|
 
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| ORIGINAL ARTICLE |
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| Year : 2023 | Volume
: 15
| Issue : 1 | Page : 47-58 |
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Metabolite fingerprinting and profiling of selected medicinal plants using nuclear magnetic resonance
Manas Ranjan Sahoo, Marakanam Srinivasan Umashankar
Department of Pharmaceutics, SRM College of Pharmacy, SRM Institute of Science and Technology, Kattankulathur, Tamil Nadu, India
| Date of Submission | 19-Oct-2022 |
| Date of Decision | 14-Jan-2023 |
| Date of Acceptance | 27-Jan-2023 |
| Date of Web Publication | 31-Mar-2023 |
Correspondence Address:
Marakanam Srinivasan Umashankar
Department of Pharmaceutics, SRM College of Pharmacy, SRM Institute of Science and Technology, Kattankulathur - 603 203, Tamil Nadu
India
 Source of Support: None, Conflict of Interest: None
DOI: 10.4103/ajprhc.ajprhc_93_22
Background: Medicinal herbs are well known for their therapeutic effects and are traditionally used in the treatment and prevention of numerous diseases. Further plant-derived natural products have also been a valuable source of lead compounds for drug discovery and development. As the bioactivity of natural extracts is due to synergism between hundreds of metabolites present in the plant extract, the complete metabolomic analysis can be used for the quality control of phytomedicine. The 1H-nuclear magnetic resonance (NMR) fingerprint of the herbal extract can be used as a promising approach for comprehensive analysis of secondary metabolites to obtain a holistic view. The 1H-NMR spectroscopy has the advantage that it requires little quantity of samples and simple sample preparation method. Objectives: The study aimed to study the secondary metabolites of seven selected herbs, namely Abies webbiana, Cuminum cyminum, Elettaria cardamomum, Zingiber officinale, Glycyrrhiza glabra, Piper longum, and Terminalia chebula. Materials and Methods: The secondary metabolites of the herbal extracts were studied by recording the 1H-NMR spectra using NMR spectrometer in suitable solvent. Results: The putative metabolites that have been identified were 4-methoxy quercetin, luteolin, cuminaldehyde, 1,8-cineole, elettarins, gingerol, shogaol, glycyrrhizin, liquiritigenin, glabridin, betulinic acid, oleanolic acid, arabinogalactan, chebulagic acid, and gallic acid. Conclusion: The study highlighted the potential of NMR metabolomics in the study of plant extracts and their application for the quality control of phytomedicine.
Keywords: 1H-nuclear magnetic resonance, medicinal plants, metabolomics, phytochemical profiling, secondary metabolites
How to cite this article:
Sahoo MR, Umashankar MS. Metabolite fingerprinting and profiling of selected medicinal plants using nuclear magnetic resonance. Asian J Pharm Res Health Care 2023;15:47-58 |
How to cite this URL:
Sahoo MR, Umashankar MS. Metabolite fingerprinting and profiling of selected medicinal plants using nuclear magnetic resonance. Asian J Pharm Res Health Care [serial online] 2023 [cited 2023 Jun 8];15:47-58. Available from: http://www.ajprhc.com/text.asp?2023/15/1/47/373382 |
| Introduction | |  |
Long before the invention of modern medicine and synthetically prepared drugs, herbal medicines have been used for the treatment and prevention of various diseases. Based on the empirical knowledge of generations, as a part of traditions, natural medicines have evolved independently across different parts of the globe. Nowadays, traditional herbal medicines have been a part of global health care. It has been estimated that 65%–80% of the world population uses traditional herbal medicine to treat or prevent various diseases.[1] As per a recent report by the WHO, 80% of the population in developing countries such as Africa and Asia uses herbal medicines for primary health care.[2] Herbal medicines are used in human health care for the prevention and treatment of various diseases, as well as to improve and restore overall health. The strong trust in herbal medicine is due to accumulating experiences and observations of longstanding traditional uses that have been passed down from generation to generation. From health-care industry perspective, the global share of herbal medicines in 2016 was estimated to be more than USD 72 billion and is expected to increase to USD $111 billion by 2023.[3],[4],[5],[6] Within India, the annual trade in herbal medicines has been estimated to be US $10 billion and with an annual export of 1.1 billion.[7] Therefore, due to the emerging demand and growth of botanical medicines, there has been an increase in the efforts to monitor and regulate herbal drugs and botanical medicines.
