The starch content was analyzed using the Total Starch assay (Megazyme International, Wicklow, Ireland) following the protocol described previously [30 (link)]. The method is based on enzymatic degradation of starch to glucose monomers by α-amylase and amyloglucosidase enzymes and measuring glucose monomers in a spectrophotometric-based assay for quantification against a D-glucose calibration control series at a wavelength of 510 nm.
Total starch assay
Manufactured by Megazyme
Sourced in Ireland
The Total Starch assay is a laboratory kit designed to quantify the total starch content in a variety of food, feed, and industrial samples. The assay utilizes enzymatic hydrolysis to break down starch into glucose, which is then measured colorimetrically. The kit provides a reliable and accurate method for determining the total starch concentration in a sample.
Automatically generated - may contain errors
Lab products found in correlation
Lysing matrix e, by MP Biomedicals (1 mentions)
Sucrose assay kit, by Merck Group (1 mentions)
Pullulanase, by Merck Group (1 mentions)
Amyloglucosidase, by Merck Group (1 mentions)
Trypsin, by Merck Group (1 mentions)
Pepsin, by Merck Group (1 mentions)
α amylase, by Merck Group (1 mentions)
β glucan assay kit, by Megazyme (1 mentions)
Sulfuric acid, by Merck Group (1 mentions)
5 protocols using total starch assay
1
Quantitative Starch Determination
2
Quantifying Starch in Microalgae Biomass
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The starch content of S. obliquus was analyzed by enzymatic degradation of starch to glucose using the thermostable α-amylase and amyloglucosidase enzymes from the Total Starch assay (Megazyme International, Wicklow, Ireland) using the protocol as described by de Winter et al.[16 (link)]. The following modifications to the protocol were made. Around 10 mg of biomass was transferred to bead beating tubes (Lysing Matrix E; MP Biomedicals) and lyophilized overnight. Freeze-dried cells were disrupted by bead beating in the presence of 80% ethanol. Starch was converted to glucose using α-amylase and amyloglucosidase enzymes. Subsequently glucose was coloured and absorbance was measured against a D-glucose calibration control series at a wavelength of 510 nm.
3
Leaf Starch and Sucrose Dynamics
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Enzymatic assays were carried out on extracts from three fully expanded leaves from three separate plants. Tissue was excised and flash‐frozen in liquid nitrogen every 2–4 h throughout the photoperiod and stored at −80 °C before analysis. Starch was estimated using a total starch assay (Megazyme, Wicklow, Eire, Method E). Sucrose measurements were collected using a sucrose assay kit (Sigma, Poole, UK). Three biological replicates were assayed at each time point and each measurement was replicated three times. Technical replicates were averaged and a two‐way anova was performed on the data (P < 0.05) using SPSS (IBM).
4
Characterization of Whole Quinoa Flour
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Three colored (red:black:white = 1:1:1) whole quinoa flour (WQ) produced in the year of 2019 were purchased from Sanjiang Fertile Soil Co., Ltd. (Qinghai, China). Total starch assay, glucose oxidase/peroxidase (GOPOD), dietary fiber, and β-glucan assay kits were provided by Megazyme International Ireland Ltd. (Bray, Ireland). Pullulanase (EC 3.2.1.41), amyloglucosidase (EC 3.2.1.3), α-amylase (EC 3.2.1.1), pepsin (EC 3.4.23.1), and trypsin (EC 3.4.4.4) were provided by Sigma Chemical Co. (St. Louis, MO, USA). Other chemical reagents obtained the standards of the analysis grade.
Hydrochloric acid-potassium chloride with pH = 2.0 was used to dissolve pepsin (10.0 mg/mL), then the miscible liquid was stored at 4 °C. Suitable amounts of α-amylase and amyloglucosidase were dissolved in 0.2 M sodium acetate buffer (pH = 5.6, mixed with 40 mM calcium chloride), and the concentrations were 290 and 60 U/mL, respectively.
Hydrochloric acid-potassium chloride with pH = 2.0 was used to dissolve pepsin (10.0 mg/mL), then the miscible liquid was stored at 4 °C. Suitable amounts of α-amylase and amyloglucosidase were dissolved in 0.2 M sodium acetate buffer (pH = 5.6, mixed with 40 mM calcium chloride), and the concentrations were 290 and 60 U/mL, respectively.
5
Characterization of Brewer's Spent Grain Biomass
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The raw material used in this work was the brewer’s spent grain kindly supplied by San Miguel S.A, from the industrial brewery located in Burgos (Spain). This raw material was first preconditioned, as soon as obtained, by washing it with water until uncolored washing water was observed and drying it in an air convection oven (45 °C) until reaching a final moisture content of 8% (w/w).
The dry BSG was milled in a SM100 mill (Retsch, Haan, Germany) to obtain a particle size lower than 0.5 mm. Biomass characterization was performed according to the NREL protocols [15 (link)]. Carbohydrates were quantified by high-performance liquid chromatography (HPLC) with a Bio-Rad Aminex-HPX-87 H column (Hercules, CA, USA) and an Agilent Technologies (Waldbronn, Germany) refractive index detector (RID), maintained at 40 °C. The mobile phase was 0.005 M sulfuric acid from Merck (Darmstadt, Germany). Megazyme Total Starch Assay and β-Glucan Assay (Wicklow, Ireland) were followed to determine starch and β-glucans content in the BSG. Protein in the raw material was estimated from the nitrogen content present in the samples as measured by the elemental analysis and considering a nitrogen factor of 6.25 according to the amino acid profile of the protein fraction of the BSG [7 (link)].
The dry BSG was milled in a SM100 mill (Retsch, Haan, Germany) to obtain a particle size lower than 0.5 mm. Biomass characterization was performed according to the NREL protocols [15 (link)]. Carbohydrates were quantified by high-performance liquid chromatography (HPLC) with a Bio-Rad Aminex-HPX-87 H column (Hercules, CA, USA) and an Agilent Technologies (Waldbronn, Germany) refractive index detector (RID), maintained at 40 °C. The mobile phase was 0.005 M sulfuric acid from Merck (Darmstadt, Germany). Megazyme Total Starch Assay and β-Glucan Assay (Wicklow, Ireland) were followed to determine starch and β-glucans content in the BSG. Protein in the raw material was estimated from the nitrogen content present in the samples as measured by the elemental analysis and considering a nitrogen factor of 6.25 according to the amino acid profile of the protein fraction of the BSG [7 (link)].
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