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DETERMINATION OF PHENYLEPHRINE HYDROCHLORIDE IN

PHARMACEUTICAL PREPARATIONS USING SPECTROPHOTOMETRIC METHOD

Article  in  Asian Journal of Pharmaceutical and Clinical Research · April 2019

DOI: 10.22159/ajpcr.2019.v12i5.32339

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 Online - 2455-3891
Vol 12, Issue 5, 2019 Print - 0974-2441
Research Article

DETERMINATION OF PHENYLEPHRINE HYDROCHLORIDE IN PHARMACEUTICAL

PREPARATIONS USING SPECTROPHOTOMETRIC METHOD

WASAN A AL-UZRI*
Department of Chemistry, College of Science, University of Baghdad, Jadriyah, Baghdad, Iraq. Email: wasanuzri67@[Link]
Received: 01 February 2019, Revised and Accepted: 09 April 2019

ABSTRACT

Objective: A simple and sensitive spectrophotometric method has been presented for the determination of phenylephrine hydrochloride by coupling
reaction with diazotized sulfacetamide sodium.

Methods: The method is based on the diazotization reaction of sulfacetamide sodium with sodium nitrite in the presence of hydrochloric acid to form
diazonium salt, which is coupled with the drug in alkaline medium to form azo dye, showing absorption maxima at 425 nm.

Results: Calibration plot was linear over the concentration range of 2–24 µg/mL and detection limit of 0.278 μg/mL with a correlation coefficient of
0.9929. All different chemical and physical experimental parameters affecting on the development and stability of the colored product were carefully
studied.

Conclusions: The proposed method was successfully applied to the determination of phenylephrine in nasal drops with good precision and sensitivity.

Keywords: Phenylephrine hydrochloride, Spectrophotometric determination, Sulfacetamide sodium, Diazotization and coupling.

© 2019 The Authors. Published by Innovare Academic Sciences Pvt Ltd. This is an open access article under the CC BY license ([Link]
org/licenses/by/4. 0/) DOI: [Link]

INTRODUCTION Materials and reagents

The reagent grade materials were used throughout this work. PHP,
Phenylephrine hydrochloride (PHP), [(R)-1-(3-hydroxyphenyl).2-
the working standard, was supplied by the State Company for Drug
(methylamino) ethanol hydrochloride], is a white crystalline
Industries and Medical Appliances (SDI), Samarra, Iraq. Pharmaceutical
powder, freely soluble in water, melts at 143°C [1,2] and its chemical
formulations (Nasophrine Nasal Drops [0.25%], SDI, Samarra, Iraq,
structure is:
and Vibrocil Nasal Drops [2.5 mg], Novartis Consumer Health, SA,
Switzerland) were obtained from local markets. Sulfacetamide sodium
(SDI, Samarra, Iraq), sodium nitrite (Merck), hydrochloric acid (HCl)
(BDH), and sodium hydroxide (NaOH) (BDH) were used.

