Analytical Method Validation for Determination of Protein and Related Protein Content in Filgrastim by Reversed-Phase High Performance Liquid Chromatography

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Surintra Lahnwong
Saiwarul Jadoonkittinan
Supaporn Phumiamorn

Abstract

      Filgrastim is used for a treatment of patients with low leukopenia. A content of protein is important for an efficacy of product and related proteins may cause side effects. The objective of this study was to validate an analysis of protein and related protein content in both oxidised and reduced forms by reversed-phase high performance liquid chromatography using European Pharmacopoeia condition. The result showed that this method was specific to filgrastims. The linearity was found between the peak area and the filgrastim concentration of 0.1 - 0.7 mg/ml (R2 = 0.9998). The percent of relative standard deviation of repeatability and intermediate precision were 0.92 and 0.65, respectively. The percent recovery of accuracy was 102.45. Moreover, the limit of quantitation and the limit of detection were 0.0250 and 0.0125 mg/ml, respectively. This method also showed good robustness. In conclusion, this method is precise and suitable to measure the content of protein and related protein in filgrastims for the quality control of the products.

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Section
Laboratory Findings

References

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