Antioxidant, Biomolecule Oxidation Protective Activities of Nardostachys jatamansi DC and Its Phytochemical Analysis by RP-HPLC and GC-MS
Abstract
:1. Introduction
2. Experimental Section
2.1. Chemicals and Reagents
2.2. Plant Material and Preparation of the Extract
2.3. Metabolite Profile of N. jatamansi
2.3.1. Reversed Phase-HPLC Analysis of the 70% Ethanol Fraction
2.3.2. Gas Chromatography and Mass Spectrometry Analysis (GC-MS-HT-TOF) of the Hexane Fraction
2.4. Phytochemical Screening of the Plant Extract
2.4.1. Determination of Total Phenolic Content
2.4.2. Determination of Total Flavonoid Content
2.5. In Vitro Free Radical Scavenging and Antioxidant Activities
2.5.1. DPPH Radical Scavenging Activity
2.5.2. ABTS Radical Cation Decolorization Assay
2.5.3. Metal Chelating
2.5.4. Superoxide Radical Scavenging Activity
2.5.5. Ferric-Reducing Antioxidant Power Assay
2.6. Biomolecules Oxidation Assays
2.6.1. AAPH-Induced Plasmid Nick Assay
2.6.2. Protein Oxidation
2.6.3. Thiobarbituric Acid-Reactive Substances Assay
2.6.4. Estimation of Reactive Oxygen Species
2.6.5. Estimation of Protein Carbonyls
2.7. Cytoprotective Effects
LDH Leakage Assay
2.8. Statistical Analysis
3. Results and Discussion
3.1. Metabolite Profile of N. jatamansi
3.1.1. Reversed Phase-HPLC Analysis of Phenolic Compounds
No. | Polyphenols | RT (min) | Concentration (mg/g) |
---|---|---|---|
1 | Gallic acid | 9.88 | 0.18 |
2 | Catechin | 18.16 | 4.37 |
3 | Chlorogenic acid | 22.27 | 19.90 |
4 | Homovanillin | 22.60 | 32.02 |
5 | Epicatechin | 24.09 | 4.23 |
6 | Rutin hydrate | 37.17 | 0.08 |
7 | Quercetin-3-rhamnoside | 40.56 | 7.13 |
3.1.2. GC-MS Analysis
No. | RT (min) | Compound Name | Chemical Formula | Mass | Area (%) | Hit |
---|---|---|---|---|---|---|
1 | 9.62 | Dodecane | C12H26 | 170.204 | 1.615 | 1 |
2 | 11.22 | 2-Furanmethanol, tetrahydro-5-methyl-trans | C6H12O | 116.084 | 0.031 | 1 |
3 | 13.98 | Linalool | C10H18O | 154.135 | 0.362 | 1 |
4 | 15.30 | Benzenemethanol, a-methyl-propanoate | C11H14O2 | 178.099 | 0.705 | 1 |
5 | 15.68 | 1H-Cyclopropa(a)naphthalene, 1a,2,3,5,6,7,7a,7b-octahydro-1,1,7,7a-tetramethyl-(1aR-(1aa,7a,7aa,7ba)) | C15H24 | 204.188 | 1.061 | 1 |
6 | 16.02 | a-Muurolene | C15H24 | 204.188 | 14.071 | 1 |
7 | 16.39 | 1H-Cycloprop(e)azulene,1a,2,3,4,4a,5,6,7b-octahydro-1,1,4,7-tetramethyl-(1aR-(1aa,4a,4aa,7ba)) | C15H24 | 204.188 | 9.184 | 5 |
8 | 16.69 | l-calamenene | C15H22 | 202.172 | 0.065 | 1 |
9 | 17.11 | Α-ionone | C13H20O | 192.151 | 0.643 | 1 |
10 | 17.41 | Bicyclo(3.3.1)nonan-2-one,1-methyl-9-(1-methylethylidene) | C13H20O | 192.151 | 8.329 | 1 |
11 | 17.63 | Neoisolongifolene,8,9-dehydro | C15H22 | 202.172 | 2.002 | 1 |
12 | 17.81 | (−)-à-Panasinsen | C15H24 | 204.188 | 1.716 | 1 |
13 | 18.13 | Neoisolongifolene,8,9-dehydro | C15H22 | 202.172 | 10.86 | 1 |
14 | 18.96 | Bicyclo(2.2.2)octa-2,5-diene, 1,2,3,6-tetramethyl | C12H18 | 162.141 | 3.643 | 1 |
15 | 19.15 | Isolongifolene,4,5,9,10-dehydro | C15H20 | 200.157 | 1.24 | 1 |
