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Spectrophotometric determination of metronidazole benzoate in

pharmaceutical dosage forms

Article  in  International Journal for Pharmaceutical Research Scholars · January 2021

DOI: 10.31838/ijpr/2020.12.03.497

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 ISSN 0975-2366
DOI:https://doi.org/10.31838/ijpr/2020.12.03.497
Research Article

Spectrophotometric determination of metronidazole

benzoate in pharmaceutical dosage forms
DHURGHAM QASIM SHAHEED 1, QASIM A. BADER 2*, ALAA ABDULHASAN ABBAS 3
1,2
Department of Pharmaceutics and Pharmaceutical Chemistry, College of Pharmacy, University of
Alkafeel, Iraq
3
Department of Pharmacy, Al-Safwa University College, Iraq
*Corresponding Author:
Email: qasim.allawi@alkafeel.edu.iq
Received: 06.07.20, Revised: 23.08.20, Accepted: 07.09.20

ABSTRACT:
Background: Metronidazole is a routinely prescribed and heavily utilized antibiotic. This drug has been
manufactured in many pharmaceutical dosage forms to treat various and serious infections. Hence, the
determination of this vital drug is of an established importance. The UV-Visible protocol is commonly utilized
machine for drugs in both active form and in pharmaceutical preparations. The standard addition procedure
combined with UV visible instrument has shown a well-defined protocol to be followed and applied for
metronidazole quantification procedures.
Methods: Three of the widely available pharmaceutical dosage forms of metronidazole were chosen and
analysed. These pharmaceutical dosage forms are tablets, vials, and oral suspensions. The samples were
handled with modified USP guidance and reasonably diluted and inoculated with the metronidazole standard
solutions. The UV readings were observed and the standard curves were plotted.
Results: The observed curves show well-fitted straight lines for the all three pharmaceutical preparations (i.e.
tablets, vials, and suspensions). The coefficients of regressions were found 0.9982, 0.9993, 0.9996, for tablets,
vials, and suspensions, respectively. Consequently, the contents percentages for the metronidazole tablets, vial,
suspensions were recorded as 100.6%, 102.4%, and 99.5%, respectively.
Conclusion: It can be concluded that this protocol is fit for the quantification purposes for the assessment of
percent contents of metronidazole in various pharmaceutical preparations.
Keywords:Standard addition method, Metronidazole benzoate, Metronidazole tablets, Metronidazole vials,
Metronidazole suspensions.

INTRODUCTION:
Metronidazole is a well-known member of the 5-
Nitroimidazole group. Metronidazole is classified
as antibiotic, antiprotozoal, and amebicide that
has a considerable activity against anaerobic
bacteria alongside specified parasites.
Nitroimidazoles including metronidazole, were
investigated for their antiparasitic activity due to
observation of the effects of azomycin antibiotic
[1-4]. Thereafter, the effect of metronidazole
against intestinal amoebiasis specifically
Entamoeba histolytica, was considered a
revolutionary finding [5, 6]
Chemically, this antibiotic (i.e. metronidazole) has
nomenclature of 2-methyl-5-nitroimidazole-1-
ethanol and the structure can be seen in figure (1)
that shows these functional groups [7].

Fig.1: The chemical structure of metronidazole (molecular formula; C6H9N3O3)

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 Dhurgham Qasim Shaheed et al/ Spectrophotometric determination of metronidazole benzoate in
pharmaceutical dosage forms

The chemical synthesis of metronidazole can be nitroimidazole (compound 2) that in turn, is

seen in figure (2) and illustrated in tree major obtained from the nitration of 2-Methylimidazole
steps; the metronidazole (compound 3) can be (compound 1) [8-10].
obtained via the alkylation of 2-methyl-4(5)-

Fig.2: Reveals schematic pathways of the chemical syntheses of metronidazole.

Metronidazole has been analysed in pure and

many pharmaceutical dosage forms as METHODS AND MATERIALS
mentioned in many articles [11-13]. UV-Visible Materials:
spectroscopy results were comparable to HPLC Metronidazole was purchased from Wuxi Hexia
[14, 15] and has been employed as robust, fast, Chemical Company, Wuxi, Jiangsu Province,
and easy to use technique [16-18]. The authors in China.
this study aimed to utilize a standard addition The analysed pharmaceuticals were purchased
technique to quantify metronidazole in from local pharmacy and these are tabulated in
pharmaceutical preparation using the classical table (1)
UV-Vis spectroscopy.

Table 1: shows the detailed information for the analysed pharmaceuticals.

