The Use of Surrogate Reference Standards in Quantitative HPLC

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MAY, 2010
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The possibilities of using surrogate reference standards in quantitative high performance liquid chromatography (HPLC) have been explored. A quantitative reverse phase-High Performance Liquid Chromatography method has been developed to determine a constant, K with a surrogate reference that makes it possible to analyse prednisolone in a formulation in the absence of pure prednisolone as reference. Indometacin, Naproxen and Piroxicam were considered as surrogate reference standards. The chromatographic apparatus consisted of Shimadzu LC-6A Liquid Chromatograph-pump, Hichrom H5ODS column, Applied Biosystems 783 programmable Absorbance UV detector at 254nm and Shimadzu CR501 Chromatopac-Integrator as a recorder. Elution was isocratic with a mobile phase consisting of methanol and phosphate buffer (pH=5.8) in a ratio 1:1. The mean retention times for both drug and surrogates were; prednisolone: (4.30 ± 0.7 min), indometacin: (9.79 ± 0.3 min), naproxen: (2.25 ± 0.6 min) and piroxicam: (1.26 ± 0.3 min). The average K constant obtained for the surrogate reference standards were, Indometacin: 1.8323, Naproxen: 1.6925 and Piroxicam: 1.3550. These constants were used in the analysis of four samples of prednisolone tablets. The percentage content obtained for each sample was then compared with that obtained using the BP 2007 method. The results showed that the surrogate reference standard can be used for the analysis of prednisolone without the use of pure prednisolone powder as reference.
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Thesis Submitted In Partial Fulfilment of the Requirements for the Award of a Degree of Master of Science Pharmaceutical Analysis and Quality Control, .
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