Contamination of milk with minute amounts of hexanal results in oxidized flavor. We have developed a trace-level procedure for aliphatic aldehydes, including hexanal, using 1,3-cyclohexandione as a fluorescent labeling agent in HPLC. However, measurement of trace amounts of hexanal with the reported method of acetic acid, 1,3-cyclohexanedione and ammonium acetate resulted in a higher quantitative value than expected. Upon investigation, we found an inseparable byproduct having the same retention time as fluorescently labelled hexanal. To solve this issue, the amount of byproduct was reduced significantly by replacing the acetic acid with formic, citric, or sulfuric acid. To confirm the precision of the modified analytical procedure, we validated the labelling method using sulfuric acid for milk, yogurt, and milk coffee. We accurately quantified several aldehydes, including hexanal, generated from milk irradiated with 2000 lux LED light with the improved analytical procedure.
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