16 | 19.76 | trans-Nerolidol | C15H26O | 222.198 | 4.781 | 1 |
17 | 19.88 | para-methoxyphenylpiperazine | C11H16N2O | 192.1263 | 5.576 | 1 |
18 | 20.2 | Cyclolongifolene oxide, dehydro | C15H22O | 218.167 | 1.368 | 1 |
19 | 21.51 | 1(2H)-Naphthalenone,octahydro-4a,8a-dimethyl-7-(1-ethylethyl)-, (4aR-(4aa,7a,8aa)) | C15H26O | 222.198 | 0.329 | 1 |
20 | 22.05 | 2-tetradecenal | C14H26O | 210.1984 | 4.873 | 1 |
21 | 30.1 | Palmitic acid | C16H32O2 | 256.24 | 1.537 | 1 |
22 | 31.64 | Oleic acid | C18H34O2 | 282.256 | 5.499 | 1 |
23 | 32.86 | 2,8,9-Trioxa-5-aza-1-silabicyclo(3.3.3)undecane,1-methoxy | C7H15NO4Si | 205.077 | 0.165 | 1 |
24 | 37.22 | Tridecanoic acid, methyl ester | C14H28O2 | 228.209 | 0.625 | 1 |
25 | 39.71 | Heptacosane | C27H56 | 380.438 | 2.664 | 1 |
3.2. In Vitro Antioxidant and Free Radical Scavenging Activities
Assay | Ethanolic Extract (NJE) | Hexane Extract (NJH) |
---|---|---|
Total polyphenolic content | 53.06 ± 2.2 mg GAE/g of extract | 13.87 ± 1.3 mg GAE/g of extract |
Total flavonoids | 25.303 ± 0.9 mg CE/g of extract | 4.58 ± 0.3 mg GAE/g of extract |
DPPH radical scavenging assay (IC50) | 222.22 ± 7.4 μg/mL | 432.68 ± 13.7 μg/mL |
Metal chelation (IC50) | 948 ± 21.1 μg/mL | 1211 ± 27.8 μg/mL |
ABTS (IC50) | 13.90 ± 0.5 μg/mL | 23.57 ± 1.4 μg/mL |
Superoxide (IC50) | 113.81 ± 4.2 μg/mL | 255.72 ± 9.7 μg/mL |
Anti-lipid peroxidation (IC50) | 465.11 ± 14.3 μg/mL (brain) | 587.53 ± 17.6 μg/mL (brain) |
539.08 ± 18.9 μg/mL (liver) | 685.15 ± 13.4 μg/mL (liver) | |
Ferric-reducing antioxidant power | 12.3 ± 0.43 mg FeSO4E/g of extract | 45.62 ± 1.34 mg FeSO4E/g of extract |
Polyphenol and Flavonoid Contents
3.3. Protective Properties against Biomolecules Oxidation
3.3.1. NJE Inhibits AAPH-Induced DNA Damage
3.3.2. NJE Inhibits Protein Oxidation
3.3.3. Inhibition of Lipid Peroxidation
3.3.4. Inhibition of Reactive Oxygen Species
3.3.5. NJE Inhibits Protein Carbonyl Content
3.4. Cytotoxicity of NJE by the Plasma Membrane Leakage Assay
4. Discussion
5. Conclusions
Acknowledgments
Author Contributions
Conflicts of Interest
References
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Razack, S.; Kumar, K.H.; Nallamuthu, I.; Naika, M.; Khanum, F. Antioxidant, Biomolecule Oxidation Protective Activities of Nardostachys jatamansi DC and Its Phytochemical Analysis by RP-HPLC and GC-MS. Antioxidants 2015, 4, 185-203. https://doi.org/10.3390/antiox4010185
Razack S, Kumar KH, Nallamuthu I, Naika M, Khanum F. Antioxidant, Biomolecule Oxidation Protective Activities of Nardostachys jatamansi DC and Its Phytochemical Analysis by RP-HPLC and GC-MS. Antioxidants. 2015; 4(1):185-203. https://doi.org/10.3390/antiox4010185
Chicago/Turabian StyleRazack, Sakina, Kandikattu Hemanth Kumar, Ilaiyaraja Nallamuthu, Mahadeva Naika, and Farhath Khanum. 2015. "Antioxidant, Biomolecule Oxidation Protective Activities of Nardostachys jatamansi DC and Its Phytochemical Analysis by RP-HPLC and GC-MS" Antioxidants 4, no. 1: 185-203. https://doi.org/10.3390/antiox4010185