Identification of Dipeptidyl Peptidase-4 and α-Amylase Inhibitors from Melicope glabra (Blume) T. G. Hartley (Rutaceae) Using Liquid Chromatography Tandem Mass Spectrometry, In Vitro and In Silico Methods
Abstract
:1. Introduction
2. Results and Discussion
2.1. Extraction Yield
2.2. Dipeptidyl Peptidase-4 Inhibitory Activity
2.3. α-Amylase Inhibitory Activity
2.4. Phytochemical Profiling of Active Fraction Using LC-MS/MS
2.5. In Silico Inhibitory Analysis of Identified Compounds
3. Materials and Methods
3.1. Chemicals and Materials
3.2. Plant Materials
3.3. Preparation of Extracts and Fractionation
3.4. Determination of Extraction Yield
3.5. Antidiabetic Assays
3.5.1. DPP-4 Inhibitory Assay
3.5.2. α-Amylase Inhibitory Assay
3.6. LC-MS/MS Analysis
3.7. In Silico Inhibitory Analysis of Identified Compounds
3.8. Statistical Analysis
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Sample Availability
References
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Plant Part | Extracts | Extraction Yield |
---|---|---|
(%) | ||
Leaves | Hexane | 2.03 ± 0.31 c |
Chloroform | 2.79 ± 0.19 d | |
Methanol | 7.30 ± 1.10 e | |
Stem bark | Hexane | 0.3 ± 0.16 a |
Chloroform | 0.7 ± 0.03 b | |
Methanol | 2.13 ± 0.25 c |
Plant Part | Extracts | IC50 (μg/mL) | |
---|---|---|---|
DPP-4 | α-Amylase | ||
Leaves | Hexane | 1623.60 ± 121.61 g | 4230.12 ± 324.76 h |
Chloroform | 169.40 ± 9.30 c | 303.64 ± 10.10 c | |
Methanol | 1086.48 ± 142.69 f | 2488.13 ± 231.54 g | |
Stem bark | Hexane | 8408.36 ± 102.23 i | 5447.01 ± 243.16 i |
Chloroform | 332.31 ± 10.07 d | 975.80 ± 17.10 e | |
Methanol | 4992.33 ± 0.84 h | 3946.12 ± 143.21 h | |
Fractions | |||
CF1 | 1711.06 ± 70.32 g | 8663.12 ± 110.75 j | |
CF2 | 619.31 ± 9.21 e | 253.30 ± 19.21 b | |
CF3 | 313.18 ± 20.92 d | 531.44 ± 11.38 d | |
CF4 | 128.35 ± 12.77 b | 170.19 ± 20.66 a | |
CF5 | 711.42 ± 10.26 e | 1817.83 ± 209.42 f | |
Sitagliptin | 0.01 ± 0.01 a | - | |
Acarbose | - | 188.60 ± 14.31 a |
Compound | RT (min) | Identification | Molecular Formula | Observed Neutral Mass (Da) | Mass Error (mDa) | Precursor ion [M−H] (m/z) | Major Fragments (m/z) |
---|---|---|---|---|---|---|---|
Flavonoid | |||||||
5 | 13.82 | 4′,5,6,7-tetramethoxy-flavone | C19H18O6 | 342.1103 | 0.8 | 341.1032 | 311.0925 |
6 | 14.07 | Isorhamnetin | C16H12O7 | 316.0583 | −0.8 | 315.0503 | 300.0270, 272.0280 |
7 | 14.22 | quercetagetin-3,4′-dimethyl ether | C17H14O8 | 346.0689 | 0.4 | 345.0617 | 330.0380, 315.0416, 164.9826 |
8 | 14.91 | 5,3′,4′-trihydroxy-6,7-dimethoxy-flavone | C17H14O7 | 330.0740 | 0.4 | 329.0662 | 314.0427, 299.0192, 271.0244 |
Cinnamic acid derivative | |||||||
3 | 11.91 | methyl 3,4,5-trimethoxycinnamate | C13H16O5 | 252.0998 | −0.4 | 251.0923 | 193.0506, 179.0352 |
Glucoside | |||||||
4 | 13.04 | Renifolin | C18H24O7 | 352.1522 | 0.4 | 351.1452 | 199.0990 |
Lactone | |||||||
2 | 11.63 | swermirin | C10H10O4 | 194.0579 | −0.4 | 193.0506 | 179.0352, 149.0611 |
