Jasmine Chaudhary et al.

IRJP 2011, 2 (12), 81-83

INTERNATIONAL RESEARCH JOURNAL OF PHARMACY ISSN 2230 – 8407

Available online www.irjponline.com Review Article

SIMULTANEOUS ESTIMATION OF MULTICOMPONENT FORMULATIONS BY

UV-VISIBLE SPECTROSCOPY: AN OVERVIEW
Jasmine Chaudhary*, Akash Jain, Vipin saini
M.M. College of Pharmacy, M.M.University, Mullana, Ambala
Article Received on: 14/10/11 Revised on: 20/11/11 Approved for publication: 29/12/11

*Email: [email protected]
ABSTRACT
UV-visible spectroscopy, a simple, rapid, precise and highly accurate method for quantitative estimation is in great use now a day. The basic principle behind this technique is
that the amount of light absorbed is proportional to the concentration of analyte. Simultaneous equation is applicable for the estimation of those drugs where the spectra of drugs
overlap properly whereas multi-component analysis can be applied on any degree of spectral overlap provided that two or more spectra are not similar exactly. Quantitative
estimation is necessary before introduction of any drug into the market as either concentration is more in formulation can cause toxicity problem or if concentration is found
less, then formulation may not be effective in prescribed dose.
Keywords: Simultaneous equation, Isoabsorptive, Derivative, Multicomponent Mode, Area under Curve

INTRODUCTION Ø A proper analytical procedure for the drug may not be available
Analytical chemistry is the branch of science which is useful in all in the literature due to patent regulations.
fields of science and medicine due to its versatile applications. It Ø Analytical methods for the estimation of drug in combination
deals basically with the two aspects of chemical characterization i.e. with other drugs may not be available.
qualitative (what it is) and quantitative (how much it is).1 The Ø The existing analytical procedures may require expensive
qualitative analysis reveals the chemical identity of the sample while reagents and solvents. It may also involve cumbersome
quantitative analysis gives the amount of one or more components extraction and separation procedure and these may not be
present in numerical terms. reliable.1, 5
The basic criterion behind these methods is the measurement of The UV-Visible spectroscopic methods for estimation of drugs are
some property which is proportional to amount of analyte in sample. highlighted in this review.
Depending on the property to be measured, these methods are Simultaneous Equation Method6, 7, 8
classified as classical methods such as gravimetry, volumetry, Consider a multicomponent system consisting of two components X
titrimetry, etc and systemic or instrumental methods like and Y, each of which absorbs at the λmax of the other, λ1 being the
refractometry, colorimetry, absorptimetry etc.1, 2 wavelength of maximum absorbance of X (λmax) and λ2 being the
In present era, market is flooded with various combinations in wavelength of maximum absorbance of Y (λmax) (Fig.1.1.)
dosage forms and the number is increasing day by day. 3 These In such cases, it can be possible to determine both the components
multi-components formulations due to greater patient acceptability, by simultaneous equation method. The information required is:
increased potency, multiple action, fewer side effects and quicker Ø The absorptivities of X at λ1 and λ2, ax1 and ax2 respectively.
relief are gaining interest.4 Therefore, it is desired that these Ø The absorptivities of Y at λ1 and λ2, ay1 and ay2 respectively.
formulations meet all the standards related to their quality, safety & Ø The absorbance of the diluted sample at λ1and λ2, A1 and A2
efficacy and this can only be possible if they are analyzed by respectively.
different methods. The motto behind this quantitative estimation is Ø cx and cy be the concentrations of X and Y respectively in the
to ensure that whether a particular drug contains the same amount of diluted sample.
drug as mentioned because if the dose given will be high then it will Thus the absorbance of the mixture at λ1 and λ2 may be expressed as
cause over dosage side effects & if it is less then the patient will not follows:
get the required dose. For the estimation of multi-component A1 = aX1bcX + aY1bcY …………………At λ1 …… (1)
formulation, the instrumental techniques like Spectrophotometric, A2 = aX2bcX + aY2bcY ………………….At λ2 …… (2)
HPLC, GLC, HPTLC etc are employed due to their inherent For measurements in 1 cm cell, b = 1, therefore,
advantages viz. avoid time consuming extraction and separation, Cx = A2ay1 – A1ay2/ ax2ay1 – ax1ay2
economical in the sense that use of expensive regents is minimized, Cy = A1ax2 – A2ax1/ ax2ay1 – ax1ay2
equally accurate and precise. These methods are based upon the Using the above two equations the concentration of component X
measurement of specific and nonspecific physical properties of the and component Y in the sample mixture can be determined.
substances. The Absorption Ratio Method: Isoabsorptive Point Method6
Sometimes the dosage form in addition to the main drugs known to This method is a modification of the simultaneous equations
contain other substances which potentially interfere in the assay and method. According to this method, the ratio of absorbance at any
if not corrected may impart a systemic error to the assay. The need two wavelengths for a substance, which obeys Beer's law, is
to develop new methods to analyze the drugs simultaneously and constant value independent of concentration and path length. This
without interferences is a basic need. Thus, it becomes necessary to constant is termed as “Hufner’s Quotient’ or Q-value. This method
develop new analytical methods for such drugs for which no involves the measurement of absorbance at two wavelengths, one
analytical method is still available for estimation. In brief the being the λ max of one of the components (λ2) and the other being a
reasons for the development of newer methods of drugs analysis are: wavelength of equal absorptivity of the two components (λ1), called
Ø The drug or drug combination may not be official in any as Iso-absorptive point (Fig. 1.2.).5
pharmacopoeias.

