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Simultaneous Determination of
Tinidazole and Furazolidone in
Suspension by HPTLC and HPLC
a

N. M. Tendolkar , B. S. Desai , J. S. Gaudh & V.

M. Shinde

Analytical Laboratory , The Institute of Science ,

15 Madam Cama Road, Bombay, 400 032, India
Published online: 23 Aug 2006.

To cite this article: N. M. Tendolkar , B. S. Desai , J. S. Gaudh & V. M. Shinde (1995)

Simultaneous Determination of Tinidazole and Furazolidone in Suspension by HPTLC
and HPLC, Analytical Letters, 28:9, 1641-1653, DOI: 10.1080/00032719508002771
To link to this article: http://dx.doi.org/10.1080/00032719508002771

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ANALYTICAL LETTERS, 28(9), 1641-1653 (1995)

SIMULTANEOUS DETERMINATION OF TINIDAZOLE AND

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FURAZOLIDONE IN SUSPENSION BY HPTLC AND HPLC

KEY WORDS : Tinidazole,

N,M.Tendolkar,

Furazolidone, HPTLC, HPLC

B. S-Desai.

J. S. Gaudh

and V. M. Shlnde*

Analytical Laboratory,
The I n s t i t u t e o f Science, 15, Madam Cama Road,
Bombay 400 032, India.

ABSTRACT
High

performance

thin

layer

chromatographic

and h i g h performance Ii q u i d chromatographic

(HPTLC)

(HPLC)

methods

were developed f o r t h e simultaneous determination of Tinidazole

and Furazol idone i n suspension.
In
and

the

method

the

plate

as

mobile

phase.

Furazol idone
concentration

of
60F

R f values obtained were 0.63

and

Tinidazole

range

10-50

Tinidazole

254
(9:l:0.1

chIoroform:rnethanol:ammonia

using

evaluation was done a t 335 nm.

the

separation

Furazolidone was c a r r i e d out on s i l i c a gel

glass
for

HPTLC

respectively.

HPTLC
vlv)

and 0.79

Densitometric

L i n e a r i t y was obtained w i t h i n
yglml

and

3.5-17.5

pglml

for

Tinidazole and Furazol idone respectively.

The second method i s based on h i g h performance l i q u i d
chromatography on a r e v e r s e d phase column
1641
Copyright 0 1995 by Marcel Dekker, Inc.

(p

Bondapak C18)

1642

TENDOLKAR ET AL.

using

mobile

phase

comprised

for

Downloaded by [University of Chicago Library] at 09:15 16 November 2014

of

acid.

Tinidazole
1.5

Retention

and

mllmin.

was

obtained

and

10.5-63

Furazolidone r e s p e c t i v e l y a t a f l o w

rate

was

the
for

were

done

5.24

at

335

concentration
Tinidazole

and

w i t h dil.
min

within

times

7.82

Detection

pglml

: acetonitrite

water

v l v ) adjusted t o p H = 3.0

t r i e t h y l a m i n e (80:20:0.1
phosphoric

of

nm.

range

and

Linearity

30-180

Pglml

Furazol idone

resp.

INTRODUCTION

su Iphony I) e t h y l I -2-methy I-5-

[ 1 [ 2- ( E t h y I

T i n idazole
nitro-1 H-imidazolel

i s a antiprotozoal and i t has a n t i m i c r o b i a l

action and i s used i n the treatment o f susceptible protozoal

infections.
Furazolidone [ 3 - [ [ (5-Nitro-2-furanyl )methylenel

amino]-

i s a n i t r o f u r a n d e r i v a t i v e w i t h antiprotozoal

2-oxazolidinone]
activity.

It i s a c t i v e against t h e protozoan g i a r d i a intestinal i s

and against a range o f enteric bacteria.

Tinidazole

is

i s o f f i c i a l i n 1.P

official

, 6.P

in

1.P'

and U.S.P

and

J.P

HPLC17-18, G.C1',

TL?

the

simultaneous

yet

reported.

