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AAMJ / Vol. 3 / Issue 6 / November – December 2017
A A M J
Anveshana Ayurveda Medical Journal
www.aamj.in
ISSN: 2395-4159
Research Article
Stability study and Evaluation of Ayurvedic formulation-
Anoac cream
Gajanan B. Bhagwat1 Swapna S Kadam2 Madhuri Hivarale3 Ashwin Porwal4
A b s t r a c t
Stability testing of drug product is important in order to know the quality and shelf life of the
product. Objectives of this study was to investigate and ensure quality of the ANOAC Cream
preparation. Accelerated stability (Temperature 400C±2, Relative Humidity (RH): 75%±5) of the
cream preparation was evaluated through different parameters like physicochemical and micro-
bial study. The stability study was carried out at time interval of 0, 1, and 3 and 6 months,
during this time period variation was seen in physicochemical characters and other properties.
No change was observed in appearance, colour and odour of ointment in the time period of 6
months. According to accelerated stability study the Anoac cream is stable for the period of 6
months when stored at Temperature 400C±2, Relative Humidity (RH): 75%±5.
Keywords: Herbal ointment, Stability testing, Anoac cream, evaluation of herbal cream
1 R&D Manager, 2Consultant, Healing Hands Clinical Research & Development, 3Quality Control
analyst, 4 Head of Institution, Healing Hands Herbs, Research & Development, Pune, Maharash-
tra, India.
CORRESPONDING AUTHOR
Dr. Swapna S Kadam
Consultant, Healing Hands Clinical Research & Development
111, Managlmurti complex, first floor,
NR. Hirabaug Ganpati, Tilak road,
Pune, Maharashtra 411002.
Email- swapnahh@gmail.com
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Gajanan
et.al,.
: Stability study and Evaluation of Ayurvedic formulation- Anoac cream
AAMJ / Vol. 3 / Issue 6 / November – December 2017 1596
INTRODUCTION
yurvedic formulations are proved to be effective
in many health aliments. Even in modern era the
popularity of the herbal medicines is increasing
day by day. Most of the herbal medicines contain multi-
ple active agents. Due to the complex nature of herbal
formulations, it becomes difficult to apply standard qual-
ity control parameters. The quality of Ayurvedic product
is questioned due to lack of standard measures. Various
factors like air, light, temperature, humidity affects shelf
life of Ayurvedic formulations. Stability is interpreted as
length of time under specific conditions and storage that
a product will remain within the pre-defined limits for all
its important characteristics (ICH Guidelines; 1993).[i]
Evaluation of Stability of a formulation is essential not
only during production of a drug, but also to evaluate the
storage condition and shelf life of existing formulations
over a period of time. Stability report is an essential com-
ponent of the documents for marketing and registration
of Ayurvedic formulations in many countries. When the
Ayurvedic products are subjected to stability studies, the
dossier of an Ayurvedic product becomes more scientific
and acceptable. [ii].
Quality of active substances or pharmaceutical drug var-
ies with time because of different factors like physical and
chemical properties of substance and environmental fac-
tors like light, humidity, temperature. A pharmaceutical
product may undergo changes like its consistency, mois-
ture content, pH etc. Stability of product can also be af-
fected by microbiological changes. The main objective
of stability study is well being of patient who is suffering
from disease for which drug has been designed. Stability
study ensures product quality and safety, as degradation
of active ingredients may form toxic compounds. Apart
from this, Real time stability study data also provide in-
formation like shelf life of newly designed drug and its
storage conditions.[iii]
Stability testing can be accelerated stability testing and
real time stability testing. In accelerated study product is
stored under stress condition i.e. at different temperature
and humidity conditions. During this period its physico-
chemical and microbiological conditions are monitored
at predetermined time interval.
The present stability study was conducted to investigate
and ensure quality of the ANOAC Cream preparation
which is Ayurvedic proprietary polyherbal formulation
MATERIALS & METHODS
Test Drug:
Anoac Ointment is an Ayurvedic proprietary polyherbal
formulation in Ointment form which comprises Kasisadi
oil, Jaytadi Oil, Neem Oil, Tankan Powder, Sphatik
Bhasma, Yashad Bhasma, Shuddha Kapoor (camphor)
and Menthol.
