1.3.2.4. Comparison of different novel extraction methods
1.3.3. Extraction of trypsin inhibitors
1.3.4. Extraction of saponins
1.3.5. Response surface methodology for optimising extraction conditions
1.4. Spectrophotometry method for quantification of bioactive compounds
1.4.1. Principle of spectrophotometry measurement
1.4.2. Devices and mechanism for spectrophotometry measurement
1.5. Oxidation and antioxidant activity of medicinal plants
1.6. Cancer and anticancer activity of medicinal plants
1.7. Experimental rationale
1.8. Hypotheses, Aims and Objectives
Chapter 2: MATERIALS AND METHODS
2.1. Gac seeds
2.1.1. Drying of the Gac seeds
2.1.2. De-coating of the Gac seeds
2.1.3. Grinding of the Gac seeds
2.1.4. Freeze-drying of the Gac seed powder
2.2. Optimisation the extraction of oil
2.2.1. Supercritical carbon dioxide extraction of Gac seed oil
2.2.1.1. Extraction procedure
2.2.1.2. Optimisation process
2.2.2. Soxhlet extraction of Gac seed oil
2.3. Optimisation the extraction of trypsin inhibitors
2.3.1. MAE procedure
2.3.2. UAE procedure
2.3.3. Optimised conventional extraction
2.3.2.1. Extraction procedure
2.3.2.2. Optimisation process
2.4. Optimised MAE for extraction of Gac seed saponins
2.4.1. Extraction procedure
2.4.2. Optimisation process
2.5. Evaporation of solvents from extracts
2.6. Analysing methods
2.6.1. Oil characterising
2.6.1.1. Specific gravity
2.6.1.2. Refractive index
2.6.1.3. Slip melting point
2.6.1.4. Iodine value
2.6.1.5. Saponification value
2.6.1.6. Free fatty acid
2.6.1.7. Peroxide value
2.6.1.8. Unsaponifiable matter
2.6.1.9. Moisture and volatile matter
2.6.1.10. Insoluble impurities
2.6.2. Oil antioxidant activity
2.6.3. Oil colour measurement
2.6.4. Determination of trypsin inhibiting activity using spectrometric method
2.6.4.1. Principle of the assay
2.6.4.2. Determination of trypsin inhibiting activity of Gac seed extracts
2.6.5. Determination of protein content
2.6.5.1. Principle of the Lowry assay
2.6.5.2. Building the standard curve
2.6.5.3. Determination of soluble protein content
2.6.6. Determination of total saponin content
2.6.6.1. Principle of the vanillin-sulfuric acid assay for TSC analysis
2.6.6.2. Building the standard curve of aescin
2.6.6.3. Determination of total saponin content
2.6.7. Determination of total phenolic content (TPC)
2.6.7.2. Building the standard curve for TPC assay
2.6.7.3. Determination of TPC
2.6.8. Determination of total antioxidant capacity of Gac seed extracts
2.6.8.1. The 2,2′-azino-bis-3-ethylbenzthiazoline-6-sulphonic (ABTS) acid assay
2.6.8.2. The 1,1-diphenyl-2-pycryl-hydrazyl (DPPH) assay
2.6.10.3. The Ferric reducing antioxidant power (FRAP) assay
2.6.9. Determination of anticancer activity
2.7. Statistical analysis
Chapter 3: RESULTS
3.1. Synopsis of research result papers
3.2. Extraction of oil – the research papers
3.2.1. Optimisation of process parameters for supercritical carbon dioxide extraction of oil from Gac seed kernel powder
3.2.2. Physicochemical properties of Gac (Momordica cochinchinensis (Lour.) Spreng) seeds and their oil extracted by supercritical carbon dioxide and Soxhlet methods
Introduction
Materials and Methods
Materials
Chemicals
Gac Seed Kernels
Proximate Analysis of the Gac Seed Kernels
Preparation of Gac Seed Kernel Powder for Oil Extraction
Methods for the Extraction of Gac Seed Oil
SC-CO2 Extraction
Soxhlet Extraction
Oil Yield Calculation
Methods for Characterisation of Oils
Determination of Physicochemical Indices
Colour Measurement
Determination of Antioxidant Activity
Statistical Analyses
Results and Discussion
Characteristics of Gac Seeds
Oil Yield
Characterisation of the Extracted Oils
Colour Characterisation
Conclusions
References
3.3. Extraction of trypsin inhibitors – the research papers
3.3.1. Effect of solvents and extraction methods on recovery of bioactive compounds from defatted Gac (Momordica cochinchinensis Spreng) seeds
Introduction (1)
Materials and Methods (1)
Materials
Solvents, Reagents, and Chemicals
Gac Seeds
Methods
Extraction Methods
Determination of Trypsin Inhibitor Activity (TIA)
Determination of Total Saponin Content (TSC)
Determination of Total Phenolic Content (TPC)
Determination of Antioxidant Capacity
Determination of Total Solids
Statistical Analyses
Results and Discussion (1)
Effect of Extraction Methods on the Trypsin Inhibitor Yield
Effect of Extraction Methods on the Total Saponin Content (TSC)
Effect of Extraction Methods on Total Phenolic Content (TPC)
Effect of Extraction Methods on Total Solids and Antioxidant Capacity
Correlations between Bioactive Compounds and Total Solids and Antioxidant Activity in the Extracts
Conclusions (1)
References (1)
3.3. Extraction of trypsin inhibitors – the research papers
3.3.2. Optimised extraction of trypsin inhibitors from defatted Gac (Momordica cochinchinensis Spreng) seeds for production of a trypsin inhibitor‐enriched freeze-dried powder.
