Chemometric profiling and anti-arthritic activity of aerial parts of Glinus oppositifolius (L.) Aug. DC.

ETHNOPHARMACOLOGICAL RELEVANCE: Glinus oppositifolius (L.) Aug. DC. belongs to the family Molluginaceae, an annual prostrate herb traditionally used to treat inflammations, arthritis, malarial, wounds, fevers, diarrhoea, cancer, stomach discomfort, jaundice, and intestinal parasites. However, the anti-arthritic activity of the aerial part has still not been reported. AIM OF THE STUDY: To investigate the antioxidant and anti-arthritic activity of G. oppositifolius in Complete Freund's Adjuvant (CFA) induced rats. MATERIALS AND METHODS: The dried aerial parts of this plant material were defatted with n-hexane and extracted by methanol using a soxhlet apparatus. The in vitro anti-arthritic activity of methanolic extract of G. oppositifolius (MEGO) was evaluated in protein denaturation, membrane stabilization, and inhibition of proteinase assay at 25, 50, 100, 200, and 400 µg/ml concentrations. Female Wistar rats were immunized sub-dermally into the right hind paw with 0.1 ml of CFA. Rats were administered with MEGO at doses of 200 and 400 mg/kg once daily for fourteen days after arthritis induction. Assessment of arthritis was performed by measuring paw diameter, arthritic index, arthritic score, body weight, organ weight, and hematological and biochemical parameters, followed by the analysis of pro-inflammatory cytokines such as tumor necrosis factor-alpha (TNF-α), interleukin 6 (IL-6), interleukin-1-beta (IL-1ß), cyclooxygenase-2 (COX-2), interleukin 13 (IL-13) and interleukin 10 (IL-10) and histopathological study. In vivo antioxidant effect was investigated in enzymatic assays. The presence of phytoconstituents was analyzed by Gas Chromatography-Mass Spectrometry (GC-MS) and Liquid Chromatography-Mass Spectrometry (LC-MS), respectively. In silico molecular docking study of the compounds was carried out against COX-2, IL-1ß, IL-6, and TNF-α using AutoDock 4.2 and BIOVIA-Discovery Studio Visualizer software. RESULTS: MEGO's in vitro anti-arthritic activity showed dose-dependent inhibition of protein denaturation, membrane stabilization, and proteinase inhibition, followed by significant in vivo anti-arthritic activity. The rats treated with MEGO showed tremendous potential in managing arthritis-like symptoms by restoring hematological, biochemical, and histological changes in CFA-induced rats. MEGO (200 and 400 mg/kg) showed a significant alleviation in the levels of hyper expressed inflammatory mediators (TNF-α, IL-1ß, and IL-6) and oxidative stress (SOD, CAT, GSH, and LPO) in CFA-induced rats. Spergulagenin-A as identified by LC-MS analysis, exhibited the highest binding affinity against COX-2 (-8.6), IL-1ß (7.2 kcal/mol), IL-6 (-7.4 kcal/mol), and TNF-α (-6.5 kcal/mol). CONCLUSIONS: Provided with the comprehensive investigation, methanolic extract of G. oppositifolius against arthritic-like condition is a proof of concept that revalidates its ethnic claim. The presence of Spergulagenin-A might be responsible for the anti-arthritic activity.

The chemometrics analysis and integrated pharmacology approach to decipher the effect and mechanism between raw and processed cistanche tubulosa.

