The research work was carried out for establishing a new Ultra Violet (UV)— Visible spectroscopy and Reverse phase-Ultra Fast Liquid Chromatography (RP-UFLC) method for the analysis and quantification of a biosimilar drug, Filgrastim. Filgrastim or recombinant methionyl granulocyte colony stimulating factor (rGCSF) is a glycoprotein. It has a biological action essential for proliferation and differentiation of hematopoetic and progenitor cells. The UV and RP-UFLC work was carried on a Shimadzu UV1800 Spectrophotometer and Shimadzu Prominence LC-20AD UFLC systems, respectively. The λmax of filgrastim was found to be 215 nm. The correlation coefficient by UV spectroscopy was found to be 0.9994 for the concentration range of 1 to 3 μg/ml in double distilled water. The Reverse phase UFLC was done by using Phenomenex C4 (25 cm × 0.46 cm internal diameter) 15 μ, 300 A° analytical column. The optimized mobile phase for binary elution was Acetonitrile and double distilled water (80:20) with a flow rate of 1 ml/min. The retention time of drug was at 3.2 min. It was observed that the response of the detector was linear in the range of 5 - 15 μg/ml with correlation coefficient value of 0.999. After developing the methods, it was assured for the intended use by validation of the analytical parameters like linearity, accuracy, precision, limit of detection, limit of quantitation, ruggedness and robustness. The results of all the parameters for both the methods were found to be within the acceptance criteria as per the International Council for Harmonisation (ICH) guidelines.
References
[1]
Alebouyeh, M., Tahzibi, A., Yaghoobzadeh, S., Zahedy, E.T., Kiumarsi, S., Soltanabad, M.H., Shahbazi, S. and Amini, H. (2016) Rapid Formulation Assessment of Filgrastim Therapeutics by a Thermal Stress Test. Biologicals, 44, 150-156. https://doi.org/10.1016/j.biologicals.2016.03.001
[2]
European Directorate for the Quality of Medicines (2009) European Pharmacopoeia 6.3: Filgrastim Concentrated Solution. Stationery Office, Strasbourg, 4142-4144.
[3]
Kubota, N., Orita, T., Hattori, K., Oheda, M., Ochi, N. and Yamazaki, T. (1990) Structural Characterization of Natural and Recombinant Human Granulocyte Colony-Stimulating Factors. The Journal of Biochemistry, 107, 486-492. https://doi.org/10.1093/oxfordjournals.jbchem.a123072
[4]
Pearlman, R. and Wang, Y.J. (2002) Formulation, Characterization, and Stability of Protein Drugs: Case Histories. Plenum Press, New York. https://doi.org/10.1007/b112935
[5]
Dalmora, S.L., D’Avila, F.B., da Silva, L.M., Bergamo, A.C. and Zimmermann, E.S. (2009) Development and Validation of a Capillary Zone Electrophoresis Method for Assessment of Recombinant Human Granulocyte Colony-Stimulating Factor in Pharmaceutical Formulations and Its Correlation with Liquid Chromatography Methods and Bioassay. Journal of Chromatography B, 877, 2471-2476. https://doi.org/10.1016/j.jchromb.2009.06.011
[6]
Sörgel, F., Schwebig, A., Holzmann, J., Prasch, S., Singh, P. and Kinzig, M. (2015) Comparability of Biosimilar Filgrastim with Originator Filgrastim: Protein Characterization, Pharmacodynamics, and Pharmacokinetics. BioDrugs, 29, 123-131. https://doi.org/10.1007/s40259-015-0124-7
[7]
Vemula, S., Vemula, S., Dedaniya, A., Kante, R.K. and Ronda, S.R. (2015) Characterization of Recombinant Human Granulocyte Colony-Stimulating Factor Expression by FT-IR Spectroscopy: Studies on Thermal Induction and Media Formulation on the Stability of the Protein Secondary Structure. Preparative Biochemistry and Biotechnology, 46, 586-595. https://doi.org/10.1080/10826068.2015.1084933
[8]
Anderlini, P. and Champlin, R.E. (2007) Biologic and Molecular Effects of Granulocyte Colony-Stimulating Factor in Healthy Individuals: Recent Findings and Current Challenges. Blood, 111, 1767-1772. https://doi.org/10.1182/blood-2007-07-097543
[9]
Kocherlakota, P. and La Gamma, E.F. (1998) Preliminary Report: rhG-CSF May Reduce the Incidence of Neonatal Sepsis in Prolonged Preeclampsia-Associated Neutropenia. Pediatrics, 102, 1107-1111. https://doi.org/10.1542/peds.102.5.1107
Nupur, N., Singh, S.K., Narula, G. and Rathore, A.S. (2016) Assessing Analytical Comparability of Biosimilars: GCSF as a Case Study. Journal of Chromatography B. 1032, 165-171. https://doi.org/10.1016/j.jchromb.2016.05.027
[12]
Levy, M.J., Gucinski, A.C., Sommers, C.D., Ghasriani, H., Wang, B., Keire, D.A. and Boyne, M.T. (2014) Analytical Techniques and Bioactivity Assays to Compare the Structure and Function of Filgrastim (Granulocyte-Colony Stimulating Factor) Therapeutics from Different Manufacturers. Analytical and Bioanalytical Chemistry, 406, 6559-6567. https://doi.org/10.1007/s00216-013-7469-x
[13]
Batkovska-Borozanova, I., Tonic-Ribarska, J. and Trajkovic-Jolevska, S. (2010) New High Performance Liquid Chromatography Method for Analysis of Filgrastim in Pharmaceutical Formulations. Webmedcentral Pharmaceutical Sciences, 1, Article ID: WMC001117.
