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Formulation and Evaluation of Parenteral Drug Edaravone

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 International Journal for
Pharmaceutical Research
Scholars (IJPRS)
V-3, I-4, 2014 ISSN No: 2277 - 7873
RESEARCH ARTICLE

Formulation and Evaluation of Parenteral Drug Edaravone

Singh Atul Pratap*, Eswari TS, Gurusharan, Verma V
Department of Pharmaceutics, IIMT College of Medical Science Meerut, India.
Manuscript No: IJPRS/V3/I4/00431, Received On: 16/11/2014, Accepted On: 20/11/2014

ABSTRACT
Edaravone have best property of neurological recovery and it is a kind of cerebral protective agent (free
redical scavenger) also it acts as an antioxidant. The present study was under taken with an intension to
develop a stable and effective parenteral formulation containing the drug Edaravone. Solubility analysis
of the drug Edaravone performs soluble in CCl4 and insoluble in water. So, various effects of various co-
solvents in the solubility of edaravone have been evaluated. Edaravone was tried with co solvents such
as CCl4, ethanol, methanol active ethyl acetate and water. The drug was made in to injection formulation
for administering as an infusion. Various batches of Edaravone injection formulation were prepared in
order to assess the influence of heat, light, atmospheric oxygen and antioxidant on the stability of the
drug. The formulation were also subjected to accelerated stability test out of all trials. Formulation
containing all the ingredients like L-cystein hydrochloride monohydrate, NaCl, sodium bi sulphate,
phosphoric acid, NaOH pellets and water for injection was found to be more stable and passed test C2
satisfactorily.
KEYWORDS
Edaravone, Formulation, Antioxidant, Evaluation, Stability.
INTRODUCTION
Edaravone (3-methyl-1-phenyl-2-pyrazolin-5- Although the neuroprotective effects of
one) is a potent free radical scavenger which has edaravone on different models of neurotoxicity
been shown to provide neuroprotection against have been described, to the best of our
cerebral ischemia-reperfusion injuries in knowledge, there is no report on the protective
experimental animal models7. Moreover, the effect of edaravone against induced
clinical efficacy of edaravone has been neurotoxicity up to now.
demonstrated in patients with acute brain
infarction and it was approved in 2001 by the Parenteral formulations are widely used
health authorities in Japan as a neuroprotective especially when an immediate psychological
agent for the treatment of acute cerebral response is needed in life threatening emergency
infarction. Recently, several studies also conditions and for administering those drug that
demonstrated the in vivo efficacy of edaravone are destroyed by digestive secretions8.
in amyotrophic lateral sclerosis, traumatic brain The aim of the present study is to formulate and
injury and Parkinson’s disease. evaluate the parenteral dosage form containing
*Address for Correspondence:
Edaravone. The objectives of the study are, to
Singh Atul Pratap study the solubility behavior of the drug in
Department of Pharmaceutics, IIMT College Medical Science,
Meerut, India.
different solvents, to develop an analytical
E-Mail Id: atulsingh2206876@gmail.com method for assay of Edaravone, to design and
formulate a stable parenteral formulation of
© Copyright reserved by IJPRS 134
 Formulation and Evaluation of Parenteral
Drug Edaravone
Edaravone, to evaluate prepared parenteral Effect of Oxygen on Drug
formulations of Edaravone.
1% of Edaravone in 0.0.1N NaOH is filled into
MATERIALS AND METHODS vials and placed at 30°C and 40°C. One group is
Edaravone is gift sample taken from UCB India purged and another group is sealed with air.
Pvt. Ltd Mumbai and other all excipients Solutions are observed for colour change and
provided from the Akums Pharmaceuticals Ltd drug content. The data are given in Table 5.