Furazolidone

G . c 1 2-1 3

methods

S i m i l a r l y spectrophotometric

14-16

methods are r e p o r t e d f o r Furazol idone b u t

determination
The

S p e c t r o p h ~ t o m e t i c ~ - ~ , HPLC8-11,
a r e r e p o r t e d f o r Tinidazole.

of

simultaneous

both

these

analysis

of

drugs

is

not

Tinidazole

and

TINIDAZOLE AND FURAZOLIDONE

Furazolidone

1643

i s required for

t h e purpose o f q u a l i t y control

of these drugs.
In

this

communication

we

propose

HPTLC

and

HPLC

methods f o r the simultaneous determination o f b o t h t h e drugs

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i n suspension form.

EXPERIMENTAL

Instrumentation

HPTLC System :Camag

Scanner

IV

Linomat

sample

II (Densitometer),

applicator,

Cats 3.15

trough chamber, Merck 6OFz5,,

v.

Camag

software,

TLC

Camag t w i n

HPTLC glass plates.

HPLC System :Perkin

LC

290

UV

Elmer

lsocratic

detector,

LC

Perkin

Rheodyne 7125 v a l v e injector,

250

Elmer

pump,
PE

Perkin

Nelson

Bondapak C,*

Elmer

Integrator,

column (Waters,

India).

Materials
Authentic

samples

of

k i n d l y donated by Kopran ( I )
were procured from market.

Tinidazole and

Furazol idone were

Ltd. Suspension ( T i n i - F ) samples

TENDOLKAR ET AL.

1644
Reagents
Chromatographic
chloroform,

grade

triethylamine

water,

methanol

a c e t o n i t r i le,
and

analytical

were

accurate1y

(E.Merck, I n d i a )

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reagent grade ammonia were used.

Standard solution
Tinidazole (1OOmg) and F u r a t o l idone (35mg)
weighed

into

acetonitrile

ml

100

volumetric

w i t h ultrasonication

and

flask,

dissolved

diluted

to

volume

in
with

acetonitri le.

Procedure f o r HPTLC

Calibration
C a l i b r a t i o n solutions
volumes

(0.5-2.5

ml)

of

p r e p a r e d by

were

standard

solution

taking

in

50

varying

ml

flasks

and d i l u t e d u p t o t h e mark w i t h acetonitrile.

10 microlitres

of

60F254 HPTLC

these

glass

solutions

plate

in

were
mm

applied
width

on

using

silicagel
sample

applicator.

The

p l a t e was developed u p t o 60 mm i n a t w i n trough chamber

containing

chloroform

methanol

ammonia

( 9 : l :0.1

vlv)

as a mobile phase.
Densitometric

evaluation

was

done

and absorbance was measured a t 335 nm.

by

TLC

scanner

TINIDAZOLE AND FURAZOLIDONE

Calibration
of

drugs

vs.

curves

1645

were

concentration

constructed by
of

drugs

for

p l o t t i n g areas
both

Tinidazole

an

accurately

and Furazol idone.

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Sample preparation
After
weighed
of

determining

amount

equivalent

Furazol idone)

flask,

ml

50

mass

of

was

per

to

millilitre,

100 mg of

transferred

acetonitrile

into

was

Tinidazole
ml

100

added,

(35 mg

calibrated

ultrasonicated

for

5 minutes and f i n a l l y d i l u t e d up t o t h e mark w i t h acetonitrile.

The

resulting

no.1

(solution A).
Then

solution

ml

of

was

filtered

solution A

through

was

Whatman paper

pipetted

into a

50

ml

c a l i b r a t e d f l a s k and d i l u t e d up t o t h e mark w i t h acetonitrile.

10

microlitres

of

the

final

s i l i c a gel 6OFz5,, HPTLC glass

developed,

dried,

scanned

solution

p l a t e and

and

peak

was
the

areas

applied
plate

were

on
was

recorded

as r e p o r t e d i n c a l i b r a t i o n procedure.