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 Jasmine Chaudhary et al. IRJP 2011, 2 (12), 81-83

The concentration of each component can be calculated by mathematical equation:

Cx = (Qm – Qy / Qx – Qy)* A/ a1
Cy = (Qm – Qx / Qy – Qx)* A/ a2
Where, Cx and Cy = concentration of x and y respectively,
A = Absorbance of sample at isoabsorptive wavelength,
a1 and a2 = Absorptivity of x and y respectively at isoabsorptive wavelength,
Qm = Absorbance of sample solution at λ max of one of the components (λ2)
Absorbance of sample solution at isoabsorptive wavelength
Qx = Absorptivity of x at λ max of one of the components (λ2)
Absorptivity of x at isoabsorptive wavelength
Qy = Absorptivity of y at λ max of one of the components (λ2)
Absorptivity of y at isoabsorptive wavelength

Derivative Spectroscopic Method6 absorptivities (E′M, E′N) at one wavelength and the other two
Derivative spectrophotometry involves the conversion of a normal absorptivities (E′′M, E′′N) at the other wavelength can be determined.
spectrum (fundamental, zeroth order or D spectrum) to its first, The absorbance of the mixture A’ and A’’ are experimentally
second or higher derivative spectrum by differentiating absorbance determinable and thus from the above two equations the
of a sample with respect to wavelength λ for higher accuracy concentration of the individual constituents CM and CN can be
(Fig.1.3.).4 readily calculated. This relationship is valid if Beer’s law is followed
[A] = f (λ): zero order and both the components behave independently of one another.
[dA/dλ ] = f (λ): first order Choosing wavelengths at which the differences in molar
[d2A/d λ 2] = f (λ): second order absorptivities are large, leads to attain greater accuracy in this
The strong positive & negative bands with maximum and minimum analysis.
at same wavelength of an absorption band as inflection point in Area Under Curve Method6-9
absorbance band governs the odd (first & third) derivative spectrum This method also utilizes two wavelength ranges. From the overlain
whereas the strong positive & negative band with minimum or spectra of both drugs the area under curve is determined at both the
maximum at same wavelength as λmax of absorbance band governs selected analytical wavelength ranges. Within the above selected
the even (second & fourth) derivative spectrum.10 wavelength ranges, the area under curve was determined for both the
Number of bands = Derivative order + 1 drugs and analysis was performed using “Cramer’s Rule” and
The amplitude (D) is directly proportional to the concentration of “Matrix Method”.
analyte provided Beer’s law is obeyed by D° spectrum. Consider a binary mixture consisting of two components M and N.
In first order derivative spectroscopy, zero crossing point for both From the two spectra (Fig.1.5. and Fig.1.6.) following information
drugs is found and the wavelengths are selected in a manner such are obtained:
M
that at the zero crossing of one drug, the other drug should show v AUC λ1 – λ2: area under curve for component M at the
substantial absorbance. wavelength range λ1 – λ2.
Advantages: M
v AUC λ3 – λ4: area under curve for component M at the
• It enhances resolution permitting identification of analyte with wavelength range λ3 – λ4.
close λmax. N
v AUC λ1 – λ2: area under curve for component N at the
• It eliminates baseline shift effect arising from instrument or