Plant 3rd Sidkul, Haridwar. Formulation Development
List of ingredients used is given in Table 1. In these process general all ingredients mix in
the tank step by step and after it injection
Preformulation Studies2,3,4,5,6
prepared the methodology is given bellow:
Solubility Studies of Edaravone in Different
Table 1: Concentration of different
Solvents (Saturation Solubility Method)
ingredients used in various trial
Excess of drug was added to different solvents formulations
in 10 ml Stoppard volumetric flasks. Then Drug
was made to dissolve in the solvent by placing Name of F1 F2 F3
the volumetric flask in the shaker bath at 25° C ingredients (gm) (gm) (gm)
for 6 hours. The volumetric flasks were then
placed at room temperature for 24 hours. The Edaravone 1.553 1.553 1.553
solutions were filtered and appropriate dilutions
were made to measure absorbance at 244nm L-Cystein
0.5 0.5 0.5
using UV visible spectrophotometer, and CCl4 Hydrochloride
as blank. The data are given in Table 2.
NaCl 6.75 6.75 6.75
Effect of Temperature on Stability of Drug
1% Edaravone solution in 0.0.1N NaOH is filled Sodium
1.0 1.0 1.0
into vials. The ampoule were sealed and placed bisulphate
at refrigeration, room-temperature, 50°C, 75°C
and 95°C for 1 week and observed for colour Phosphoric Acid - 39.00 39.00
change and crystal growth. The samples placed
at refrigeration and room temperature served as NaOH pellets 16.00 16.00 -
controls. The data are given in Table 3.
Light Stability of Drug Water For
q.s q.s q.s
Injection
1% of Edaravone solution in 0.0.1N NaOH is
filled in to 20ml glass vials (amber and clear). Active material has been taken equivalent to its
Also samples of drug substance are placed in an 100%. Assay on as is basis considering the
open Petri dish to expose a large surface. Drug minimum.
and dilutions placed in a light-resistant amber
Assay – NLT 99% (ODB) and maximum LOD
colored glass vials, foil wrapped and in a
NMT 0.5%w/w # this quantity is taken only for
cardboard box as controls. This is carried out for
pH adjustment.
4 weeks with weekly examinations for visible
colour change or precipitation in solution in Calculation
clear vials, the compound can be considered as Required Quantity of Edaravone
potentially light sensitive and should be handled
accordingly. The data are given in Table 4. Averages: 2% “Q” indicates the %
“P” indicates assay LOD NMT 0.5%w/w
NLT 99% (ODB) Note: - During
formulation required
© Copyright reserved by IJPRS 135
 Formulation and Evaluation of Parenteral
quantity of active  Drug Edaravone
Electronic 100ml further dilute
Sample Preparation
shall be taken weighing balance the 10ml of this
equivalent to100% solution to 100ml Dilute 2ml of sample
 HPLC with the same. to 100ml with the
Assay on as is
 pH meter mobile phase.
basic considering
the estimated assay Apparatus Required: Chromatographic
and %LOD% water System
contents  Volumetric flask
-100ml,1000ml Column :5
The Detection :2
 Measuring
data cylinder Flow Rate :1
are  Clean and dry Injection Volume : 20
give beaker
Procedure
n in  Bulb pipette –
Separately inject
2ml,10ml
Tabl equal volume
e 6. Reagent Required: (20µl) of diluents,
six replicate
Post  Methanol HPLC
standard
Grade
For preparations and
 Sodium Acetate two injection of
mul sample preparation
Edaravone Working
atio Standard in to the
chromatograph,
n Sodium Acetate record the
Eval Buffer Preparation chromatogram and
uati Dissolve 13.61g of measure the
Sodium acetate in responses for
ons major peaks.
sufficient H2O to
Ass produce 1000 ml, System suitability
ay adjust the pH 5.0 parameter
with HCl solution or Tailing factor: NMT
Con
NaOH solution 2.0%
tent
Mobile Phase Relative standard
of Preparation deviation: NMT 2.0%
Eda Mix sodium acetate Calculation
ravo buffer and methanol
(HPLC grade) in the
ne ratio of 30:70 filter
Instr and degas.
