Expanding resolution of metalloprotein separations from soybean seeds using 2D-HPLC-ICP-MS and SDS-PAGE as a third dimension

Atomic Spectrometry Update: review of advances in elemental speciation

“…The 201 Hg : 202 Hg isotope ratio for the MeHg peak in the resultant GC chromatogram of 0.45 was comparable with the natural ratio of 0.44. Subsequently, MeHg was purged from the distillate onto Tenax traps, desorbed at 200 C and swept with He gas to the diphenyl (5%)-polydimethyl siloxane (95%) capillary GC column (15 Â 0.32 mm ID, 0.25 mm lm thickness) via a transfer line heated to 60 C. The GC column outlet was inserted directly into the torch injector with, unusually, no heated transfer line being used.…”

Recent update on methodologies for extraction and analysis of soybean seed proteins

“…Size exclusion (first dimension) and anion exchange chromatography (second dimension) followed by separation using SDS‐PAGE (third dimension) showed different bands with improved resolution. After digestion and purification of these bands, the metal‐associated proteins were identified as 3‐lipoxygenase A chain complex, 13‐( S )‐hydroperoxy‐9( Z) ,11( E )‐octadecadienoic acid, β ‐amylase and seed lipoxygenase 1 and 2 …”

“…Table provides multiple references of various MS methods (matrix‐assisted laser desorption/ionization (MALDI), Electrospray ionization (ESI), MS/MS) used for detection of proteins from soybeans separated by LC and capillary electrophoresis. Either purified proteins based on gel electrophoresis or a mixture of proteins can be identified using the above‐mentioned methods . A two‐step approach is used to detect and identify the peptides from samples.…”

“…used MALDI‐MS/MS as a fast screening tool for the identification of stable soybean‐derived tryptic markers even if the proteins were subjected to various changes on the molecular level through several reactions typically occurring during food processing. Mataveli and Arruda used ICP‐MS for the identification of different metalloproteins ( β ‐amylase, β ‐conglycinin, lipoxygenase) from soybean seed. Blazek and Caldwell investigated and compared the application of SDS gel capillary electrophoresis (equipped with UV detection) and microfluidic lab‐on‐a‐chip technology to quantify relative amounts of storage proteins (7S and 11S) from 20 soybean cultivars.…”

“…Either purified proteins based on gel electrophoresis or a mixture of proteins can be identified using the above-mentioned methods. 67,70 A two-step approach is used to detect and identify the peptides from samples. In MS, a precursor ion is detected from the full MS scan for quantification.…”