Simultaneous Determination of Aspirin, Paracetamol, Caffeine in Tablets

Alano, Alesha O., Figueroa, Jasmin Marciel S., Hermosura, Sarah C. & Mendoza, Tria B. (Pharmacy 4 –
2)

College of Pharmacy, De La Salle Medical and Health Sciences Institute

ABSTRACT

Aspirin, caffeine, and phenacetin in synthetic mixtures were subjected to quantification using
Ultraviolet and Visible Spectrophotometry. Stock solutions were prepared. Factors such as temperature,
nature of solvent, and pH of the solutions were controlled which caused a bathochromic shift. The
percentage label claim for aspirin, caffeine, and phenacetin were found to be 58.94%, 43.34%, and 0%,
respectively. This did not conform to the USP requirement where they should be 90 – 110%.

Keywords: synthetic mixture, aspirin, Beer-Lambert Law, caffeine, phenacetin, UV spectroscopy

INTRODUCTION

Absorption Spectroscopic method of analysis based on Ultraviolet (UV) and Visible Radiation is
considered as one of the most widely used and powerful tools for quantitative analysis. It can determine
concentrations of colored compounds in various chemicals. (Skoog, 2007). Absorption of visible and UV
radiation is associated with excitation of electrons, in both atoms and molecules, from lower to higher
energy levels.

Many analgesic tablets contain paracetamol, aspirin, and caffeine as active components. These
as combined are applied in Ultraviolet-Visible Spectrophotometry to quantify the concentration of each
drug component in the mixture. Aspirin, a salicylate drug, is used to relieve pain, reduce a high
temperature (fever), and reduce swelling. It has a stronger anti-inflammatory action than paracetamol.
Paracetamol, on the other hand, cause less gastric irritation and less toxic effects to blood than aspirin.
Caffeine, a central nervous system stimulant, is used to relax muscle concentrations in blood vessels to
improve blood flow during headaches. (Drugs.com, 2018)

Figure 1. The structures of the different drugs. Aspirin, Caffeine, and Paracetamol, respectively.

The absorbance of different species is guided by the Beer - Lambert Law, which is the linear
relationship between absorbance and concentration the absorbing species. The general Beer - Lambert
law is usually written as A=Ɛbc, where A is the absorbance, Ɛ is the Molar Absorptivity, b is the
pathlength, and c is for the concentration. To determine the absorbance in a mixture, the formula used is
A = Ɛ1b[C1] + Ɛ2b[C2] + Ɛ3b[C3]+…+ Ɛnb[Cn], where [C] is the concentration of one component. (Skoog,
2007)

The objective of this study is to determine the working wavelength for quantification of each drug in a
ternary mixture, calculate the molar absorptivity in the determined ‫ג‬max, and solve for the concentration of
each component using elimination - substitution technique of solving system equations and using matrix
solution technique.
METHODOLOGY
Preparation of Standard Stock Solution
10mg was weighed and dissolved in 50mL 0.5N Sodium Hydroxide. The solution was further diluted to
100 mL with 0.5N Sodium Hydroxide.

Preparation of Standard Calibration Solutions

2.50 mL from the stock solution was then further diluted to 25 mL with 0.5N Sodium Hydroxide. This
contained 10 µg/mL concentration.

Determination of Working Wavelength

For each of the standard solutions, their maximum absorbance were then determined from 210 to 380
nm.

Preparation of Sample Solution

The sample containing aspirin, phenacetin and caffeine was weighed to about 100mg of aspirin. This was
then dissolved to 50 mL with 0.5N Sodium Hydroxide. The solution was then further diluted to 100 mL
with 0.5 N Sodium Hydroxide. 100 µL from the solution was then further diluted to 10 mL with the same
solvent, 0.5N Sodium Hydroxide. The absorbance was measured in three wavelengths identified by the
standard.