Procedure f o r HPLC
Cal ibration
Calibration

solutions

were

volumes o f standard solution (1.5-9

prepared by taking

varying

m l ) i n s i x 50 ml standard

flasks and d i l u t e d u p t o t h e mark w i t h mobile phase.

TENDOLKAR ET AL.

1646
microlitres

20

of

the

above

solutions

were

injected

into chromatograph.
The

mobile

triethylamine

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dilute

0.22

mllmin.

consisted

acid.

of

adjusted

vlv)

(80:20:0.1

phosphoric

through
1.5

phase

water:acetonitrile:
to

This

was

filtered

Whatman paper.

Flow

rate

with

an

average

operating

pH

and
was

pressure

with

3.0

degassed
kept

at

2200

psi

and the UV response was monitored a t 335 nm.

Calibration

curves

areas o f drugs vs.

were

constructed

by

plotting

peak

concentration.

Sample p r e p a r a t i o n

3 m l o f solution A ( p r e p a r e d as above1 was t r a n s f e r r e d

into

50

ml

calibrated

t h e mobile phase.
into

flask

and

diluted

to

the

mark

with

20 m i c r o l i t r e s o f t h i s solution was injected

chromatograph

and

peak

areas

were

recorded

for

both

t h e drugs as i n t h e c a l i b r a t i o n experiment.

Recovery Studies
To
and

to

study
check

formulation,

the

the

accuracy,

interference

recovery

reproducibility,
from

experiments

excepients

were

carried

precision

used
out

in the
by

the

standard a d d i t i o n method f o r b o t h HPTLC and HPLC methods.

TINIDAZOLE AND FURAZOLIDONE

Known amount

of

standard

1647

Tinidazole

and

Furazol idone were

added t o t h e f i x e d amount o f sample solution a t f i v e concentra-

% recovery

t i o n l e v e l s and from t o t a l amount o f d r u g found,

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was calculated.

RESULTS AND DISCUSSION

HPTLC :To c a r r y out separation o f Tinidazole and Furazol idone

by

TLC,

various

mobile

phase m i x t u r e s

s i l i c a g e l 60F254 HPTLC glass plate.

methano1:ammonia
t h e drugs.

vlv)

(9:l:O:l

R f values obtained

were

tried

on

A m i x t u r e o f chloroform:

gave
were

good

0.63

resolution

and

0.79

of
for

Furazolidone and Tinidazole respectively.

Scanning of

the

p l a t e was done a t 335 nm as a t t h i s

wavelength b o t h Tinidazole and Furazol idone show good response

and t h e r e was no

interference o f t h e

bands

of

inactive

ingredients.
For
were

obtained

and 3.5-17.5
The

quantitative
within

application,
the

linear

concentration

calibration
range

10-50

graphs
yglml

F g / m l f o r Tinidazole and Furazol idone respectively.

calibration

curves

regression equations.

could

be

represented

by

following

TENDOLKAR ET AL.

1648
Table 1
Determination

of

Tinidazole

and

by

F u r a z o l idone

HPTLC

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L a b e l c l a i m :- T i n i d a z o l e (100 m g / 5 m l )
F u r a z o l i d o n e (35 mg/5 m l )

101.72

34.01

100.02

34.25

99.43

34.21

99.38

34.03

97.96

34.81

100.26

34.69

98.43

34.98

(Tinidazole)
(Furazol
These

= 33.5860

idone)
equations

= 51.8000
were

23.000

(r=0.999)

X + 7.0800
used

for

(r=0.998)

direct

evaluation

of

drugs.
Results o f t h e r e p l i c a t e a n a l y s i s and r e c o v e r y e x p e r i m e n t
o f suspension a r e t a b u l a t e d i n T a b l e 1 and T a b l e 2 r e s p e c t i v e l y .

TINIDAZOLE AND FURAZOLIDONE

1649

Table 2
Result of the r e c o v e r y experiment by HPTLC.

