A simple and fast procedure to determine 3-nitropropanoic acid and 3-nitropropanol in freeze dried Canadian milkvetch (Astragalus canadensis)
Citation
Liu, H., Shao, S., Schellenberg, M. (2017). A simple and fast procedure to determine 3-nitropropanoic acid and 3-nitropropanol in freeze dried Canadian milkvetch (Astragalus canadensis). Toxins, [online] 9(7), http://dx.doi.org/10.3390/toxins9070204
Plain language summary
Canadian milkvetch (Astragalus canadensis) is a North American plant species from the legume family. Parts of this plant is fatally poisonous to livestock due to the naturally occurring notrotoxins. These compounds cause nitrite oxidization of hemoglobin and inhibition of cellular metabolism leading to pulmonary emphysema in sheep and spinal cord demyelination in cattle or even death of the animals. To determine the toxicity of the plant, it is very important to develop an analytical method capable of separating and quantifying these toxic compounds. In this study we have successfully developed an extraction procedure followed by high performance liquid chromatography analysis to simultaneously analyze notrotoxins. The developed method has proven to be simple and efficient. Since there is increasing interest in using A. canadensis as forage, this developed method provides an important tool for the determination of the toxicity of Canadian milkvetch from different genomic backgrounds and/or growing conditions.
Abstract
Canadian milkvetch (Astragalus canadensis) is a North American plant species in the legume family and some of this plant is fatally poisonous to livestock. The poisoning is attributed to the natural occurrence of notrotoxins, i.e., 3-nitropropanoic acid and 3-nitropropanol, present as aglycones and conjugated forms in the plant. Those compounds cause nitrite oxidization of hemoglobin and inhibition of cellular metabolism. To determine the toxicity of the plant, it is very important to develop an analytical method for the contents of the compounds in the plant. In this study, we have successfully developed an extraction procedure followed by HPLC-UV analysis to simultaneously analyze notrotoxins. The aglycones could be released from its conjugated forms in the freeze dried plant and extracted by water at room temperature. An HPLC-UV method using a Phenomenex Kinetex 2.6 µ F5 100 Å 100 × 4.6 mm column with pH 3.5 phosphonate buffer as mobile phase have been developed and validated for the detection of the two compounds at 210 nm. This developed procedure for the analysis of 3-nitropropanoic acid and 3-nitropropanol has proven simple and efficient and it has been successfully applied for batch sample analysis.