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Experimental design for chemometrics-based evaluation of bioactivity of 21 GTE panels representing multicomponent systems.
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Although understanding their chemical composition is vital for accurately predicting the bioactivity of multicomponent drugs, nutraceuticals, and foods, no analytical approach exists to easily predict the bioactivity of multicomponent systems from complex behaviors of multiple coexisting factors. We herein represent a metabolic profiling (MP) strat...
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... screening-driven MALDI-MS-MP technique for quality evaluation of diverse GTEs with different properties. Herein, we aimed to develop a MALDI-MS system suitable for the chemometrics-driven evaluation of the bioactivity of diverse GTE panels (Fig. 1). In MALDI-MS, the matrix preparation, including the selection of the matrix and solvent, is a critical step to ensure efficient ionization of the analyte, because the detection of the analyte is completely dependent on these conditions 11 . Because little is known about which matrices can simultaneously ionize multiple compounds from ...
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... mixture was spotted onto a stainless steel MALDI sample plate and analyzed by MALDI-TOF-MS in the negative ionization mode. All the resulting spectral data were processed using an in-house script ( Supplementary Fig. S1) that simultaneously detects authentic phytochemical peaks in their basic, deprotonated ion form [M-H] − while effectively excluding various matrix background peaks. The obtained information on the intensity of all the peaks was converted to a heatmap to visually compare the extent of each ionization (Fig. 2A and Supplementary Table S1). ...
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... however, MP based on the MALDI-MS datasets (Fig. 3B) showed a better performance than that based on LC-MS. As shown in Supplementary Fig. S10, LC-MS data was not able to construct appropriate OPLS models using datasets excluding components with higher VIP values (>1) (Supplementary Table S6). In contrast, MALDI-MS data was not greatly affected by the exclusion of such components (Supplementary Tables S3 and S7) although its predictive performance was slightly lowered. ...
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... we attempted to apply this established technique to crude herbal extracts. The appropriate linearity was obtained for the representative GTE components (EGCG, EGC, ECG, and EC) at the standard level (Supplementary Fig. S11A). These compounds were detected in a dilution series of the representative GTE (YB1) (Supplementary Fig. S11B). ...
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... appropriate linearity was obtained for the representative GTE components (EGCG, EGC, ECG, and EC) at the standard level (Supplementary Fig. S11A). These compounds were detected in a dilution series of the representative GTE (YB1) (Supplementary Fig. S11B). A 10-fold dilution of the GTE stock solution was used in all experiments under the condition ensuring the linearity of four compounds. ...
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... 10-fold dilution of the GTE stock solution was used in all experiments under the condition ensuring the linearity of four compounds. MS data of the representative GTE (YB1) sample were acquired on a different day ( Supplementary Fig. S11C), and there were no significant changes in relative spectral patterns. We were also able to stably construct preferable OPLS models using such MS data ( Supplementary Fig. S11D). ...
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... data of the representative GTE (YB1) sample were acquired on a different day ( Supplementary Fig. S11C), and there were no significant changes in relative spectral patterns. We were also able to stably construct preferable OPLS models using such MS data ( Supplementary Fig. S11D). These results ensure the robustness of our proposed MALDI-MS-MP system. ...
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... proposed MALDI-MS procedures (sample preparation, matrix selection, peak detection and alignment, and multivariate statistical analysis) contribute to the construction of standardization of crude herbal extracts for successfully performing MP and the screening of chemical combination. These results allowed us to overcome the drawbacks of the conventional MP technique ( Supplementary Fig. S8), and may enable a rapid and simple high-throughput MP of crude samples for the evaluation of their bioactivity (Supplementary Fig. S12) as well as various applications, including the quality assessment, breeding, screening, and monitoring of low-molecular-weight chemicals. In addition, further chemometric research will open new avenues for investigating the potential relationship between the bioactivity of crude extracts and their multiple coexisting factors, and for determining effective chemical combinations for bioactivity prediction. ...
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... the GTE experiments, GTE samples (1/10 dilution in water) were subjected to MALDI-MS analysis using 1,5-DAN or 9-AA as the matrix under the same spotting and measuring conditions as for the phytochemicals. Then, peak picking and alignment, noise reduction, and deisotoping of the obtained spectral data were performed using an in-house script ( Supplementary Fig. 1S), and the resultant data were subjected to multivariate statistical analysis. Phytochemical peaks were assigned by MS/MS analysis or by searching for their precise masses using the MassBank metabolite databases. ...
