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IOSR Journal of Pharmacy and Biological Sciences (IOSRJPBS)
ISSN: 2278-3008 Volume 2, Issue 2 (July-August 2012), PP 46-48
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Spectrophotometric Estimation of Flucloaxacillin in Pure Drug
and Pharmaceutical Dosage Formulation
C.M.Bhaskar Reddy 1, G.V.Subba Reddy, 2
1
(Associate professor of chemistry, HOD of chemistry , Dept of humanities &basic sciences , Samskruti
college of engineering & Technology (Affiliated to JNT University Hyderabad,)
Kondapur, Ghatkesar, Hyderabad, Rangareddy (Dist), pin-501301, A. P , India
2
(Associate professor of chemistry ,Dept of Chemistry Jntu college of Engineering, Pulivendula . Kadapa
(Dist), pin-516390, A.P, India
Abstract : A simple and sensitive spectrophotometric method has been described for the assay of Flucloxacillin
either in pure form or in pharmaceutical solid dosage form. Absorption maxima of Flucloxacillin in methanol
were found to be at 220 nm. Beer’s law is obeyed in the range 5-70 μg/mL. Result of percentage recovery
and placebo interference shows that the method was not affected by the presence of common excipients. The
percentages assay of Flucloxacillin in tablet was more than 99%. The method was validated by determining its
sensitivity, accuracy and precision which proves suitability of the developed method for the routine estimation
of Flucloxacillin in bulk and solid dosage form.
Keywords: Flucloxacillin , methanol, UV spectrophotometer, validation
I.
Introduction
.Flucloxacillin [1] is chemically (2S, 5R, 6R)-6-({[3-(2-chloro-6-fluorophenyl)-5-methylisoxazole -4yl] carbonyl} amino)-3, 3-dimethyl-7-oxo-4-thia-1-azabicyclo [3.2.0] heptane-2-carboxylic acid which is a
narrow spectrum beta lactam antibiotic of the penicillin class. It is used to treat infections caused by susceptible
gram-positive bacteria [1] FLUCLOXACILLIN chemical formula is C19H17ClFN3O5S [2]
.FLUCLOXACILLIN is stable against hydrolysis by a variety of beta-lactamases, including penicillinases, and
cephalosporinases and extended spectrum beta-lactamases. By binding to specific penicillin-binding proteins
(PBPs) located inside the bacterial cell wall, flucloxacillin inhibits the third and last stage of bacterial cellwall
synthesis. Cell lysis is then mediated by bacterial cell wall autolytic enzymes such asautolysins; it is possible
that flucloxacillin interferes with an autolysin inhibitor.. Literature survey revealed that FLUCLOXACILLIN
was estimated either individually or in presence of amoxicillin trihydrate by using RP-HPLC method[3,4]
Flucloxacillin (INN) or floxacillin (USAN) is a narrow-spectrum beta-lactam antibiotic of the penicillin class
Unlike other penicillins, flucloxacillin has activity against beta-lactamase-producing organisms such as
Staphylococcus aureus[5] Flucloxacillin has similar pharmacokinetics, antibacterial activity, and indications to
dicloxacillin, and the two agents are considered interchangeable. It is believed to have higher incidence of
severe hepatic adverse effects than dicloxacillin, but a lower incidence of renal adverse effects.[6]
Flucloxacillin is contraindicated in those with a previous history of allergy to penicillins, cephalosporins, or
carbapenems. It should also not be used in the eye, or administered to those with a history of cholestatic
hepatitis associated with the use of dicloxacillin or flucloxacillin.[6] It should be used with caution in the
elderly; patients with renal impairment, where a reduced dose is required; and those with hepatic impairment,
due to the risk of cholestatic hepatitis.[6] Flucloxacillin is indicated for the treatment of infections caused by
susceptible bacteria. Specific approved indications include
Staphylococcal skin infections and cellulitis –
including impetigo, otitis externa, folliculitis, boils, carbuncles, and mastitis :[6][7] Common adverse drug
reactions (ADRs) associated with the use of flucloxacillin include: diarrhoea, nausea, rash, urticaria, pain and
inflammation at injection site, superinfection (including candidiasis), allergy, and transient increases in liver
enzymes and bilirubin.[6] eeks.[6][7] Rarely, cholestatic jaundice (also referred to as cholestatic hepatitis) has
been associated with flucloxacillin therapy. The reaction may occur up to several weeks after treatment has
stopped, and takes weeks to resolve. The estimated incidence is 1 in 15,000 exposures, and is more frequent in
people >55 years, females, and those with treatment longer than 2 w eeks.[6][7] Fluclox was determined
spectrophotometrically in pharmaceutical preparations using some nitrophenols as chromogenic reagents [8].
