Acid-Catalyzed Isomerization of Carbonyls-2,4- dinitrophenylhydrazone in Mainstream Smoke of Heat-Not-Burn Tobacco Product for HPLC Analysis

Abstract

Carbonyls (such as acrolein, acetaldehyde, and formaldehyde) are the critical type of carcinogens and toxicants contained within the heat-not-burn (HNB) tobacco products. Using HNB products can have negative effects on human health; therefore, it is important to measure carbonyl contents within the HNB mainstream smoke. Typically, the 2,4-dinitrophenylhydrazine (DNPH) approach involves forming the 2,4-dinitrophenylhydrazone derivatives, which is the most extensively adopted approach to qualitatively and quantitatively analyze carbonyl compounds. However, the approach can result in analytical error because 2,4-dinitrophenylhydrazones contains the E-stereoisomer as well as the Z-stereoisomer. Only an E-isomers exists in the purified carbonyls-2,4-dinitrophenylhydrazone, but when acid is added, the E-isomer and Z-isomer can be observed. For propionaldehyde-, acetaldehyde-, crotonaldehyde-, acrolein-, and 2-butanone-2,4-dinitrophenylhydrazones, their equilibrium Z/E isomer ratios are 0.143, 0.309, 0.093, 0.028, and 0.154. In the case of adding trace water into hydrazone derivatives dissolved within the acetonitrile solution, the derivative contents decrease, whereas the free DNPH content increases. Therefore, catalytic acid should be added in the low content. To determine carbonyls-2,4-dinitrophenylhydrazones through HPLC, the optimal approach is adding phosphoric acid into the samples and the standard reference solution to form the 0.02–1.0% acid solution.