Method Development for Reversed-Phase Separations

To avoid disposing of acetonitrile as hazardous waste, you can hydrolyze it to sodium acetate and flush it down the drain.23 

Mobile phase CH,CN/H20 for neutral analytes CH,CN/aqueous buffer 1 for ionic analytes 5 vol% CH, CN in H20 to 100% CH1CN for gradient elution 

Step 2 Optimize the separation with methanol/buffer to generate chromatogram B. 

Step 3 Optimize the separation with tetrahydrofuran/buffer to generate chromatogram C. Step 4 Mix the solvents used in A. B. and C, one pair at a time, in 1 1 proportion, to generate chromatograms D, E, and F. 

Step 5 Construct a 1 1 1 mixture of the solvents for A, B, and C to generate chromatogram G. Step 6 If some of the results A through G are almost good enough, select the two best points and mix the solvents to obtain points between those two. 

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