ETHNOPHARMACOLOGICAL RELEVANCE: Cistanche tubulosa (CT) is the dried fleshy stem with scaly leaves of Cistanche tubiflora (Schenk) Wight, which has the effects of tonifying the kidney-yang, benefiting the vital essence and blood, and moisturizing the intestines and laxatives. There are differences in the activity of CT before and after processing, but the mechanism of processing is not clear. AIM OF THE STUDY: The study aimed to compare the strength of action of CT before and after yellow-wine processing in the treatment of constipation and kidney yang deficiency and to identify the active ingredients responsible for the differences in activity before and after yellow-wine processing. MATERIALS AND METHODS: This study established the fingerprints of CT and PCT using HPLC to identify their shared components. Then efficacy of KYDS and FC were carried out to compare the differences between CT and PCT in terms of efficacy. Next, this study established the spectrum-effect relationship between the shared chemical components and the medical effects of CT and PCT using the gray correlation analysis and entropy methods. Ultimately, the activity of the analyzed chemical components was verified using the zebrafish model. RESULTS: CT was more effective than PCT in promoting intestinal peristalsis, regulating gastrointestinal hormone levels, and thus treating FC. PCT was more effective than CT in improving the level of hormone indexes of the hypothalamus-pituitary-target gland axis, replenishing blood, and enhancing immunity. Through the analysis of the spectrum-effect relationship, it was finally found that 5, 6, 12 (tubuloside A), and 13 (isoacteoside) might be more closely related to the activity of tonifying kidney yang, and peaks 9, 10, and 11 (acteoside) are more closely associated with the treatment of constipation, and peaks 3 (salidroside), 4, 1, 2 (geniposidic acid), and 8 (echinacoside) were associated with both kidney yang tonic and treatment of constipation. At the same time, an activity verification experiment showed that echinacoside, geniposidic acid, and salidroside were effective in the treatment of FC and KYDS, while acteoside was very effective in the treatment of FC, and tubuloside A was significant in supplementing the blood, which validated the spectrum-effect relationship analysis. CONCLUSION: This study proved that the raw CT had a better laxative effect, while the yellow-wine processed CT had a better kidney-yang tonic effect; moreover, spectrum-effect relationships were established to analyze the chemical components leading to changes in the activity of CT before and after yellow-wine processing.

Fast Discrimination and Quantification Analysis of Atractylodis rhizoma Using NIR Spectroscopy Coupled with Chemometrics Tools.

In this study, near-infrared (NIR) spectroscopy and high-performance liquid chromatography (HPLC) combined with chemometrics tools were applied for quick discrimination and quantitative analysis of different varieties and origins of Atractylodis rhizoma samples. Based on NIR data, orthogonal partial least squares discriminant analysis (OPLS-DA) and K-nearest neighbor (KNN) models achieved greater than 90% discriminant accuracy of the three species and two origins of Atractylodis rhizoma. Moreover, the contents of three active ingredients (atractyloxin, atractylone, and ß-eudesmol) in Atractylodis rhizoma were simultaneously determined by HPLC. There are significant differences in the content of the three components in the samples of Atractylodis rhizoma from different varieties and origins. Then, partial least squares regression (PLSR) models for the prediction of atractyloxin, atractylone, and ß-eudesmol content were successfully established. The complete Atractylodis rhizoma spectra gave rise to good predictions of atractyloxin, atractylone, and ß-eudesmol content with R2 values of 0.9642, 0.9588, and 0.9812, respectively. Based on the results of this present research, it can be concluded that NIR is a great nondestructive alternative to be applied as a rapid classification system by the drug industry.

Characterization and classification of oleogels and edible oil using vibrational spectroscopy in tandem with one-class and multiclass chemometric methods.

Oleogel represents a promising healthier alternative to act as a substitute for conventional fat in various food products. Oil selection is a crucial factor in determining the technological properties and applications of oleogels due to their distinct fatty acid composition, molecular weight, and thermal properties, as well as the presence of antioxidants and oxidative stability. Hence, the relevance of monitoring oleogel properties by non-destructive, eco-friendly, portable, fast, and effective techniques is a relevant task and constitutes an advance in the evaluation of oleogels quality. Thus, the present study aims to classify oleogels rapidly and reliably, without the use of chemicals, comparing two handheld near infrared (NIR) spectrometers and one portable Raman device. Furthermore, two different multivariate methods are compared for oleogel classification according to oil type. Three types of oleogels were prepared, containing 95 % oil (sunflower, soy, olive) and 5 % beeswax as a structuring agent, melted at 90 °C. Polarized light microscopy (PLM) images were acquired, and fatty acid composition, peroxide index and free fatty acid content were determined using official methods. A total of 240 oleogel and 92 oil spectra were obtained for each instrument. After spectra pretreatment, Principal Component Analysis (PCA) was performed, and two classification methods were investigated. The Data Driven - Soft Independent Modelling of Class Analogy (DD-SIMCA) and Partial Least Squares Discriminant Analysis (PLS-DA) models demonstrated 95 % to 100 % of accuracy for the external test set. In conclusion, the use of vibrational spectroscopy using handheld and portable instruments in tandem with chemometrics showed to be an efficient alternative for classifying oils and oleogels and could be extended to other food samples. Although the classification of vegetable oils by NIR is widely used and known, this work proposes the classification of different types of oil in oleogel matrices, which has not yet been explored in the literature.