Currently, the quality control practice of the herbal product includes the estimation of extractable matter, thin-layer chromatography, and standardization of the herbal extract with respect to its active marker or analytical marker constituents.[7] But this does not represent the complete spectrum of the chemical complexity of the phytochemical profile of the herbal constituent, as the biological activity of the herbal extracts is a result of whole phytochemical constituents rather than due to single constituents.[8] Hence, the complete fingerprint of the metabolites representing all types of phytochemicals of specific extract of a plant can be used for its standardization. This method uses multidisciplinary approaches of complementary analytical tools such as chromatography separation techniques, Fourier transform infrared spectroscopy, gas chromatography-mass spectrometry, liquid chromatography-mass spectrometry, nuclear magnetic resonance (NMR), and high-performance thin-layer chromatography.[9] In recent times, the NMR has emerged as a powerful tool for the study of plant metabolomics that provide preliminary structural information about its primary and secondary metabolites. The metabolic profiling of the plant secondary metabolite and fingerprinting is the key process in the quality control of herbal drugs. Therefore, NMR and FTIR spectra can be used for fingerprint development as well as to find out the phytochemical consistency between herbal extracts through identification of diagnostic peaks of the metabolites or markers in the herbal samples. Further, these experiments are rapid testing techniques that take very less time as compared to other methods. In the present study, we have used 1H-NMR and 13C-NMR-based fingerprinting techniques to identify the possible metabolites on the basis of the diagnostic peaks with the reported metabolites as per the literature. NMR spectroscopy is an essential analytical tool used for the identification and quantification of several molecules in natural samples such as food materials, plant extracts, herbal remedies, and biofluids.[10],[11] Furthermore, the NMR technique is nondestructive and does not require any complex sample preparation procedures. The herbs selected for the NMR-based characterization are Abies webbiana, Cuminum cyminum, Elettaria cardamomum, Zingiber officinale, Glycyrrhiza glabra, Piper longum, and Terminalia chebula. These herbs are well known in Ayurveda for various therapeutic indications and used in various Ayurvedic formulations.
| Materials and Methods | | |
For metabolite profiling, the herbs selected were Abies webbiana, Cuminum cyminum, Elettaria cardamomum, Zingiber officinale, Glycyrrhiza glabra, Piper longum, Terminalia chebula, and Curcuma longa. The herbs were extracted on the basis of the nature of phytochemical constituents. The herbs Glycyrrhiza glabra and Terminalia chebula were extracted with polar aqueous and chloroform solvent, whereas the herbs Abies webbiana, Cuminum cyminum, Elettaria cardamomum, and Zingiber officinale that are rich in phenolics and oleoresin were extracted with the ethyl alcohol. The samples were dissolved in CDCl3, D2O, and DMSO-d6. After the preparation of the extracts, the NMR analysis was carried out to obtain 1H-NMR and 13C-NMR spectrum. The NMR spectrum was recorded using a 400 MHZ Bruker Avance-400 spectrometer.
| Results and Discussion | | |
1H-nuclear magnetic resonance and 13C-nuclear magnetic resonance fingerprint of ethyl alcohol extract of Abies webbiana
The NMR spectra of both 1H [Figure 1] and 13C-NMR [Figure 2] showed prominent peaks in the aliphatic region as compared to the aromatic regions. | Figure 1: 1H-NMR spectrum alcohol extract of Abies webbiana. NMR: Nuclear magnetic resonance
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 | Figure 2: 1C NMR spectrum alcohol extract of Abies webbiana. NMR: Nuclear magnetic resonance
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1H-NMR (400 MHZ; DMSO-d6): 9.12, 6.97, 6.66, 5.32, 4.90, 4.86, 4.50, 4.42, 4.27, 3.82, 3.76, 3.64, 3.17, 3.04, 2.50, 2.06, 1.98, 1.53, 1.22, and 0.84.
