Biomedical & Pharmacology Journal Vol.

7(2), 659-663 (2014)

Hepatoprotective Effect of Sarvakalp Kwath Against Carbon

Tetrachloride Induced Hepatic Injury in Albino Rats
A. YADAV1* and S. KUMAR2

1
Department of Pharmacology, MES Medical College, Perinthalmanna, Kerala, India.
2
Department of Pharmacology, Kalinga Institute of Medical Sciences, Bhubaneswar, Odisha, India.
*Corresponding author E-mail: dr.ashishyadav@gmail.com

http://dx.doi.org/10.13005/bpj/538

(Received: September 30, 2014; accepted: November 04, 2014)

ABSTRACT

Modern medicine, even in the modern era, lacks effective medications in its armamentarium
which could be useful to treat or prevent hepatic ailments. However, Indian Ayurvedic system
claims to have many indigenous plants and herbal preparations which provide therapeutic benefit
in the treatment of liver diseases. One of such herbal preparations claiming to be of benefit in liver
diseases is Sarvakalp Kwath by Divya Pharmacy, Haridwar. This study was undertaken to explore
the hepatoprotective effects of Sarvakalp Kwath in animal model using albino rats. The animals
were administered aqueous extract of the preparation in the dose of 120 mg/kg for seven days
followed by carbon tetrachloride (CCl4) challenge. Animals were then sacrificed. Hepatic damage
and protection from the damage was assessed by biochemical parameters viz. alanine
transaminase, alkaline phosphatase, total bilirubin and albumin as well as histological studies. It
was observed that aqueous extract of Sarvakalp Kwath offered significant hepatoprotection as
reflected by improvement in all biochemical and histological parameters as compared to saline
controls. In conclusion, Sarvakalp Kwath offers significant hepatoprotection as is observed in this
study. Further studies are warranted to establish its therapeutic efficacy in humans.

Key words: Hepatoprotective, Sarvakalp, Kwath, CCl4, Divya Pharmacy, Ayurveda

INTRODUCTION the liver problems. Various indigenous plants have

been scientifically screened for hepatoprotective
Liver is the largest gland of the human activity and quite a few have been found to possess
body and also the seat of metabolism for significant activity in this regard1,2,3,4,5. Also, there
endogenous as well as xenobiotics. The fact that are some formulations which have been
the liver is constantly exposed to xenobiotics, which established as hepatoprotective agents both in
also include harmful entities, makes it vulnerable experimental as well as clinical studies. An
to ailment. Although, despite having a remarkable outstanding example of one such formulation is
reserve and regeneration capacity, continued insult Liv.52, a product of The Himalaya Drug Company6.
to the liver by toxins, alcohol, infectious agents or
cardiac decompensation result in liver disease. One of such formulations to claim
hepatoprotective action is “Sarvkalp Kwath” (Divya
Despite gigantic strides have been taken Pharmacy, Haridwar). Up till now, no scientific study
in the field of modern medicine, unfortunately, no has been performed to evaluate these claims.
significant drug has yet been developed for liver Therefore, this study was undertaken to explore
diseases. the hepatoprotective activity of this formulation in
animal models with experimentally induced
The indigenous system of Ayurvedic hepatotoxicity.
medicine claims to have a wealth of solutions for
660 YADAV & KUMAR, Biomed. & Pharmacol. J., Vol. 7(2), 659-663 (2014)

