High-throughput measurement methodologies for developing nutrient-dense crops

With the development of nutrient-dense crops comes the need for analytical methodologies to enable rapid and accurate analysis of the micronutrients of interest. The analysis of provitamin A carotenoids (pVACs) and the minerals iron (Fe) and zinc (Zn) are the focus of this chapter with the considerations and commonly employed methods discussed. When analyzing samples there are various considerations to minimise analyte degradation (in the case of provitamin A) and reduce possible contamination from external sources (for Fe and Zn). Spectroscopic and chromatographic analyses are the most common analysis approaches utilised when screening for carotenoids. Spectroscopic analyses including near-infrared spectroscopy (NIRS) and iCheck are rapid and require minimal samples preparation and provide fast analysis times. The carotenoids present in the sample is dependent on the crop analyzed and resulting number and concentration of carotenoids present will impact the final decision on suitable analysis techniques. For example, in crops with high concentrations of non-pVACs, chromatographic analysis is necessary in order to accurately quantify the micronutrients. This process is able to accurately identify and quantify individual carotenoids, but requires extensive sample preparation and often long chromatographic separation analysis. When analyzing the minerals Fe and Zn, these same techniques are not suitable, but it is still important to ensure careful sample preparation to deliver accurate analytical results. Degradation of these micronutrients is not a concern, however, possible contamination from soil/ dust/ insects can lead to inaccurate results. Commonly employed analysis such as atomic absorption spectroscopy (AAS) and Inductively Coupled Plasma-Optical Emission Spectrometry ICP-OES or Inductively Coupled Plasma-Mass spectrometry (ICP-MS) require sample digestion prior to analysis and highly pure reagents and gases. These techniques are able to analyze multiple elements and have high accuracy and sensitivity but require specialised facilities and highly trained staff. The use of high-throughput analyses to complement these high-accuracy methods include colorimetric and X-ray flourescence (XRF) technologies. These approaches enable much higher throughput with simple sample preparation and enable screening for micronutrient concentration without the need for specialised facilities. Keywords: Screening, Analysis, Carotenoid, Provitamin A, Iron, Zinc, Micronutrient analysis