INTRODUCTION:

Pharmaceutical formulation with combinations of drugs have shown an increasing trend to counteract the symptoms specific to one drug and formulation and hence analytical chemist will have to accept the challenge of developing reliable and easy simultaneous estimation methods because it does not required manual individual calculations and gives marginally better result.

Paracetamol [N-(4-hydroxy phenyl) acetamide] is one of the common drugs used in the world. Paracetamol is used worldwide for its analgesic and antipyretic actions. It has the spectrum of action similar to that of NSAIDs and resembles particularly the COX-2 selective inhibitors. Paracetamol on average a weaker analgesic than NSAIDs or COX-2 selective inhibitors but is often preferred because of its better tolerance it has central and peripheral inhibitor effects. Paracetamol are reduced by inhibitors of many endogenous neurotransmitter systems including serotogenic, opioid and cannabinoid system.

Caffeine- [1,3,7-trimethyl-1,3, dihydro-1H-purine-2,5-dione. (OR)1,3,7-trimethyl xanthine]. Caffeine is the most widely used central nervous system (CNS) stimulant in the world. It has numerous pharmacological and physiological effects, including cardiovascular, respiratory, renal and smooth muscle effects, as well as effect on mood, memory, alertness and physical and cognitive performance.

EXPERIMENTAL:

Materials:

A UV -VIS Spectrophotometer, model Shimadzu 1700 series was employed with spectral with a pair of matched quartz cells of 10mm optical path length. Paracetamol and Caffeine was obtained as a gift sample from Zim Laboratories, Nagpur. Methanol was procured from S.D five chem. Limited, Mumbai. Brands Combiflam^plus (650mg Paracetamol and 50mg Caffeine) and Novalgin nu (650mg Paracetamol and 50mg Caffeine) was obtained from Sanofi India limited. All other chemicals and reagents were of analytical reagent grade.

Preparation of standard solutions:

Accurately weighed quantities (100mg) of Paracetamol and Caffeine.Both drugs were prepared separately dissolve in 20ml of distilled and volume was made upto 100ml.

Selection of analytical wavelength:

Aliquot portion of stock solution of Paracetamol and Caffeine were diluted with distilled water to obtain 10µg/ml. The solution were scanned in the range of 200-400 in 1cm cell against solvent blank. The overlain spectrum of Paracetamol and Caffeine is shown in fig. from the overlain spectra the wavelength selected for estimation were 243nm as ƛmax for Paracetamol, 273nm as ƛmax for Caffeine.

Fig - Overlain spectra of Paracetamol and Caffeine

ƛmax for Paracetamol = 243nm, and ƛmax for Caffeine = 273nm.

Analysis of laboratory mixture proposed by simultaneous estimation method:

In order to feasibility of the proposed method for simultaneous estimation of Paracetamol and Caffeine in pharmaceutical formulation. The proposed method was tried for the estimation of drug in standard laboratory mixture. The standard solution of PARA and CAF (10µg/ml) were scanned separately in the range of 200-400nm against distilled water as blank and wavelength of maximum absorbance were determined. Absorptivity coefficients of two drugs were determined at both wavelengths and two simultaneous equations were formed. (Table-1).

Table-1 Result of % Estimation of laboratory mixture

Sr. No	Weight of pure drug (g)		Absorbance at nm		% Estimation
1.	0.65	0.050	1.876	2.305	100.04	100.23
2.	0.64	0.051	1.917	2.396	98.039	98.522
3.	0.64	0.050	1.936	2.345	98.039	100.23
4.	0.65	0.050	1.995	2.998	100.04	100.23
5.	0.64	0.051	2.302	2.282	98.039	98.522
Mean					98.839	99.54
±SD					0.9802	0.8367
S.E.					0.4901	0.4185
C.V.					0.2040	0.2390
R.SD.					0.9971	0.8405

Table -2 Result of % estimation in drug formulation

Sr. No.	Weight of drug (g)	Absorbance at nm				% Estimation
Sr. No.	Weight of drug (g)	234	243	255	273	PARA	CAF
1.	0.8092	1.633	1.635	1.986	0.987	100.49	99.87
2.	0.8095	1.789	1.968	1.756	1.003	100.80	101.27
3.	0.8096	1.698	1.789	1.896	0.954	101.95	99.85
4.	0.8098	2.030	1.888	1.669	0.669	99.87	101.71
5.	0.8099	1.988	1.963	1.596	0.968	101.09	101.67
Mean						100.84	100.87
±SD						0.3072	0.3783
SE						0.1536	0.1891
CV						0.0030	0.00375
RSD						0.3046	0.3750