RESULTS AND DISCUSSION

Table 1. Molar absorptivities and final concentration of aspirin, phenacetin, and caffeine at different λmax
Solution Final Concentration Wavelength Absorbance Absorptivity

SS10 ASA 6.9384x-5 mol/L λ1 245 0.199 2868.0964

λ2 271 -0.047 -677.3896

λ3 302 0.170 2450.1326

SS10 ACP 5.8030x10-5 mol/L λ1 245 0.498 8581.7681

λ2 271 0.057 1121.3850

λ3 302 -0.070 -1206.2726

SS10 CAF 5.4071x10-5 mol/L λ1 245 0.285 5270.8476

λ2 271 0.482 8914.2054

λ3 302 -0.047 -869.2275

Table 2. Mean and standard deviation of the absorbances of the different trials for the synthetic mixture of
aspirin, phenacetin, and caffeine.
Wavelength Absorbance

Trial 1 Trial 2 Trial 3 Mean Standard

Deviation

245 0.328 0.364 0.353 0.348 0.0184

271 0.044 0.080 0.082 0.069 0.02139

302 0.101 0.126 0.124 0.117 0.01389

The wavelengths of aspirin, phenacetin and caffeine were absorbed at 302 nm, 245 nm and 271
nm, respectively. The molar absorptivity for each compound at each wavelength were computed using
the Beer-Lambert’s Law while the concentrations of each compound in the mixture were computed
through Matrix Resolution Technique. In order to determine mean drug contents, aspirin 250mg,
phenacetin 205mg, and caffeine 45mg were tested. Their percentage label claims were 58.94%, -67.54%
or 0% and 43.34%, respectively. Based on the obtained results, these three did not conform to the
requirement of the USP, which should be 90 – 100%.

CONCLUSION AND RECOMMENDATION

Based on the computed percentage label claim, the sample did not conform with the official
requirement of the USP for all of the three components—aspirin, caffeine, and phenacetin which should
be 90 – 100%. Dilution of the caffeine may have interfered as to why its percentage label claimed to be
0%. It is then recommended to observe proper dilution techniques, as well as, more trials be performed in
order to obtain an accurate result. The small amounts weighed produced higher percentage of error,
which signifies the recommendation of performing serial dilution; higher concentration of stock solution is
also recommended to avoid such errors.

REFERENCES:

Drugs.com. (2018). Aspirin. Retrieved from

https://www.drugs.com/search.php?searchterm=aspirin on September 22, 2018.
Drugs.com. (2018). Caffeine. Retrieved from
https://www.drugs.com/search.php?searchterm=caffeine on September 22, 2018.
Drugs.com. (2018). Paracetamol. Retrieved from
https://www.drugs.com/search.php?searchterm=paracetamol on September 22,
2018.
Murtaza, G., et.al. (2011, January 18). Development of a UV-spectrophotometric method for the
simultaneous determination of aspirin and paracetamol tablets. Research gate. Retrieved from
https://www.researchgate.net/publication/233860462_Development_of_a_UV-
spectrophotometric_method_forthe_simultaneous_determination_of_aspirin_andparacetamol_in_
tablets on September 22, 2018.
Skoog, D.A., Holler, F.J. & Neiman, T.A. (2007). Principles of instrumental analysis, 6th
ed. Philadelphia: Harcourt Brace & Co.
APPENDIX

FINAL CONCENTRATION
For ASA:
𝑤𝑡 𝑜𝑓 𝑡ℎ𝑒 𝑠𝑎𝑚𝑝𝑙𝑒 𝑎𝑙𝑖𝑞𝑢𝑜𝑡 𝑚𝑜𝑙 1000 𝑚𝐿
𝑥 𝑥 𝑥
𝑣𝑜𝑙𝑢𝑚𝑒 𝑜𝑓 𝑡ℎ𝑒 𝑠𝑡𝑜𝑐𝑘 𝑠𝑜𝑙 𝑣𝑜𝑙𝑢𝑚𝑒 𝑜𝑓 𝑡ℎ𝑒 𝑐𝑎𝑙𝑖𝑏𝑟𝑎𝑡𝑖𝑜𝑛 𝑠𝑜𝑙 𝑚𝑜𝑙𝑒𝑐𝑢𝑙𝑎𝑟 𝑤𝑒𝑖𝑔ℎ𝑡 1 𝐿
;.;=>? @ >.? AB = ACD =;;; AB
𝑥 𝑥 𝑥 = 6.9384x-5 mol/L ASA
=;; AB >? AB =E;.=?FG = B

For ACP:
;.;=;H @
>.? AB = ACD =;;; AB
𝑥 𝑥 𝑥 = 5.8030x10-5 mol/L ACP
=;; AB >? AB =FG.>=G @ = B