[14]
Souza, L., Boone, T., Gabrilove, J., Lai, P., Zsebo, K., Murdock, D., et al. (1986) Recombinant Human Granulocyte Colony-Stimulating Factor: Effects on Normal and Leukemic Myeloid Cells. Science, 232, 61-65. https://doi.org/10.1126/science.2420009
[15]
Chu, J.W., Yin, J., Wang, D.I.C. and Trout, B.L. (2004) Molecular Dynamics Simulations and Oxidation Rates of Methionine Residues of Granulocyte Colony-Stimulating Factor at Different pH Values. Biochemistry, 43, 1019-1029. https://doi.org/10.1021/bi0356000
[16]
Faraji, F., Mofid, M.R., Babaeipour, V., Divsalar, A. and Abolghasemi Dehaghani, S. (2010) The Structural Characterization of Recombinant Human Granulocyte Colony Stimulating Factor. International Journal of Environmental Science and Development, 1, 15-19. https://doi.org/10.7763/IJESD.2010.V1.4
[17]
Halim, L.A., Márquez, M., Maas-Bakker, R.F., Castaneda-Hernández, G., Jiskoot, W. and Schellekens, H. (2018) Quality Comparison of Biosimilar and Copy Filgrastim Products with the Innovator Product. Pharmaceutical Research, 35, Article No. 226. https://doi.org/10.1007/s11095-018-2491-5
[18]
Choi, C., Yoo, B.W., Kim, C.O., Hong, T., Jin, B.H., Seo, K.S., Jang, J.Y. and Park, M.S. (2018) Comparison of Biosimilar Filgrastim with a Reference Product: Pharmacokinetics, Pharmacodynamics, and Safety Profiles in Healthy Volunteers. Drug Design, Development and Therapy, 12, 2381-2387. https://doi.org/10.2147/DDDT.S158277
[19]
Sörgel, F., Lerch, H. and Lauber, T. (2010) Physicochemical and Biologic Comparability of a Biosimilar Granulocyte Colony-Stimulating Factor with Its Reference Product. BioDrugs, 24, 347-357. https://doi.org/10.2165/11585100-000000000-00000
[20]
Magalhaes, V., Mantovani, M., Caruso, C., Facchini, F., Pascon, R. and Cagnacci, P. (2016) Physicochemical and Biological Comparison of the First Brazilian Biosimilar Filgrastim with Its Reference Product. Biosimilars, 6, 45-60. https://doi.org/10.2147/BS.S107898
[21]
Luiz Dalmora, S., Maria Krug Masiero, S., Renato de Oliveira, P., da Silva Sangoi, M. and Brum Junior, L. (2006) Validation of an RP-LC Method and Assessment of rhG-CSF in Pharmaceutical Formulations by Liquid Chromatography and Biological Assay. Journal of Liquid Chromatography & Related Technologies, 29, 1753-1767. https://doi.org/10.1080/10826070600716900
[22]
Codevilla, C.F., Brum, L., de Oliveira, P.R., Dolman, C., Rafferty, B. and Dalmora, S.L. (2004) Validation of an SEC-HPLC Method for the Analysis of rhG-CSF in Pharmaceutical Formulations. Journal of Liquid Chromatography & Related Technologies, 27, 2689-2698. https://doi.org/10.1081/JLC-200029121
[23]
Atole, D.M. and Rajput, H.H. (2018) Ultraviolet Spectroscopy and Its Pharmaceutical Applications—A Brief Review. Asian Journal of Pharmaceutical and Clinical Research, 11, 59-66. https://doi.org/10.22159/ajpcr.2018.v11i2.21361