9 | 15.15 | 2-methoxy-5-acetoxy-fruranogermacr-1(10)-en-6-one | C18H24O5 | 320.1624 | 0.7 | 319.1554 | 305.1400, 179.0356 |
Coumarin | |||||||
1 | 10.20 | trans-decursidinol | C14H14O5 | 262.0841 | −0.2 | 261.0765 | 203.0349, 159.0460 |
Compounds | DPP-4 | α-Amylase | ||||
---|---|---|---|---|---|---|
Binding Affinity (kcal/mol) | H-Bond | Hydrophobic | Binding Affinity (kcal/mol) | H-Bond | Hydrophobic | |
1 | −7.7 | Glu206, Arg125, Ser630, Glu205 | Glu205, Tyr666 | −8.2 | Glu233, Asp300 | Ala198, Leu162, His201 |
2 | −5.4 | Lys122, Asp739 | His740, Arg125 | −6.4 | His299, Asp197, Tyr62, Asp300 | Ala198, Trp58 |
3 | −5.6 | Arg125, Tyr547, Arg669, Val 207, Tyr662 | Tyr666, Glu205 | −6.0 | His299, Tyr62, Thr163 | Trp59 |
4 | −7.8 | Glu206, Glu205, Asn710, Arg125, Ser630 | Tyr547, Phe357 | −8.2 | Asp300, His299, Asp197, Glu233, His305 | Trp59, Leu165 |
5 | −7.7 | Arg125, Ser630, His740, Asn710, Tyr547, Val207 | Glu205, Phe357 | −8.4 | His299, Gln63, Asp300, Glu233 | Tyr62, Trp59 |
6 | −7.8 | Glu206, Arg358, Tyr547 | Phe357, Tyr666 | −8.0 | Lys200, Tyr151, Asp197 | Ile235, Ala198, His201, Leu162, Lys200 |
7 | −7.9 | Ser209, Arg125, Tyr631 | Glu205, Glu206, Tyr666, Phe662 | −8.6 | Thr163, Glu233, Gln63, Arg195 | Leu162, Leu165, Asp197, Asp300 |
8 | −8.1 | Glu206, Arg125, Asn710, His740, Ser630 | Glu205 | −8.1 | Glu233, Asp197, Asp300 | Trp59, Leu165 |
9 | −6.3 | His126, Arg125 | Glu205, Tyr666, Phe357 | −7.5 | Gln63, Asp197 | Leu162, His299, Tyr62, His305, Trp59, Trp58 |
sitagliptin | −8.6 | Ser209, Arg125, Arg358, Glu205, Glu206, Tyr 662 | Phe357, Ser630, Tyr666, Val207, His740 | - | - | - |
acarbose | - | - | - | −6.9 | His305, Glu233, Tyr62, Asp197, Asp300, Thr163 | - |
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Quek, A.; Kassim, N.K.; Ismail, A.; Latif, M.A.M.; Shaari, K.; Tan, D.C.; Lim, P.C. Identification of Dipeptidyl Peptidase-4 and α-Amylase Inhibitors from Melicope glabra (Blume) T. G. Hartley (Rutaceae) Using Liquid Chromatography Tandem Mass Spectrometry, In Vitro and In Silico Methods. Molecules 2021, 26, 1. https://doi.org/10.3390/molecules26010001
Quek A, Kassim NK, Ismail A, Latif MAM, Shaari K, Tan DC, Lim PC. Identification of Dipeptidyl Peptidase-4 and α-Amylase Inhibitors from Melicope glabra (Blume) T. G. Hartley (Rutaceae) Using Liquid Chromatography Tandem Mass Spectrometry, In Vitro and In Silico Methods. Molecules. 2021; 26(1):1. https://doi.org/10.3390/molecules26010001
Chicago/Turabian StyleQuek, Alexandra, Nur Kartinee Kassim, Amin Ismail, Muhammad Alif Mohammad Latif, Khozirah Shaari, Dai Chuan Tan, and Pei Cee Lim. 2021. "Identification of Dipeptidyl Peptidase-4 and α-Amylase Inhibitors from Melicope glabra (Blume) T. G. Hartley (Rutaceae) Using Liquid Chromatography Tandem Mass Spectrometry, In Vitro and In Silico Methods" Molecules 26, no. 1: 1. https://doi.org/10.3390/molecules26010001