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... statistical analysis. The datasets of the 21 GTEs were subjected to multivariate statistical analysis to identify similarity/dissimilarity among the samples (149 (1,5-DAN) or 18 (9-AA) distinct m/z peaks). We conducted an unsupervised multivariate principle component analysis (PCA) and a supervised multivariate OPLS analysis using SIMCA-P+ ver.12 (Umetrics, Umea, Sweden). ...
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... LC-MS and GC-MS have been the techniques most commonly used for the MP of medicinal herbs, agricultural products, and foods for quality evaluation. Previously, we showed for the first time the useful- ness of LC-MS-MP for evaluating the apoptosis-inducing activity of GTEs and for screening anti-cancer com- pounds or synergetic sensitizers 20 . In this study, we also performed conventional LC-MS-MP for the same GTE samples used in the main experiment ( Supplementary Fig. S9 and Supplementary Table 5S). LC-MS detected a greater number of peaks (507) than did MALDI-MS (149), and MP based on the composition profiles from LC-MS successfully achieved various quality evaluations, such as distinguishing the similarity and dissimilarity among cultivars and picking seasons ( Supplementary Fig. S9A), and the prediction of ORAC and polyphenol content ( Supplementary Fig. S9B,C). Interestingly, however, MP based on the MALDI-MS datasets (Fig. 3B) showed a better performance than that based on LC-MS. As shown in Supplementary Fig. S10, LC-MS data was not able to construct appropriate OPLS models using datasets excluding components with higher VIP values (>1) (Supplementary Table S6). In contrast, MALDI-MS data was not greatly affected by the exclusion of such compo- nents (Supplementary Tables S3 and S7) although its predictive performance was slightly lowered. These results suggest that MALDI-MS datasets include more number of peaks correlated with ORAC values or polyphenol contents compared to LC-MS datasets, and such peaks may coordinately contribute to the better performance of MALDI-MS datasets. The lowered performance of LC-MS datasets may be due to the low-correlative ability (predictability) of components with VIP values (<1) to ORAC values or polyphenol contents. These findings endorse the quality of the composition profiles obtained from MALDI-MS measurements and their applicability to the rapid and simple non-targeted MP of medicinal herbs, agricultural products, and ...
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... summary, we have established an effective strategy for evaluating a measure of the bioactivity, namely the antioxidant activity, of diverse GTE panels by non-targeted MP-based chemometric analy- sis using a high-throughput analytical system, MALDI-MS. This approach also enabled us to easily extract a bioactivity-predictive chemical combination from multicomponent information. Our proposed MALDI-MS procedures (sample preparation, matrix selection, peak detection and alignment, and multivariate statistical anal- ysis) contribute to the construction of standardization of crude herbal extracts for successfully performing MP and the screening of chemical combination. These results allowed us to overcome the drawbacks of the conven- tional MP technique ( Supplementary Fig. S8), and may enable a rapid and simple high-throughput MP of crude samples for the evaluation of their bioactivity (Supplementary Fig. S12) as well as various applications, including the quality assessment, breeding, screening, and monitoring of low-molecular-weight chemicals. In addition, further chemometric research will open new avenues for investigating the potential relationship between the bioactivity of crude extracts and their multiple coexisting factors, and for determining effective chemical com- binations for bioactivity prediction. This may contribute to the discovery of new scientific data helpful for the development of multicomponent botanical drugs and dietary supplements, herbal medicines, functional foods, and combinations of foods/beverages optimized to promote health and reduce the risk of ...
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... we have reported the performance of basic MALDI-MS procedures, including the repeatabil- ity, sensitivity, and linearity of detection of low-molecular-weight metabolites [6][7][8]22 . Here we attempted to apply this established technique to crude herbal extracts. The appropriate linearity was obtained for the represent- ative GTE components (EGCG, EGC, ECG, and EC) at the standard level (Supplementary Fig. S11A). These compounds were detected in a dilution series of the representative GTE (YB1) (Supplementary Fig. S11B). A 10-fold dilution of the GTE stock solution was used in all experiments under the condition ensuring the lin- earity of four compounds. MS data of the representative GTE (YB1) sample were acquired on a different day ( Supplementary Fig. S11C), and there were no significant changes in relative spectral patterns. We were also able to stably construct preferable OPLS models using such MS data ( Supplementary Fig. S11D). These results ensure the robustness of our proposed MALDI-MS-MP system. The present technique is effective for performing the high-throughput first screening of bioactive compounds and combinations from crude herbal samples at the ini- tial step of research. However, this MS technique is still in vitro cell-free system, and further its technical validity would be reinforced through some sort of comparison with in vitro cellular systems and/or in vivo systems. In our research groups, some MALDI-MS-MP researches, using other bioactivities and medicinal herbal extracts, bacterial samples, and animal/human body fluids, are in progress. These attempts may also contribute to the eval- uation of technical validity and further improvement of our proposed MS ...