Preparation of solutions
It belongs to a group of drugs named sympathomimetics [3]. It PHP stock standard solution (1000 μg/mL) was prepared by
stimulates alpha receptors in certain areas of the body. It is used
dissolving 0.100 g of pure PHP in distilled water and made up to
locally, as decongestant, for non-specific and allergic conjunctivitis,
100 mL volumetric flask with distilled water. Working standard
sinusitis, and nasopharyngitis [4]. Phenylephrine nasal drops are
solutions were prepared by suitable dilution of the stock standard
used for treating symptoms such as runny nose, sneezing, itching of
solution with distilled water.
the nose, and throat [5]. PHP is normally used to increase the blood
pressure in unstable patients with hypotension, especially resulting
Sodium nitrite solution (3.9×10−3 M) was prepared by dissolving
from septic shock [5]. Various methods reported in literature for
0.0673 g of sodium nitrite in distilled water and diluting to the mark in
analysis of phenylephrine hydrochloride. Examples of these methods
250 mL volumetric flask.
are conductometric titration [6], voltammetry [7-9], thin-layer
chromatography [10], high-performance liquid chromatography
HCl solution (0.5 M) was prepared by diluting 10.88 mL of 11.49 M of
(HPLC) [11-14], flow injection [15-17], and fluorescence [18]. Among
concentrated HCl with distilled water in 250 mL volumetric flask.
the different techniques, the most popular and simple method for rapid
and trace analysis of drugs is spectrophotometry [19-26].
Sulfacetamide sodium solution (0.1%) was prepared by dissolving
0.1 g of sulfacetamide sodium in distilled water and diluting to 100 mL
In this work, a spectrophotometric method for estimation of
volumetric flask with the same solvent.
phenylephrine was described. The method was based on coupling
reaction between diazotized sulfacetamide sodium with the medicine
NaOH solution (2 M) was prepared by dissolving 20 g of NaOH with
in alkaline medium to form a yellow water-soluble azo dye measured
distilled water in 250 mL volumetric flask.
at 425 nm. This method has been successfully applied for the
determination of phenylephrine in nasal drops.
General procedure for calibration
About 2 mL of 0.1% sulfacetamide sodium was transferred into a series
METHODS
of 25 mL calibrated flask. To this solution, equimolar of sodium nitrite
Apparatus solution (3.9×10−3 M) was added and the acidity was adjusted with 1 mL
An optima spectrophotometer ultraviolet-visible (Japan) double beam of 0.5 M HCl solution. The solution was shaken thoroughly. Then, an aliquot
with 1 cm quartz cells was used in all absorbance measurements. of a standard solution (500 μg/mL) containing 0.1–1.2  mL of PHP was
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Asian J Pharm Clin Res, Vol 12, Issue 5, 2019, 1-5

transferred into this series of 25 mL calibrated flasks and 1 mL of 2 M shows that 2 mL of sulfacetamide sodium (0.1%) was enough to obtain
NaOH solution was added, and the contents were diluted to the mark with a maximum absorbance.
distilled water and mixed well. After 15 min, the absorbance of the colored
azo dye was measured at 425 nm against the corresponding reagent blank. Effect of the volume of NaOH (2 M)
For the optimization of conditions and in all subsequent experiments, 1 mL The effect of different volumes (0.5–3 mL) of NaOH was examined on
of 500 µg/mL of PHP in a final volume of 25 mL was used. the maximum absorbance of the formed product. Fig. 4 shows that 1 mL
of NaOH (2 M) was enough to obtain a maximum absorbance.
Procedure for PHP in nasal drops
The contents of three bottles of nasal drops were mixed. An aliquot Effect of reaction time
corresponding to 50 mg of PHP was diluted to 50 mL with distilled water The stability of the product was studied for 180 min following the
in a volumetric flask to obtain 500 μg/mL of PHP. Further, appropriate mixing of the reagents. The colored product developed rapidly after
solutions of pharmaceutical preparations were made by simple dilution mixing and attained maximum absorbance about 15 min at room
with distilled water. temperature. The color was stable for a period of 180 min.

RESULTS AND DISCUSSION Structures of the products

The stoichiometry of the reaction between PHP and diazotized
Determination of absorption maximum
sulfacetamide sodium was investigated under the recommended
An aqueous solution of PHP is reacted with diazotized sulfacetamide
optimum conditions using continuous variation method. The result
sodium in alkaline medium giving yellow dye which became stable
obtained in Fig. 5 shows that a 1:2 azo dye was formed between PHP
after 15 min and has a maximum absorption at 425 nm. Fig. 1 shows
and diazotized sulfacetamide sodium.
the spectra of the product formed.
A reaction subsequent based on the above result is shown in Scheme
Optimization of the experimental conditions
(1) [22].
The effects of various parameters on the absorption intensity of the
formed product were optimized. Determination of stability constant and Gibbs free energy of the
reaction
Effect of the volume of HCl (0.5 M)
The apparent stability constant was calculated by comparing the
The effect of different volumes (0.3–3 mL) of HCl was examined on the
absorbance of a solution containing 1 mL of PHP (1×10−3 M) and 2 mL
maximum absorbance of the formed product. Fig. 2 shows that 1 mL of
of diazotized sulfacetamide sodium (1×10−3 M) (AS) with that of a
HCl (0.5 M) was enough to obtain a maximum absorbance.
solution containing a 5-fold excess of diazotized sulfacetamide sodium
(Am) and according to analytical procedure. The average stability
Effect of the volume of sulfacetamide sodium (0.1%)
constant was (K) = 4.399 × 107 L2 mol-2 where (K = [1-α]/4 α3 C2;
The effect of different volumes (0.5–4 mL) of sulfacetamide sodium was
α = [Am − As]/Am) [27]. This indicates a stable reaction product. The
examined on the maximum absorbance of the formed azo dye. Fig. 3
Gibbs free energy (ΔG) of this reaction was calculated adopting the
following equation: ΔG= −2.303RTlogK where, R is the universal
gas constant (8.314 J mole−1 deg−1), T is the absolute temperature
(273+25°C), and K is the stability constant of the reaction. The value of

