Abstract Biofortification is a sustainable and cost‐effective strategy that uses plant breeding and agronomic approaches to improve the nutrient content of staple crops consumed by vulnerable populations. The approach requires high‐throughput phenotyping to effectively identify and develop nutrient‐rich genotypes. This study aimed to develop a multielement, nondestructive method to quantify calcium (Ca), manganese (Mn), iron (Fe), copper (Cu), and zinc (Zn) in whole seed wheat ( Triticum aestivum L.) samples using a benchtop energy dispersive x‐ray fluorescence (EDXRF) spectrometer. Grain samples from 29 and 41 wheat genotypes were used for the EDXRF calibration and validation, respectively. A microwave plasma–atomic emission spectrometer (MP‐AES) provided the analyte reference values for each sample. The EDXRF calibration showed moderate to high correlation with MP‐AES values for Ca, Mn, Cu, and Zn, while Fe exhibited a weak correlation. The limits of quantification (mg kg −1 ) were 103.9 for Ca, 8.5 for Mn, 3.5 for Fe, 4.7 for Zn, and 1.0 for Cu—all below the observed analyte range in wheat grain. The method is suitable for use in early generation selection, as indicated by standard errors of prediction (mg kg −1 ) of 36.4 for Ca, 3.3 for Mn, 2.5 for Fe, 0.3 for Cu, and 1.5 for Zn. This study builds upon previous nondestructive EDXRF methods by introducing additional elements that can be reliably phenotyped in wheat, supporting broader use in biofortification programs.
Klarquist et al. (2026) studied this question.