Journal of Biotechnology and Bioengineering

Volume 4, Issue 1, 2020, PP 20-25

ISSN 2637-5362

New Spectrophotometric Methods for Determination of

Albendazole in Presence of Cerium as Oxidant and Both Indigo
Carmine and Alizarin Red Dyes in Bulk and Dosage Forms
Oday T. Alia&b, Khaled M. Elgendy a, Monir Z. Saada, Wafaa S. Hassanc, Mahmoud M. Sebaiyd*
a
Department of Chemistry, Faculty of Science, Zagazig University, Zagazig, Egypt
b
Ministry of Education, Iraq
c
Department of Analytical Chemistry, Faculty of Pharmacy, Zagazig University, Zagazig, Egypt
d
Department of Medicinal Chemistry, Faculty of Pharmacy, Zagazig University, Zagazig, Egypt
*Corresponding Author: Mahmoud M.Sebaiy, Department of Medicinal Chemistry, Faculty of
Pharmacy, Zagazig University, Zagazig, Egypt, Email: sebaiym@gmail.com

ABSTRACT
Novel spectrophotometric methods have been developed for the determination of albendazole in bulk, tablet
and suspension dosage forms. Solution of albendazole in methanol was oxidized using known excess ceric
ammonium nitrate (Ce4+) then the un-reacted amount of Ce4+ was determined using two different dyes,
indigo carmine and alizarin red using universal buffer. The reaction showed maximum absorbance at 610
nm for indigo carmine and at 401 nm for alizarin red. Different factors were studied such as type of buffer,
dye volume, oxidant volume, time, temperature, organic solvents, and sequence addition. It was found that
Beer's law is obeyed in the range of 1.32-7.95 µg mL-1 for albendazole with both dyes with molar
absorptivity of 4.69× 103 L.mol-1.cm-1 for indigo carmine and 97.53× 103 L.mol-1.cm-1 for alizarin red.
Also the methods showed a high degree of sensitivity in respect of limit of detection which was reported to
be as low as 0.24 and 0.35 µg mL-1 in case of indigo carmine and alizarin red, respectively. By applying
statistical treatment, it was found that the methods were accurate and precise with low standard deviation
values. The methods were then applied for the determination of albendazole in tablet and suspensions with
satisfactory results.
Keywords: spectrophotometric; albendazole; ceric ammonium nitrate; indigo carmine; alizarin red.

as a result, the pharmacokinetic studies were

INTRODUCTION done using ABZSO as an active metabolite and
Albendazole (ABZ), chemically known as methyl- ABZSO2 as an inactive metabolite [4-8]. As such,
5-(propylthio)-2-benzimidazole carbamate [1] the therapeutic importance of this compound
(Fig. 1), is widely used as an anthelmintic with a justifies research to develop analytical methods
broad spectrum of activity [2]. ABZ is an inactive for its determination in bulk form, pharmaceu-
moiety but it readily gets metabolized to an tical formulations and in biological samples.
active metabolite, albendazole sulfoxide (ABZSO) Several techniques can be found in the literature
which is then further metabolized to an inactive for monitoring ABZ therapy such as UV-vis
metabolite, albendazole sulfone (ABZSO2) [3]. spectrophotometry [9-12], spectroflourimetry
As ABZ is metabolized extensively, therefore, the [13], HPLC [14], HPTLC [15], LC/MS [16],
plasma concentration of ABZ is also very low, capillary electrophoresis [17] and voltammetry [18].