MATERIALS AND METHODS A dose limit of 2000 mg/kg of the test compound
was administered in five healthy female adult Wistar
Study Drug rats. Rats were fasted overnight from food, but not
Sarvakalp Kwath is a proprietary water, prior to dosing and weighed before the extract
preparation of Divya Pharmacy, Haridwar. It is was administered orally. A dose of 2000 mg/kg was
available as a crude powder and is claimed to be given to one animal, and this rat was observed for
beneficial for liver disorders. Each 5g powder mortality and clinical signs (behaviours: unusual
contains: aggressiveness, unusual vocalisation,
· Boerhavia diffusa 2.50g restlessness, sedation and somnolence;
· Phyllanthus urinaria 1.25g movements: twitch, tremor, ataxia, catatonia,
· Solanum nigrum 1.25g paralysis, convulsion, fasciculation, prostration and
unusual locomotion) for the first hour, then hourly
The instructions for consumption are as follows for 3 h and, finally periodically until 48 h. If the animal
Boil 5-10g powder in 400 ml water till only survived, then four additional animals were given
100 ml volume of the mixture remains. Filter and the dose 2000 mg/kg sequentially at 48 h intervals.
consume. All of the experimental animals were maintained
under close observation for 14 days, and the
Sarva Kalp Kwath was procured from number of rats that died within the study period
Divya Pharmacy authorized vendor. Working on the was noted. The LD50 was predicted to be greater
same lines as instructions on the Sarvakalp Kwath than 2000 mg/kg if three or more rats survived7.
pack (which leads to the preparation of an aqueous
extract), aqueous extract of the preparation was Study Groups
obtained using Soxhlet extraction apparatus These animals were divided into three
followed by evaporation of the extract over water groups of six animals each.
bath to obtain a solid product which served as the
test compound. Group-I: This group was given normal saline, once
daily, per orally for seven days followed by 1 ml/kg
Experimental Animals intraperitoneal injection of normal saline.
The study was undertaken after due Group-II: This group was given normal saline, once
permission of the institutional animal ethics daily, per orally for seven days followed by 1 ml/kg
committee. of a 50% v/v solution of Carbon tetrachloride (Nice
Chemicals Pvt. Ltd., Cochin) in olive oil
Wistar strain albino rats of either sex and intraperitoneally.
weighing 150-200g were utilized for the main study Group-III: This group received the test compound in
while acute toxicity study was carried out in female dose of 120 mg/kg, once daily, per orally for seven
rats only of the same strain and same weight range. days followed by carbon tetrachloride
intraperitoneally as in Group-II.
The animals were housed in cages under
controlled conditions of temperature (25°C) and Per oral administration of normal saline or
alternating 12 hour cycle of light and darkness and the test compound were carried out by gavage
the animals had free access to standard rat pellet method with animals fasted 3-4 hours prior and 1
diet (Lipton India Ltd.) and tap water ad libitum. hour after administration to ensure proper
absorption. Intraperitoneal normal saline (Group I)
After one week of acclimatization, the or carbon tetrachloride (Groups II, and III) dose was
animals were considered suitable for study. given concomitantly with the last (seventh day)
corresponding oral dose of the group (normal saline
Acute Toxicity Study for groups I and II; test compound for group III). Food
The acute toxicity study was performed was completely restricted after the corresponding
according to the Organisation for Economic Co- intraperitoneal administration in each group.
operation and Development (OECD) guideline 425. Animals of all the groups were fasted for 24 hours
 YADAV & KUMAR, Biomed. & Pharmacol. J., Vol. 7(2), 659-663 (2014) 661

(during which duration water remained freely RESULTS

available) after which they were sacrificed under
Ketamine (75 mg/kg) and Diazepam (10 mg/kg) No symptoms or signs of toxicity or death
anaesthesia given intraperitoneally8. were observed in the experimental rats in the acute
toxicity study. Hence the LD50 of aqueous extract
Blood was collected from the of Sarvakalp Kwath was considered to be greater
anaesthetized animals from retro-orbital plexus. than 2000 mg/kg.
After blood collection, the animals were sacrificed
to obtain the liver. In this study, Group I provided the baseline
values of the studied parameters viz. normal
Biological Study Parameters biochemical parameters (Table 1) and normal
After a standing time of half an hour, the histological study (Fig 1). In Group II, which was
collected blood was centrifuged at 2500 rpm for 10 challenged with CCl4 without any hepatoprotection,
min. The serum so obtained was used to estimate extensive hepatic injury was observed. With the
the biochemical study parameters viz. Alanine exception of albumin, all the biochemical
transaminase (ALT), Alkaline phosphatase (ALP), parameters were significantly raised. The
Total bilirubin and Albumin which were all estimated histological study of Group II confirmed hepatic
spectrophotometrically via commercially available damage wherein classical centrizonal hepatic
reagent kits based on established methods9,10,11,12. necrosis was observed (Fig 2). The Group III, which
was challenged with CCl 4 after one week
The liver was excised from the animals administration of Sarvakalp Kwath, showed
and washed with the normal saline. About one cm evidence of hepatic injury as compared to Group I.
piece was cut and fixed in 10% neutral formalin for However, the extent of derangement of biochemical
12-24 hours. It was then dehydrated and cleared parameters was significantly lower as compared to
with ethanol and xylene respectively followed by Group II and this was statistically significant with p-
embedding in paraffin wax from which blocks were value less than 0.01 (Table 1). The histological study
prepared. Sections of 5m thickness were taken from of Group III showed a remarkable level of
the blocks using a microtome 13. These were hepatoprotection potential of Sarvakalp Kwath from
processed in alcohol-xylene series and were the CCl4 assault (Fig 3). No necrosis was observed
stained with Harris haematoxylin and eosin stain14 and the extent of liver injury was limited to fatty
and subjected to histopathological examination. changes in the hepatocytes. Normal architecture of
the liver was largely preserved.
Statistics
Mean ± SD was calculated for each group DISCUSSION
to observe the general trend of the group. Wilcoxon
Rank Sum Test was applied to test the significance Liver has a pivotal role in regulation of
of the results. P-values were estimated by referring several vital body functions such as metabolism,
to appropriate tables. storage, secretion etc. Liver diseases are common

