Biochemistry

Antioxidant Activity Assays

• DPPH assay: Measures the ability of compounds to scavenge free radicals using a purple-colored DPPH solution.

• ABTS assay: Similar to DPPH but uses ABTS radicals, producing a green-blue color change.

• FRAP assay: Determines the reducing power of antioxidants by converting ferric ions (Fe³⁺) to ferrous ions (Fe²⁺).

 

Antidiabetic Assays

• α-Amylase inhibition: Checks how well a sample can slow down the breakdown of starch into sugars.

• α-Glucosidase inhibition: Tests the ability of compounds to reduce glucose release from carbohydrates, important in controlling blood sugar.

 

Cholesterol Assimilation by Microbes

Studies how beneficial microbes (like probiotics) can absorb or break down cholesterol, reducing its levels in the body.

 

Anti-inflammatory Assays

• NO inhibition: Measures suppression of nitric oxide production in cells, linked to inflammation.

• COX inhibition: Tests whether compounds block cyclooxygenase enzymes, reducing inflammation and pain.

 

Oxalate Degradation Enzyme Assay

Analyzes enzymes that can break down oxalates, which are linked to kidney stone formation.

 

Enzyme Kinetics and Inhibition Studies

Measures the speed of enzyme reactions and how inhibitors affect them (Michaelis-Menten kinetics).

 

Lipid Peroxidation Assay (MDA Content)

Determines oxidative damage in lipids by measuring malondialdehyde (MDA), a marker of lipid peroxidation.

 

Vitamin and Mineral Analysis

• HPLC (High-Performance Liquid Chromatography): Used for separating and quantifying vitamins.

• AAS (Atomic Absorption Spectroscopy): Used to detect and measure mineral content (iron, calcium, zinc, etc.).

 

Phytochemical Estimations

• Phenolic content: Total phenols in a sample, linked to antioxidant activity.

• Flavonoid content: Compounds with antioxidant, anti-inflammatory properties.

• Tannin content: Plant compounds with astringent and protective effects.

 

Proximate Composition Analysis

Measures nutritional composition:

• Moisture (water content)

• Ash (mineral content)

• Fat

• Carbohydrates

• Protein

 

Protein Extraction and Ammonium Sulfate Precipitation

Methods for isolating proteins from biological samples and concentrating them by “salting out” with ammonium sulfate.

 

Protein Purification

• Dialysis: Removes small molecules and salts.

• Ultrafiltration: Separates proteins based on size.

• Chromatography: Further purification using affinity, ion-exchange, or gel filtration methods.

 

Protein Quantification

• Bradford assay: Uses dye binding for quick protein measurement.

• Lowry assay: More sensitive, based on copper-protein reactions.

• BCA assay: Uses bicinchoninic acid, suitable for micro-quantities.

 

SDS-PAGE Analysis

Separates proteins based on size (molecular weight) using an electric field in a polyacrylamide gel.

 

 Western Blotting

Detects specific proteins using antibodies after separation by SDS-PAGE.

 

Chromatography Methods

• TLC (Thin Layer Chromatography): Separates small molecules on a coated plate.

• Column Chromatography: Separates compounds using a stationary phase packed in a column.

 

Immunoassays

• ELISA (Enzyme-Linked Immunosorbent Assay): Detects and quantifies proteins, hormones, or antigens.

• Competitive ELISA: Measures concentration of an analyte by competition with a known antigen.

 

Spectroscopic Techniques

• FTIR (Fourier Transform Infrared Spectroscopy): Identifies functional groups in compounds.

• UV-Vis spectroscopy: Measures absorbance of compounds in ultraviolet/visible light.

• Fluorescence spectroscopy: Detects molecules that emit light when excited, highly sensitive for biomolecules.