COMBINED MIXTURE DESIGN-DOE AND ION PAIR REAGENT: A SYNERGISTIC MODEL FOR THE SEPARATION OF MULTIPLE AMINO ACIDS IN RP-HPLCAbstract
The aim of the present study is to apply the Design of Experiments (DoE) to develop an assay method for the separation of amino acids by RP-HPLC. The method used in this study is the DoE Combined- Randomized method. Design of Experiments allows interpreting the results with better outcomes and enhanced understanding process. Ion pair reagent- Octane 1 Sulphonic acid sodium salt monohydrate of 0.01M at pH 2.2 is used along with Methanol and Tetra-hydro Furan. The column used is INETSIL C8, with 5µ Particle size and 4.6×250 mm column dimensions. The Solvent composition and ion pair concentration is evaluated as variables in the DoE. The wavelength for all amino acids is 205 nm. Conclusively DoE is an efficient tool for the separation of amino acids. Method development was established, and the design is validated. The proposed method has adequate reproducibility and accuracy for the estimation of the amino acids in the routine analysis.
P. S. Krishna *, P. S. Babu, K. S. Babu and A. Viswanath
Department of Pharmaceutical Analysis, Vignan Pharmacy College, Vadlamudi, Guntur, Andhra Pradesh, India.
06 May 2020
16 September 2020
21 September 2020
01 May 2021