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Cyanogenic Glycoside Analysis in American Elderberry.

Molecules (Basel, Switzerland) (2021-04-04)
Michael K Appenteng, Ritter Krueger, Mitch C Johnson, Harrison Ingold, Richard Bell, Andrew L Thomas, C Michael Greenlief
ABSTRACT

Cyanogenic glycosides (CNGs) are naturally occurring plant molecules (nitrogenous plant secondary metabolites) which consist of an aglycone and a sugar moiety. Hydrogen cyanide (HCN) is released from these compounds following enzymatic hydrolysis causing potential toxicity issues. The presence of CNGs in American elderberry (AE) fruit, Sambucus nigra (subsp. canadensis), is uncertain. A sensitive, reproducible and robust LC-MS/MS method was developed and optimized for accurate identification and quantification of the intact glycoside. A complimentary picrate paper test method was modified to determine the total cyanogenic potential (TCP). TCP analysis was performed using a camera-phone and UV-Vis spectrophotometry. A method validation was conducted and the developed methods were successfully applied to the assessment of TCP and quantification of intact CNGs in different tissues of AE samples. Results showed no quantifiable trace of CNGs in commercial AE juice. Levels of CNGs found in various fruit tissues of AE cultivars studied ranged from between 0.12 and 6.38 µg/g. In pressed juice samples, the concentration range measured was 0.29-2.36 µg/mL and in seeds the levels were 0.12-2.38 µg/g. TCP was highest in the stems and green berries. Concentration levels in all tissues were generally low and at a level that poses no threat to consumers of fresh and processed AE products.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Prunasin, ≥90% (LC/MS-ELSD)
Supelco
Picric acid solution, 100 μg/mL in acetonitrile, PESTANAL®, analytical standard
Sigma-Aldrich
Amygdalin, ≥99% (HPLC), from apricot kernels
Sigma-Aldrich
α-Hydroxyisobutyronitrile β-D-glucopyranoside, ≥97% (HPLC)