An Efficient HPLC Approach to Quantify Kunitz Trypsin Inhibitor in Soybean Seeds

• DOI: 10.2135/cropsci2018.01.0061
• Publication Date: 2018-06-07T17:36:20Z
• Authors: M. Luciana Rosso; Chao Shang; Edgar Correa; Bo Zhang
• Keywords: Repeatability; Trypsin inhibitor; Glycine soja; Sodium acetate

Trypsin inhibitor (TI), an important antinutritional factor present in soybean [ Glycine max (L.) Merr.] seeds, prevents animal protein digestibility. Accurately determining seed TI concentration is essential for screening breeding lines to select genotypes with low TI. A colorimetric assay widely used measure activity seeds. This bioassay time consuming, expensive, and has repeatability issues. study developed a high‐performance liquid chromatography (HPLC) method as high‐throughput, less more reliable quantify Kunitz trypsin (KTI), the major TI, We extracted KTI using sodium acetate buffer, separated on Poros R2/H perfusion column detected at 220 nm. The HPLC was compared two popular enzymatic bioassays 100 lines. For bioassays, HCl or NaOH extractants determined according Kakade et al. (1974). from ranged 0.52 12.15 mg g −1 average of 5.25 limit detection 0.05 . recovery spiked samples 82.33%. data both were strongly correlated ( r = 0.82 0.80, p ≤ 0.0001). CVs (66.20% HPLC, 18.84 32.55% bioassays) suggest that capable detecting wider range quantities providing better sensitivity quantifying samples.