No. Dosage Trade name Strength Batch no. Expiry
form date
1 Tablet METROSULE-500 500 mg/tablet G / 25 / 2080DJ0609H 07 -
2022
2 Suspension Flazole 200 200 mg/5mL AB0204 12 -
2021
3 Vial Pezole 500mg/100mL 9NSEX50006 02 -
2022

Instruments: The flask was sonicated for five minutes to assure

UV-6100PC DOUBLE BEAM fill dissolution. This led to the final clear solution
SPECTROPHOTOMETER, EMGLAB GmbH metronidazole standard with a concentration of
Germany, (model: UV-6100PC 10 mg/mL.
spectrophotometer). Digital Ultrasonic Cleaner,
1.5 L, CODYSON, China, (Model: CD-4820). λmax selection:
Micropipette; (0.5-10) L, (10-100) L, and (100- An exact volume of 2.5 mL of metronidazole
1000) L, Dragon Lab, Shanghai, China. Four standard solution was transferred into a 10-mL
digits electric sensitive balance (Model: OHAUS - tube and the volume was completed with distilled
AV114 Adventurer Pro, Switzerland). water. The later solution was scanned and the
maximal absorption was recorded, accordingly.
Methodologies: Metrosule-500® tablets:
Standard Solution: Twenty Metrosule-500® tablets were weighed and
An accurately weighed 1g of metronidazole was entirely crashed to get a homogenized fine
transferred into a 100-mL volumetric flask. The powder. A suitable portion of that powder was
volume was filled up to mark by distilled water. transferred into 100-mL volumetric flask with 30
mL distilled water. This mixture was sonicated for

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 Dhurgham Qasim Shaheed et al/ Spectrophotometric determination of metronidazole benzoate in
pharmaceutical dosage forms

5 minutes, shaken mechanically for 0.5 hour, and
the volume was completed to the mark with
distilled water to get an equivalent concentration
of 10 mg/mL. Thereafter, this flask was let to
settle down and 100 µL of the supernatant was
further diluted up to 100 mL mixed well to get a
clear sample solution. Eventually, this sample
solution was aliquoted to six tubes of ten millilitres
portions to be utilized in the "standard addition"
curve as revealed below.
Pezole® vial:
A metronidazole injection of 5 mg/mL called
Pezole® vial was analysed. A portion of 5mL of
Pezole® vial is transferred into 25 mL volumetric
flask and filled with distilled water up to the mark
and mixed. Then, 1 mL is taken from this solution
and was diluted up to 100-mL with distilled water.
This solution was mixed well and was aliquoted to
six tubes of ten millilitres portions to be utilized in
the "standard addition" curve as revealed below.
Flazole® 200 suspension:
An exact volume of 250 µL was allocated in 100
mL volumetric flask and the volume was
completed with distilled water up to the mark.
This flask was sonicated for three minutes to
assure dissolving of the suspended particles of the
metronidazole suspension. Thereafter, 10 mL of
this solution was transferred into 100 mL
volumetric flask and filled up with distilled water.
This solution was mixed well and was aliquoted to
six tubes of ten millilitres portions to be utilized in
the "standard addition" curve as revealed below.
Standard Addition Curve:
For each series the above six tubes of 10-mL of
metronidazole sample preparations, spikes of
(0.0, 10, 20, 30, 40, and 50) µL of
metronidazole standard solution, were
inoculated. These tubes were mixed with the
added standards. This leads to observed final
standard concentrations of (0, 10, 20, 30, 40,
and 50) µg/mL, respectively. Then, for each
sample preparation, triplicates were precisely
prepared. Finally, all the prepared samples were
submitted to the UV-Vis light at 320 nm and the
absorbed portion to be measured at room
temperature. The experiments were performed at
the scientific labs of the College of Pharmacy.
RESULTS:
Metronidazole λmax:
The scanned standard solution of metronidazole
shows λmax of 320 nm. This can be seen in
figure (3) neatly presented:

Fig.3:This figure presents the maximal absorption of metronidazole standard solution as 320 nm
was found as the λmax.

Metronidazole Standard Addition Calibration
Curve Observed with UV-Vis Machine:
This curve was plotted for each drug sample
resulting in observing of three distinct calibration
curves, one for each pharmaceutical preparation.
These are; metronidazole tablets (shown in figure
4), metronidazole vial (shown in figure 5), and
metronidazole oral suspension (shown in figure
6).