Fig. 1: Absorption spectra of the azo dye (20 µg/mL) of

phenylephrine hydrochloride against reagent blank and blank
against distilled water Fig. 3: Effect of the volume of sulfacetamide sodium (0.1%) for the
determination of phenylephrine hydrochloride (20 µg/mL)

Fig. 2: Effect of the volume of hydrochloric acid (0.5 M) for the Fig. 4: Effect of the volume of NaOH (2 M) for the determination of
determination of phenylephrine hydrochloride (20 µg/mL) phenylephrine hydrochloride (20 µg/mL)

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Asian J Pharm Clin Res, Vol 12, Issue 5, 2019, 1-5

Scheme 1: Proposed mechanism of the reaction between phenylephrine hydrochloride and diazotized sulfacetamide sodium

Table 1: Determination of 20 μg/mL of PHP in the presence of excipients

Excipient 200 µg/ml Conc. of phenylephrine, 20 µg/mL Erel.* (%) Rec.* (%)

(Found*)
Lactose 19.917 −0.415 99.585
Starch 20.028 0.140 100.140
Talc 19.945 −0.275 99.725
Sodium chloride 20.012 0.060 100.060
Magnesium stearate 19.945 −0.275 99.725
Polyvinylpyrrolidone 19.986 0.275 100.275
*Average of four determinations, Erel.: Relative error, Rec.: Recovery

Table 2: Analytical data obtained from the determination of PHP ΔG was found to be −43.612 kJ/mole. The negative value of ΔG refers to
hydrochloride the spontaneity of the reaction.

Parameter Value Interferences

λmax (nm) 425 The extent of interfering by some excipients which often accompanied
Beer’s law limits (µg/mL) 2–24 pharmaceutical preparations was studied by measuring the absorbance
Regression equation Y=0.0169X+0.0084 of solutions containing 20 µg/mL of PHP and excess amounts (10-fold
Sandell’s sensitivity (µg/mL) 5.917×10−5 excess) of each excipient, none of these substances interfered seriously
Molar absorptivity (L mol−1 cm−1) 3.442×103 (Table 1).
Correlation coefficient (R2) 0.9929
LOD (µg/mL) 0.278 Analytical characteristics of spectrophotometric method
Stability (min) 180 Calibration graph (Fig. 6) was obtained after optimized all the reaction
Molar ratio (D:R) 1:2 conditions mentioned previously and a series of standard solutions
Color Yellow were analyzed in triplicates to test the linearity. The molar absorptivity
LOD: Limit of detection (ε), the Sandell’s sensitivity (S), the intercept (b), and the slope (a)
were determined and are included in Table 2. The limit of detection
was determined by taking the ratio of the standard deviation (SD) of
Table 3: Accuracy and precision for the proposed method the blank with respect to water and the slope of the calibration curve
multiplied by the factor three [28].
Amount of PHP Recovery % Erel. % RSD %
hydrochloride (µg/mL) The accuracy and precision of the proposed method were
tested by analyzing five replicate of phenylephrine by proposed
Present Found
spectrophotometric method for two different concentrations of
12.00 11.771 98.092 −1.908 1.266 phenylephrine. The values of relative SD relative standard deviation%
16.00 15.953 99.706 −0.294 0.373 and relative error % are summarized in Table 3. These values indicated
Erel.: Relative error, RSD: Relative standard deviation the high accuracy and precision of the proposed method.

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Asian J Pharm Clin Res, Vol 12, Issue 5, 2019, 1-5

AUTHOR CONTRIBUTION

Wasan A. Al-Uzri: The idea of research, preparation of reagents and

solutions with execution of experiments, data interpretation, and
manuscript writing.

CONFLICTS OF INTEREST

There are no conflicts of interest.

Fig. 5: Continuous variation plot REFERENCES

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