13C-NMR (100 MHZ; DMSO-d6): 155.58,132.90,129. 48, 115.4, 98.5, 97.3, 92.6, 82.7, 77.2, 75.2, 74.1, 73.5, 72.8, 72.6, 72.4, 71.1, 71.0, 70.4, 70.7, 69.6, 68.2, 65.6, 63.5, 61.6, 57.8, 56.5, 31.1, 29.4, 24.06, and 19.00.
The peaks at δH of 9.12, 6.97 6.66, 3.32, and 2.50 and δC at 155.58, 132.7,115.45, 98.51,92.62 found to reported for the compound 4-methoxy quercetin as per the published literatures.[12]
1H-nuclear magnetic resonance and 13C-nuclear magnetic resonance fingerprint of ethyl alcohol extract Cuminum cyminum
Both the 1H-NMR [Figure 3] and 13C-NMR [Figure 4] showed the characteristic peaks in the region of aromatic region and aliphatic regions. | Figure 3: 1H-NMR spectra of Cuminum cyminum). NMR: Nuclear magnetic resonance
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 | Figure 4: 13C-NMR spectra of Cuminum cyminum. NMR: Nuclear magnetic resonance
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1H-NMR (DMSO-d6, 400MHz): 7.24, 7.21, 7.17, 6.99, 6.97, 6.93, 6.90, 6.89, 6.85, 6.69, 6.66, 6.04, 5.98, 5.96, 5.32, 3.51, 3.36, 2.95, 2.50, 2.27, 2.26, 2.13, 1.98, 1.60, 1.59, 1.47, 1.23, 0.84, and 0.83.
13C-NMR (DMSO-d6, 100MHz): 164.69, 48.40, 148.21, 142.2, 39.4, 138.1, 126.1, 122.9, 121.2, 108.9, 105.9, 101.7, 46.5, 24.6, and 20.6.
The peaks at δH at 6.99, 6.85, 6.69, and 6.66 ppm and δC at 164.69, 105.9,122.9, and 126.1 ppm were found to belong to the compound apigenin (5, 7, 4′-trihydroxyflavone) as per the reported literature. 7.24, 7.21, 6.89, and 6.66 and 13C-NMR signals at 164.69, 101.75, 105.94, 121.21, 142.27 and 148.40 ppm corresponds to the luteolin. The peaks at δH of 7·79, 7.34, 2.95, and 1.23 ppm have been reported for the compound cuminaldehyde as per the published literature values.[13],[14]
1H-nuclear magnetic resonance and 13C-nuclear magnetic resonance fingerprint of ethyl alcohol extract of Elettaria cardamomum
The NMR spectra of both 1H-NMR [Figure 5] and 13C-NMR [Figure 6] showed most of the peaks in aliphatic region. On the basis of the diagnostic peaks, [Table 1] 1,8-cineol, α-terpinyl acetate, and tetranorlabdane diterpenoids (elettarins) were identified by comparing with the published literature values.[15],[16],[17] | Figure 5: 1H-NMR of Elettaria cardamomum. NMR: Nuclear magnetic resonance
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 | Figure 6: 13C-NMR of Elettaria cardamomum. NMR: Nuclear magnetic resonance
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 | Table 1: 1H-nuclear magnetic resonance and 13C-nuclear magnetic resonance characteristic signals of identified metabolites in extract of Elettaria cardamomum (δ in ppm)[15],[16],[17]
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1H-NMR (DMSO-d6, 400MHz): 5.34, 5.31, 5.17, 4.90, 4.88, 4.27, 4.25, 3.83, 3.79, 3.76, 3.58, 3.55, 3.40, 3.27, 3.24, 3.12, 3.04, 2.50, 2.25, 1.94, 1.87, 1.60, 1.47, 1.37, 1.35, 1.22, and 0.85.
13C-NMR (DMSO-d6, 100MHz): 178.12, 170.30, 133.7, 120.7, 102.4, 98.5, 97.3, 84.5, 82.2, 77.1, 76.2, 75.7, 73.4, 75.24, 73.49, 73.27, 70.7, 70.31, 69.61, 68.20, 63.47, 63.38, 61.6, 53.65, 40.26, 39.88, 39.68, 39.47, 39.26, 32.78, 31.74, 30.84, 29.48, 29.15, and 26.24.