3528| International Journal of Pharmaceutical Research | Jul - Sep 2020 | Vol 12 | Issue 3
 Dhurgham Qasim Shaheed et al/ Spectrophotometric determination of metronidazole benzoate in
pharmaceutical dosage forms

Metronidazole Tablets Standard Addition Curve

1.2
Absorbance at 320 nm

0.8

0.6

0.4 y = 0.0168x + 0.169

R² = 0.9982
0.2

0
-15 -10 -5 0 5 10 15 20 25 30 35 40 45 50 55
(Standard - Sample concentration) (µg/mL)

Fig.4: Reveals the metronidazole standard addition calibration curve observed with UV-Vis
machine. The intersection with the x-axis of the standard curve represents the diluted
concentration of the sampled metronidazole tablets.

Metronidazole vial Standard Addition Curve

1.2

1
Absorbance at 320 nm

0.8

0.6

0.4 y = 0.0162x + 0.1658

R² = 0.9993

0.2

0
-15 -10 -5 0 5 - Sample
(Standard 10 15 20 25 (µg/mL)
concentration) 30 35 40 45 50 55

Fig.5: Reveals the metronidazole standard addition calibration curve observed with UV-Vis
machine. The intersection with the x-axis of the standard curve represents the diluted
concentration of the sampled metronidazole vial.

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 Dhurgham Qasim Shaheed et al/ Spectrophotometric determination of metronidazole benzoate in
pharmaceutical dosage forms

Metronidazole Suspension Standard Addition Curve

1.2
Absorbance at 320 nm

0.8

0.6

0.4
y = 0.0172x + 0.1712
0.2 R² = 0.9996

0
-15 -10 -5 0 5 10 15 20 25 30 35 40 45 50
(Standard - Sample concentration) (µg/mL)

Fig.6: Reveals the metronidazole standard addition calibration curve observed with UV-Vis
machine. The intersection with the x-axis of the standard curve represents the diluted
concentration of the sampled metronidazole suspension.

The analyses of the observed data of the given curves gave the parameters shown in table (2).

Table 2: Shows some statistics gained from the standard addition curves for the analyses of
metronidazole in different pharmaceutical preparations.
No. Item Metrosule- Pezole® vial Flazole® 200
®
500 tablets suspension
1 Intercept with the Y-axis 0.169 0.1658 0.1712
2 Standard Deviation (SD) 0.001713 0.002502 0.00229
3 Coefficient of Variation (CV) 0.231895 0.650654 0.536289
4 Regression coefficient (R2) 0.9982 0.9993 0.9996
5 Slope 0.0168 0.0162 0.0172
6 Limit of detection (LOD = 3SD) 0.005138 0.007505 0.006871
7 Limit of quantification (LOQ = 10SD) 0.017128 0.025018 0.022904
8 Dilution-factor 50,000 500 4,000
9 Stated amount or concentration 500 mg / 5 mg / 1 mL 200 mg / 5 mL
tab
10 Found amount or concentration 503 mg / 5.12 mg / 1 mL 199 mg / 5 mL
tab
11 Percent content of metronidazole 100.6 % 102.4 % 99.5 %

DISCUSSION: 110.0% stated in the USP [30]. Moreover, the

Metronidazole is a worldwide routinely and
extensively prescribed antibiotic. It is indicated in
variant conditions and for various causative
organisms those commonly lead to an
inflammation [19]. Therefore, it is required to find
a rapid, inexpensive and easy to perform daily
utilized method of determination of such
antibiotic. The UV-Vis has been approved for such
purpose [20-23]. The observed lambda maximum
was found to be 320 nm. This is agreed with
many researches [24-28] and as far as possible
from the solvent (water) cut off value (190 nm)
[29]. The content percentage of metronidazole
tablets was found as 100.6%. The found
percentage obeys the allowed range of 90.0% -
percent content of metronidazole vial was found
to be 102.4%. This percent is falling within the
accepted stated range of 90%-110% as allowed
in the USP [31]. The last pharmaceutical
preparation is the metronidazole oral suspension.
This analyte has a percent content of 99.5%. This
percent is perfect with the knowledge of the
accepted allowance of 98.5% - 101.0% as stated
within the USP [32].
CONCLUSION
It has been concluded that the standard addition
method is fit for purpose regarding the
determination of percent contents of

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 Dhurgham Qasim Shaheed et al/ Spectrophotometric determination of metronidazole benzoate in
pharmaceutical dosage forms
metronidazole in various pharmaceutical
preparations.
CONFLICT OF INTEREST:
"None are declared".
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