ume Standard Preparation
= mg/ml
nt Dissolve accurately
weighed 30 mg of Acceptance Criteria
Req edaravone working For finished products-
uire standard in 50 ml of 90% to 110%
mobile phase, mix
d: For bulk products -98%
and make the volume
© Copyright reserved by IJPRS 136
 Formulation and Evaluation of Parenteral
to 105% filtersDrug
of Edaravone
0.22, 10 ml of the each
RESULTS AND
0.45, 1.2 and prepared
Sterilization DISCUSSION
1.5 micrometers formulations C2 were
Studies
were placed kept at different FT-IR Spectrum of
The injection successively temperatures/ Pure Edaravone
samples were and tested conditions such as
taken in glass whether the refrigeration, room
syringe, the injection sample temperature, 37°C,
membrane could pass 40°C, 45°C and
filter holder through these under light. At
was attached membrane or regular time intervals
to the not. The data the samples were
syringe. A are given in examined for pH
prefilter of Table 7. changes for six
1.5 weeks using a digital
micrometers Sterility Testing
pH meter. The data
was placed in Direct Transfer are given in Table 14.
this holder, Method Figure 1: FT-
Clarity
after which IR spectrum of
10 ml of the pure
Aliquots of the from point of view of
formulations were Edaravone
samples are safety of the patient
placed at
transferred being treated with and Table 2:
refrigeration, room
aseptically into to get a safe and Solubility
temperature, 37°C,
fluid thioglycolate maximum therapeutic profile of
40°C and 45°C for
medium and response of the drug. Edaravone in
six weeks and
soybean casein The provision of different
observed for colour
digest medium. rapid means of solvents
change or turbidity.
The inoculated quality control, which
The data are given in
thioglycolate ensures that no
Table 16. Descriptive Par
medium is unexpected changes
incubated at 32°C in the stored product % Drug Content Term requir
and soybean casein are occurred like: The drug content of
digest samples at Crystal growth, pH the formulations C2 Very soluble L
22°C for 7 days. changes, Clarity and were determined by
Likewise negative % Drug content. following the same Freely soluble Fr
and positive Crystal Growth procedures as
controls are mentioned in assay. Soluble Fro
prepared. The data 10 ml of the each
The estimates were
are given in Table prepared formulations Sparingly soluble Fro
done at intervals of
13. C2 were placed at
one week upto six
refrigeration, room Slightly soluble From
Stability Studies weeks. The data are
temperature, 37°C,
given in Table 17. Very slightly
For any 40°C and 45°C From
pharmaceutical respectively for six soluble
dosage form weeks and observed
Insoluble or
stability of the for crystal growth.
practically 100
prepared The data are given in
insoluble
formulation is a Table 15.
very basic and Soluble in CCl4,
pH Changes
important factor,
© Copyright reserved by IJPRS 137
 Formulation and Evaluation of Parenteral
freely Stability
DrugEvaluation
Edaravone
various tests and size
soluble in Various stress results are and
methanol, tests are discussed in the filterab
ethanol, performed on following section. ility of
slightly solid and the
soluble in Table 6: Drug
solution sample formul
acetone or in conte
to establish the ation
ethyl acetate nt of
effect of heat, vario of
and in light and Edarav
us
soluble in oxygen on the one
form
water. drug substance ulati
stability. on Filter pore
Formulation
trials size (µm)
Heat Stability l
e conta
Table 3: Heat 0.22
a ining
stability profile Edar 0.45
r C1
of Edaravone avon 1.2
, 1.5
e
- 0.22
Temperature (C)
T Drug content 0.45
C2
u Formulation 1.2
(mg/ml)
Refrigeration r 1.5
b C1 1.55234 0.22
Room i 0.45
Temperature d C2 1.50133 C3
1.2
i C3 1.49532 1.5
40 t
y *Each value is an +Injection passes
50 average of three though.-Injection
E determinants does not pass
75 f through
f Table 7: Filter
+ Color change – e pore All the formulation was
No color change c found to be easily
t subjected for
Light Stability
Table 4: Light o + Color change – No
stability study of f color change
Edaravone
Formulation
O
Withdrawal Development
x
week y A stable parenteral
1 g formulation of water
2 e insoluble drug
n edaravone was
3 formulated after
Table 5: Test for color performing trials
4 change after a week
Temperature Air sealed Perged with various solvent.
