Analgesic, Anti-Inflammatory, Cytotoxic Activity Screening and UPLC-PDA-ESI-MS Metabolites Determination of Bioactive Fractions of Kleinia pendula
Abstract
:1. Introduction
2. Results
2.1. Acute Oral Toxicity Studies
2.2. Analgesic Activity of Fractions of Kleinia pendula
2.3. Anti-Inflammatory Activity of Fractions of Kleinia Pendula
2.4. Cytotoxic Assay
2.5. Characterization of Metabolites from the Bioactive Fractions Using UPLC-PDA-ESI-MS
3. Discussion
4. Materials and Methods
4.1. Plant Material
4.2. Preparation of Extract and Fractions
4.3. Animals
4.4. Acute Toxicity Studies
4.5. Measurement of Analgesic Activity
4.6. Measurement of Anti-Inflammatory Activity
4.7. Cell Culture
4.8. Cytotoxicity Studies
4.9. UPLC-PDA-ESI-MS Analysis of the Active Fractions
4.10. Statistical Methods
5. Conclusions
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Sample Availability: Samples of the compounds are not available from the authors. |
Fractions/Extract | IC50 (μg/mL) | ||
---|---|---|---|
MCF-7 (µg) | HepG2 (µg) | HCT-116 (µg) | |
Methanol extract | 3.2 ± 0.6 | 3.17 ± 0.69 | 4.8 ± 0.6 |
Hexane fraction | 0.07 ± 0.03 | 0.19 ± 0.02 | 0.11 ± 0.01 |
Chloroform fraction | 0.13 ± 0.07 | 0.24 ± 0.03 | 0.19 ± 0.15 |
Ethyl acetate fraction | 90.22 ± 18.6 | ≥100 | 28.1 ± 5.3 |
Butanol fraction | ≥100 | ≥100 | ≥100 |
Water fraction | ≥100 | ≥100 | ≥100 |
Doxorubicin | 0.014 ± 0.008 | 0.0065 ± 0.005 | 0.013 ± 0.0005 |
Peak | Retention Time | Identified Compd. | UV-Vis (λ Max) | [M − H]− (m/z) | Fragment Ions (m/z) | Percentage (%) | Ref. |
---|---|---|---|---|---|---|---|
Kp1 | 4.2 | Quinic acid | 323 | 191 | 178, 173, 148, 110 | 0.8 | [15]. |
Kp2 | 6.2 | Protocatechuic acid | 320 | 153 | 113, 105 | 0.5 | [16]. |
Kp3 | 20.48 | Di-caffeoyl quinic acid | 246, 310 | 515 | 191 | 5.7 | [15,17] |
Kp4 | 21.32 | Chlorogenic acid | 246, 310 | 353 | 191 | 1.21 | [17] |
Kp5 | 21.98 | Feruloyl-quinic acid | 247,310 | 367 | 191 | 0.62 | [15] |
Kp6 | 22.9 | Isorhamnetin-3-O-glucoside-7-O-rhamnoside | 230, 262 | 623 | 315, 153 | 3.2 | [18] |
Kp7 | 23.09 | Feruloyl-quinic acid | 247,310 | 367 | 191 | 1.32 | [15] |
Kp8 | 24.6 | Trihydroxyphenethyl-O-rhamnopyranosyl-(1-6)-4-O-caffeoyl-glucopyranoside | 232, 282 | 621 | 487, 469 | 3.4 | [19] |
Kp9 | 25.72 | Sinapic acid hexoside | 310 | 385 | 223 | 4.82 | [20] |
Kp10 | 25.85 | Di-caffeoyl-hexuronide derivative | 328 | 710 | 355, 135, 113 | 2.5 | [21] |
Kp11 | 26.27 | Chlorogenic acid | 246, 310 | 353 | 191 | 4.95 | [17] |
Kp12 | 28.86 | Feruloyl-quinic acid | 333 | 367 | 191 | 5.1 | [15] |
Kp13 | 29.22 | Sulphate conjugate of dimethyl gallic acid | 260 | 277 | 197,163 | 1.6 | [22,23] |
Kp14 | 30.62 | Di-methylgallic acid derivative | 267 | 291 | 155 | 0.7 | [22,23] |
Kp15 | 35.9 | Coumaroyl-shikimic acid | 225 | 319 | 155 | 0.92 | [15] |
Kp16 | 38.9 | Apigenin-6,8-di-C-glucoside | 246, 310 | 593 | 297,135 | 1.6 | [22,23] |
Kp17 | 40.21 | Formylipolamiidic acid | Undetected | 419 | 401, 257, 155 | 0.9 | [22,23] |
Kp18 | 42.82 | Procyanidin B3 | 330 | 579 | 453, 127 | 0.5 | [24,25] |
Peak | Retention Time | Identified Compd. | UV-Vis (λ Max) | [M − H]− (m/z) | Fragment Ions (m/z) | Percentage (%) | Ref. |
---|---|---|---|---|---|---|---|
