Journal: Journal of Agricultural and Food Chemistry

Article Title: Potential of Quantitative α‑Amylase or Trypsin Inhibition by Refined and Whole Wheat and Einkorn Using High-Performance Thin-Layer Chromatography–NanoGIT versus Conventional Spectrophotometry

doi: 10.1021/acs.jafc.5c14009

Figure Lengend Snippet: Inhibition of amylolysis by different acarbose amounts (1–5 μg/band): (A) HPTLC–nanoGIT (amylolysis inhibition)–FLD chromatogram and (B) percentage deviation in intensity (Δ%) of released maltotriose ( Mal3 ), maltose ( Mal ), and glucose ( Glc ) in contrast to the NC (α-amylase, 2.5 μg/band and soluble starch, 4 μg/band) and for acarbose (2 μg/band) oversprayed by α-amylase (2.5 μg/band) in contrast to α-amylase (2.5 μg/band); analyzed on HPTLC plate silica gel 60 with acetonitrile/water/2-propanol/acetone (12:3:4:1, V / V / V / V ) up to 70 mm, derivatized with p -aminobenzoic acid reagent, detected at FLD 366 nm, and quantified after densitometric fluorescence measurement at 366/>400 nm.

Article Snippet: The plate, except the application zone, was covered with a cut HPTLC plate (layer upward, 20 cm × 8.5 cm) and wetted with a 0.1 M sodium chloride solution by piezoelectrical spraying (2.5 mL, yellow nozzle, level 6, Derivatizer, CAMAG).

Techniques: Inhibition, High Performance Thin Layer Chromatography, Starch, Fluorescence

Journal: Journal of Agricultural and Food Chemistry

Article Title: Potential of Quantitative α‑Amylase or Trypsin Inhibition by Refined and Whole Wheat and Einkorn Using High-Performance Thin-Layer Chromatography–NanoGIT versus Conventional Spectrophotometry

doi: 10.1021/acs.jafc.5c14009

Figure Lengend Snippet: Saccharide removal from α-amylase (5 mg/mL) over a 2–12 h dialysis period: HPTLC–nanoGIT (amylolysis inhibition)–FLD/Vis chromatograms (2–8 h on plate 1 and 10-12 h on plate 2) and respective graphs of filtered α-amylase (2.5 μg/band, −) overspotted with soluble starch (4.0 μg/band, +) versus amylolysis to prove the activity of the purified α-amylase; analyzed on HPTLC plate silica gel 60 with acetonitrile/water/2-propanol 3:1:1 ( V / V / V ) up to 70 mm, derivatized via a reagent sequence first with the p -aminobenzoic acid reagent ( p -ABA), detected at FLD 366 nm, and quantified after densitometric fluorescence measurement at 366/>400 nm, followed (on the same plate) by the 2-naphthol reagent, detected at Vis (remission-transmission mode), and quantified after densitometric absorbance measurement at 500 nm.

Article Snippet: The plate, except the application zone, was covered with a cut HPTLC plate (layer upward, 20 cm × 8.5 cm) and wetted with a 0.1 M sodium chloride solution by piezoelectrical spraying (2.5 mL, yellow nozzle, level 6, Derivatizer, CAMAG).

Techniques: High Performance Thin Layer Chromatography, Inhibition, Starch, Activity Assay, Purification, Sequencing, Fluorescence, Transmission Assay

Journal: Journal of Agricultural and Food Chemistry

Article Title: Potential of Quantitative α‑Amylase or Trypsin Inhibition by Refined and Whole Wheat and Einkorn Using High-Performance Thin-Layer Chromatography–NanoGIT versus Conventional Spectrophotometry

doi: 10.1021/acs.jafc.5c14009

Figure Lengend Snippet: In-vial trypsin inhibition assay (7 μL/band), followed by HPTLC analysis: HPTLC–Vis chromatograms showing inhibition of proteolysis (marked +, framed region of interest) by three flour extracts (100 and 150 μL each) versus corresponding non-proteolyzed flour extracts (150 μL, marked −), proteolyzed casein ( NC , 1:100), proteolyzed trypsin inhibitor ( PC 1:2 and 1:3 , mixed with trypsin 1:2 and 1:3 in a vial, 10 min preincubated, and 30 min incubated at 37 °C), TI, and trypsin; analyzed on HPTLC plate silica gel 60 with 2-butanol/pyridine/ammonia (25%)/water 10:17:5:13 ( V / V / V / V ) up to 50 mm, derivatized with ninhydrin reagent, and detected at Vis (remission–transmission mode).

Article Snippet: The plate, except the application zone, was covered with a cut HPTLC plate (layer upward, 20 cm × 8.5 cm) and wetted with a 0.1 M sodium chloride solution by piezoelectrical spraying (2.5 mL, yellow nozzle, level 6, Derivatizer, CAMAG).

Techniques: Inhibition, High Performance Thin Layer Chromatography, Incubation, Transmission Assay

Journal: Journal of Agricultural and Food Chemistry

Article Title: Potential of Quantitative α‑Amylase or Trypsin Inhibition by Refined and Whole Wheat and Einkorn Using High-Performance Thin-Layer Chromatography–NanoGIT versus Conventional Spectrophotometry

doi: 10.1021/acs.jafc.5c14009

Figure Lengend Snippet: Qualitative all-in-one trypsin inhibition assay: HPTLC–nanoGIT (proteolysis inhibition)–Vis chromatograms showing the inhibition of the proteolysis (marked +, framed region of interest) by three flour extracts (4 and 7 μL/band) versus corresponding non-proteolyzed flour extracts (marked −) and casein (2 μL/band, 20 mg/mL), maximal proteolysis ( NC , 2-fold determination, casein/trypsin E/S 1:50, 30 min incubation), maximal inhibition ( PC , trypsin/trypsin inhibitor E/I 1:2.5) versus trypsin inhibitor ( TI , 1 μL/band, 0.5 mg/mL), and trypsin (4 μL/band, 0.2 mg/mL); analyzed on HPTLC plate silica gel 60 with 2-butanol/pyridine/ammonia (25%)/water 10:17:5:13 ( V / V / V / V ) up to 50 mm, derivatized with the ninhydrin reagent, and detected at Vis (remission–transmission mode).

Article Snippet: The plate, except the application zone, was covered with a cut HPTLC plate (layer upward, 20 cm × 8.5 cm) and wetted with a 0.1 M sodium chloride solution by piezoelectrical spraying (2.5 mL, yellow nozzle, level 6, Derivatizer, CAMAG).

Techniques: Inhibition, High Performance Thin Layer Chromatography, Incubation, Transmission Assay