Chemical Profiling and Therapeutic Evaluation of Standardized Hydroalcoholic Extracts of Terminalia chebula Fruits Collected from Different Locations in Manipur against Colorectal Cancer
Abstract
:1. Introduction
2. Results
2.1. Yield of the Extract
2.2. GC-MS Analysis of Hydroalcoholic Extracts of T. chebula Fruits
2.3. High-Performance Thin-Layer Chromatography (HPTLC)
2.4. Total Phenolic, Total Flavonoid, and Antioxidant Activity of T. chebula Extracts
2.5. MTT Assay
2.6. Effect of T. chebula Extracts on Colony-Forming Capacity of HCT 116 Cells
2.7. Cytotoxic Activity of TYH on HCT 116 Colon Cancer Cell
2.8. Effect of TYH on Migration of HCT 116 Cells
2.9. Analyses for Apoptosis
2.9.1. Effect of TYH on the Morphology of HCT 116 Cells
2.9.2. Apoptotic Effect of TYH Observed through DAPI Staining
2.9.3. TYH Activates Caspase-3 in HCT 116 Cells
3. Discussion
4. Materials and Methods
4.1. Chemical Reagents
4.2. Collection of Samples
4.3. Preparation of Extracts
4.4. Determination of Total Phenolic Content
4.5. Determination of Total Flavonoid Content
4.6. Maintenance of Cell
4.7. Qualitative Phytochemical Screening
4.8. GC-MS Analysis
4.9. Quantitative Estimation of Gallic Acid and Ellagic Acid by HPTLC
4.10. Antioxidant Activity
4.10.1. DPPH Free Radical Scavenging Assay
4.10.2. ABTS Free Radical Scavenging Assay
4.11. Cell Viability Assay
4.12. Colony-Forming Assay
4.13. Test for Cytotoxicity Using LDH Assay
activity/Maximum LDH activity − Spontaneous LDHactivity) × 100
4.14. Effect on Cell Migration
4.15. Giemsa Staining
4.16. DAPI Staining
4.17. Caspase-3 Activity Assay
5. Statistical Analysis
6. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
Sample Availability
Abbreviations
References
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Sample | Collection Site with GPS Coordinates | Yield Percentage (w/w) | Gallic Acid Content (µg/mg) as Determined by HPTLC | Ellagic Acid Content (µg/mg) as Determined by HPTLC |
---|---|---|---|---|
TCH | Chandel, N 24°25′35.7″ E94°00′47.7″ | 25.2 | 22.44 ± 0.056 | 2.652 ± 0.07 |
TSH | Sagolband, N 24°48′06″ E93°55′16″ | 28.7 | 21.61 ± 0.183 | 3.597 ± 0.145 |
TKH | Kakching, N 24°30′01.9″ E93°58′27.1″ | 30.3 | 18.275 ± 0.272 | 3.081 ± 0.089 |
TYH | Yumnam Huidrom, N 24°39′46″ E93°54′14″ | 28.7 | 19.629 ± 0.055 | 11.265 ± 0.089 |
Sl.No. | Name of the Compounds | % Content in TCH | % Content in TKH | % Content in TYH | % Content in TSH |
---|---|---|---|---|---|
1 | 1,2,3-Benzenetriol | 20.95 | 26.53 | 43.56 | 40.41 |
2 | 2-Cyclopenten-1-one | 19.35 | 12.79 | 9.07 | 14.66 |
3 | Catechol | 12.82 | 6.85 | NA | 12.47 |
4 | 5-Hydroxymethylfurfural | 6.04 | 11.26 | 4.03 | NA |
5 | 1-Hexyl-2-nitrocyclohexane | 4.13 | 2.76 | NA | 2.11 |
6 | 1-Undecanol | 3.15 | 1.88 | NA | 1.74 |
7 | Phenol | 3.27 | 0.37 | 5.40 | NA |
8 | Hexadecanamide | 1.95 | 1.31 | NA | 1.56 |
7 | 4H-Pyran-4-one, 2,3-dihydro-3,5-dihydroxy-6-methyl | 1.74 | 1.68 | NA | 0.74 |
8 | 2,6-Difluorobenzoic acid, 4-nitrophenyl ester | 1.72 | 3.05 | NA | NA |
9 | 1-(1′-Pyrrolidinyl)-2-butanone | 1.33 | NA | NA | NA |
