Antioxidant and Anti-Inflammatory Activities of Endemic Plants of the Australian Wet Tropics
Abstract
:1. Introduction
2. Results
2.1. Crude Extraction and Extract Yield
2.2. Major Classes of Phytochemical Analysis
2.3. Phenolic and Flavonoid Content
2.4. DPPH Free Radical Scavenging Activity
2.5. Ferric Reducing Antioxidant Power (FRAP) Activity
2.6. Correlation of Pseudo-IC50 Values of Antioxidant Activities with TPC and TFC
2.7. Cell Viability Assay
2.8. Anti-Inflammatory Activity
3. Discussion
4. Materials and Methods
4.1. Plant Collection and Reagents
4.2. Extract Preparation
4.3. Qualitative Screening for Major Classes of Phytochemicals
- Alkaloid test: Dragendorf’s reagent (1 mL) was added to the sample, and the formation of orange precipitate confirmed the presence of alkaloid.
- Steroid test: chloroform (2 mL) was added to the sample, followed by a few drops of acetic anhydride. Next, the reaction mixture was boiled in a water bath and cooled in the ice water. Concentrated sulphuric acid (1 mL) was then added, and the formation of a brown ring at the junction and change of the upper layer into green confirmed the presence of steroid.
- Terpenoid test: chloroform (2 mL) was added to the sample, followed by a few drops of concentrated sulphuric acid, and the formation of reddish-brown coloration at the interface confirmed the presence of terpenoid.
- Tannin test: a few drops of 10% aqueous ferric chloride was added to the sample, and the formation of a black precipitate or blue-green coloration confirmed the presence of tannin.
- Saponin test: distilled water was added and the sample was shaken for few minutes. The presence of foam/frothing that persisted after warming in the water bath (37 °C) for 5 min confirmed the presence of saponin.
- Anthraquinone glycoside test: 5% sulphuric acid (1 mL) and chloroform (2 mL) were added to the sample, followed by shaking the lower layer with dilute ammonia. Rose pink to red colour of the ammoniacal layer confirmed a positive test result.
- Cardiac glycoside test: glacial acetic acid (2 mL) was added to the sample, followed by a few drops of 10% ferric chloride and then 5% sulphuric acid (1 mL). The formation of a brown ring at the interface confirmed the presence of cardiac glycoside.
4.4. Quantitative Phytochemical Estimation
4.4.1. Total Phenolics Content (TPC)
4.4.2. Total Flavonoids Content (TFC)
4.5. Antioxidant Activity Assays
4.5.1. 2,2-Diphenyl-1-picrylhydrazyl (DPPH) Assay
4.5.2. Ferric Reducing Antioxidant Power (FRAP) Assay
4.6. Calculation of Pseudo-IC50 of Antioxidant Activities
4.7. In Vitro Anti-Inflammatory Activity and Quantification
4.7.1. PBMCs Culture and Sample Treatment
4.7.2. Determination of Cell Viability
4.7.3. Quantification of Proinflammatory Cytokines
4.8. Statistical Analysis
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Plants (Family) | Specimen Voucher Number | Abbreviation | Extract Yield (%) |
---|---|---|---|
