Application of total error concept in the analytical method validation for the assay of essential amino acids by precolumn derivatization
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Date
2020-05
Journal Title
Journal ISSN
Volume Title
Publisher
Open Science Publishers LLP
Abstract
A pre-Ultraperformance Liquid Chromatography (UPLC) column derivatization procedure was developed for thesimultaneous quantification of essential amino acids (EAAs) in the solid oral dosage pharmaceutical formulation.This analytical procedure has been validated with the help of the concept of total error. The total error approach (thecombination of systematic and random error) is a decision-making tool for ensuring the performance of the method.Fluorenylmethyloxycarbonyl chloride was used as a derivatization reagent. The amino acid derivatives were separatedon a C18 column (internal diameter 2.1 × 100 mm, 1.6 µm) by gradient elution with 0.1% trifluoroacetic acid andacetonitrile:water (90:10, v/v), respectively, as mobile phase A and B. About 10 EAAs could be detected at 265 nm in35 minutes with a flow rate of 0.25 ml/min. The linearity range of each amino acid was between 0.1 and 1.0 mg/ml.The accuracy and risk profiles were considered acceptable across the range. The precolumn derivatization procedureand the concept of the validation of total error could be used as an appropriate strategy to demonstrate the suitabilityof the analytical procedure for the separation and evaluation of EAA in solid oral dosage formulations.
Description
Keywords
Essential amino acids, pre-column derivatization, total error, UPLC, accuracy profile.
Citation
Aasodi Ramachandra Reddy, Murugan V, Premakumari KB. Application of total error concept in the analytical method validation for the assay of essential amino acids by precolumn derivatization. Journal of Applied Pharmaceutical Science. 2020 May; 2020 May: 031-042