Many laboratories are struggling to find alternatives for HPLC separations that use acetonitrile for the organic. While there are other options, it can become difficult to find the right conditions, and method revalidation might also be required.
The automotive industry commonly performs the analysis of DNPH-aldehydes and ketones using a custom-designed HPLC Column (Restek Allure-AK), but this separation uses acetonitrile. Recently at Restek, Ty Kahler and Randy Romesberg developed a method to separate the "CARB" list of compounds using methanol. Using a Restek Ultra-C18 column they were able to obtain complete separation in less than 15 minutes. The chromatogram is the result which was performed under the following conditions: Column - Ultra-C18 (cat # 9174525) 4.6 mm i.d. X 200 mm, Flow=1mL/min,temp=50°C, 365nm bandwidth 8, Reference 600/8, gradient:
Time %B (MeOH) 0 70 10 75 11 90 13 100 15 100