Antioxidant Activities of Different Peanut (Arachis hypogaea L.) Market Types by Spectrophotometric Techniques Combined with Chemometrics.

Peanut is rich in oil and protein and has a large content of bioactive constituents consisting of tocopherols, phytosterols, and so on. Generally, Virginia, Spanish, Valencia and Runner market types are grown of peanut. In this study, it is aimed to determine the antioxidant activity, total phenolic content and total flavonoid content of peanuts from four different market types, for the first time, and group them with principal component analysis (PCA) and hierarchical cluster analysis (HCA). For PCA, PC1 and PC2 explained 87.655 % of the total variation and, according to the HCA of peanut samples, two main groups were determined. The total phenolic content changed 1.556 to 2.899 mg GAE/g. The lowest value have seen at Spanish merket type to determine the antioxidant activities of peanut samples were maked FRAP and DPPH assay, the lowest FRAP value (8.136 µmol FeSO47H2O/g sample) was seen at Valencia market type, the highest (14.004 µmol FeSO47H2O/g sample) was seen at Virginia market type. It was determined that the total flavonoid, total phenolic content, and antioxidant activities of the Virginia, Valencia, Spanish, and Runner market types included in the study were different from each other, and the Virginia market type showed superior characteristics compared to the others. The results obtained suggest that Virginia market type may be preferred more especially in peanut cultivation for food uses. It is thought that this study can be a source for future studies by eliminating a deficiency in the literature.

[Quality evaluation of salt-fired Eucommiae Cortex based on HPLC fingerprint, multi-component content determination, and chemometrics].

This study established an HPLC fingerprint and multi-component content determination method for salt-fired Eucommiae Cortex, and evaluated the quality of salt-fired Eucommiae Cortex from different sources using fingerprint similarity evaluation, cluster analysis(CA), principal component analysis(PCA), and orthogonal partial least square discriminate analysis(OPLS-DA). HPLC was launched on a Cosmosil 5C_(18)-MS-Ⅱ column(4.6 mm×250 mm, 5 µm) by gradient elution with a mobile phase of methanol-0.2% phosphoric acid aqueous solution at a flow rate of 1.0 mL·min~(-1), detection wavelength of 238 nm, column temperature of 30 ℃, and an injection volume of 10 µL. The results of fingerprint similarity evaluation for 20 batches of salt-fired Eucommiae Cortex indicated that, except for batch S3 with a similarity of 0.893, the similarity of the other 19 batches was of ≥ 0.919, suggesting good similarity. Fourteen common peaks were calibrated and seven common peaks were identified including geniposidic acid. The mass fractions of geniposidic acid, chlorogenic acid, geniposide, genipin, pinoresinol diglucoside, liriodendrin, and pinoresinol-4-O-ß-D-glucopyranoside were 0.062 0%-0.426 9%, 0.024 9%-0.116 5%, 0.009 5%-0.052 9%, 0.005 5%-0.034 8%, 0.115 9%-0.317 8%, 0.016 4%-0.108 8%, and 0.026 4%-0.039 8%, respectively. Using CA, PCA, and OPLS-DA, the 20 batches of salt-fired Eucommiae Cortex were classified into three categories. Additionally, through the analysis of variable importance in projection(VIP) under OPLS-DA, two differential quality markers, geniposidic acid and chlorogenic acid, were identified. The established HPLC fingerprint and multi-component content determination method is stable and reliable, providing a reference for quality control of salt-fired Eucommiae Cortex.

Distinguishment of different varieties of rhubarb based on UPLC fingerprints and chemometrics.