1H-nuclear magnetic resonance and 13C-nuclear magnetic resonance fingerprint of chloroform extract of Zingiber officinale
The spectra of both 1H-NMR [Figure 7] and 13C-NMR [Figure 8] showed the peaks in the region of the aliphatic and aromatic regions. The spectra indicate the presence of the phytochemicals with carbonyl functional groups and the presence of oxygenated carbons and aromatic compounds. | Figure 7: 1H-NMR chloroform extract of Zingiber officinale. NMR: Nuclear magnetic resonance
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 | Figure 8: 1H-NMR chloroform extract of Zingiber officinale. NMR: Nuclear magnetic resonance
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1H-NMR (CDCl3, 400MHz): δH ppm 7.26, 6.83, 6.70, 6.69, 6.67, 6.11, 5.35, 5.09, 2.86, 2.85, 2.73, 2.33, 2.31, 2.20, 2.18, 2.05, 2.00, 1.68, 1.60, 1.44, 1.25, and 0.81.
13C-NMR (CDCl3, 400MHz): δC ppm 199.9, 147.9, 146.4, 146.4, 143.8, 133.2, 132.6, 130.3, 120.8, 120.7, 114.4, 114.33, 111.1, 111.01, 67.71, 55.8, 49.3, 45.4, 41.9, 36.4, 32.4, 31.9, 31.7, 31.5, 29.8, 29.7, 29.6, 29.3, 29.2, 29.11, and 29.1.
The 1H-NMR spectrum indicated aromatic proton signals at δH 6.83, 6.67, and 6.11, methine proton at 5.35, methoxy protons at δH 3.86, and methylene protons in the range of δH 1.2–1.6 and δH 2.1–2.8. The spectrum exhibited a signal at δH 0.88 owing to the methyl group. The 13C-NMR spectrum exhibited the presence of methyl groups δc 13.96, 14.03, 14.08, 14.13, and 17.69, several methylene protons at δc 24.76, 25.13, 25.73, 25.64, 29.11, 29.17, 29.28, 29.33, 29.37, 29.61, 29.71, 29.88, 31.34, 31.53, 31.73, 31.93, 32.46, 36.42, 41.98, 45.44, and 49.34, methoxy group at δc 55.88, oxymethine group at δc 67.71. 76.73, double-bond carbon with signal at δc 143.8 and 120.8, aromatic carbons at 111.1, 111.0, 114.33, 114.42, 120.4, 130.3, 130.64, 132.6, 132.2, 143.8, 146.41, 146.47, and 147.98, and carbonyl carbon at δc 208.0 and 199.93. On the basis of comparison of the spectral data with that of the reported value, gingerol and shogaol classes of compounds were identified in the ginger extract by comparing with the published literature values.[18],[19],[20]
1H-nuclear magnetic resonance and 13C-nuclear magnetic resonance fingerprint of Glycyrrhiza glabra aqueous extract
The 1H-NMR [Figure 9] spectra distinctly showed the presence of methyl groups, oxymethine group at δ 3.37, and protons of glucuronic acid in the δ range of 4 to 5.5. The 13C-NMR [Figure 10] showed the presence of oxymethine carbon at 92.11, olefinic carbon at δ 129.95, and oxygenated carbons. Further, a Liebermann–Burchard reaction revealed the presence of a triterpenoid skeleton. The diagnostic peaks of the NMR spectra were in good agreement with that of the reported literature value of glycyrrhizin.[21],[22],[23],[24],[25] The characteristic NMR data of glycyrrhizin are presented in below [Table 2]. | Figure 9: 1H-NMR spectrum of Glycyrrhiza glabra in D2O. NMR: Nuclear magnetic resonance
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 | Figure 10: 13C-NMR spectrum of Glycyrrhiza glabra in D2O. NMR: Nuclear magnetic resonance
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 | Table 2: 1H-nuclear magnetic resonance and 13C-nuclear magnetic resonance characteristic signals of identified metabolites in aqueous extract of Glycyrrhiza glabra
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1H-NMR (400MHz, D2O): δppm: 7.03, 6.71, 5.29, 4.69, 4.08, 3.95, 3.69, 3.61, 3.55, 3.47, 3.45, 3.44, 3.42, 3.37, 3.34, 3.07, 2.91, 1.90, 1.88, 1.09, 0.82, and 0.63.