The organanoleptic characteristics were examined and
Accelerated stability study was conducted as per ICH
guideline Q1. A (R2)3 to evaluate the ointment.Storage
condition were: Temperature: 40 °C ± 2, Relative Humid-
ity (RH): 75 % ± 5.The change was observed during 6
months for accelerated stability.
The following parameters were considered for evalua-
tion
.
Color and appearance: accurately weighed 5 gram
Anoac ointment was taken into watch glasses and exam-
ined for the color by naked eye after placing against
white background in white tube light.[iv,v]
Odour: 2 gram Anoac ointment was smelled. [iv,v]
pH of the Cream: 0.5 g of accurately weighed Anoac
cream was dissolved in 50.0 ml of distilled water and
its pH was measured. Caliberation of pH meter was
done using standard buffer solution.[iv,v,vi]
Viscosity:
The viscosity of ointment was measured by using a
Brookfield Viscometer. Brookfield digital viscometer
(model DVE-LV, Brookfield Engineering Laboratory, INC.,
USA) was used to measure the viscosity (in cps) of the
prepared ointment formulations as such that is in semi-
solid state. The spindle (spindle code S 64) was rotated
at 0.5, 1, 2, 3, 4, 5 and 10 rpm. The reading, near to
100% torque was noted. Samples were measured at 30
± 1°C.[iv.v.vii]
Thermal Stability: Thermal stability was determined by
placing the formulation into the humidity chamber con-
trolled at 75% RH and temperature 40 ± 2oC. [iv,v,vi]
Homogeneity: The formulations were tested for the ho-
mogeneity by touch as well as by visual appearance.
The appearance of the cream was judged by its color,
pearlscence and roughness and graded. [iv,v,viii]
Spreadability:
Spreadability is a term which indicates the area to which
the cream easily spreads on when applied to the skin.
The spreading ability of cream determines its therapeutic
efficacy. It is expressed as time in seconds taken to slip
off by two slides where the formulation is sandwiched. A
special instrument is used to determine it. To calculate
spreadaibility following formula is used.[iv,v,ix]
Spreadability= m × l/t
m = weight tied to the upper slide (30g)
A
Gajanan
et.al,.
: Stability study and Evaluation of Ayurvedic formulation- Anoac cream
AAMJ / Vol. 3 / Issue 6 / November – December 2017 1597
l =length of glass slide (5cm)
t =time taken in
It is expressed as second cm/ gm.
After feel: Fixed amount of cream was applied and emol-
liency, slipperiness and amount of residue left after the
application was checked.[iv,v]
Type of Smear: A certain amount of formulation was
applied on the dorsum of skin of human volunteer and
observed. The type of film or smear formed on the skin
after application of the cream was checked.[iv,v]
Removal: The ease of wipe out of the applied cream
was examined by washing the area of application with
tap water.[iv,v]
Degradation of product: product was examined for any
degradation or oil separation occurred during study pe-
riod under accelerated conditions.[iv,v]
Test for Microbial Growth:
The formulated creams were inoculated on the plates of
Muller Hilton agar media by streak plate method. The
plates were placed in the incubator and are incubated at
37 °C for 24 hours. After the incubation period, plates
were taken out and check the microbial growth by com-
paring it with the control.[iv,v,ix]
Patch Test:
About 1-3gm of test drug was applied to the sensitive
part of the skin (skin behind ears) with the help of piece
of fabric or funnel. The cream to be tested was applied
to an area of 1sq.m. of the skin. Control patches were
also applied. The site of patch is inspected after 24
hrs.[iv,v,ix]
RESULTS AND DISCUSSION
Anoac cream is semisolid, buff colored ointment with
characteristic odor and smooth texture. Composition of
contents is as per table number 1. The nature of the Ayur-
vedic formulation indicated by its colour while taste and
odour of the formulation are very susceptible indicators.[x]
Three batches were tested for quality control, all the pa-
rameters found within permissible limit and complying
with standards as per API Part I Vol I n II [xi] as shown in
table number 2.
Viscosity of creams is inversely proportional to rate of
shear (rpm) and the results are showed in table no. 3.
Results of microbial load of Anoac ointment was comply-
ing with Ayurvedic drug standards. Table no 4 gives the
result of accelerated stability study.