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... we have reported the performance of basic MALDI-MS procedures, including the repeatabil- ity, sensitivity, and linearity of detection of low-molecular-weight metabolites [6][7][8]22 . Here we attempted to apply this established technique to crude herbal extracts. The appropriate linearity was obtained for the represent- ative GTE components (EGCG, EGC, ECG, and EC) at the standard level (Supplementary Fig. S11A). These compounds were detected in a dilution series of the representative GTE (YB1) (Supplementary Fig. S11B). A 10-fold dilution of the GTE stock solution was used in all experiments under the condition ensuring the lin- earity of four compounds. MS data of the representative GTE (YB1) sample were acquired on a different day ( Supplementary Fig. S11C), and there were no significant changes in relative spectral patterns. We were also able to stably construct preferable OPLS models using such MS data ( Supplementary Fig. S11D). These results ensure the robustness of our proposed MALDI-MS-MP system. The present technique is effective for performing the high-throughput first screening of bioactive compounds and combinations from crude herbal samples at the ini- tial step of research. However, this MS technique is still in vitro cell-free system, and further its technical validity would be reinforced through some sort of comparison with in vitro cellular systems and/or in vivo systems. In our research groups, some MALDI-MS-MP researches, using other bioactivities and medicinal herbal extracts, bacterial samples, and animal/human body fluids, are in progress. These attempts may also contribute to the eval- uation of technical validity and further improvement of our proposed MS ...
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... we have reported the performance of basic MALDI-MS procedures, including the repeatabil- ity, sensitivity, and linearity of detection of low-molecular-weight metabolites [6][7][8]22 . Here we attempted to apply this established technique to crude herbal extracts. The appropriate linearity was obtained for the represent- ative GTE components (EGCG, EGC, ECG, and EC) at the standard level (Supplementary Fig. S11A). These compounds were detected in a dilution series of the representative GTE (YB1) (Supplementary Fig. S11B). A 10-fold dilution of the GTE stock solution was used in all experiments under the condition ensuring the lin- earity of four compounds. MS data of the representative GTE (YB1) sample were acquired on a different day ( Supplementary Fig. S11C), and there were no significant changes in relative spectral patterns. We were also able to stably construct preferable OPLS models using such MS data ( Supplementary Fig. S11D). These results ensure the robustness of our proposed MALDI-MS-MP system. The present technique is effective for performing the high-throughput first screening of bioactive compounds and combinations from crude herbal samples at the ini- tial step of research. However, this MS technique is still in vitro cell-free system, and further its technical validity would be reinforced through some sort of comparison with in vitro cellular systems and/or in vivo systems. In our research groups, some MALDI-MS-MP researches, using other bioactivities and medicinal herbal extracts, bacterial samples, and animal/human body fluids, are in progress. These attempts may also contribute to the eval- uation of technical validity and further improvement of our proposed MS ...
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... we have reported the performance of basic MALDI-MS procedures, including the repeatabil- ity, sensitivity, and linearity of detection of low-molecular-weight metabolites [6][7][8]22 . Here we attempted to apply this established technique to crude herbal extracts. The appropriate linearity was obtained for the represent- ative GTE components (EGCG, EGC, ECG, and EC) at the standard level (Supplementary Fig. S11A). These compounds were detected in a dilution series of the representative GTE (YB1) (Supplementary Fig. S11B). A 10-fold dilution of the GTE stock solution was used in all experiments under the condition ensuring the lin- earity of four compounds. MS data of the representative GTE (YB1) sample were acquired on a different day ( Supplementary Fig. S11C), and there were no significant changes in relative spectral patterns. We were also able to stably construct preferable OPLS models using such MS data ( Supplementary Fig. S11D). These results ensure the robustness of our proposed MALDI-MS-MP system. The present technique is effective for performing the high-throughput first screening of bioactive compounds and combinations from crude herbal samples at the ini- tial step of research. However, this MS technique is still in vitro cell-free system, and further its technical validity would be reinforced through some sort of comparison with in vitro cellular systems and/or in vivo systems. In our research groups, some MALDI-MS-MP researches, using other bioactivities and medicinal herbal extracts, bacterial samples, and animal/human body fluids, are in progress. These attempts may also contribute to the eval- uation of technical validity and further improvement of our proposed MS ...
