A Validated Ecofriendly Chromatographic Method for Determination of Myasthenia Gravis Combined Medications in Spiked Human Plasma.

Recently, the main interest of the analytical chemistry researchers is the development of green analytical methods to minimize the harmful effect on the environment and the natural life. So, a RP-HPLC method was developed and assessed regarding its greenness criteria using three greenness assessment tools: analytical eco-scale, analytical greenness metric approach, and green analytical procedure index. This method aims to separate and quantitavely determine three co-administered drugs namely pyridostigmine bromide (PYR), 6-mercaptopurine (MRC) and prednisolone (PRD) in their tertiary mixture and spiked human plasma. These drugs are co-administered to manage myasthenia gravis autoimmune disease. The separation was done using C 18 column and gradient elution of a mixture of 0.1% H 3 PO 4 aqueous solution (pH 2.3) and methanol. The flow rate was adjusted to 1 mL/min and detection was done at 254 (for PYR and PRD) and at 330 nm (for MRC). The lower limits of quantitation (LLOQ) were 15, 2, and 5 μg/mL for PYR, MER and PRD, respectively. Linear correlations were obtained and found to be near 1. In addition, the proposed method was validated according to the FDA instructions, and the results proved its success to determine the three studied drugs in their tertiary mixture and spiked human plasma.