SOP TRANSACTIONS ON ANALYTICAL CHEMISTRY

VOLUME 1, NUMBER 1, JUNE 2014

SOP TRANSACTIONS ON ANALYTICAL CHEMISTRY

Quantitative Analysis of Paracetamol,

Acetylcysteine and Guaifenesin in
Commercial Cold Medicines by UV-Vis
Spectroscopy
UK
TUNCA*
Aysel KUC
Sakarya University, Faculty of Arts and Sciences, Department of Chemistry, Sakarya, 54187, Turkey
*Corresponding author: ayselk@[Link]

Abstract:
Paracetamol (acetaminophen) is a painkiller and antipyretic agent. Acetylcysteine is a mucolytic
agents and it is N-acetylated derivative of L-Cysteine, which is a natural amino acid. Guaifenesin
(glyceryl guaiacolate) is an expectorant agent. All these three agents are active ingredients of
several cold and pain killer medicines sold in Turkish market today (Parol, Asist Plus, Cinetix,
Mucoplus and Vicks Vapo Expectorant). A spectroscopic quantitative determination method not
requiring a derivation process has been developed successfully for all these agents and their
amounts in these trade preparations were determined. Finally, bioanalytical method validation
was performed for the developed method and main validation parameters were determined.
Keywords:
Paracetamol; Acetylcysteine; Guaifenesin; UV-Vis. Spectroscopy

1. INTRODUCTION
Paracetamol (P) has a painkiller effect on headaches, toothaches, joint aches, middle ear aches, aches
stem from cold, flue, migraine, neuralgia, lumbago, sinusitis, medical operations or injuries [15].
Acetylcysteine (A) decreases the density and sliminess of mucus accumulated in the breath system,
transforms mucus into a viscous state like water and solves mucus. It also helps excretion of bronchial
secretions and regulates lung functions by making breathing easier [610]. On the other hand, Guaifenesin
(G) enables efficient coughing by decreasing the sliminess of the secretions in the respiratory system and
the mucus density and lowers the chest congestions by extracting mucus out easily [1114] (See Figure
1).
In the literature, there are several studies on quantitative analysis of these three active ingredients in
trade preparations by UV-spectroscopy [15, 7, 9, 12, 13]. On the other hand, in the literature there is
no study combining the analysis of these three active ingredients together. The method developed in
this study does not require a derivation process and it is also practical, easy and economical for UV
spectroscopy analysis of these three active ingredients.

40

Quantitative Analysis of Paracetamol, Acetylcysteine and Guaifenesin in Commercial Cold Medicines by UV-Vis
Spectroscopy

Figure 1.

Molecular formulas for Paracetamol (N-(4-hydroxyphenyl) ethanamide) (C8 H9 NO2 ) (1), Acetylcysteine ((2R)-2-acetamido-3-sulfanyl propanoic acid) (C5 H9 NO3 S) (2) and Guaifenesin ((RS)-3-(2methoxyphenoxy) propane-1,2-diol) (C10 H14 O4 ) (3)

Figure 2.

Calibration curve for P standards prepared in methanol

2. MATERIAL AND METHOD

Thermo Scientific brand UV-Vis. Spectrophotometer device connected to a Samsung brand computer
was used in metricconverterProductID1 cm1 cm quartz cells for UV spectroscopic studies.
Pure, certificated P, A and G standards, 500 mg Parol (tablet), 600 mg Asist Plus (sachet), 900 mg
Cinetix (Effervescent tablet), 1200 mg Mucoplus (Effervescent tablet) and 200 mg Vicks Vapo Expectorant
(syrup) trade preparation forms and analytical grade methyl alcohol (Merck) were obtained from trade
sources. Deionized water obtained from deionizer system (Milli-Q, Millipore-advantage 10) in our lab
was used only for Acetylsystein standard solutions and all its trade sample solutions.

