INTRODUCTION:

Meropenem is chemically (4R, 5S, 6s) – 3- [ (2S, 5S)5- (Di methyl carbamoyl) pyrrolidin-2-yl] sulfanyl – 6 – (1- hydroxy ethyl) – 4 – methyl – 7 –oxo – 1- aza bicyclo [3.2.0] hept -2 ene-2-carboxylic acid. It is a broad–spectrum carbapenem antibiotic and is active against Gram-positive and Gram – negative bacteria, exerts its action by penetrating bacterial cells readily and interfering with the synthesis of vital cell wall components, which leads to cell death. A very few physico-chemical methods appeared in the literature for the determination of Meropenem in serum and in pharmaceutical formulations, which include spectrophotometric^1-8, HPLC^9-14, liquid chromatographic^15-17 and electrophoresis^18-20 techniques. The analytically important functional groups of Meropenem are not fully exploited for designing suitable spectrophotometric methods for its determination. Hence the author made an attempt and succeeded in developing this sensitive, precise and accurate spectrophotometric method based on the formation of an ion-association complex^21-30 between the drug and acidic dye methyl orange.

MATERIALS AND METHODS:

Apparatus: All spectral and absorbance measurements were made on a Techcomp UV – 2301 UV-Visible spectrophotometer with 1 cm matched quartz cells.

Chemicals and reagents:

All chemicals used were of analytical grade. Meropenem (pharmaceutical grade) was obtained from local pharmaceutical laboratory and commercial formulations were procured from the market. Aqueous solutions of methyl orange (0.1% w/v) and 0.1M HCl were prepared.

Preparation of standard solution:

About 100mg of Meropenem was accurately weighed and transferred into a 100ml volumetric flask and diluted to volume with methanol to get the stock solution (1mg /ml). From this, suitable dilutions were made to obtain a final working concentration of 250 µg/ml.

Preparation of sample solution:

Accurately weighed formulation powder equivalent to 100mg of Meropenem was transferred to a 100ml volumetric flask. About 20ml of methanol was added and sonicated for 10min. finally made up the volume with methanol and mixed thoroughly. The resulting solution was filtered through a Whatman filter paper. From this, suitable dilutions were made to obtain the working concentration of 250µg/ml.

Procedure for Calibration Curve:

Into a series of 250ml separating funnels containing aliquots of standard drug solution ranging from 0.5 to 5.0ml (250µg/ml) were transferred. To that 1.0ml of HCl and 2.0ml of methyl orange solution were added and the total volume of the aqueous phase was made up to 15ml with distilled water. About 10ml of dichloromethane was added to each funnel and the contents were shaken for few min. and two phases were allowed to separate and the absorbance of the dichloromethane layer was measured at 400nm against the corresponding reagent blank (Fig. 1). The amount of drug present in the unknown sample was measured from the calibration curve (Fig. 2). For comparing accuracy of the results obtained by the proposed method, the author has developed a simple and sensitive UV-spectrophotometric method (Fig. 3 and 4) and adapted it as a reference method.

Fig. 1: Absorption spectrum of the proposed method

Fig. 2: Calibration curve of proposed method

Fig. 3: Absorption spectrum of UV reference method

Fig. 4: Calibration curve of UV reference method

RESULTS:

The proposed method is based on the formation of an ion-association complex due to the reaction of protonated form of Meropenem with acidic dye methyl orange, which was a pale yellow colored chromogen extracted with dichloro methane from the aqueous phase. Optical characteristics such as absorption maximum, Beer’s law limits, molar absorptivity, sandell’s sensitivity are summarized in table-1. The relative standard deviation and confidence limits were considered satisfactorily for this method. To find out the suitability of the proposed method for the assay of formulations, different dosage forms containing selected drug were analyzed by proposed method and the reference method. The results obtained from the proposed and reference methods were compared statistically by the t-test and F-test and are summarized in table-2. In order to check the accuracy of the method, recovery experiments were performed by analyzing each pharmaceutical formulation in the first instance for the active ingredients by the proposed method. A known amount of pure drug was then added to each of the previously analyzed formulation and the total amount of the drug was once again determined by the proposed method and percent recovery values obtained are listed in table-2.

Table 1: Optical characteristics of the proposed method for Meropenem

S. No.	Parameter	Proposed Method
1	λ max, nm	400
2	Beer’s law limits (µg/ml)	10 – 30
3	Molar absorptivity (l mol^-1 cm^-1)	0.2439×10³
4	Sandell’s sensitivity (µg cm^-2)	1.5723
5	Regression equation (Y= mX+b)
	Slope (m)	0.0052
	Intercept (b)	0.0090
6	Correlation coefficient (r)	0.9998
7	Relative Standard Deviation (%)*	0.6818
8	% Range of error (confidence limits)
	0.05 level	± 0.8482
	0.01 level	± 1.4039
* Mean of five determinations

Table 2: Assay of Meropenem in commercial formulations by proposed method

Formu lation	Labeled amount mg/vial	Proposed Method			Reference Method	% Recovery**
Formu lation	Labeled amount mg/vial	Amount found*	t-Value	F-Value	Reference Method	% Recovery**
A	500	499.86 ± 0.152	0.3120	3.7019	499.89 ± 0.079	98.79 ± 0.783
B	500	499.93 ± 0.069	0.4583	4.1185	499.91 ± 0.034	98.97 ± 0.057
C	500	499.96 ± 0.046	0.6875	1.0929	499.94 ± 0.044	99.86 ± 0.070

* Mean ± standard deviation of five determinations. The t-test and F-test values refer to

comparison of the proposed method with the reference method. Theoretical values

at 95% confidence limits, t=2.306, F=6.39.

**Mean ± standard deviation of three determinations.

DISCUSSION:

The characteristics of the proposed method summarized in table -1 clearly indicate that the reagent methyl orange in acidic medium can be successfully employed for the determination of Meropenem. The low standard deviation and high percent recovery values shows that precision and accuracy of the method is satisfactory. The results obtained in the determination of Meropenem in commercial formulations listed in table-2 are very close to that of manufacturers specifications.

CONCLUSION:

The proposed method is simple, accurate, sensitive and is suitable for the determination of Meropenem in pharmaceutical formulations without interference from the excipients and also for quality control in the pharmaceutical laboratories.

ACKNOWLEDGEMENTS:

I would like to acknowledge the contribution of my research supervisor Prof. K. Saraswathi with whose help I was able to incorporate this work in my thesis. I tender grateful thanks to the Management and Principal, R.V.R. and J.C. College of Engineering, Guntur for their encouragement to me for doing this work.

CONFLICT OF INTEREST:

The authors declare no conflict of interest.

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Received on 17.03.2022 Modified on 21.10.2022

Research J. Pharm. and Tech 2023; 16(9):4148-4150.

DOI: 10.52711/0974-360X.2023.00679