Alterations of the Benzoxazinoid Profiles of Uninjured Maize Seedlings During Freezing, Storage, and Lyophilization

Benzoxazinoids are highly studied compounds due to their biological activity and presence in several cereals. They include compound classes such as hydroxamic acids and lactams and usually occur as inactive glucosides in unstressed plants. Injury to the plant causes enzymatic hydrolysis of the inactive glucosides to the biologically active hydroxamic acid and lactam aglucones. The hydroxamic acids further undergo spontaneous hydrolysis to benzoxazolinones in aqueous solution. Extraction methods that do not cause immediate inactivation of enzymes result in accumulation of aglucones in samples. Using HPLC-MS to profile benzoxazinoids in maize seedlings subjected to several sample preparation techniques, we have found that hydroxamic acid aglucones and benzoxazolinones are present in uninjured maize seedlings, but that the benxozazinoid profile varies depending on sample treatment, potentially underrepresenting the glucoside content and overrepresenting the aglucone and benzoxazolinone content.