Figure1. Chemical structures of albendazole (ABZ), indigo carmine (INC) and alizarin red (ALR)

Journal of Biotechnology and Bioengineering V4 ● 11 ● 2020 20

New Spectrophotometric Methods for Determination of Albendazole in Presence of Cerium as Oxidant
and Both Indigo Carmine and Alizarin Red Dyes in Bulk and Dosage Forms

To our knowledge, no spectrophotometric method oxidation reaction of ABZ with a known excess
for the determination of ABZ using cerium as an of ceric ammonium nitrate (Ce4+) followed by a
oxidant has yet been reported despite the versatility, subsequent determination of the residual oxidant
simplicity and reliability of the technique in by reacting it with a measured amount of the
chemical analysis. The requirements of pharma- dyes, indigo carmine (INC) at 610 nm or
ceutical quality control are more severe than in alizarin red (ALR) at 401 nm.
other fields; drug control requires excelled
EXPERIMENTAL
accuracy, specificity and precision. Further,
because active components are often present in Apparatus
low amounts in pharmaceutical formulations, All absorbance measurements were made with a
the methods must be very sensitive. These Systronics (model 6800 UV / VIS digital
requirements are fulfilled by the two methods spectrophotometer JENWAY) equipped with 1
presented in this paper which are based on an cm matched quartz cells.

Materials
Name Formula Mol.Wt Supplier
Albendazole C12H15N3O2S 265.34mg ̸ mol EIPICO
ceric ammonium nitrate (NH4)2Ce(NO3)6 548.26 GLOBAL chemie
Indigo carmine C16H8N2Na2O8S2 466.34 SDFCL
Alizarin red C14H8O4 240.21 SDFCL
Methanol CH3OH 32.04 SIGMA
Ethanol CH3CH2OH 46.06 SIGMA
Propanol CH3CH2CH2OH 60.096 SIGMA
DMF HCON(CH3)2 73.095 SIGMA
Acetone C3H6O 58.08 SIGMA
Formaldehyde CH2O 30.026 SIGMA
Ethylene glycol C2H6O2 62.068 SIGMA

 A stock solution of 1 × 10-3 M ABZ was Phosphate Buffer Solution

prepared by dissolving an exact weight in A series of universal buffer solutions, covering
100 mL measuring flask and completed to the pH range from (7 to 10) were prepared as
the mark by diluted methanol. recommended by Lurie [21].
 Stock solutions of 1 × 10-3 M INC dye, ALR Pharmaceutical Preparations
dye, and Ce4+ were prepared by dissolving an Drug Trade Name company Formulation
exact weight in 100 mL measuring flasks and Alzental® 200mg EIPICO Tablets
completing to the mark by distilled water. ABZ Alzental® 20mg/mL EIPICO suspension
Bendax® 20mg/mL SIGMA suspension
Buffers
General Procedure
Universal Buffer Solutions
In 10 mL volumetric flask, accurate volume of
A series of universal buffer solutions, covering ABZ (1×10-3 M) and 2 mL Ce4+ were mixed for
the pH range from (2 to 10) were prepared as 10 minutes in presence of universal buffer (pH 9
recommended by Britton [19]. for INC dye or pH 10 for ALR dye). 1.25 mL of
Borate Buffer Solutions INC dye followed by 1 mL ethanol or 1 mL of
ALR dye followed by 1 mL propanol was then
A series of borate buffer solutions, covering the added with shaking for 1 minute before completing
pH range from (7 to 9) were prepared as to the mark with distilled water. Using
recommended by Bower and Bates [20]. spectrophotometer, the un-reacted INC dye or
ALR dye was measured at 610 nm, and 401 nm,
Acetate Buffer Solution respectively, using a reagent blank similarly
A series of acetate buffer solutions, covering the prepared without ABZ. The concentration of
pH range from (3 to 6) were prepared. ABZ was then determined form calibration