Amount o f
drug
present
mg/5 m l

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Drug
name

T i n idazole

Amount o f
standard
added
m g / 5 ml

Total
Amount
% Recovery
recovered*
mg/5 m l

100
100
100
100
100

0.00
20.00
40.00
60.00
80.00

99.48
118.24
140.77
157.89
179.33

99.48
98.53
100.55
98.68
99.63

35
35
35
35
35

0.00
7.00
14.00
21.00
28.00

34.58
41.92
49.51
55.02
62.77

98.80
99.81
101.04
98.25
99.63

...................................
Mean Recovery :99.37
................................................................
Furazol idone

* .. Average

o f t h r e e experiments.

HPLC :In

order

to

effect

the

separation

of

components by r e v e r s e d phase HPLC system,

column as
of
The

stationary

phase and

mobile

water: acetoni tri le: tr i e t h y lami ne (80:20 :0.1

mobile

phase

composition

was

the

two

drugs

a Bondapak

C18

phase comprising
v / v 1 was used.

optimised

and

pH

for

TENDOLKAR ET AL.

1650
Table 3

Determination

of

Downloaded by [University of Chicago Library] at 09:15 16 November 2014

L a b e l c l a i m :-

Tinidazole

and

Furazol idone by

Tinidazole ( 1 0 0 m g / 5 m l )
Furazolidone (35 mg/5 m l )

Samp Ie
no.

Furazol idone
Content obtained
(mgl5 ml)

T i n idazole
Content obtained
(mg15 m l )

99.11

34.54

100.54

34.23

100.18

34.18

98.35

34.40

98.28

35.01

100.90

34.49

99.89

34.64

optimum r e s o l u t i o n was adjusted t o 3.0

acid.
were
flow
7.82

Under
well

the

described

defined,

r a t e of

HPLC

1.5

resolved

using d i l u t e p h o s p h o r i c

conditions,
and

free

the

analyte

peaks

from

tailing.

At

m l l m i n t h e retention times were 5.24

min f o r Tinidazole and Furazol idone.

and

TINIDAZOLE AND FURAZOLIDONE

1651

Table 4
Results o f t h e r e c o v e r y experiment by HPLC.

Amount of
drug
present
m g / 5 ml

Downloaded by [University of Chicago Library] at 09:15 16 November 2014

Drug
name

Amount o f
standard
added
m g / 5 ml

Total
% Recovery
Amount
recovered*
m g / 5 ml

0.00
20.00
40.00
60.00
80.00

100.08
120.45
139.41
1 58.82
178.40

................................................................
100
100
100
100
100

Tinidazole

100.08
100.38
99.58
99.26
99.11

Mean Recovery :0.00

7.00
14.00
21.00
28.00

35
35
35
35
35

Furazol idone

99.68
99.00
101.62
98.94
99.86
100.13

34.65
42.68
48.48
55.92
63.08

___________________---_-------Mean Recovery :-

99.91

:- Average o f t h r e e experiments.

The

optimum

wavelength

for

detection

was

335

nm

a t w h i c h good detector response was obtained.

Linearity

was

30-180 p g l r n l and

obtained

10.5-63

in

the

concentration

Fglrnl respectively.

range

The c a l i b r a t i o n

curves could be represented by following regression equation.

( T i n idazole)

(Furazolidone)

= 23.0323

X + 18.650

= 41.3796

X + 15.0636

(r=0.999)
(r=0.999)

TENDOLKAR ET AL.

1652
These
drugs.
ment

equations

used

for

direct

evaluation

of

Results o f t h e r e p l i c a t e analysis and r e c o v e r y e x p e r i of

suspension

are

tabulated

i n Table 3

Anchrom

Enterprises

and

Table

respectively

Downloaded by [University of Chicago Library] at 09:15 16 November 2014

were

ACKNOWLEDGEMENT
Authors

are

grateful

to

for

providing

HPTLC f a c i l i t i e s .

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Received December 1 2 , 1994
Accepted March 28, 1995