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... of GTEs. We prepared 21 distinct hot-water extracts from the leaves (Table 1) of 7 Japanese green tea cultivars (Camellia sinensis L. and C. sinensis x C. taliensis), which were cropped during 3 different pick- ing seasons (7 April-7 May, 28 May-11 June, and 4-17 July). Six cultivars (Camellia sinensis L.) were purchased from retail tea stores, and one cultivar (C. sinensis x C. taliensis), Sunrouge (SR), was kindly donated by Nippon Paper Industries Co., Ltd (Tokyo, Japan). A fine powder of the dried leaf (30 mg) from each cultivar was added to 1.5 mL boiling water for 10 min. The extract was centrifuged at 15,000 × g for 10 min, and the supernatant was subjected to further analyses. MALDI-MS analysis. Individual phytochemical standard compounds were dissolved in water or MeOH, diluted to give graded concentrations (100 ppm), and mixed with a matrix (9-AA, 1,5-DAN, Nor-Ho, or harmine)/100% MeOH or acetone solution (10 mg/mL) at a ratio of 1:1 (v/v). The sample (0.5 μL) was spotted onto the ground-steel MALDI plate and air-dried. Four spots were deposited from each individual sample and their data averaged for the subsequent data analyses. A MALDI-TOF-MS (AXIMA Performance, Shimadzu, Japan) was used for all the analyses. Each mass spectrum was acquired with 5 laser shots. For each sample spot, 121 spectra were mean-centered. All spectrometric data were processed and analysed using the Shimadzu Biotech Launchpad software. The ionization of each phytochemical was confirmed by a deprotonated ion peak [M-H] − . In the GTE experiments, GTE samples (1/10 dilution in water) were subjected to MALDI-MS analysis using 1,5-DAN or 9-AA as the matrix under the same spotting and measuring conditions as for the phytochemicals. Then, peak picking and alignment, noise reduction, and deisotoping of the obtained spectral data were performed using an in-house script ( Supplementary Fig. 1S), and the resultant data were subjected to multivariate statistical analysis. Phytochemical peaks were assigned by MS/MS analysis or by searching for their precise masses using the MassBank metabolite ...
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... statistical analysis. The datasets of the 21 GTEs were subjected to multivariate statistical analysis to identify similarity/dissimilarity among the samples (149 (1,5-DAN) or 18 (9-AA) distinct m/z peaks). We conducted an unsupervised multivariate principle component analysis (PCA) and a supervised multivariate OPLS analysis using SIMCA-P+ ver.12 (Umetrics, Umea, Sweden). PCA models are depicted as score plots and consist of two synthetic variables: principal component (PC) 1 (accounting for the greatest proportion of the total variance) and PC2 (accounting for the second greatest proportion of the total variance orthogonal to PC1). These plots display intrinsic groups of samples based on their spectral variations. This analysis attempts to explain the original features of the samples as far as possible based on a ratio of the sum of the percentages of PC1 and ...
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... screening-driven MALDI-MS-MP technique for quality evaluation of diverse GTEs with different properties. Herein, we aimed to develop a MALDI-MS system suitable for the chemomet- rics-driven evaluation of the bioactivity of diverse GTE panels (Fig. 1). In MALDI-MS, the matrix preparation, including the selection of the matrix and solvent, is a critical step to ensure efficient ionization of the analyte, because the detection of the analyte is completely dependent on these conditions 11 . Because little is known about which matrices can simultaneously ionize multiple compounds from herbal extracts, including various phyto- chemicals, we first screened the optimum matrix for detecting approximately 70 representative phytochemicals Table S1) among four potentially high-performing matrices previously reported 7,12,13 . A solution of each matrix in 100% methanol (MeOH) or acetone was mixed with an equal volume of a phytochemical solu- tion. This mixture was spotted onto a stainless steel MALDI sample plate and analyzed by MALDI-TOF-MS in the negative ionization mode. All the resulting spectral data were processed using an in-house script ( Supplementary Fig. S1) that simultaneously detects authentic phytochemical peaks in their basic, deprotonated ion form [M-H] − while effectively excluding various matrix background peaks. The obtained information on the intensity of all the peaks was converted to a heatmap to visually compare the extent of each ionization (Fig. 2A and Supplementary Table S1). The detectability of the phytochemicals was differed among the four matrices, namely 9-AA, 1,5-diami- nonaphthalene (1,5-DAN), norharmane (nor-Ho), and harmine. Based on the ionization rate and the number of phytochemicals successfully ionized, the best ionization performance was achieved by 1,5-DAN, and the ace- tone ( Fig. 2A) provide a better solvent than MeOH (Supplementary Fig. S2). The ionization efficiency of the phytochemicals varied widely depending on their chemical structures (Supplementary Fig. S3). Furthermore, 1,5-DAN obtained the greatest number of peaks (138 following background subtraction) from the crude aqueous GTEs, many more than were detected by 9-AA (Fig. 2B). In fact, while 9-AA is the most commonly used matrix in non-targeted metabolomic analysis 6,8 , it showed the worst detection performance in this study ( Fig. 2A and Supplementary