Analysis of formulation by proposed method:

20 tablet were weighed and ground to fine powder. An accurately weighed powder equivalent to 650mg of Paracetamol and 50mg of Caffeine was transferred to a 100ml of volumetric flask containing 10ml methanol and 50ml of distilled water and ultrasonicated for about 15min, the volume was made up to the mark with distilled water. The solution was filtrate through whatman filter paper no. 41. The absorbance’s of resulting solution was measured at 234nm, 243nm, 255nm, 273nm in 1cm cell against solvent blank. Percent label claim was calculated using following formula.

% Label claim = [(cx or cy)/Wm× L] × d × w × 100

Where, C = cx or cy =concentration of paracetamol and caffeine in g/100ml

D = Dilution factor

W =Average of sample of tablet

Wm =Weight of sample taken.

L =label claim of sample taken

Average weight of drug formulation =0.8090 gm

Validation parameter:

Accuracy:

Accuracy of an analytical method is the closeness of the test result obtained by analytical method to the true value conventional value.

Precision:

“Precision of an analytical method is a degree of agreement among individual test results, when method is applied repeatedly to multiple sampling of a homogenous sample. Precision may be a measure of either the degree of reproducibility or repeatability of the analytical method under normal operating conditions.

Recovery:

Recovery study was done by standard addition method:

Preparation of standard solution:

Accurately weighed quantities of pure paracetamol and caffeine were dissolved in 20ml of HCL (0.1N) in separate 100ml volumetric flask volume was made up to the mark with HCL (0.1N) to obtained concentration of 10µg/ml of each drug

Preparation of sample solution:

Accurately weighted quantity of pre-analysed tablet powder equivalent to average weight was taken in 100ml volumetric flask, to it 20ml of HCL (0.1 N) was added and shaken for 15 minutes and volume was adjusted to the mark with of HCL (0.1N). the solution was filtered through whatman filter paper No.41.the aliquot portion of filtrate were further diluted with of HCL (0.1N) the solutions were analyzed as per the procedure describe above.

The % recovery was then calculated by using formula –

%Recovery =A/B+C×100

Where,

A=total drug estimated

B=Amount of drug found in pre -analyzed basis.

C=amount of pure drug added

Table -3 Result and statistical data for recovery study

Sr. No.	Wt of tablet powder	Wt of pure drug (g)		Absorbance at nm				% Recovery
Sr. No.	Wt of tablet powder	PARA	CAF	234	243	255	273	PARA	CAF
1.	0.6503	0.6010	0.049	0.381	0.269	0.348	0.691	100.04	100.20
2.	0.6500	0.6100	0.050	0.353	0.248	0.354	0.657	98.68	100.04
3.	0.6510	0.6120	0.049	0.644	0.448	0.637	1.198	98.38	99.895
Mean								99.033	100.04
±SD								0.3230	0.2778
SE								0.2284	0.1964
CV								0.0032	0.0027
RSD								0.3261	0.2776

RESULT AND DISCUSSION:

Selection of solvent:

During preliminary studies it was observed that Paracetamol were sparingly soluble in water. Caffeine is soluble in freely soluble in boiling water and slightly in ethanol but, ƛ max 243 Paracetamol, ƛ max Caffeine was observed for Paracetamol and Caffeine respectively. However, all two drugs have solubility in water and the overlain spectra of Paracetamol and Caffeine, suggested that there is wide scope of development of several UV analytical methods. The results of recovery study were found to be within the prescribed limit of 98-102% showing recovery for all developed UV spectrophotometric methods. The methods were found to be precise with respective S.D. and R.S.D. values for all the UV spectrophotometric methods. In the specificity study. The sample was exposed to different stress conditions like alkali, acid, heat and oxide. The study showed degradation Paracetamol under oxide, but the exact nature of degradation of drug could not be found out. The low RSD value of inter day and intraday variation revealed that proposed method is robust and rugged. The method is linear as correlation co-efficient were more than 0.999.

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Received on 06.01.2021 Modified on 13.04.2021

Asian J. Pharm. Ana. 2021; 11(3):191-194.

DOI: 10.52711/2231-5675.2021.00033