Table 1. Calibration Curve Values Obtained for P, A and G (n=6)

Concentration
Ranges (g/mL)

Regression Equations

Regression Coeffi- Relative Standard Decients (r)

viation* (RSD%)

P(5-25g/mL)

y=0.0040x-0.0086

0.9986

1.49-5.92

A(10-40 g/mL)

y=0.0453x+0.3296

0.9859

0.78-2.55

G(5-25 g/mL)
y=0.0027x+0.1342
0.9794
1.77-2.32
*RSD: (100xSD/X) (Relative Standard Deviation: 100xStandart Deviation/Average Value)
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SOP TRANSACTIONS ON ANALYTICAL CHEMISTRY

Figure 3.

Calibration curve for A standards prepared in deionized water

Figure 4.

Calibration curve for G standards prepared in methanol

3. EXPERIMENTAL
A serial of standard solutions was prepared from the stock solutions containing P, A and G to develop a
new quantitative method by UV spectroscopy for trade medicines containing these agents and then the
calibration curves were obtained. After that, the analysis method for these active ingredients was applied
to several trade preparations and their recoveries were tested. The method developed was validated using

Table 2. Precision Values Obtained for P, A and G (n=6)

Selected Concentra- Intra-day Precision Values Inter-day Precision Values
tions (g/mL)
(RSD%)
(RSD%)
P

42

4.08

15

5.19

4.00
4.77

20

1.49

1.46

10

1.15

2.09

20

2.55

2.73

30

1.95

2.20

1.77

2.07

15

2.32

4.83

20

1.90

2.72

Quantitative Analysis of Paracetamol, Acetylcysteine and Guaifenesin in Commercial Cold Medicines by UV-Vis
Spectroscopy

Table 3.

Average Recovery Data for P, A and G Obtained from Trade Preparations (n=6)
P 500 mg
A 600 mg (Asist Plus A 900 mg (Cinetix Ef- A 1200 mg
(Parol tablet) Effervescent tablet)
fervescent tablet)
(Mucoplus sa(15 g/mL) (25 g/mL)
(25 g/mL)
chet)
(25 g/mL)

G 200 mg/15 mL
(Vicks Vapo Expectorant syrup)
(15 g/mL)

5.002

1.541

1.576

1.583

0.200

4.989

1.478

1.577

1.509

0.202

5.002

1.553

1.549

1.565

0.203

5.002

1.544

1.537

1.568

0.193

5.002

1.485

1.498

1.511

0.202

4.989

1.581

1.562

1.488

0.205

Average
Abs. (Xort )

4.998

1.527

1.550

1.537

0.201

Standard
Deviation (SD)

0.0067

0.0390

0.0300

0.0390

0.0042

Relative Standard De- 0.13

viation (RSD)

2.55

1.92

2.56

2.07

Found Concentration
(g/mL)

15.00

25.20

25.58

25.36

17.02

Recovery*
(%)

103.00

100.80

102.30

101.45

106.80

Relative Error*
+3.00
+0.79
+2.26
+1.43
+6.80
(%)
*Recovery: (Found amount/Added amount)x100, Relative Error (Accuracy): (Fount amount-Added Amount/ Added Amount)x100

all validation parameters such as specificity, sensitivity, precision, accuracy, linearity and repeatability
[15].

3.1 Preparation of Standard and Trade Sample Solutions

100 g/mL main stock solutions were prepared in deionized water from pure A standard and in
methanol from pure P and G standards. After that standard solutions at concentration ranges of 5-25
g/mL, 10-40 g/mL and 5-25 g/mL were prepared from these stock solutions by diluting with water
and methanol in fixed volumes for P, A and G respectively.
A 15 g/mL sample solution was prepared in methanol media from Parol tablet containing 500 mg P.
25 g/mL sample solutions were prepared in water media from 900 mg Cinetix and 1200 mg Mucoplus
efervesan tablets and 600 mg Asist Plus sachet preparation containing A active ingredient. A 15 g/mL
trade sample solution was prepared in methanol media from Vicks Vapo Expectorant syrup containing
200 mg G in 15 mL.

