Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2015 Sep;23(4):444-52.
doi: 10.1016/j.jsps.2015.01.016. Epub 2015 Jan 16.

Optimization of HPLC method for determination of cefixime using 2-thiophenecarboxaldehyde as derivatizing reagent: A new approach

Madan Lal Maheshwari  1 , Ayaz Ali Memon  2 , Shahabuddin Memon  2 , Fakhar-Un-Nisa Memon  2 , Ubed Ur Rahman Mughal  1 , Abdullah Dayo  1 , Naheed Memon  1 , Mohammed Ali Ghoto  1 , M Khan Leghari  3
Affiliations

Optimization of HPLC method for determination of cefixime using 2-thiophenecarboxaldehyde as derivatizing reagent: A new approach

Madan Lal Maheshwari et al. Saudi Pharm J. 2015 Sep.

Abstract

The determination of cefixime 1 has clinical and analytical importance due to its broad spectrum antimicrobial activity and stability. Cefixime is a significant member of orally active third generation cephalosporin and has excellent activity against many pathogens. It is for first time that we have developed a new HPLC-DAD method for analysis of imine derivative 3 of cefixime by using reflux method at 100 °C for 50 min without any buffer solution. 2 Thiophenecarboxaldehyde (2TCA) 2 was used first time as a derivatizing reagent for cefixime drug. Furthermore, separation of three components, i.e. drug (cefixime), reagent (2TCA) and derivative was carried out using kromasil 100 C-18 (15 mm × 0.46 mm, 5 μm) column with isocratic elution of methanol: 0.1% aqueous formic acid (70:30 v/v) with flow rate of 1 ml min(-) (1) at retention time of 1.8, 2.4 and 3.3 min, respectively; while, total run time was 5 min. The developed method was repeatable with a relative standard deviation (RSD) of 0.81-1.88% for imine derivative. The limit of detection and quantification of imine derivative 3 were obtained within the range of 0.132-0.401 μg ml(-) (1) and compared with cefixime drug as 0.30-0.90 μg ml(-1), respectively. However, the formation of imine derivative 3 was confirmed by comparing peak height, retention time and spectral changes. The method is rapid, simple, very stable and accurate for the separation and determination of imine derivative 3 of cefixime 1.

Keywords: 2-Thiophenecarboxaldehyde; Cefixime; Derivatization; HPLC–DAD; Reflux method.

PubMed Disclaimer

Figures

Figure 1
Figure 1
Verification (retention time and UV spectra) and separation of methanolic solution of (a) cefixime (drug) and (b) 2TCA (reagent) by HPLC–DAD.
Figure 2
Figure 2
Comparison of UV spectra of imine derivative, cefixime and 2TCA reagent by diode array detector.
Figure 3
Figure 3
(a) High pressure liquid chromatography separation of three components including cefixime 1 (drug), 2TCA (reagent) 2 and imine derivative 3, (b) high pressure liquid chromatography separation after internal addition of two components including cefixime 1, and 2TCA reagent 2 with imine derivative 3.
Figure 4
Figure 4
HPLC chromatogram of extracted imine derivative peak and their UV spectra at λmax of 349 nm.
Figure 5
Figure 5
Comparison of UV spectra (at λmax 349 nm) of imine derivative before and after extraction from a mixture by n-hexane solvent by DAD detector.
Figure 6
Figure 6
HPLC chromatograph of linear calibration of imine derivative within the concentration range of 1–50 μg mL−1.
Scheme 1
Scheme 1
A representative diagram for general reaction procedure of Schiffs base reaction.
Scheme 2
Scheme 2
Synthetic route for the derivatization of cefixime using 2TCA as derivatizing reagent.
See this image and copyright information in PMC

References

    1. Abass K., Zafar I., Mohammad I.K., Khalid J., Abad K., Lateef A., Yasar S., Fazli N. Simultaneous determination of cefdinir and cefixime in human plasma by RP-HPLC/UV detection method: method development, optimization, validation, and its application to pharmaceutical study. J. Chromatog. B. 2011;879(24):2423–2429. - PubMed
    1. Adegoke O.A., Quadri M.O. Novel spectrophotometric determinations of some cephalosporins following azo dye formation with p-dimethylaminobenzaldehyde. Arab. J. Chem. 2012
    1. Adegoke O.A., Umoh O.E. A new approach to the spectrophotometric determination of metronidazole and tinidazole using p-dimethylaminobenzaldehyde. Acta Pharm. 2009;59(4):407–419. - PubMed
    1. Agbaba D., Eric S., Karljikovic-Rajic K., Valdimirov S., Zivanov-stakic D. Spectrophotometric determination of certain cephalosporins using ferihydroxamate method. Spectrosc. Lett. 1997;30:309–319.
    1. Ahemed O.A. Simultaneous determination of ofloxacin and cefixime in tablet formulation using capillary electrophoresis. J. Liq. Chromatogr. Relat. Technol. 2013;36(19):2687–2697.

LinkOut - more resources