Rhubarb, a widely used traditional Chinese medicine (TCM), is primarily used for purging in practice. It is derived from the dried roots and rhizomes of R. tanguticum Maxim. ex Balf. (RT), Rheum officinale Baill. (RO) and R. palmatum L. (RP). To date, although the three varieties of rhubarb have been used as the same medicine in clinical, studies have found that they have different chemical compositions and pharmacological effects. To ensure the stability of rhubarb for clinical use, a simple and effective method should be built to compare and discriminate three varieties of rhubarb. Here, ultra-performance liquid chromatography-diode array detection (UPLC-DAD) fingerprints combined with chemometric methods were developed to evaluate and discriminate 29 batches of rhubarb. Similarity evaluation, hierarchical cluster analysis (HCA) and principal component analysis (PCA) showed that the chemical constituents of the three varieties of rhubarb were significantly different, and the three varieties could be effectively distinguished. Finally, all the 14 common peaks were identified by ultra-performance liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry (UPLC-Q-TOF-MS). In this research, the developed UPLC fingerprints offer a simple, reliable and specific approach for distinguishing different varieties of rhubarb. This research aims to promote the scientific and appropriate clinical application of rhubarb from three varieties.

Integration of untargeted and pseudotargeted metabolomics reveals specific markers for authentication and adulteration detection of Fritillariae Bulbus using tandem mass spectrometry and chemometrics.

Authentication and adulteration detection of closely related herbal medicines is a thorny issue in the quality control and market standardization of traditional Chinese medicine. Taking Fritillariae Bulbus (FB) as a case study, we herein proposed a three-step strategy that integrates mass spectrometry-based metabolomics and multivariate statistical analysis to identify specific markers, thereby accurately identifying FBs and determining the adulteration level. First, an ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry-based untargeted metabolomics method was employed to profile steroid alkaloids in five sorts of FB and screen potential differential markers. Then, the reliability of the screened markers was further verified by the distribution in different FB groups acquired from ultra-high performance liquid chromatography triple quadrupole mass spectrometry-based pseudotargeted metabolomics analysis. As a result, a total of 16 compounds were screened out to be the specific markers, which were successfully applied to distinguish five FBs by using discriminant analysis model. Besides, partial least squares regression models based on specific markers allowed accurate prediction of three sets of adulterated FBs. All the models afforded good linearity and good predictive ability with regression coefficient of prediction (R2p) > 0.9 and root mean square error of prediction (RMSEP) < 0.1. The reliable results of discriminant and quantitative analysis revealed that this proposed strategy could be potentially used to identify specific markers, which contributes to rapid chemical discrimination and adulteration detection of herbal medicines with close genetic relationship.

Rapid identification of Radix Astragali origin by using fluorescence probe combined with chemometrics.

Fluorescent probes for metal ion recognition can be divided into selective probes, weakly selective probes, and non-selective probes roughly. Weakly selective probes are not often used for quantitative analysis of metal ions due to their overlapping spectra resulting from simultaneous interactions with multiple metal ions. Conversely, the different metal ions contained in herbal medicine extracts from different geographical origins will produce corresponding fluorescence fingerprint profiles after interaction with weakly selective fluorescence probes. The performance can be used in the study of origin tracing of food or Chinese herbal medicine. Weakly selective fluorescent probes of benzimidazole derivatives have been synthesized and attempted to be used in the origin tracing of Radix Astragali in this work. Radix Astragali from different origins will produce different fluorescence fingerprint spectra due to the difference of metal ions and content in combination with the probe. Excitation-emission matrix (EEM) fluorescence spectroscopy in conjunction with N-way partial least squares discriminant analysis (N-PLS-DA), and unfolded partial least squares discriminant analysis (U-PLS-DA) were used to identify the origin of 150 Radix Astragali samples from five geographical origins. The prediction results showed that the correct recognition rates of the U-PLS-DA model and N-PLS-DA model are 95.92% and 93.88%, respectively. In comparison, the results of U-PLS-DA are slightly better than those of N-PLS-DA. These findings indicate that EEM fluorescence spectroscopy based on weakly selective fluorescent probes combined with multi-way chemometrics provides a good idea for the origin tracing of traditional Chinese medicine.

Polar licit and illicit ingredients in dietary supplements: chemometric optimization of extraction and HILIC-MS/MS analysis.