In this accelerated stability study, Temperature: 40°C ±
2, Relative Humidity (RH): 75% ± 5 was maintained up
to 6 months. The product was analyzed on 0, 1, 3 and
6 months. No change was observed in color, odour and
spreadability of formulation up to storage of 6 months at
accelerated condition (Table 4).
pH of the Cream: The pH of the cream was found to be
in range of 6.1 to 6.6 which is good for skin. pH was
nearer to the ph required for skin.
Viscosity: The viscosity of cream was in the range of
14950 – 236400 cps. This shows that the cream is eas-
ily spreadable by small amounts of shear.
Homogeneity: formulation produced uniform distribution
of extracts in cream. This was confirmed by visual ap-
pearance and by touch.
Appearance: On keeping formulation for long time, no
change in colour of cream was observed.
After feel: On application of fixed amount of cream,
emolliency, slipperiness and amount of residue left was
found.
Type of smear: the type of smear, on application of
cream, formed on the skin was non greasy
Removal: The cream applied on skin was easily removed
by washing with tap water.
Anoac cream is a formulation with smoothening, healing
and anti-inflammatory functions used for anal conditions.
The excipients of the formulation contains oils which has
got anti-infective and healing properties .The essential
oils have been used for the treatment of various ailments
externally and internally, described in the pharmaco-
poeia, traditional systems of medicine and reported in
folk medicine.[xii] The quality of drug product should be
maintained under the various conditions that pharmaceu-
ticals encounter, during production, storage in ware-
houses, transportation and storage in hospital and com-
munity pharmacies, as well as in the home.[xiii]
In accelerated conditions Anoac cream found to be sta-
ble for six months and no significant variations seen dur-
ing study period.
The pH was found to be suitable for topical application.
It was in range of 6.2 to 6.8 which is nearer to skin. The
viscosity, spreadiability, after feel results showed that it
is suitable for topical application. It was found to be non-
greasy and no irritation or adverse effects like redness,
edema seen during Patch Test.
The current accelerated stability study was carried out as
per ICH guidelines[i], investigations revealed no signifi-
cant physically observable variations.
CONCLUSION
The stability studies of the various parameters like visual
appearance, nature, pH of the formulations showed that
Gajanan
et.al,.
: Stability study and Evaluation of Ayurvedic formulation- Anoac cream
AAMJ / Vol. 3 / Issue 6 / November – December 2017 1598
there was no significant variation after six months of the
study period. There were no signs of microbial growth
after incubation period of 24 hours at 370 C and having
more antimicrobial property as compare to standard.
From the results we can also say that the product can be
stored up to six months.
ΛΛΛΛ
Acknowledgement –
Authors would like to thank Dr. Snehal Porwal, Healing
Hands & Herbs, Pune for valuable guidance and encour-
agement. We would also like to thank all the team of
healing hands research & development.
REFERENCES
i. ICH Guidelines: Stability testing of New Drug Substances
and Products, 27th October 1993.
ii. Neeraj PT, PA02.21 Stability study in ayurvedic products,
Ancient Science of Life 2013; 32 (s2):66).
iii. Anonymous. ICH Harmonised Tripartite Guideline. Stability
testing of new drug substances and products – Q1A (R2).
2003 Feb.
iv. Indian Pharmacopoeia: Ministry of Health and Family Wel-
fare,Government of India. Published by the Indian Pharma-
copoeial commission: Ghaziabad; 2010.
v. United States Pharmacopoeia. The US pharmacopoeial con-
vention. 35th edn. Inc.12601 Twinbrook Parkway, Rockville
MD20852; 2007.
vi. Sheraz MA, Ahmed S, Ahmad I, Shaikh RH, Vaid FHM, Iqbal
K.Formulation and stability of ascorbic acid in topical prep-
arations. Syst Rev Pharm 2011;2:86-90.
vii. Sujith S Nair, Molly Mathew, Sreena K.Formulation and Eval-
uation of Herbal Cream containing
Curcuma longa
.
http://www.ijpcsonline.com/files/22-261.pdf
viii. Singhal M, Khanna S, Nasa A. Cosmeceuticals for the skin.
Asian J Pharm Clin Res 2011;4:1-6.
ix. Akash S, Mali, Karekar P, Yadav AV. Formulation and eval-
uation of multipurpose herbal cream. Int J Sci Res
2015;4:1495-8.
x. Gajanan B. Bhagwat, Swapna S Kadam, Madhuri Hivarale.
