Development and optimization of Assay method of Filgrastim injection by Reverse Phase- High Performance Liquid Chromatography

Development and optimization of Assay method of Filgrastim injection by Reverse Phase- High Performance Liquid Chromatography

  • Shaily Tyagi Shriram institute for Industrial Research
  • Ruchi Tyagi, Dr. Shriram institute for Industrial Research
  • Rajavelly Prem, Dr. Shriram institute for Industrial Research
  • Ajeet Kumar Agrawal, Dr. Shriram Institute for industrial research
  • M L Agarwal, Dr. Shriram institute for Industrial Research
  • K M Chacko, Dr. Shriram institute for Industrial Research
Keywords: RP-HPLC method, PDA detector, ICH, Validation, Filgrastim

Abstract

Filgrastim is a human granulocyte colony stimulating factor which belongs to the family of cytokines. The goal of this study was to develop and validate a simple and rapid RP-HPLC method for determination of Filgrastim in protein formulations (Injection). The experiment was carried out by RP-HPLC using YMC-Pack C4 (150 X 4.6mm ID S-5 µm, 12nm column). The HPLC was operated at gradient mode using mobile phase composed of solvent A i.e. Water: Trifluoroacetic Acid (1000:1) and solvent B Water: Trifluoroacetic acid: ACN (100:1:899 and at 60°C and UV detection was set 215 nm. Under the proposed conditions the retention time of Filgrastim was about 21 minutes. Linearity was performed using six concentration of Filgrastim solutions in the range of 35-225 µg/ml and the value of correlation co-efficient was found to be R2 =0.99 that shows good linearity curve. The RSD value for repeatability and reproducibility were 0.1%, 0.23% respectively. The obtained recovery value of 96.6 % indicate that the proposed method is quantitative and accurate.

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Published
2018-10-02
How to Cite
Tyagi, S., Tyagi, R., Prem, R., Agrawal, A., Agarwal, M., & Chacko, K. (2018). Development and optimization of Assay method of Filgrastim injection by Reverse Phase- High Performance Liquid Chromatography. The Pharma Research, 19(02), 1-7. Retrieved from http://www.s-epub.in/ejournal/index.php/tphres/article/view/31
Section
Research