Fig. S2). To examine the potential of 1,5-DAN as a matrix for the acquisition of the chemical compositions and the subsequent quality evaluation of diverse GTEs, MALDI-MS measurements and the sub- sequent multivariate statistical analysis were performed using 21 distinct GTEs from 7 representative Japanese green tea cultivars (Camellia sinensis L. and C. sinensis x C. taliensis) cropped during 3 different picking seasons (Table 1 and Supplementary Table S2). The score plot of the principal component analysis (PCA), an unsuper- vised multivariate statistical analysis, showed clear clusters, one consisting of the Sunrouge (SR) cultivar (C. sin- ensis x C. taliensis), and the other consisting of the remaining cultivars (Camellia sinensis L.) (Fig. 2C). The cluster separation of the cultivars was observed along the PC2 axis (1,5-DAN, right panel). Regardless of whether the SR cultivar was included or excluded (leaving 21 or 18 GTEs, respectively), clusters related to the picking season could be observed along PC1 (1,5-DAN, left panel). These results strongly suggest that the compositional differ- ences among the GTEs can account for the different cultivars and picking seasons. In contrast, no such cluster formation was observed in the MALDI-MS-MP results when using 9-AA as a matrix (Fig. 2C, right panel). These results show that phytochemical-based matrix screening is an effective strategy for selecting the optimal matrix, in this case 1,5-DAN, for the analysis of the chemical compositions of diverse GTEs and their quality ...
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... screening-driven MALDI-MS-MP technique for quality evaluation of diverse GTEs with different properties. Herein, we aimed to develop a MALDI-MS system suitable for the chemomet- rics-driven evaluation of the bioactivity of diverse GTE panels (Fig. 1). In MALDI-MS, the matrix preparation, including the selection of the matrix and solvent, is a critical step to ensure efficient ionization of the analyte, because the detection of the analyte is completely dependent on these conditions 11 . Because little is known about which matrices can simultaneously ionize multiple compounds from herbal extracts, including various phyto- chemicals, we first screened the optimum matrix for detecting approximately 70 representative phytochemicals Table S1) among four potentially high-performing matrices previously reported 7,12,13 . A solution of each matrix in 100% methanol (MeOH) or acetone was mixed with an equal volume of a phytochemical solu- tion. This mixture was spotted onto a stainless steel MALDI sample plate and analyzed by MALDI-TOF-MS in the negative ionization mode. All the resulting spectral data were processed using an in-house script ( Supplementary Fig. S1) that simultaneously detects authentic phytochemical peaks in their basic, deprotonated ion form [M-H] − while effectively excluding various matrix background peaks. The obtained information on the intensity of all the peaks was converted to a heatmap to visually compare the extent of each ionization (Fig. 2A and Supplementary Table S1). The detectability of the phytochemicals was differed among the four matrices, namely 9-AA, 1,5-diami- nonaphthalene (1,5-DAN), norharmane (nor-Ho), and harmine. Based on the ionization rate and the number of phytochemicals successfully ionized, the best ionization performance was achieved by 1,5-DAN, and the ace- tone ( Fig. 2A) provide a better solvent than MeOH (Supplementary Fig. S2). The ionization efficiency of the phytochemicals varied widely depending on their chemical structures (Supplementary Fig. S3). Furthermore, 1,5-DAN obtained the greatest number of peaks (138 following background subtraction) from the crude aqueous GTEs, many more than were detected by 9-AA (Fig. 2B). In fact, while 9-AA is the most commonly used matrix in non-targeted metabolomic analysis 6,8 , it showed the worst detection performance in this study ( Fig. 2A and Supplementary Fig. S2). To examine the potential of 1,5-DAN as a matrix for the acquisition of the chemical compositions and the subsequent quality evaluation of diverse GTEs, MALDI-MS measurements and the sub- sequent multivariate statistical analysis were performed using 21 distinct GTEs from 7 representative Japanese green tea cultivars (Camellia sinensis L. and C. sinensis x C. taliensis) cropped during 3 different picking seasons (Table 1 and Supplementary Table S2). The score plot of the principal component analysis (PCA), an unsuper- vised multivariate statistical analysis, showed clear clusters, one consisting of the Sunrouge (SR) cultivar (C. sin- ensis x C. taliensis), and the other consisting of the remaining cultivars (Camellia sinensis L.) (Fig. 2C). The cluster separation of the cultivars was observed along the PC2 axis (1,5-DAN, right panel). Regardless of whether the SR cultivar was included or excluded (leaving 21 or 18 GTEs, respectively), clusters related to the picking season could be observed along PC1 (1,5-DAN, left panel). These results strongly suggest that the compositional differ- ences among the GTEs can account for the different cultivars and picking seasons. In contrast, no such cluster formation was observed in the MALDI-MS-MP results when using 9-AA as a matrix (Fig. 2C, right panel). These results show that phytochemical-based matrix screening is an effective strategy for selecting the optimal matrix, in this case 1,5-DAN, for the analysis of the chemical compositions of diverse GTEs and their quality ...
