Abdelhamid_2022_J.AOAC.Int__

BACKGROUND: Prednisolone, 6-mercaptopurine and pyridostigmine bromide are co-administered together to treat a neuromuscular autoimmune disease called myasthenia gravis. Prednisolone and 6-mercaptopurine are immunosuppressant drugs. 6-mercaptopurine is the active form of the pro-drug azathioprine. Pyridostigmine bromide is cholinesterase enzyme inhibitor. Recently, the green characteristics are taken into account by analysts when they develop new methods. Objective: The spectra of the 3 compounds and plasma are highly overlapped, so this study aims to remove this overlapping and determine the 3 components quantitavely in raw powders and spiked human plasma using green methods. METHODS: Two multivariate updated chemometric models called principle component analysis and partial least squares were developed. Three greenness assessment tools: the eco-scale, the analytical greenness metric approach, and the green analytical method index, were used to evaluate the greenness behavior of the generated models. RESULTS: The two models were verified in accordance with FDA requirements, and the results were within acceptable parameters. In addition they are relatively green in accordance to the mentioned greenness evaluation tools. CONCLUSION: The developed models succeeded to predict the proposed drugs in their tertiary combinations and spiked human plasma with satisfied precision, accuracy, and good greenness behavior. HIGHLIGHTS: Two ecologically evaluated multivariate spectrophotometric methods were developed for determination of pyridostigmine bromide, 6-mercaptopurine, and prednisolone in spiked human plasma.