wavelength range λ1 – λ2.
sample handling. N
• It eliminates scattering effects thus helpful for analyte present in v AUC λ3 – λ4: area under curve for component N at the
turbid solution.11 wavelength range λ3 – λ4.
Multicomponent Mode Method The total area under the curve of a mixture at a particular
This method requires two wavelengths. One wavelength is selected wavelength range is equal to the sum of area under curve of the
such that one drug shows maximum absorbance while other drug individual components at same wavelength range. The area under
shows considerable absorbance. The second wavelength is selected curve of the mixture containing component M and N can be given as
such that other drug shows maximum absorbance while the first one follows:
shows considerable absorbance. AUC λ1– λ2 = AUCMλ1– λ2 + AUCNλ1 – λ2 ------------------------------- (1)
M N
Consider a mixture consisting of two components M and N where AUC λ3 – λ4 = AUC λ3 – λ4 + AUC λ3 – λ4 ------------------------------- (2)
X1 nm and X2 nm are the maximum absorbance of component M and Now the above equation can also be written as follows:
M N
N respectively (Fig.1.4.) AUC λ1– λ2 = X λ1 – λ2bCM + X λ1 – λ2bCN ------------------------------ (3)
M N
The absorbance of mixture containing components M and N at AUC λ3– λ4 = X λ3 – λ4bCM + X λ3 – λ4bCN- ----------------------------- (4)
wavelength X1 and X2 may be expressed as follows, Where, X λ1 – λ2 = AUC λ1 – λ2/ Conc. in g/l
A′ = E′MBCM + E′NBCN------------------------------at X1 X λ3 – λ4 = AUC λ3 – λ4/ Conc. in g/l
A′′ = E′′MBCM + E′′NBCN-----------------------------at X2
Using individual standard solution of M and N, the two

By applying “Cramers Rule” and “Matrix Method”, the concentration of component M and component N can be determined as follows:
N N N M N M
CM = X λ1– λ2 AUC λ3 – λ4 – X λ3 – λ4 AUC λ1– λ2 / X λ1– λ2 X λ3 – λ4 – X λ3 – λ4 X λ1– λ2 ----------------- (5)
M M N M N M
CN = X λ1– λ2 AUC λ3 – λ4 – X λ3 – λ4 AUC λ1– λ2 / X λ1 – λ2 X λ3 – λ4 – X λ3 – λ4 X λ1– λ2 --------------- (6)

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 Jasmine Chaudhary et al. IRJP 2011, 2 (12), 81-83

From the study, it has been found that so many combinations have Ofloxacin and Ornidazole in Tablet Dosage Form. Asian J. Research
Chem.2009; 2(1): 60-62.
been successfully estimated by UV-Visible spectroscopic methods
like combinations of Diclofenac sodium and tizanidine10,
Pantoprazole & domperidone11, Dexibuprofen & Paracetamol12,
Enalapril maleate & Amlodipine besylate13, Sulphamethaxole &
Trimethoprin14, Rabeprazole and Itopride15, Ondansetron and
Paracetamol16, Ofloxacin and Satranidazole17, Nebivolol and
Hydrochlorothiazide18, Metronidazole & Amoxicillin19, Norfloxacin
& Ornidazole20, Ofloxacin & Ornidazole21etc.
CONCLUSION
Fig: 1.1. Overlain spectra of component X, Y and Mixture containing X & Y
It can be concluded that UV-visible spectroscopy can be effectively
used for estimation of many drug combinations for which no method
of estimation has been reported so far and it is simple, less time
consuming, accurate and highly sensitive.
ACKNOWLEDGMENT
The authors are very thankful to the management of
Maharishi Markandeshwer University, Mullana (Ambala) for
constant support.
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INTERNATIONAL RESEARCH JOURNAL OF PHARMACY, 2(12), 2011