Many dietary supplements claim the ability to enhance sports performance and to improve the fitness of the consumers. Occasionally, along with legal ingredients, illicit compounds may be added without being labelled, leading to unintended doping. Hence, the aim of this study was to develop an analytical method to determine a set of 12 polar (logDpH=7 from -2.0 to +0.3) compounds including diuretics, stimulants, ß2-agonists, methylxanthines, and sweeteners. Hydrophilic interaction liquid chromatography was chosen as separation strategy, coupled with tandem mass spectrometry. The instrumental method was optimized using a two-step design of experiments (DoE). Firstly, a Plackett-Burman (PB) DoE was performed to identify the more influencing variables affecting peak areas and chromatographic resolution among temperature, water percentage in the mobile phase, and flow rate, as well as type and concentration of buffers. Secondly, a D-optimal DoE was set, considering only the most significant variables from the PB-DoE results, achieving a deeper understanding of the retention mechanism. Sample processing by salt-assisted liquid-liquid extraction was studied through DoE as well, and the whole method showed recoveries in the range 40-107% and procedural precision ≤11% for all analytes. Finally, it was applied to real samples, in which the four methylxanthines and two artificial sweeteners were detected and quantified in the range of 0.02-192 mg g-1. These values were compared to the quantities declared on the DS labels, when possible. Furthermore, a sequence of MS/MS scans allowed detection of a signal in one of the samples, structurally similar to the ß2-agonist clenbuterol.

A multiplex metabolomic approach for quality control of Spirulina supplement and its allied microalgae (Amphora & Chlorella) assisted by chemometrics and molecular networking.

Microalgae species are of economic importance regarded as "green gold" being rich in bioactive compounds. Spirulina and Chlorella are the most popular microalgal species and are marketed as healthy food supplements. At the same time, Amphora holds potential as a source of healthy lipids and essential fatty acids. Yet, there are considerable variations in their reported chemical composition, and less is known about their compositional differences. A multiplexed metabolomic approach was adopted for the quality control (QC) of Spirulina supplements and to compare its constitutive metabolome to Chlorella and Amphora. The adopted protocol comprised gas chromatography-mass spectrometry (GC-MS), ultra-high performance liquid chromatography coupled with high-resolution tandem mass spectrometry (UPLC-HRMS/MS), and ultraviolet-visible spectrophotometry (UV/Vis) for mapping their primary and secondary metabolome. Interestingly, UPLC-HRMS/MS analysis delineated the abundance of fatty acids in Amphora versus glycolipids enrichment in Spirulina, and porphyrins were the main pigments identified in Spirulina, with scarce occurrence in Chlorella. Orthogonal projections to latent structures discriminant analysis (OPLS-DA) analysis of GC-MS data set revealed palmitic acid, 3-mannobiose, and glyceryl-glycoside as being most enriched in Spirulina, versus sucrose and leucine in Chlorella and Amphora, respectively. Despite being of low discriminatory potential, UV/Vis OPLS-DA modeling showed that Spirulina was distinguished with the UV absorbances of carotenoids and chlorophyll pigments, as indicated by its OPLS-DA derived S-plot. Our study provides a QC approach for the analysis of the microalgal species and poses alternative spectral and compositional markers for their discrimination.

Chemical and biological characterization of Melaleuca subulata (Cheel) Craven leaves' volatile constituents supported by chemometric analysis and molecular docking.

BACKGROUND: The genus Melaleuca (Myrtaceae) comprises dozens of essential oil (EO)-rich species that are appreciated worldwide for their various medicinal values. Additionally, they are renowned in traditional medicine for their antimicrobial, antifungal, and other skin-related activities. The current study investigated the chemical profile and skin-related activities of volatile constituents derived from M. subulata (Cheel) Craven (Synonym Callistemon subulatus) leaves cultivated in Egypt for the first time. METHODS: The volatile components were extracted using hydrodistillation (HD), headspace (HS), and supercritical fluid (SF). GC/MS and Kovat's retention indices were implemented to identify the volatile compounds, while the variations among the components were assessed using Principal Component Analysis and Hierarchical Cluster Analysis. The radical scavenging activity was assessed using 2,2-diphenyl-1-picrylhydrazyl (DPPH), oxygen radical absorbance capacity (ORAC) and ß-carotene assays. Moreover, the anti-aging effect was evaluated using anti-elastase, and anti-collagenase, while the antimicrobial potential was deduced from the agar diffusion and broth microdilution assays. Lastly, the molecular docking study was executed using C-docker protocol in Discovery Studio 4.5 to rationalize the binding affinity with targeted enzymes. RESULTS: The SF extraction approach offered the highest EO yield, being 0.75%. According to the GC/MS analysis, monoterpene hydrocarbons were the most abundant volatile class in the HD oil sample (54.95%), with α-pinene being the most copious component (35.17%). On the contrary, the HS and SF volatile constituents were pioneered with oxygenated monoterpenes (72.01 and 36.41%) with eucalyptol and isopulegone being the most recognized components, representing 67.75 and 23.46%, respectively. The chemometric analysis showed segregate clustering of the three extraction methods with α-pinene, eucalyptol, and isopulegone serving as the main discriminating phytomarkers. Concerning the bioactivity context, both SF and HD-EOs exhibited antioxidant effects in terms of ORAC and ß-carotene bleaching. The HD-EO displayed potent anti-tyrosinase activity, whereas the SF-EO exhibited significant anti-elastase properties. Moreover, SF-EO shows selective activity against gram-positive skin pathogens, especially S. aureus. Ultimately, molecular docking revealed binding scores for the volatile constituents; analogous to those of the docked reference drugs. CONCLUSIONS: M. subulata leaves constitute bioactive volatile components that may be indorsed as bioactive hits for managing skin aging and infection, though further in vivo studies are recommended.