- (0C) vials vials Thus prepared
C 25 + - formulations were
© Copyright
35 reserved by IJPRS
+ + 138
 Formulation and Evaluation of Parenteral
passing through the microbial
Drug Edaravone n %
entire pore size growth (except c
filter and the positive o
hence 0.22 m control) at the l
pore size end of o
filter was incubation r
selected to period, c
filter all the indicating all h
prepared the a
formulations formulations n
separately. were sterile and g
thus all the e
None of the formulations
formulations are subjected to Formulation 5ºC
showed further C1 -
turbidity or evaluations.
signs of C2 -
Post Formulation C2 - C3 -
Studies
C3 - + Color change – No
color change
Scale up Studies
Effect of Different +Turbidity, -Clear.
Assay of the
Temperature on Effect of Different
formulations
Crystal Growth Temperature on
Table 8: Effect of different Color Change
=
temperature Table 10: Effect
on crystal o 1
growth f .
d 5
Formulation RT if 0
C1 f 1
e 3
C2 r 3
C3 e
n m
+crystal growth,- t g
crystal growth t /
e m
Effect of Different m
Temperature on l
p
Clarity e =
ble 9: Effect of different r
temperature a 1
on clarity t 0
u 0
Formulation RT r .
e 0
C1 - 8
o
8
© Copyright reserved by IJPRS 139
 Formulation and Evaluation of Parenteral
Acceptance Drug Edaravone el Crystal growth
Criteria Filter poreer Table 15: Crystal
Formulation
For finished size(µm) growth of
at
products- 90% Withdrawal
to formulation
Formulation
110% 37ºC 40ºC 45ºC 0.22 e C2 at
week
0 - - - 0.45 d different
For bulk
1 - C2 - - temperatur
pro 1.2 St es
2 - - -
duct C2 3 - - - 1.5 a
4 - - - bi
s +Injection
5 - - -
-98 6 passes
- through,-
- - lit
Injection does
% y
not pass through
to St
The results of
105 filterability u
show that both di
%
the formulation
Tab of Edaravone es
le passes through p
all the four
11: membrane H
Dru filters. Hence c
they can be
g h
sterilized by
cont filtration a
ent Direct Transfer n
Method g
of
Table 13: The es
C2
growth of
bacteria in Table 14: pH
soya bean changes
Formulation of
casein digest
medium and formulat
fluid ion C2 at
C2 different
thioglycollate
Table 12: medium after temperat
filter pore ures
seven days
size and Withdrawal
filterability Soyabean- Formulation week
of the casein digest
Formulation 0
formulation medium
s of (SCDM) 1
Edaravone 2
C2 - C2 3
4
-clear, +Turbid 5
6
A cc
© Copyright reserved by IJPRS 140
 Formulation and Evaluation of Parenteral
All the Drugperatu
Edaravone Te ., chapter 26
formulations were res sti in the
clear at different ng Theory and
temperatures. Sample % Drug content
withdrawal No crystal growth Practice of Industrial
Drug Content (week) 37 ºC was Pharmacy. page-902.
Table17: 1 100.003 observed 2. Krishna, G.,
Per the formulations at Hodnick, W. F.,
2 100.023 different
cen Lang, W., Lin,
t temperatures. X., Karra, S.,
3 100.018
dru Clarity Studies Mao, J., &
g 4 100.011 Almassian, B.
Table16: Clarity of
con 5 099.998 (2001).
for
tent Pharmaceutical
6 100.006 mul
of development
atio
for n and
mul C2a manufacturing
ACKNOWLEDGEM
atio t of a parenteral
ENTS
n diff formulation of
C2 Special thanks to a novel
eren
at akums antitumor
t
diff tem agent,
ere pharmaceutical pera VNP40101M.
nt haridwar for ture AAPS
tem providing labs, s PharmSciTech,
equipments and 2(3), 39-47.
API Withdrawal
Formulation 3. Nahar, M., &
(Edaravone) week
with 0 Jain, N. K.
excipients. 1 (2006).
Author 2 Formulation
thankfull to C2 3 and evaluation
Mr. S. N. 4 of saquinavir
+ Crystal Tripathi for 5 injection.
growth,- his 6 Indian Journal
no crystal continuous of
+Turbid,-clear Pharmaceutica
growth assistance
for research. l Sciences,
68(5), 608-14.
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s and Health
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© Copyright reserved by IJPRS 141
 Formulation and Evaluation of Parenteral
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© Copyright reserved by IJPRS 142

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