13C-NMR (100 MHz, D2O): δppm: 192.11, 183.15, 168.18, 129.95, 115.23, 103.61, 91.11, 82.61, 81.30, 76.32, 73.91, 72.49, 72.33, 72.06, 71.60, 71.00, 70.46, 69.75, 69.14, 62.30, 61.26, 61.18, 60.03, 59.66, 46.02, 28.92, and 23.70.
1H-nuclear magnetic resonance and 13C-nuclear magnetic resonance fingerprint of chloroform extract of Glycyrrhiza glabra
The 1H-NMR and 13C-NMR spectra of the chloroform fraction of Glycyrrhiza glabra are given in below [Figure 11] and [Figure 12]. The 1H-NMR [Figure 11] and 13C-NMR [Figure 12] spectra showed the signals in the aliphatic and aromatic regions. On the basis of the appearance of diagnostic peaks from the spectra and its comparison with the reported literature, the compounds such as liquiritigenin, glabridin, betulinic acid, and oleanolic acid were identified in the chloroform fraction of Glycyrrhiza glabra. The characteristic peaks of these compounds are presented in [Table 3].[26],[27] | Figure 11: 1H-NMR spectra of chloroform fraction of Glycyrrhiza glabra. NMR: Nuclear magnetic resonance
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 | Figure 12: 13C NMR spectra of chloroform fraction of Glycyrrhiza glabra. NMR: Nuclear magnetic resonance
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 | Table 3: 1H-nuclear magnetic resonance and 13C-nuclear magnetic resonance characteristic signals of identified metabolites in chloroform fraction of Glycyrrhiza glabra
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1H-NMR (400MHz, CDCl3) δH ppm: 7.85, 6.82, 6.78, 6.64, 6.36, 5.55, 5.35, 5.33, 5.23, 5.02, 4.35, 3.96, 3.85, 3.74, 3.45, 3.38, 2.82, 2.34, 2.05, 2.02, 1.77, 1.72, 1.68, 1.60, 1.41, and 1.29.
13C-NMR (100MHz, CDCl3) δC ppm: 192.5, 178.2, 161.10, 154.74, 151.75, 149.74, 129.23, 128.94, 127.99, 125.40, 121.40, 121.11, 119.11, 116.99, 115.80, 114.76, 114.56, 109.92, 108.68, 107.75, 55.33, 31.94, 31.61, 30.54, 29.72, 29.38, 29.27, 29.15, 27.73, 27.52, 27.22, 25.83, 24.75, 22.71, 17.92, and 14.14.
1H- and 13C-nuclear magnetic resonance fingerprint of Piper longum chloroform extract
1H-NMR (CDCl3 400MHz): δH ppm 7.43, 7.39, 7.27, 6.97, 6.89, 6.87, 6.78, 6.76, 6.74, 6.45, 6.41, 5.97, 5.92, 5.80, 5.76, 5.34, 3.63, 3.52, 3.39, 3.38, 3.16, 3.13, 2.33, 2.31, 2.17, 2.13, 2.12, 2.01, 1.65, 1.59, 1.58, 1.28, 1.25, 0.92, and 0.91.
13C-NMR (CDCl3 100 MHz): δC ppm 172.32, 166.48, 165.42, 148.20, 148.14, 142.59, 141.20, 139.22, 138.31, 135.31, 131.00, 129.32, 128.26, 126.66, 125.33, 122.52, 119.98, 108.49, 108.19, 105.67, 105.50, 105.38, 101.28, 101.22, 100.89, 76.76, 46.94, 32.95, 32.84, 31.92, 29.70, 29.65, 29.35, 29.31, 19.19, 19.15, 18.98, 28.81, 28.62, 26.91, 26.72, 25.62, 14.65, 22.69, 22.14, 20.14, 14.12, and 12.01.
