration of T.
chebula to aged rats prevented the depletion of SOD, CAT, GPx activities and GSH, vitamin C
and E contents. Also, the level of MDA content was decreased in heart tissues. The results of the
present study show that T. chebula aqueous extract modulates the activities of antioxidants and
lipid peroxidation through the management of oxidant/antioxidant imbalance in rat heart tissues.

Withania somnifera

Possible neuroprotective effect of Withania somnifera root extract against 3-nitropropionic

acid-induced behavioral, biochemical, and mitochondrial dysfunction in an animal model of
Huntington's disease
J Med Food. 2009 Jun;12 (3):591-600, Kumar P, Kumar A. University Institute of Pharmaceutical
Sciences, Panjab University, Chandigarh, India.
Huntington's disease (HD) is a neurodegenerative disorder that results from the destruction of
neurons in the basal ganglia, and oxidative stress has been implicated in its pathogenesis. 3-
Nitropropionic acid (3-NP), a potent neurotoxin, has been reported to induce oxidative/nitrosative
stress and causes neurobehavioral and biochemical changes that mimic HD in humans.

Neuroprotective effects of Withania somnifera on 6-hydroxydopamine induced

Parkinsonism in rats.
Hum Exp Toxicol. 2005 Mar;24(3):137-47, Ahmad M, Saleem S, Ahmad AS, Ansari MA,
Yousuf S, Hoda MN, Islam F.

6-Hydroxydopamine (6-OHDA) is one of the most widely used rat models for Parkinson's
disease. There is ample evidence in the literature that 6-OHDA elicits its toxic manifestations
through oxidant stress. In the present study, we evaluated the anti-parkinsonian effects of
Withania somnifera extract, which has been reported to have potent anti-oxidant, anti-
peroxidative and free radical quenching properties in various diseased conditions.W. somnifera
extract was found to reverse all the parameters significantly in a dose-dependent manner. Thus,
the study demonstrates that the extract of W. somnifera may be helpful in protecting the neuronal
injury in Parkinson's disease.

The neuroprotective effect of Withania somnifera root extract in MPTP-intoxicated mice: an

analysis of behavioral and biochemical variables.
Cell Mol Biol Lett. 2007; 12(4):473-81. Epub 2007 Apr 6.
Sankar SR, Manivasagam T, Krishnamurti A, Ramanathan M.

We studied the influence of Withania somnifera (Ws) root extract (100 mg/kg body weight) on
parkinsonism induced by 1-methyl 4-phenyl 1,2,3,6-tetrahydropyridine (MPTP; i.p, 20 mg/kg
body weight for 4 days), via the analysis of behavioral features and the oxidant-antioxidant
imbalance in the midbrain of mice. The results indicated that at least part of the chronic stress-
induced pathology may be due to oxidative stress, which is mitigated by Ws. Further studies are
needed to assess the precise mechanism to support the clinical use of the plant as an
antiparkinsonic drug.

Withania somnifera Improves Semen Quality in Stress-Related Male Fertility

Evid Based Complement Alternat Med. 2009 Sep 29.

Stress has been reported to be a causative factor for male infertility. Withania somnifera has been
documented in Ayurveda and Unani medicine system for its stress-combating properties.

Antistressor effect of Withania somnifera

Journal of Ethnopharmacology, Volume 64, Issue 1, 1 January 1998, Pages 91-93
Withania somnifera is an Indian medicinal plant used widely in the treatment of many clinical
conditions in India. Its antistressor properties have been investigated in this study using adult
Wistar strain albino rats and cold-water swimming stress test. The results indicate that the drug
treated animals show better stress tolerance.

Neuroprotective effects of Withania somnifera Dunn. in hippocampal sub-regions of female

albino rat.
Phytother Res. 2001 Sep;15 (6):544-8
The neuroprotective effects of W. somnifera were studied on stressed adult female Swiss albino
rats. Experimental rats were subjected to immobilization stress for 14 h and were treated with a
root powder extract of W. somnifera available as Stresscom capsules (Dabur India Ltd). Control
rats were maintained in completely, non stressed conditions. Thionin stained serial coronal
sections (7 microm) of brain passing through the hippocampal region of stressed rats (E(1) group)
demonstrated 85% degenerating cells (dark cells and pyknotic cells) in the CA(2) and CA(3) sub-
areas. Treatment with W. somnifera root powder extract significantly reduced (80%) the number
of degenerating cells in both the areas. The study thus demonstrates the antistress neuroprotective
effects of W. somnifera.

PHYSICO-CHEMICAL QUALITY SPECIFICATIONS

Parameter Unit Standard

Organoleptic
Description - Clear, dark brown liquid without frothing and
significant sedimentation; with astringent taste
Chemical
Total solids % w/v 20.0 – 30.0
Specific gravity - 1.05 – 1.15
Total reducing sugar %w/v 10.0 – 25.0
Non reducing sugar %w/v NMT 1.0
pH - 3.5 – 5.0
Alcohol %v/v 4.0 – 11.0
Total phenolic content: % w/v 0.104 - 0.260
Assay of marker Micrograms per 9.0 – 12.0
Sigmasitosterol by HPTLC 50 ml
Methanol Absent
Aflatoxin
B1 ppm NMT 0.5
G1 ppm NMT 0.5
B2 ppm NMT 0.1
G2 ppm NMT 0.1
Heavy metal
Lead ppm NMT 10
Arsenic ppm NMT 3
Cadmium ppm NMT 0.3
Mercury ppm NMT 1
Microbiology
Total aerobic microbial count cfu/g NMT 105
Total yeast and mould cfu/g NMT 103
Staphylococcus aureus cfu/g Absent
Salmonella cfu/g Absent
Pseudomonas aeruginosa cfu/g Absent
Escherichia coli cfu/g Absent
PHYTOCHEMICAL FINGERPRINTING BY HIGH PERFORMANCE THIN LAYER
CHROMATOGRAPHY.

HPTLC of non-volatile methanol fraction (using in-house developed HPTLC conditions)

HPTLC fingerprint of non-volatile methanol fraction of arishtam showing the presence of
marker molecule

Calibration graph for standard levels

HPTLC of volatile toluene fraction (using in-house developed HPTLC conditions)

TLC plate views

Derivatised TLC plate view of non volatile TLC plate view at 254 nm (a) of
fraction volatile fraction and its derivatised
view (b) at visible light and 366 nm (c)