The effects of extraction techniques and quantitative determination of oxalates in Nerium oleander and feeds

Shrubs represent the most affordable and accessible form of feed that livestock can rely on to acquire both essential and non-essential elements of life. In addition to their inherent toxins, they contain endogenous substances commonly referred to as 'antinutritive factors' (ANFs) that often interfere with the utilisation of nutrients. Their abundance may lead to severe clinical trauma. Hence, the objective of the study was to investigate the effects of different extraction techniques on Nerium oleander L. and animal feeds as well as to quantify oxalates. Organic (hexane, acetone and methanol) sequential and aqueous (infusion and decoction) extractions were explored. Qualitative and quantitative analyses were conducted to determine the presence of various phytochemicals and oxalate contents as putative ANFs, respectively. The results showed higher extraction yields of 22.6% and 43.1% in the decoction and infusion of N. oleander, respectively. The quantification methods were validated for linearity, accuracy and precision. Oxalate contents of 6.76 +/- 0.245 (0.65%) mg/g and 5.74 +/- 0.236 mg/g dry weight (0.55%) were obtained in N. oleander and feeds, respectively. This difference was statistically significant with p < 0.05. Percentage recoveries of 98.5 (percent relative standard deviation [% RSD] = 2.3), 85.7 (% RSD = 1.03) and 80.3 (% RSD = 1.22) at 76%, 95% and 112% fortifications were obtained, respectively. Relative standard deviation for precision was 0.99% and 1.13% at 0.33 mg and 0.39 mg fortifications, respectively, while reproducibility showed 2.21% RSD. Therefore, these methods can be used to provide a valuable basis for qualitative determination of ANFs, particularly in shrub foliage.