10 | Phosphonic acid, (p-hydroxyphenyl)- | NA | 3.82 | NA | 4.64 |
11 | Sucrose | 0.22 | 2.27 | NA | NA |
12 | 3-Furaldehyde | 0.68 | 1.59 | 0.89 | 0.50 |
13 | 2-Cyclohexen-1-one | NA | 1.49 | NA | 1.22 |
14 | Cyclobutaneethanol, á-methylene | 0.92 | 1.40 | 0.93 | NA |
15 | Benzeneacetaldehyde | 0.89 | 1.13 | 1.56 | NA |
16 | 2,3,4,5-Tetrahydropyridazine | NA | NA | 1.65 | NA |
17 | 1,2-Benzenediol, mono(methylcarbamate) | NA | 0.19 | 1.57 | NA |
18 | 2,5-Furandione, 3-methyl- | NA | NA | 1.18 | 0.18 |
19 | 2,6,10,14-Tetramethylpentade can-6-ol | NA | NA | 1.06 | NA |
20 | S-Methyl 2-methylpropanethioate | NA | NA | NA | 1.26 |
Sample | DPPH Assay IC50 (µg/mL) | ABTS Assay IC50 (µg/mL) | TPC (GAEmg/g ext) | TFC (QEmg/g ext.) |
---|---|---|---|---|
TCH | 23.00 ± 3.11 | 17.81 ± 1.21 | 286.76 ± 0.35 | 8.98 ± 0.35 |
TKH | 19.85 ± 0.93 | 12.35 ± 0.92 | 295.35 ± 0.98 | 8.10 ± 0.87 |
TYH | 12.16 ± 0.42 | 7.80 ± 0.23 | 304.56 ± 1.23 | 7.65 ± 0.45 |
TSH | 24.42 ± 1.78 | 13.45 ± 0.28 | 270.65 ± 0.78 | 7.95 ± 0.79 |
Trolox(TC) | 18.01 ± 0.44 | 10.15 ± 0.24 | - | - |
Ascorbic Acid(AC) | 3.46 ± 0.12 | 2.19 ± 0.003 | - | - |
Sample/Drugs | IC50 (µg/mL) Values of the T chebula Extracts against HCT 116 Cells |
---|---|
TSH | 133.58 ± 2.0 |
TCH | 172.05 ± 2.0 |
TYH | 52.42 ± 0.87 |
TKH | 94.07 ± 1.70 |
Cisplatin | 24.38 ± 1.26 |
5-FU | 12.75 ± 1.03 |
Sl.No | Sample (Place of Collection) | Voucher Specimen No. | Latitude Longitude | Elevation(m) |
---|---|---|---|---|
1 | TCH (Chandel, Manipur) | IBSD/M-283-A | N 24°25′35.7″ E94°00′47.7″ | 817 |
2 | TKH (Kakching Manipur) | IBSD/M-283-C | N 24°30′01.9″ E93°58′27.1″ | 778 |
3 | TSH (Sagolband, Imphal, Manipur) | IBSD/M-283-B | N 24°48′06″ E93°55′16′ | 780 |
4 | TYH (Yumnam Huidrom, Imphal, Manipur) | IBSD/M-283-D | N 24°39′46″ E93°54′14″ | 784 |
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Thoithoisana Devi, S.; Devika Chanu, K.; Singh, N.B.; Chaudhary, S.K.; Keithellakpam, O.S.; Singh, K.B.; Mukherjee, P.K.; Sharma, N. Chemical Profiling and Therapeutic Evaluation of Standardized Hydroalcoholic Extracts of Terminalia chebula Fruits Collected from Different Locations in Manipur against Colorectal Cancer. Molecules 2023, 28, 2901. https://doi.org/10.3390/molecules28072901
Thoithoisana Devi S, Devika Chanu K, Singh NB, Chaudhary SK, Keithellakpam OS, Singh KB, Mukherjee PK, Sharma N. Chemical Profiling and Therapeutic Evaluation of Standardized Hydroalcoholic Extracts of Terminalia chebula Fruits Collected from Different Locations in Manipur against Colorectal Cancer. Molecules. 2023; 28(7):2901. https://doi.org/10.3390/molecules28072901
Chicago/Turabian StyleThoithoisana Devi, Soibam, Khaidem Devika Chanu, Nameirakpam Bunindro Singh, Sushil Kumar Chaudhary, Ojit Singh Keithellakpam, Kshetrimayum Birla Singh, Pulok K. Mukherjee, and Nanaocha Sharma. 2023. "Chemical Profiling and Therapeutic Evaluation of Standardized Hydroalcoholic Extracts of Terminalia chebula Fruits Collected from Different Locations in Manipur against Colorectal Cancer" Molecules 28, no. 7: 2901. https://doi.org/10.3390/molecules28072901