The Accelerated Stability Study of Constalax Churna- An
Ayurvedic Formulation. International Journal of Ayurveda
and Pharma Research. 2017;5(7):85-90.
xi. Lohar DR. (2007). Protocol for testing of Ayurvedic, Siddha
& Unani medicines. Government of India, Department of
AYUSH, Ministry of Health & Family Welfare: Pharmacopoe-
ial laboratory for Indian medicines, Ghaziabad, 47-52
xii. J.T. Coon and E. Erenst,
Alimen. Pharmacol. Therap.
, 16,
1689 (2002).
xiii. NK Sachan, S Pushkar, AK Sachan, SK Ghosh; Thermal Sta-
bility and Drug-Excipient Compatibility Studies of Peppermint
and Caraway Oils for Formulation of Chewable Tab-
lets.Asian Journal of Chemistry 25 (11), 5930 – 5934.
https://doi.org/10.14233/ajchem.2013.14186 )
Source of Support: Nil.
Conflict of Interest: None declared
ΛΛΛΛ
TABLES
Table No 1: Composition of Anoac Cream
Sr. No.
Ingredients Name
Quantity
Percentage Wise
01
Jatyadi oil
17%
02
Kasisadi oil
31%
03
Neem oil
07%
04
Yasad Bhasma
01%
05
Spatik Bhasma
02%
06
Tankan Bhasma
01%
07
Menthol
0.3%
08
Camphor
0.7%
09
Cream Base
Q.S.
How to cite this article: Gajanan et.al,. : Stability study
and Evaluation of Ayurvedic formulation- Anoac cream.
AAMJ 2017; 6:1595 – 1599.
Gajanan
et.al,.
: Stability study and Evaluation of Ayurvedic formulation- Anoac cream
AAMJ / Vol. 3 / Issue 6 / November – December 2017 1599
Table No 2 : Quality analysis Batch wise
Batch No
AO-12
AO-13
AO-14
General Appearance
Uniform
Uniform
Uniform
Color
Buff colored
Buff colored
Buff colored
Odor
Characteristic
Characteristic
Characteristic
Spreadability
Uniform Spreading
Uniform Spreading
Uniform Spreading
pH
6.2
6.5
6.6
LOD (% w/w)
43
45
45
Homogeneity
Uniform
Uniform
Uniform
microbiological tests like TBC, Pathogens
< 100 colonies
< 100 colonies
< 100 colonies
Table No 3: Viscosity of Anoac Ointment
Batch No
Viscosity (cps) at rpm
0.5
01
02
03
04
05
10
10
212400
66600
47700
31000
24300
21600
16200
09
236400
70800
41700
33400
28200
22200
15000
08
236000
70500
41711
32900
27500
22000
14900
07
240000
60900
41277
31740
27010
22700
15300
06
237000
70811
40711
33050
28350
21900
15200
05
236100
70320
42611
33720
28000
22400
14960
04
237500
70440
41612
31200
27900
22785
14870
03
236950
60820
47202
32000
28150
22665
15100
02
237000
66600
46450
31800
28300
21400
15210
01
236000
70511
47750
32300
27500
22000
14950
Table No 4: Accelerated Stability Studies of Anoac cream as per below
Sr. No.
MONTHS/ TESTS
Initial Months
After 1 months
After 2 months
After 6 months
01
Colour
Buff colored
Buff colored
Buff colored
Buff colored
02
Appearance
Semisolid
Semisolid
Semisolid
Semisolid
03
Odour
Characteristic
Characteristic
Characteristic
Characteristics
04
PH
6.2
6.2
6.4
6.8
05
Thermal Stability
Ok
Ok
Ok
Ok
06
Homogeneity
Uniform
Uniform
Uniform
Uniform
07
Spreadability
Easy
Easy
Easy
Easy
08
After Feel
Emollient
Emollient
Emollient
Emollient
09
Type of Smear
Non greasy
Non greasy
Non greasy
Non greasy
10
Removal
Easy
Easy
Easy
Easy
11
Degradation of product
Nil
Nil
Nil
Nil
12
Microbial limit test
< 100 colonies
< 100 colonies
< 100 colonies
< 100 colonies