Table. 1: Biochemical parameters in the studied groups

ALT ALP Total Bilirubin Albumin

(IU/L) (IU/L) (mg/dl) (g/dl)

Group I (Saline control) 20.69±5.94 78.89±12.54 0.15±0.17 5.24±0.13

Group II (Saline + CCl4) 428.78±40.49^ 602.37±34.19^ 1.76±0.69^ 5.20±0.12
Group III (Test compound + CCl4) 173.28±13.04* 253.70±14.23* 0.51±0.10* 5.17±0.12

^ p<0.01 as compared to group I

* p<0.01 as compared to group II
662 YADAV & KUMAR, Biomed. & Pharmacol. J., Vol. 7(2), 659-663 (2014)

and usually come to be noticed when they are well scientific studies on hepatoprotective plants/
advanced. In spite of the tremendous advances formulations have been carried out by chemically
made in modern medicine, no effective inducing liver damage in rodents.
hepatoprotective medicine has yet emerged.
The injury of CCl4 has been ascribed to its
There are numerous plants and metabolite – tricholormethyl radical (CCl3*) which
polyherbal formulations in ayurvedic system of is generated by microsomal enzymes namely
medicine which have been claimed to have CYP2E1, CYP2B1 or CYP2B2, and possibly
hepatoprotective actions and many of these have CYP3A. This metabolite and not CCl4 per se is toxic
been proved by scientific studies as well. Most to the hepatocytes. This radical can also react with
oxygen to form the trichloromethylperoxy radical
(CCl3OO*), which initiates the chain reaction of lipid
peroxidation15. This affects the permeabilities of
mitochondrial, endoplasmic reticulum, and plasma
membranes resulting in elevated levels of
transaminases, alkaline phosphatase, bilirubin and
other biochemical parameters16. Observed albumin
levels were similar in all the groups because of the
half life of albumin is 15 to 20 days.

Based on the mechanism of injury of CCl4,

the plausible mechanism of hepatoprotective
action of Savakalp Kwath can be due to microsomal
enzyme inhibition, free radical scavenging action
Fig, 1: Group I (Saline control) – Microscopic or anti-oxidant action on lipid peroxidation or
features of liver. Normal hepatic study. combination of two or more of the above actions.
Hepatocytes and their nuclei are well visible Also it is quite possible that the hepatoprotection of
(H& E stain 400x) CV: Central Vein; Arrows: Sarvakalp Kwath may be due to a mechanism
Sinusoids which is yet to be identified.

Fig. 2: Group II (CCl4 control) – Microscopic Fig. 3: Group III (Sarvakalp Kwath) –
features of liver. Extensive necrosis apparent. Microscopic features of the liver. Centrizonal
No nucleated hepatocytes can be demarcated. hepatocytes show fatty changes. No necrosis
(H & E stain 400x) is seen. Normal hepatocytes are visible in the
Arrow: Central vein periphery of the hepatic lobule. (H & E stain
400x) CV: Central vein; N: Normal hepatocytes
 YADAV & KUMAR, Biomed. & Pharmacol. J., Vol. 7(2), 659-663 (2014) 663

CONCLUSION observed in this animal study. Although it is

reasonable to extrapolate these results to the
Sarvakalp Kwath, a proprietary product of human population for which this medicine is
Divya Pharmacy, Haridwar, offers significant intended, only clinical trials of Sarvakalp Kwath can
preventive hepatoprotection against CCl 4 as firmly establish its role in therapy of liver diseases
in humans.

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