An). isocratic ion exchange high performance liquid chromatography method was developed for the
simultaneous determination of Fluclox and amoxicillin in pharmaceutical formulations for injections [9]. A
modified spectrophotometric method was developed for the determination of some ß-lactam antibiotic s
including Fluclox and Diclox with pyrocatechol violet as chromogenic reagents [10] and potassium periodate
[11]. Diclox was determined spectrophotometrically in a binary mixture with amoxicillin [12]. Liquid
chromatographic methods [13–18] were used as quantitative tools for the determination of Fluclox and Diclox.
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Spectrophotometric Estimation Of Flucloaxacillin In Pure Drug And Pharmaceutical Dosage Formulation
High performance thin-layer chromatography was developed for determination of chloramphenicol, ampicillin,
benzylpenicillin, Diclox and erythromycin in cow’s milk [19]. Therefore, an attempt was made to develop a
simple spectrophotometric method for the estimation of the present drug in bulk and pharmaceutical dosage
formulations
II.
Experimental Section
1. 1
Instrument and apparatus
Perkin Elmer UV-Visible Spectrophotometer Lambda 25 model was used for spectral measurements
with spectral band width 1 nm; wavelength accuracy is 0.5 nm and 1 cm matched quartz cells. Glassware used
in each procedure was soaked ove r night in a mixture of chromic acid and sulphuric acid rinsed thoroughly with
methanol and dried in hot air oven.
1.2 Reagents and Materials
All chemicals were of analytical reagent grade and methanol was used to prepare solutions.
1.3 Standard drug solution
Pharmaceutical grade Flucloxacillin was kindly provided by Synchron Research Services Pvt. Ltd,
Ahmedabad, India. A stock standard solution equivalent to 1mg/mL Flucloxacillin was prepared by dissolving
50 mg of pure drug in methanol and diluting to 50 mL in calibrated flask with. Methanol
III.
Method
Different aliquots (0.0, 0.5, 1.0,…… , 7.0 mL) of 1 mg/mL Flucloxacillin solution were accurately
measured and transferred into a series of 100 mL volumetric flasks and volume made up to the mark with.
Methanol Then all dilutions were scanned between 200-400 nm against blank which shows the maximum
absorbance at 220 nm The same λ max was used for further measurement of drug. A calibration curve for
absorbance vs. concentration was plotted (Fig.2).
2.1 Assay of pharmaceutical Formulations
Twenty tablets were weighed accurately and ground into a fine powder. Powder equivalent to 100mg
of Flucloxacillin was weighed accurately and transferred into a 100 mL volumetric flask with 60 mL. The
content was shaken for 15-20 min, diluted to volume with Methanol, and filtered using a Whatman No. 42 filter
paper. First 10 mL portion of filtrate was discarded and subsequent portions were subjected to analysis.
IV.
Results And Discussion
The absorption spectrum of Flucloxacillin was measured in the range 200–400 nm against the blank
solution Methanol similarly prepared .The standard solution show maximum absorbance at λ max for each
three systems as recorded in Table 1. And the method was validated by studying the following parameters in
Table1.
FIG: 1 The structure of flucloxacillin
Systematic (IUPAC) name is
(2S, 5R, 6R)-6-({[3-(2-chloro-6-fluorophenyl)-5-methylisoxazole-4-yl]
carbonyl}amino)-3,3-dimethyl-7-oxo-4-thia-1-azabicyclo[3.2.0] heptane-2-carboxylic acid
FIG: 2 The calibration curve of flucloxacillin
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Spectrophotometric Estimation Of Flucloaxacillin In Pure Drug And Pharmaceutical Dosage Formulation
Table1:
Parameters for determination of Flucloxacillin against Methanol
Parameters
Values
λ max, nm
220
Beer’s law limit, μg /mL
5– 70
Molar absorptivity, L mol1.52x104
1cm-1
Regression equation
y=a+bc
Slope (m)
0.0297
Intercept (c)
0.01714
Correlation coefficient
0.9657
The accuracy of the above method was ascertained by comparing the results obtained with the proposed and
reference methods in the case of formulation are presented in Table 2.
Table 2: Assay and Recovery of Flucloxacillin in Pharmaceutical Formulations
Formulation
Label
Amount
% Recovery
%Recovery
Claim (mg) Found (mg)
Proposed method
Reference method
Floxapen,
40
39.63
99.07
98.56
Fluclomix,
40
39.71
99.27
99.13
As an additional check on the accuracy of these methods, recovery experiments were performed by
adding known amounts of pure drug to pre-analyzed formulation and percent recovery experiments were also
done. Recovery experiments indicated the absence of interferences from the commonly encountered
pharmaceutical additives and excipients.
V.
Conclusion
It could be concluded that the developed method for estimation of Flucloxacillin in pharmaceutical
dosage forms and in bulk is simple, sensitive, relatively precise and economical .The proposed methods are
used for the routine analysis of the drugs in the quality control.
VI.
Acknowledgment
Authors are thankful to the Principal Dr A.S, Reddy Samskruti college of Engineering & Technology,
Ghatkesar ,Kondapur , Hyderabad India, for providing research facilities for this work.
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