For demonstrating the use of the graph for the determination of OD260 from HPLC peak area, crude ODN 1b was synthesized at 0.2 µmol scale on CPG, cleaved, deprotected, and purified with trityl-on RP HPLC. One twentieth of the ODN was injected into HPLC. The area of the ODN peak was found to be 1690.6, which corresponds to an OD260 of 0.608 according to the line obtained using the UV-Vis spectrometer in Figure . Thus, the OD260 for the 0.2 µmol ODN synthesis was 12.16. To validate the result, the OD260 of the synthesis was also determined using standard method by measuring the absorbance at 260 nm on a UV-Vis spectrometer. The number obtained was 11.20. The use of the graph was further validated using ODN 1c following the same procedure for 1b. The OD260 values calculated from the line obtained using the UV-Vis spectrometer and obtained a UV-Vis spectrometer were 16.92 and 15.40, respectively.