Among the above peaks from 1H-NMR spectra [Figure 13], δH at 7.43, 7.39, 7.25, 6.95, 6.88, 6.86, 6.76, 6.74, 6.43, 6.39, 5.95, 3.63, and 1.65 ppm and δC at 165.47, 148.20, 148.14, 142.62, 138.31, 131.04, 125.37, 122.50, 119.96, 108.49, 105.67, 101.27, 46.94, 26.91, 25.62, 24.72, and 14.10 ppm. The peak at 5.95 corresponds to the dioxy-methylene protons (O-CH2-O). All these diagnostic peaks correspond to the reported NMR signals of piperine as per the published literature. Hence, piperine was identified clearly in the extract. The diagnostic peaks at δH 6.78, 2.17, and 1.25 and δC from 13C-NMR [Figure 14] at 60.80, 53.70, 178.5, 148.14, 141.70, 135.31, 105.50, 165.42, 105.38, 142.59, 128.26, 129.32, 126.66, 22.14, and 14.12 correspond to flavonoid class of compounds as per the reported literature values. The diagnostic peaks are δH 0.92, 2.01, 3.16, 5.92, 5.97, 6.74, 6.87, 6.89, and 7.36 ppm and δC ppm at 19.19, 28.62, 46.94, 101.28, 105.38, 108.49, 122.52, 126.66, 129.8, 138.31, 139.22, 141.20, 148.14, and 166.48 and correspond to the compound isopiperlongumine as per the reported literature. The diagnostic peaks are δH at 0.92, 1.28, 1.58, 1.59, 1.65, 2.17, 3.16, 5.76, 5.97, 6.41, 6.74, 6.89, and 7.27 ppm and δC at 22.14, 28.62, 28.81, 29.32, 29.35, 46.94, 100.89, 105.38, 108.19, 119.98, 122.52, 128.26, 129.32, 135.31, 141.20, 142.59, 148.14, and 166.42 ppm and correspond to brachystamide-D on comparison with that reported in the literature.[28],[29] | Figure 13: 1H-NMR of Piper longum extract. NMR: Nuclear magnetic resonance
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 | Figure 14: 13C NMR Piper longum extract. NMR: Nuclear magnetic resonance
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1H-nuclear magnetic resonance and 13C-nuclear magnetic resonance fingerprint of Terminalia chebula aqueous extract
[Figure 15] shows the 1H-NMR and 13C-NMR spectrum [Figure 16] of aqueous extract of T. chebula recorded in D2O. A number of resonances were seen in the spectrum, as expected from a complex mixture of plant extracts. Distinct signals between 2.0 to 3.0 and 6.0 to 7.0 indicates presence of proteins and presence of different signals at 5.04, 4.98, 4.69 come from anomeric protons and signals at 3.51 and 4.28 indicates oxygenated protons of carbohydrate ring. The spectral pattern of water extract showed the presence of polysaccharides, carbohydrates, amino acids, and phenolic compounds. | Figure 15: 1H-NMR Spectrum in D2O (400 MHz). NMR: Nuclear magnetic resonance
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 | Figure 16: 13C-NMR spectrum in D2O (100 MHz). NMR: Nuclear magnetic resonance
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1H-NMR (400MHz, D2O) δH ppm: 6.98, 6.92, 6.86, 6.81, 6.75, 6.62, 6.60, 6.59, 6.52, 6.31, 6.28, 6.03, 5.04, 4.98, 4.69, 4.28, 4.01, 3.90, 3.88, 3.87, 3.85, 3.80, 3.78, 3.73, 3.71, 3.60, 3.58, 3.55, 3.54, 3.52, 2.73, 2.71, 2.69, 2.53, 2.51, 2.48, 2.47, 2.02, 1.92, and 1.90.
13C-NMR (100 MHz, D2O) δC ppm: 175.90, 167.35, 144.64, 144.20, 143.62, 142.06, 139.60, 137,73, 136.45, 130.25, 118.55, 115.51, 109.72, 109.14, 79.09, 71.16, 70.74, 69.17, 66.50, 65.75, 79.09, 71.16, 70.74, 69.17, 66.50, 65.75, 44.46, 36.29, 34.11, and 30.57.