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... The CP and cumP showed a VIP score that was >2 ( Figure. 5C) and those metabolites with VIP value ≥1 could be considered as key predictors for bioactivity [74]. Similarly, in the S-plot, CP, and cumP were placed far from the origin indicating their high correlation to the measured AGI activities of pepper leaves extracts. ...
... The CP and cumP showed a VIP score that was >2 (Figure. 5C) and those metabolites with VIP value ≥1 could be considered as key predictors for bioactivity [74]. Similarly, in the S-plot, CP, and cumP were placed far from the origin indicating their high correlation to the measured AGI activities of pepper leaves extracts. ...
Metabolomics and in vitro α-glucosidase inhibitory (AGI) activities of pepper leaves were used to identify bioactive compounds and select genotypes for the management of type 2 diabetes mellitus (T2DM). Targeted metabolite analysis using UPLC-DAD-QToF-MS was employed and identified compounds that belong to flavone and hydroxycinnamic acid derivatives from extracts of pepper leaves. A total of 21 metabolites were detected from 155 samples and identified based on MS fragmentations, retention time, UV absorbance, and previous reports. Apigenin-O-(malonyl) hexoside, luteolin-O-(malonyl) hexoside, and chrysoeriol-O-(malonyl) hexoside were identified for the first time from pepper leaves. Pepper genotypes showed a huge variation in their inhibitory activity against α-glucosidase enzyme(AGE) ranging from 17% to 79%. Genotype GP38 with inhib-itory activity of 79% was found to be more potent than the positive control acarbose (70.8%.). Orthogonal partial least square (OPLS) analyses were conducted for the prediction of the AGI activities of pepper leaves based on their metabolite composition. Compounds that contributed the most to the bioactivity prediction model (VIP >1.5), showed a strong inhibitory potency. Caffeoyl-putrescine was found to show a stronger inhibitory potency (IC50 = 145 µM) compared to acarbose (IC50 = 197 µM). The chemometric procedure combined with high-throughput AGI screening was effective in selecting polyphenols of pepper leaf for T2DM management.
... Specific characteristics in each cluster facilitated the selection of two promising extracts. This technique has previously been used to analyze the connection between biological activity and phytochemical profile for selection of extracts/plants with the best biological activities [26,52,53]. ...
Reactive oxygen species (ROS) are related to several degenerative diseases. In this study, Acacia, a genus with many fast-growing species, was investigated to explore the many phytochemical compounds that are biologically active in processes dealing with ROS-related diseases. This study aimed to select extracts of Acacia heartwood on the basis of their pharmacological and phytochemical profiles and identify their bioactive compounds. Five methanolic extracts from Acacia heartwood were evaluated for their antioxidant activity using three different in vitro assays: toxicity toward Artemia salina and phenolic and polyphenolic content. Multivariate analysis was conducted to select two promising extracts and then their bioactive compounds were identified using liquid chromatography coupled with mass spectrometry. Acacia crassicarpa extracts showed the highest antioxidant activity, as well as phenolic and hydrolyzable tannin contents, but low toxicity. The A. mangium extract exhibited high flavonoid and condensed tannin content, whereas A. decurrrens had the highest toxicity with low antioxidant activity. Pearson’s correlation analysis demonstrated no correlation between antioxidant activity and toxicity. Moreover, the phytochemical profile exhibited an association with pharmacological parameters. Principal component analysis followed by cluster analysis divided the extracts into three clusters. Two heartwood extracts of A. crassicarpa and A. auriculiformis were chosen as the best extracts. Identification showed that these extracts were dominated by phenolic compounds, as well as anthraquinone and xanthone.