Alyxia orophila Domin (Myrtaceae) | D.M. Crayn 1168 (CNS 135636.1) | AO | 9.5 |
Ceratopetalum hylandii Rozefelds & R.W.Barnes (Cunoniaceae) | S.J. Worboys 1682 (CNS 148538.1) | CH | 3.5 |
Garcinia brassii C.T. White (Clusiaceae) | S.J. Worboys 1681 (CNS 148513.2) | GB | 8.2 |
Leptospermum wooroonooran F.M.Bailey (Myrtaceae) | D.M. Crayn 1172 (CNS 135635.1) | LW | 16.8 |
Linospadix apetiolatus Dowe & A.K.Irvine (Arecaceae) | D.M. Crayn 1572 (CNS 145626.1) | LA | 5.7 |
Litsea granitica B.Hyland (Lauraceae) | D.M. Crayn 1437 (CNS144467.1) | LG | 10.4 |
Polyscias willmottii (F.Muell.) Philipson (Araliaceae) | D.M. Crayn 1575 (CNS 145629.2) | PW | 16.4 |
Plants | Alkaloid | Steroid | Terpenoid | Tannin | Saponin | Anthraquinone Glycoside | Cardiac Glycoside |
---|---|---|---|---|---|---|---|
Alyxia orophila | − | ++ | + | +++ | +++ | − | + |
Ceratopetalum hylandii | +++ | ++ | +++ | +++ | − | − | +++ |
Garcinia brassii | + | + | − | +++ | +++ | + | − |
Leptospermum wooroonooran | − | + | + | +++ | − | − | − |
Linospadix apetiolatus | − | − | − | +++ | ++ | − | + |
Litsea granitica | − | ++ | +++ | +++ | + | − | − |
Polyscias willmottii | + | ++ | +++ | ++ | − | − | − |
Plants | Extract Concentration | Pseudo-IC50 (mg/mL) | |||
---|---|---|---|---|---|
25 μg/mL | 100 μg/mL | 250 μg/mL | 500 μg/mL | ||
Alyxia orophila | 0.26 ± 0.20 | 0.91 ± 0.71 | 0.93 ± 0.72 | 1.39 ± 0.23 | 24.78 |
Ceratopetalum hylandii | 0.48 ± 0.42 | 0.67 ± 0.13 | 1.78 ± 0.74 | 2.85 ± 0.45 | 9.56 |
Garcinia brassii | 0.27 ± 0.07 | 1.09 ± 0.38 | 2.02 ± 1.04 | 3.23 ± 0.23 | 8.28 |
Leptospermum wooroonooran | 0.63 ± 0.07 | 1.57 ± 0.07 | 3.48 ± 0.33 | 6.39 ± 0.25 | 4.10 |
Linospadix apetiolatus | 0.65 ± 0.47 | 0.97 ± 0.53 | 0.94 ± 0.07 | 1.90 ± 0.33 | 20.59 |
Litsea granitica | 0.99 ± 0.75 | 2.50 ± 1.00 | 5.10 ± 0.74 | 8.40 ± 0.54 | 3.19 |
Polyscias willmottii | 0.66 ± 0.29 | 1.44 ± 0.33 | 3.60 ± 0.35 | 5.80 ± 0.66 | 4.50 |
Ascorbic acid | 1.70 ± 0.25 | 6.37 ± 0.75 | 14.59 ± 1.95 | 29.59 ± 0.51 | 0.85 |
Assays | Correlation R2 | |
---|---|---|
Phenolics (TPC) | Flavonoid (TFC) | |
IC50 of DPPH radical scavenging ability | 0.4933 | 0.1698 |
IC50 of FRAP activity | 0.7947 ** | 0.4139 |
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Yeshi, K.; Ruscher, R.; Miles, K.; Crayn, D.; Liddell, M.; Wangchuk, P. Antioxidant and Anti-Inflammatory Activities of Endemic Plants of the Australian Wet Tropics. Plants 2022, 11, 2519. https://doi.org/10.3390/plants11192519
Yeshi K, Ruscher R, Miles K, Crayn D, Liddell M, Wangchuk P. Antioxidant and Anti-Inflammatory Activities of Endemic Plants of the Australian Wet Tropics. Plants. 2022; 11(19):2519. https://doi.org/10.3390/plants11192519
Chicago/Turabian StyleYeshi, Karma, Roland Ruscher, Kim Miles, Darren Crayn, Michael Liddell, and Phurpa Wangchuk. 2022. "Antioxidant and Anti-Inflammatory Activities of Endemic Plants of the Australian Wet Tropics" Plants 11, no. 19: 2519. https://doi.org/10.3390/plants11192519