3.2 Validation Tests

Six measurements were taken repeatedly during the day for three different concentrations selected
from the defined working ranges of each active ingredient. After that, three separate calibration curves
were drawn from the averages of these six measurements (n=6). The data were analyzed by linear
regression model using least squares method and the regression coefficients (r) were obtained ( Figures 24). Furthermore, quite good RSD (Relative Standard Deviation) values obtained from the measurements
of the solutions within these working ranges (See Table 1). After that intra-day and inter-day precision

43

SOP TRANSACTIONS ON ANALYTICAL CHEMISTRY

Figure 5.

Spectra obtained from all standards at concentration ranges of 5-25 g/mL, 10-40 g/mL and 5-25
g/mL for P, A and G respectively

Figure 6.

UV spectrum for 500 mg Parol trade preparation

tests were performed for three different concentrations (low, medium and high) selected from the prepared
standards and the repeatability tests were carried out for the same standards. RSD values obtained from
intra-day and inter-day precision tests showed quite good precision values (As shown in Table 2).
For precision determination, RSD values for intra-day and inter-day measurements should be < 10%
and LOQ (Limit of Quantitation) should be < 20%. For accuracy inter-day average value X should be
44

Quantitative Analysis of Paracetamol, Acetylcysteine and Guaifenesin in Commercial Cold Medicines by UV-Vis
Spectroscopy

Figure 7. UV spectra for 600 mg Asist Plus (1), 900 mg Cinetix (2) and 1200 mg Mucoplus (3) trade preparations

Figure 8.

UV spectrum for 200 mg/15 mL Vicks Vapo Expectorant trade preparation

in 15% of the real value and LOQ should not deviate more than 20%. All these conditions were
obtained. In this study, LOQ values (LOQ=10SD/m) (m, slope value in calibration curve) were 1.23
g/mL, 5.70 g/mL and 1.48 g/mL for P, A and G respectively and LOD values (Limit of Detection)
(LOD=3SD/m) were calculated as 0.37 g/mL, 1.71 g/mL and 0.45 g/mL for P, A and G respectively.
Absorbance data were measured at 287 nm, 193 nm and 284 nm for P, A and G respectively. In Figure
5, spectra obtained for all standards at concentration ranges of 5-25 g/mL, 10-40 g/mL and 5-25
45

SOP TRANSACTIONS ON ANALYTICAL CHEMISTRY

g/mL for P, A and G respectively were shown together. While UV Spectrum for P 500 mg Parol was
given in Figure 6, UV spectra for A 600, 900 and 1200 mg Asist Plus, Cinetix and Mucoplus were given
in Figure 7 and UV spectrum for G 200 mg/15 mL Vicks Vapo Expectorant was given in Figure 8.
RSD data, relative errors and recoveries obtained from six repeated measurements of analysis samples
prepared from several trade preparations were given in Table 3. It can be seen from this table that the
RSD data obtained are 2.56%, relative error data are < almost 7.00% and recovery data are quite good
(almost 100%).

4. RESULT AND DISCUSSION

In this study, UV-Vis. Spectroscopy quantitative analysis method was developed for five different
trade preparations containing P, A and G active ingredients. The developed method is an alternative for
the other spectroscopic methods in the literature since there is no need to derivate all these three active
ingredients and it is both easy to apply and economic. Furthermore quite good recoveries were obtained
in a shorter period of time and in simpler way. It was concluded that the method can solely be applied to
any trade preparations containing P, A and G active ingredients easily. All the validation parameters such
as specificity, sensitivity, precision, accuracy, linearity and repeatability were applied to test the validity
of the method and bioanalytical method validation was also performed.