... Metabolic profiling in food science enables us to evaluate the balance of components in samples, determine the differences in samples containing multi-component compounds, and predict the correlations between bioactivity/functional activity and multi-component compounds [14][15][16]. Recently, Fujimura et al. identified multi-component compounds in tea extracts, which were correlated with bioactivity, by employing non-targeted analyses using matrix-assisted laser desorption ionization-mass spectrometry [17]. Among many metabolic profiling analytical techniques, such as nuclear magnetic resonance spectroscopy [18], visible micro-Raman spectroscopy [19], Fourier-transform infrared spectroscopy [20], hyperspectral imaging [21,22], liquid chromatography (LC) [23], and combined platforms [24], MS-based techniques are particularly sensitive and selective [25]. ...
This paper reports on the chemical compounds in arabica coffee beans with a high Specialty Coffee Association (SCA) cupping score, especially those in specialty coffee beans. We investigated the relationship between the chemical compounds and cupping scores by considering 16 types of Coffea arabica (arabica coffee) beans from Guatemala (SCA cupping score of 76.5–89.0 points). Non-targeted gas chromatography-mass spectrometry-based chemometric profiling indicated that specialty beans with a high cupping score contained considerable amounts of methyl-esterified compounds (MECs), including 3-methylbutanoic acid methyl ester (3-MBM), and other fatty acid methyl esters. The effect of MECs on flavor quality was verified by spiking the coffee brew with 3-MBM, which was the top-ranked component, as obtained through a regression model associated with cupping scores. Notably, 3-MBM was responsible for the fresh-fruity aroma and cleanness of the coffee brew. Although cleanness is a significant factor for specialty beans, the identification of compounds that contribute to cleanness has not been reported in previous research. The chemometric profiling approach coupled with spiking test validation will improve the identification and characterization of 3-MBM commonly found in arabica specialty beans. Therefore, 3-MBM, either alone or together with MECs, can be used as a marker in coffee production.
... Additionally, chemometrics supply the foundation for the bourgeoning field of biochemometrics the combined studies of chemometrics and bioactive compounds. A central challenge of natural products research is assigning bioactive compounds from complex mixtures or extracts [18][19][20][21]. Bioactive compounds are increasingly valued in the field of pharmacology for their utility and variety, and the study of biochemometrics aids further development of the efficacy and verification of these compounds [22,23]. ...
This review serves to highlight the role of chemometrics and biochemometrics in recent literature as well as including a perspective on the current state of the field, as well as the future needs and possible directions. Specifically examining the analytical methods and statistical tools that are available to chemists, current applications of QTOF-MS, Orbitrap-MS, LC with PDA/UV detectors, NMR, and IMS coupled MS are detailed. Of specific interest, these techniques can be applied to botanical dietary supplement quality, efficacy, and safety. Application in natural products drug discovery, industrial quality control, experimental design, and more are also discussed.
... Further analysis of different peak profiles obtained by other LC-MS conditions will become an effective strategy for constructing more precise bioactivity-prediction models and the subsequent identification of more effective bioactive compounds. Recently, we have reported that the MP-based data-mining technique could be used for effectively identifying bioactive chemical combinations in green tea extracts 10,25 . An introduction of this chemical combination-oriented approach to the present study may open new avenues for investigating the potential relationship between the bioactivity of crude green tea extracts and their multiple coexisting ingredients, and for determining effective chemical combinations for prediction of glycolytic enzyme inhibitory activity. ...
Although the major green tea catechins can inhibit the activity of carbohydrate-hydrolyzing enzymes, there is a paucity of information describing the potential of other green tea ingredients and numerous green tea cultivars. Herein, we reveled that a green tea cultivar Sunrouge significantly suppressed the postprandial blood glucose level in mice. Unlike the most representative Japanese green tea cultivar, Yabukita, the suppression by Sunrouge was observed clearly during the initial period after oral dosing of starch. Sunrouge also strongly inhibited the carbohydrate-hydrolyzing enzymes α-glucosidase and α-amylase when compared with that of Yabukita and many other cultivars. Liquid chromatography–mass spectrometry (LC–MS)-based metabolic profiling (MP) of 42 Japanese green tea cultivars was performed. Multivariate statistical analysis enabled visualization of the differences among cultivars with respect to their ability to inhibit carbohydrate-hydrolyzing activities. Analysis of metabolites, contributing to the discrimination and prediction of the bioactivity of cultivars, showed that O-methylated catechins, epicatechin-3-O-(3-O-methyl) gallate (ECG3”Me) and epigallocatechin-3-O-(3-O-methyl) gallate (EGCG3”Me), were newly identified α-glucosidase inhibitors. Such ability was also observed in epigallocatechin-3-O-gallate (EGCG), epicatechin-3-O-gallate (ECG), delphinidin-3-O-glucoside and myricetin-3-O-glucoside. The amounts of these compounds in Sunrouge were higher than that in many other cultivars. These results suggest that Sunrouge has high potential for suppressing the elevation of the postprandial blood glucose level, and an MP approach may become a valuable strategy for evaluating the anti-hyperglycemic activity of green tea and for screening its active ingredients.