The compound chebulagic acid and gallic acid[30],[31],[32] were identified in aqueous extract on the basis of observed spectral data and comparison with the reported data from the literature. The diagonostic peaks of these compounds are presented in [Table 4]. Further another compound arabinogalactan protein polysaccharide was identified on basis of the presence of signals at 4.98, 4.28 for the sugar protons, glycosidic protons at 3.5 to 4.2, the anomeric proton at 5.2 and 5.3 and the protons of protein moiety in the range of 6.0 to 7.0 (6.28, 6.52, 6.62, 6.75, 6.81, 6.92 and 6.99) 2.0 to 3.0 (2.02, 2.47, 2.51, 2.69, 2.73 and these signals were in accordance with that of published literature.[33] | Table 4: 1H-nuclear magnetic resonance and 13C-nuclear magnetic resonance characteristic signals of identified metabolites in aqueous extract of Terminalia chebula
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1H- and 13C-nuclear magnetic resonance fingerprint of Terminalia chebula chloroform fraction
The chloroform fraction of Terminalia chebula showed both the proton and carbon signals in the region of aromatic and aliphatic regions. The 1H- and 13C-NMR are given in [Figure 17] and [Figure 18], respectively. | Figure 17: 1H-NMR spectrum in CDCl3 (400 MHz). NMR: Nuclear magnetic resonance
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 | Figure 18: 1H-NMR spectrum in CDCl3 (100 MHz). NMR: Nuclear magnetic resonance
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1H-NMR (400MHz, CDCL3) δH ppm: 7.71, 7.54, 7.52, 7.26, 5.34, 4.23, 2.36, 2.32, 2.30, 2.02, 1.63, 1.61, 1.42, 1.41, 1.33, 1.29, 1.28, 1.25, 0.94, 0.92, 0.89, and 0.88.
13C-NMR (100MHz, CDCL3) δC ppm: 167.79, 132.46, 130.89, 128.81, 114.07, 76.70, 68.17, 38.73, 33.84, 33.67, 31.94, 31.44, 30.37, 29.71, 29.37, 28.93, 23.75, 22.99, 22.70, 14.13, 14.06, and 10.97.
The compound was identified to be gallic acid and its ethyl ester was identified in the chloroform fraction on the basis of the reported literature evidence.[32],[33],[34] The details of diagnostic NMR peaks are presented in [Table 5]. | Table 5: 1H-nuclear magnetic resonance and 13C-nuclear magnetic resonance characteristic signals of identified metabolites in aqueous extract of Terminalia chebula
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| Conclusion | | |
The secondary metabolites are present in very low quantity, and the isolation of the compounds from the natural sources is a time-consuming and complicated process with repeated chromatography purification due to the complex nature of the plant extract. Metabolomic experiments offer an improved and speed route for the identification of secondary metabolites in plant extracts. The metabolomic approach helps to simultaneous explore of the phytochemical diversity of herbal samples. The NMR can be a suitable method for fingerprinting of the herbal samples that offer reproducibility to confirm the batch-to-batch consistency within a shorter time with simple sample preparation method. NMR spectroscopy provides a powerful technique for the identification of the plant metabolites. In the present work, the metabolite profiling of the medicinal plants has been successfully carried out using NMR fingerprinting. 1H-NMR and 13C-NMR spectral fingerprints of the extracts of selected medicinal plants using techniques were investigated. The key NMR spectra peaks were identified for the selected herbs. Various putative compounds were identified in various medicinal plant extracts without going for prepurification. Highly significant qualitative differences were noticed between different extracts with respect to the secondary metabolites. The NMR spectra showed a clear discrimination between all samples. Hence, NMR metabolomics could be a promising and effective, fast, and convenient technique for the quality control of herbal medicines.
Financial support and sponsorship
Nil.
Conflicts of interest
There are no conflicts of interest.
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[Figure 1], [Figure 2], [Figure 3], [Figure 4], [Figure 5], [Figure 6], [Figure 7], [Figure 8], [Figure 9], [Figure 10], [Figure 11], [Figure 12], [Figure 13], [Figure 14], [Figure 15], [Figure 16], [Figure 17], [Figure 18]
[Table 1], [Table 2], [Table 3], [Table 4], [Table 5]
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