Untitled
Chapter 3: RESULTS (2)
3.4. Extraction of saponins – the research papers
3.4.1. Improving the vanillin-sulphuric acid method for quantifying total saponins
Introduction (2)
Materials and Methods (2)
Materials
Methods
The Usual Vanillin-Sulphuric Acid TSC Assay
Solvents and the Reagent Blank
Solvents Used to Prepare Saponin (Aescin) Solutions
The Modified Vanillin-Sulphuric Acid TSC Assay
Determining the TSC of a Gac Seed Extract Using the Modified Method
Statistical Analyses
Results
The Effect of Different Solvents in the Reagent Blank
Effect of Solvents on the Aescin Standard Curve
Aescin Standard Curve for the Modified Method
Saponin Content of the Gac Seed Kernel Powder
Discussion
Effect of Common Extraction Solvents on the Vanillin-Sulphuric Acid Method
Modifications of the Vanillin-Sulphuric Acid Method
Advantages of the Modified Vanillin-Sulphuric Acid Method
Implementation of the Modified Vanillin-Sulphuric Acid Method
Conclusions (2)
References (2)
3.4. Extraction of saponins – the research papers
3.4.2. Optimisation of the microwave-assisted ethanol extraction of saponins from Gac (Momordica cochinchinensis Spreng) seeds
Introduction (3)
Materials and Methods (3)
Materials
Solvents, Reagents and Chemicals
Gac Seed Kernel Powder
Preparation of Gac Seed Kernel Powder
Preparation of Defatted Gac Seed Kernel Powder
Methods
Microwave Assisted Extraction (MAE)
Extraction of Saponins from Full-Fat and Defatted Gac Seed Kernel Powders
Extraction of Saponins from the Full-Fat Seed Kernel Powder
Verifying Optimal Conditions for Gac Seed Saponin Extraction
Analytical Methods
Determination of Total Saponin Content (TSC)
Determination of Antioxidant Capacity
Statistical Analyses
Results (1)
Effect of the Ethanol Concentration on the MAE of Saponins from Full-Fat and Defatted Gac Seed Kernel Powders
Effect of the Ethanol to Sample Ratio on the MAE of Saponins from the Full-Fat Gac Seed Kernel Powder
Effect of the Microwave Parameters on the MAE of Saponins from the Full-Fat Gac Seed Kernel Powder
Correlations between the TSC and the MAE Temperature
Verification of the Optimal MAE Conditions for the Extraction of Saponins from Full-Fat Gac Seed Kernel Powder
Discussion and Conclusions
References (3)
3.5. Anticancer and antioxidant activities of Gac seed extracts – the research paper
3.5.1. Bioactive composition, antioxidant activity, and anticancer potential of freeze-dried extracts from defatted Gac (Momordica cochinchinensis Spreng) seeds
Introduction (4)
Materials and Methods (4)
Materials
Solvents, Reagents, and Chemicals
Gac Seeds
Methods
Extraction
Freeze Drying Extracts
Determination of Extractable Yield
Determination of Dry Mass Yield
Determination of Trypsin Inhibitor Activity (TIA)
Determination of Total Saponin Content (TSC)
Determination of Total Phenolic Content (TPC)
Determination of Antioxidant Capacity
Determination of Cytotoxicity
Statistical Analyses
Results (2)
Effect of Solvent on the Extractable Yield and the Dry Mass Yield
Effect of Solvents on the Content of Bioactive Compounds
Trypsin Inhibitors
Saponins
Phenolics
Effect of Solvents on Antioxidant Activity
Effect of Extraction Solvent on Cancer Cell Viability
Correlations between Extract Yields, Bioactive Compounds, Antioxidant Activity, and Cancer Cell Viability across the FD Crude Extracts
Discussion (1)
Conclusions (3)
References (4)
Chapter 4: GENERAL DISCUSSION AND CONCLUSIONS
4.1. General discussion
4.1.1. Gac seed oil
4.1.2. Gac seed bioactives
4.1.2.1. Gac seed trypsin inhibitors
4.1.2.2. Gac seed saponins
4.1.2.3. Gac seed phenolic compounds
4.1.3. Antioxidant activity and anticancer potential of Gac seed extracts