... Considering the principle of this methodology, it is expected that MP may become an effective strategy for obtaining a comprehensive understanding of the physiological activities of multicomponent drugs and nutraceuticals. Recently, we demonstrated that the MALDI-MS-MP technique could be used to evaluate the anti-oxidant activity, oxygen radical absorbance capacity (ORAC), of diverse GTE panels based on their compositional balances, and select an effective chemical combination to predict the bioactivity [117]. ...
... Selected representative charts of the GTEs are shown, demonstrating that the ORAC can be visually estimated from the abundances of the four components. Adapted with permission from reference [117]. ...
... Selected representative charts of the GTEs are shown, demonstrating that the ORAC can be visually estimated from the abundances of the four components. Adapted with permission from reference[117]. ...
Low-molecular-weight phytochemicals have health benefits and reduce the risk of diseases, but the mechanisms underlying their activities have remained elusive because of the lack of a methodology that can easily visualize the exact behavior of such small molecules. Recently, we developed an in situ label-free imaging technique, called mass spectrometry imaging, for visualizing spatially-resolved biotransformations based on simultaneous mapping of the major bioactive green tea polyphenol and its phase II metabolites. In addition, we established a mass spectrometry-based metabolic profiling technique capable of evaluating the bioactivities of diverse green tea extracts, which contain multiple phytochemicals, by focusing on their compositional balances. This methodology allowed us to simultaneously evaluate the relative contributions of the multiple compounds present in a multicomponent system to its bioactivity. This review highlights small molecule-sensing techniques for visualizing the complex behaviors of herbal components and linking such information to an enhanced understanding of the functionalities of multicomponent medicinal herbs.
There are numerous cultivars of tea (Camellia sinensis L.), but the differences in their anti-hyperglycemic-related effects are largely unknown. The inhibition of the dipeptidyl peptidase (DPP)-IV enzyme plays an essential role in controlling hyperglycemia in diabetes by blocking the degradation of incretin hormones, which is necessary for insulin secretion. In this study, we examined the DPP-IV inhibitory activity of leaf extracts from diverse Japanese green tea cultivars. The inhibitory rates differed among tea extracts. Metabolic profiling (MP), using liquid chromatography-mass spectrometry, of all cultivars revealed compositional differences among cultivars according to their DPP-IV inhibitory capacity. Epigallocatechin-3-O-(3-O-methyl)gallate, kaempferol-3-O-rutinoside, myricetin-3-O-glucoside/galactoside, and theogallin were newly identified as DPP-IV inhibitors. The bioactivity of a tea extract was potentiated by adding these ingredients in combination. Our results show that MP is a useful approach for evaluating the DPP-IV inhibitory potency of green tea and for determining bioactivity-related ingredients and combinations.
Many bioactive peptides have been described from marine sources and much marine biomass is still not explored or utilized in products. Marine peptides can be developed into a variety of products, and there is a significant interest in the use of bioactive peptides from marine sources towards nutraceuticals or functional foods. We present here a mini review collecting the knowledge about the value chain of bioactive peptides from marine sources used in nutraceuticals and functional foods. Many reports describe bioactive peptides from marine sources, but in order to make these available to the consumers in commercial products it is important to connect the bioactivities associated with these peptides to commercial opportunities and possibilities. In this mini review, we present challenges and opportunities for the commercial use of bioactive peptides in nutraceuticals and functional food products. We start the paper by introducing approaches for isolation and identification of bioactive peptides and candidates for functional food. We further discuss market driven innovation targeted to ensure that isolated peptides and suggested products are marketable and acceptable by targeted consumers. To increase the commercial potential and ensure sustainability of the identified bioactive peptides and products, we discuss scalability, regulatory frameworks, production possibilities and the shift towards greener technologies. Finally, we discuss some commercial products from marine peptides within the functional food market. We discuss around the placement these products within the larger picture of the commercial sphere of functional food products from bioactive peptides.