References
[1] V. Rodenas, M. Garca, C. Sanchez-Pedreno, and M. Albero, Simultaneous determination of
propacetamol and paracetamol by derivative spectrophotometry, Talanta, vol. 52, no. 3, pp. 517
523, 2000.
[2] S. Behera, S. Ghanty, F. Ahmad, S. Santra, and S. Banerjee, UV-Visible Spectrophotometric Method
Development and Validation of Assay of Paracetamol Tablet Formulation, Journal of Analytical &
Bioanalytical Techniques, vol. 3, no. 6, 2012.
[3] V. Vichare, P. Mujgond, V. Tambe, and S. Dhole, Simultaneous Spectrophotometric determination
of Paracetamol and Caffeine in Tablet formulation, Int. J PharmTech Res, vol. 2, no. 4, pp. 251216,
2010.
[4] X. Feng, Q. Zhang, P. Cong, and Z. Zhu, Determination of the paracetamol degradation process
with online UV spectroscopic and multivariate curve resolution-alternating least squares methods:
comparative validation by HPLC, Analytical Methods, vol. 5, no. 19, pp. 52865293, 2013.
[5] B. R. Shrestha and R. R. Pradhananga, Spectrophotometric method for the determination of
paracetamol, Journal of Nepal Chemical Society, vol. 24, pp. 3944, 2009.
[6] H. N. Tse, L. Raiteri, K. Y. Wong, K. S. Yee, L. Y. Ng, K. Y. Wai, C. K. Loo, and M. H. Chan,
High-Dose N-Acetylcysteine in Stable COPD, Chest, vol. 144, no. 1, pp. 106118, 2013.
[7] L. Kukoc-Modun and N. Radic, Spectrophotometric Determination of N-Acetyl-L-Cysteine and N(2-Mercaptopropionyl)-Glycine in Pharmaceutical Preparations, International journal of analytical
chemistry, vol. 2011, 2011.
[8] W. T. Suarez, O. D. Pessoa-Neto, B. C. Janegitz, H. J. Vieira, R. C. Faria, and O. Fatibello-Filho,
Flow injection spectrophotometric determination of N-acetylcysteine and captopril employing
Prussian blue generation reaction, Analytical Letters, vol. 44, no. 14, pp. 23942405, 2011.
[9] M. A. Eid, Spectrophotometric determination of cysteine andN-acetylcysteine in pharmaceutical
preparations, Microchimica Acta, vol. 129, no. 1-2, pp. 9195, 1998.
46

Quantitative Analysis of Paracetamol, Acetylcysteine and Guaifenesin in Commercial Cold Medicines by UV-Vis
Spectroscopy

[10] M. G. Alvarez-Coque, M. M. Hernandez, R. V. Camanas, and C. M. Fernandez, Spectrophotometric

determination of N-acetylcysteine in drug formulations with o-phthalaldehyde and isoleucine,
Analyst, vol. 114, no. 8, pp. 975977, 1989.
[11] S. P. Reddy, K. Sudhakar Babu, N. Kumar, and Y. Sasi Sekhar, Development and validation of
Stability Indicating the RP-HPLC method for the estimation of related compounds of Guaifenesin in
pharmaceutical dosage forms, Pharmaceutical methods, vol. 2, no. 4, pp. 229234, 2011.
[12] A. A. Bankar, S. R. Lokhande, R. Sawant, and A. R. Bhagat, Spectrophotometric estimation of
guaifenesin and salbutamol in pure and tablet dosage form by using different methods,
[13] I. Bhattacharyya, S. Bhattacharyya, C. Kyal, P. Choudhury, B. Dhakal, and S. Ghosh, Estimation
and Validation of Stability Indicating UV Spectrophotometric Method for the Determination of
Guaifenesin in Presence of Its Degradant Products, Int. J. Pharm. & Pharma. Sci, vol. 5, no. 1,
pp. 262268, 2013.
[14] O. M. Abdallah, Sensitive spectrophotometric method for quantitation of guaifenesin and
dropropizine in their dosage forms, International journal of analytical chemistry, vol. 2010, 2010.
[15] I. H. T. Guideline, Q2B validation of analytical procedures: methodology, Fed. Regist, vol. 62,
1997.

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