Water and organic content tuning of mobile phase for Enantioseparation of fungicides by capillary electrochromatography using cellulose Tris(3‑chloro-4-methylphenylcarbamate) based stationary phase

• Capillary electrochromatography for enantioseparation of chiral imidazole azole fungicides. • Cellulose Tris(3-chloro-4-methylphenylcarbamate) based stationary phase • Effect of mobile phase composition on chiral separation This study investigated the effect of mobile phase composition, especially water content in a mixture of protic and non-protic organic solvents, on chiral recognition and separation efficiency using a cellulose tris(3-chloro-4-methylphenylcarbamate)-based stationary phase in the enantioseparation of selected fungicide compounds. The chiral stationary phase, coated on porous native silica particles, was packed into fused silica capillaries (250 mm x 100 µm I.D.) via the "slurry" packing method. The capillary column was utilized for the enantiomeric separation of five azole compounds through capillary electrochromatography. The primary objective was to thoroughly investigate how the composition of the mobile phase affects enantioresolution. This was achieved by systematically varying the ratios of methanol, acetonitrile, and water while keeping the buffer concentration, type, and pH constant. The role of water content was studied in relation to chiral interactions by evaluating its effect on key chromatographic parameters such as retention factor, resolution, and efficiency. By selecting an appropriate acetonitrile to methanol ratio at a fixed water content, chiral resolutions ( Rs ) ranging from approximately 2.0 to 3.5 were achieved for the tested analytes.