For CAF:
;.;=;? @ >.? AB = ACD =;;; AB
𝑥 𝑥 𝑥 = 5.4071x10-5 mol/L CAF
=;; AB >? AB =GH.=G @ = B

ABSORPTIVITY
𝐴
𝑎=
𝑏𝑐
For ASA:
λ1 245
;.=GG
𝑎= = 2868.0964 L/mol/cm of ASA
= KA (M.GNEHO=;P?ACD/B)
λ2 271
P;.;HF
𝑎= = -677.3896 L/mol/cm of ASA
= KA (M.GNEHO=;P?ACD/B)
λ3 302
;.=F;
𝑎= = 2450.1326 L/mol/cm of ASA
= KA (M.GNEHO=;P?ACD/B)
For ACP:
λ1 245
;.HGE
𝑎= = 8581.7681L/mol/cm of ACP
= KA (?.E;N;S=;P? TUV/W)
λ2 271
;.;?F
𝑎= = 1121.3850 L/mol/cm of ACP
= KA (?.E;N;S=;P? TUV/W)
λ3 302
P;.;F;
𝑎= = -1206.2726 L/mol/cm of ACP
= KA (?.E;N;S=;P? TUV/W)
For CAF:
λ1 245
;.>E?
𝑎= = 5270.8476 L/mol/cm of CAF
= KA (?.H;F=S=;P? TUV/W)
λ2 271
;.HE>
𝑎= = 8914.2054 L/mol/cm of CAF
= KA (?.H;F=S=;P? TUV/W)
λ3 302
P;.;HF
𝑎= = -869.2275 L/mol/cm of CAF
= KA (?.H;F=S=;P? TUV/W)

MEAN ABSORBANCE AT λ:
Σ(𝑇𝑟𝑖𝑎𝑙 1 + 𝑇𝑟𝑖𝑎𝑙 2 + 𝑇𝑟𝑖𝑎𝑙 3)
x=
𝑛

Mean Absorbance at λ1
Σ(0.328 + 0.364 + 0.353)
x= = 0.348
3
Mean Absorbance at λ2
Σ 0.044 + 0.080 + 0.082
x= = 0.069
3
Mean Absorbance at λ3
Σ 0.101 + 0.126 + 0.124
x= = 0.117
3

Concentration of each compound in the mixture

Concentration of compound in the

mixture
E.=FGM=;;E=O=;P? ACD =E;.=?FG @ =;;;;;; AK@ = B
CASA= 𝑥 𝑥 𝑥
B ACD = @ =;;; AB
= 14.73621375 mcg/mL ASA
PF.F>?>=MFEFMO=;P? ACD
=FG.>=G @ =;;;;;; AK@ = B
CACP= 𝑥 𝑥 𝑥
B ACD = @ =;;; AB
= -13.84505627 mcg/mL ACP
=.;;HNH;G=FO=;P? ACD
=GH.=G @ =;;;;;; AK@ = B
CCAF= 𝑥 𝑥 𝑥
B ACD = @ =;;; AB
= 1.950319627 mcg/mL CAF

Label Claim
ASA = 250 mg/tab
>?; A@ ;.= AB
LCASAmg/mL = 𝑥 = 0.025 𝑚𝑔/𝑚𝐿 or 25 mcg/mL ASA
=;; AB =; AB
ACP = 205 mg/tab
>;? A@ ;.= AB
LCACPmg/mL = 𝑥 = 0.0205 𝑚𝑔/𝑚𝐿 or 20.5 mcg/mL ACP
=;; AB =; AB
CAF = 45 mg/tab
H? A@ ;.= AB
LCCAFmg/mL = 𝑥 = 0.0045 𝑚𝑔/𝑚𝐿 or 4.5 mcg/mL CAF
=;; AB =; AB

Percentage Label Claim

dCeK.Cf ghg
%LC = x100
Bd Cf ghg
For ASA:
=H.FNM>=NF? Tij/TW
%LCASA = x100
>? Tij/TW
= 58.94% ASA
P=N.EH?;?M>F Tij/TW
%LCACP = x100
>;.? Tij/TW
= -67.54% ACP
=.G?;N=GM>F Tij/TW
%LCCAF = x100
H.? Tij/TW
= 43.34% CAF