21 Journal of Biotechnology and Bioengineering V4 ● 11 ● 2020

New Spectrophotometric Methods for Determination of Albendazole in Presence of Cerium as Oxidant
and Both Indigo Carmine and Alizarin Red Dyes in Bulk and Dosage Forms

curve previously constructed under the optimum RESULTS AND DISCUSSION

condition.
Absorption Spectra
Pharmaceutical Applications
Ce4+ has been used as an effective oxidizing
ABZ was determined in different drug samples of agent for determination of different materials
different forms and companies such as Alzental® like ABZ giving a number of oxidized products.
tablet, Alzental® and Bendax ® suspensions. A The un-reacted Ce4+ oxidizes known amount of
suitable weight or volume of each form was the dye while the remaining dye is measured
dissolved and diluted in methanol in 100 mL spectrophotometrically at corresponding maximum
volumetric flasks. 1.00 – 3.00 mL sample of wavelengths. The absorption spectra of the
each was mixed with 2 mL Ce4+ and universal remaining dye showed characteristic λ max values
buffer (pH 9 for INC dye or pH 10 for ALR
as shown in Fig. 2 at 610 for INC and at 401 nm
dye) and the methods were completed according
for ALR.
to the above mentioned procedure.

Figure2. Absorption spectra of ABZ with (A) INC dye at 610 nm and (B) ALR dye at 401 nm.

Factors Affecting the Absorbance phosphate buffer solution (pH 7-10). The
absorbance was measured with 1 mL ABZ (1 ×
Effect of Buffer Type 10-3 M) against a blank solution prepared by the
The effect of buffer on the oxidation of ABZ same way without ABZ. Universal buffer (pH 9)
with Ce4+ was studied. Different Buffers were was chosen as the optimum buffer in case of
used such as universal buffer (pH 2-10), borate INC while universal buffer (pH 10) was the
buffer (pH 7-9), acetate buffer (pH 3-6), and of optimum media for ALR as shown in Fig. 3.

Figure3. Effect of buffer pH on the absorption of ABZ with (A) INC dye at 610 nm and (B) ALR dye at 401 nm.

Effect of Oxidant Volume concentrations of dye and it was recorded that

1.25 mL is the best volume for INC while 1 mL
The effect of different volumes of Ce4+ in
is the best one for ALR.
presence of dye on the oxidation process was
investigated using 1 mL ABZ (1 × 10-3 M) and it Effect f Sequence of Addition
was found that the suitable volume of Ce4+ was
Under the above mentioned conditions, the
2 mL in case of both dyes.
effect of sequence of additions on the oxidation
Effect of Dye Concentration process of ABZ was investigated by measuring
The effect of dye concentration on the oxidation the absorbance of solution prepared by different
process was investigated by adding different sequences of addition against a blank solution

Journal of Biotechnology and Bioengineering V4 ● 11 ● 2020 22

New Spectrophotometric Methods for Determination of Albendazole in Presence of Cerium as Oxidant
and Both Indigo Carmine and Alizarin Red Dyes in Bulk and Dosage Forms

prepared by the same way without the studied was studied by measuring the absorbance at
material. The most favorable sequence was different time intervals. The results indicated
(ABZ - Ce4+ - buffer universal - dye) which showed that shaking for one minute was sufficient to
the best results for determination of ABZ while give reliable and stable values.
other sequences gave lower absorbance values. Effect of Temperature
Effect of Time The effect of temperature on the oxidation process
The time required to complete the oxidation of was studied by measuring the absorbance of a
ABZ by Ce4+ was studied at different time intervals solution containing ABZ and Ce4+ against blank
(5-50 min). The proper time for completing the solution at different temperatures (30 – 60˚C). The
oxidation process is 10 minutes for both dyes. On optimum temperature is almost at room temperature
the other hand, the effect of time after dye addition (30 ± 1 C˚) for both dyes as shown in Fig. 4.

Figure4. Effect of temperature (0C) on the absorption of ABZ and Ce4+ with (A) INC dye at 610 nm and (B)
ALR dye at 401 nm

Effect of Organic Solvents the absorbance was measured against the blank
solution. The results illustrated that the value of
The effect of organic solvents on the absorption
absorbance was improved by using ethanol as a
spectra was studied in presence of different
solvent in case of INC while it increased in prese-
organic solvents such as propanol, ethanol, DMF,
nce of propanol in case of ALR as shown in Fig. 5.
acetone, formaldehyde, and ethylene glycol and

Figure5. Effect of organic solvents on the absorption of ABZ with (A) INC dye at 610 nm and (B) ALR dye at 401 nm.