Kp19 | 3.5 | Hexahydroxydiphenoyl (HHDP)-galloylglucose | 274 | 633 | 301, 257, 229 | 5.4 | [26] |
Kp20 | 5.18 | Dihydroxy-4-methoxyl isoflavan | 226, 284 | 271 | 227, 135 | 3.75 | [27] |
Kp21 | 5.2 | Gallocatechin | 274 | 305 | 179 | 1.29 | [17] |
Kp22 | 6.2 | HHDP-galloylglucose | 274 | 633 | 301, 257, 229 | 1.3 | [26] |
Kp23 | 6.9 | Trihydroxyphenethyl-O-rhamnopyranosyl-(1-6)-4-O-caffeoyl-glucopyranoside | 232, 282 | 621 | 487, 469 | 3.4 | [19] |
Kp24 | 7.38 | unidentified | 230, 290 | 604 | 582, 462, 342 | 2.38 | [28] |
Kp25 | 9.2 | Methylretusin | 230, 283 | 297 | 281, 239 | 0.18 | [27] |
Kp26 | 9.35 | Acyl-feruloyl-4-O-caffeoyl-quinic acid | 220, 232 | 571 | 277, 191 | 0.95 | [15] |
Kp27 | 12.08 | Glycycoumarin hydroxylate glucuronide | 258 | 559 | 338 | 3.29 | [22,23] |
Kp28 | 12.65 | Sulfate conjugate of glycycoumarin | 283 | 447 | 367 | 2.01 | [22,23] |
Kp29 | 14.78 | trisgalloyl (hexahydroxydiphenoyl) glucose derivative | 275 | 907 | 765,191 | 0.72 | [29] |
Peak | Retention Time | Identified Compd. | [M − H]− (m/z) | Fragment Ions (m/z) | Percentage (%) | Ref. |
---|---|---|---|---|---|---|
Kp30 | 4.2 | Tinosposinenside B | 581 | 379, 343, 297 | 0.21 | [30] |
Kp31 | 4.93 | Amritoside A | 555 | 537, 513 | 0.42 | [30] |
Kp32 | 4.95 | Isocryptotanshinone II | 297 | 225, 211 | 0.8 | [31] |
Kp33 | 5.23 | Linolenic acid | 277 | 250, 219 | 3.5 | [31] |
Kp34 | 11.3 | Octadecadienoic acid derivative | 265 | 249, 179 | 0.2 | [31] |
Kp35 | 14.2 | Tanshinone IIB | 311 | 275, 250 | 0.73 | [32] |
Kp36 | 25.6 | Salvianolic acid G | 339 | 277, 249 | 0.35 | [33] |
Kp37 | 27.97 | Tinocrisposide | 535 | 521, 355 | 1.9 | [30] |
Kp38 | 31.43 | Tormentic acid | 487 | 469 | 4.9 | [34] |
Kp39 | 34.3 | 6,7-Dehydroroyleanone | 315 | 297, 216 | 1.87 | [35] |
Kp40 | 36.8 | Salvianolic acid D | 417 | 197, 175, 135 | 1.2 | [31] |
Kp41 | 37.80 | Dehydrated derivative of Tinosposinenside B | 419 | 297 | 3.75 | [30] |
Kp42 | 38.45 | 5-Hydroxydavisiosidec | 513 | 197 | 4.89 | [36] |
Kp43 | 39.5 | Citrostadienyl | 432 | 419 | 4.3 | [37] |
Kp44 | 40.58 | Unidentified | 319 | 305, 291, 277 | 6.75 | n.d. |
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Alfaifi, M.; Alsayari, A.; Gurusamy, N.; Louis, J.; Eldin Elbehairi, S.; Venkatesan, K.; Annadurai, S.; I. Asiri, Y.; Shati, A.; Saleh, K.; et al. Analgesic, Anti-Inflammatory, Cytotoxic Activity Screening and UPLC-PDA-ESI-MS Metabolites Determination of Bioactive Fractions of Kleinia pendula. Molecules 2020, 25, 418. https://doi.org/10.3390/molecules25020418
Alfaifi M, Alsayari A, Gurusamy N, Louis J, Eldin Elbehairi S, Venkatesan K, Annadurai S, I. Asiri Y, Shati A, Saleh K, et al. Analgesic, Anti-Inflammatory, Cytotoxic Activity Screening and UPLC-PDA-ESI-MS Metabolites Determination of Bioactive Fractions of Kleinia pendula. Molecules. 2020; 25(2):418. https://doi.org/10.3390/molecules25020418
Chicago/Turabian StyleAlfaifi, Mohammad, Abdulrhman Alsayari, Narasimman Gurusamy, Justin Louis, Serag Eldin Elbehairi, Kumar Venkatesan, Sivakumar Annadurai, Yahya I. Asiri, Ali Shati, Kamel Saleh, and et al. 2020. "Analgesic, Anti-Inflammatory, Cytotoxic Activity Screening and UPLC-PDA-ESI-MS Metabolites Determination of Bioactive Fractions of Kleinia pendula" Molecules 25, no. 2: 418. https://doi.org/10.3390/molecules25020418