Linearity Also the methods showed a high degree of

Seven different concentrations of ABZ were sensitivity in respect of limit of detection which
specified for linearity studies. The calibration was reported to be as low as 0.24 and 0.35 µg
curves obtained by plotting absorbance against mL-1 in case of INC and ALR, respectively. Other
concentration showed linearity in the concentr- parameters are reported in Table 1 indicating a
ation range of 1.23-7.95 µg mL-1 for both dyes. high degree of linearity, accuracy and precision.

23 Journal of Biotechnology and Bioengineering V4 ● 11 ● 2020

New Spectrophotometric Methods for Determination of Albendazole in Presence of Cerium as Oxidant
and Both Indigo Carmine and Alizarin Red Dyes in Bulk and Dosage Forms

Table1. Analytical merits for determination of ABZ in pure sample using the proposed spectrophotometric methods.
INC ALR

Found µg mL-1

Found µg mL-1
Taken µg mL-1

Taken µg mL-1

Recovery %
Recovery %
Parameters

1.32 1.30 98.48 1.23 1.22 99.18

1.85 1.84 99.45 1.85 1.83 98.91
2.65 2.64 99.62 3.97 3.95 99.49
3.44 3.42 99.41 4.50 4.49 99.77
3.97 3.96 99.74 5.30 5.20 98.11
4.50 4.48 99.55 6.62 6.61 99.84
7.95 7.93 99.74 7.95 7.94 99.89
Mean 99.42 99.31
±SD 0.43 0.64
±RSD 0.44 0.65
±SE 0.13 0.19
Variance 0.19 0.41
LOD (µg mL-1) 0.24 0.35
LOQ (µg mL-1) 0.72 1.07
Molar absorptivity (L.mol-1.cm-1) 4.69× 103 97.53× 103

analyzed by the proposed methods. Excipients

Analytical Applications of Pharmaceutical
and impurities did not show interference
Formulations
indicating a high degree of specificity regarding
Alzental® tablet, Alzental® and Bendax® the percentage recovery and RSD % for the
suspensions containing ABZ had been successfully methods as recorded in Table 2.
Table2. Application of the spectrophotometric methods for the determination of Alzental® tablet, Alzental® and
Bendax® suspensions dosage forms.
Recommended Found Taken
Type RSD % Recovery % Dye
(µg mL-1) (µg mL-1) (µg mL-1)
Alzental® Tablet 7.50 92.50 1.83 1.85 2
Alzental® suspension INC 3.00 98.80 9.74 9.88 10
Bendax® suspension 2.80 97.20 4.85 4.86 5
Alzental® Tablet 3.00 97.00 1.97 1.94 2
Alzental® suspension ALR 2.30 97.70 9.73 9.77 10
Bendax® suspension 0.26 98.20 4.90 4.91 5

CONCLUSION antifungal, anthelmintic and antitimor activity a

review. International Journal of Pharmacy and
The proposed methods are reported to be Pharmaceutical Sciences. 2014; 6:17-25.
simple, sensitive, precise and accurate based on [2] A. Hemphill. Treatment of echinococcosis:
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1.23-7.95 µg mL-1 for albendazole with both J.L. Del Estal, J.G. Prieto. Small intestinal
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Citation: Mahmoud M. Sebaiy, Oday T. Ali, Khaled M. Elgendy, Monir Z. Saad, Wafaa S. Hassan, "New
Spectrophotometric Methods for Determination of Albendazole in Presence of Cerium as Oxidant and Both
Indigo Carmine and Alizarin Red Dyes in Bulk and Dosage Forms", Journal of Biotechnology and
Bioengineering, 4(1), 2020, pp. 20-25.
Copyright: © 2020 Mahmoud M. Sebaiy. This is an open-access article distributed under the terms of the
Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any
medium, provided the original author and source are credited.

25 Journal of Biotechnology and Bioengineering V4 ● 11 ● 2020