Exploring the dynamics of bioactive metabolites changes in barley grains (Hordeum vulgare L.) during roasting: Insights from UPLC-QqQ-MS/MS analysis combined to chemometrics.

This investigation delves into the dynamic metabolic shifts within barley grains during the roasting process, employing UPLC-QqQ-MS/MS analysis. The complex spectrum of metabolites before and after roasting is revealed. The resulting data, unveils substantial transformations in chemical composition during roasting. A total of 62 chromatographic peaks spanning phenolic compounds, flavones, Millard Reaction Products, amino acids, lignans, vitamins, folates, and anthocyanins were annotated. Leveraging UPLC-QqQ-MS/MS analysis, we scrutinized the intricate metabolite profile before and after roasting where the roasting process was found to trigger dynamic changes across diverse metabolite classes particularly Millard Reaction Products, produced through the Maillard reaction, with dihydro-5-methyl-5H-cyclopentapyrazine, maltol and hydroxy maltol emerging as discerning markers of roasting progression. Amino acids and sugars showed degradation, while beta-glucan, a signature barley sugar, experienced notable decline. Folate derivatives witnessed pronounced reduction, aligning with the heat sensitivity of folates. Harnessing the power of multivariate data analysis, the consequences of roasting materialize through distinct clusters in PCA and OPLS-DA plots. Noteworthy, roasting duration governs the trajectory of metabolic divergence, culminating in the identification of roasting-specific markers. Epigallocatechin, procyanidin B, 10-HCO-H4 folate, and hordatine A emerge as pivotal discriminators. Orthogonal Projection to Latent Structure (OPLS) analysis linked anti-inflammatory activity with 30-min, 1-hour, and 1.5-hour roasted samples, with hordatine B in addition to some Millard Reaction Products being correlated with pro-inflammatory marker downregulation.. This study encapsulates the intricate metabolic metamorphosis ignited by roasting in barley grains, offering a holistic comprehension of their potential health-enhancing attributes. Key metabolites act as poignant indicators of these transformations, substantiating the complex interplay between roasting and the barley grain metabolome.

Influence of maturation and roasting on the quality and chemical composition of new conilon coffee cultivar by chemometrics.

Coffee is one of the most consumed beverages worldwide. Espírito Santo is the largest Brazilian producer of conilon coffee, and invested in the creation of new cultivars, such as "Conquista ES8152", launched in 2019. Therefore, the present study aimed to evaluate the effects of maturation and roasting on the chemical and sensorial composition of the new conilon coffee cultivar "Conquista ES8152". The coffee was harvested containing 3 different percentages of ripe fruits: 60%, 80%, and 100%, and roasted at 3 different degrees of roasting: light, medium, and dark, to evaluate the moisture and ash content, yield of soluble extract, volatile compound profile, chlorogenic acid and caffeine content, and sensory profile. "Conquista ES8152" coffee has a moisture content between 1.38 and 2.62%; ash between 4.34 and 4.72%; and yield between 30.7 and 35.8%. Sensory scores ranged between 75 and 80 and the majority of volatile compounds belong to the pyrazine, phenol, furan, and pyrrole groups. The content of total chlorogenic acids was drastically reduced by roasting, with values between 2.40 and 9.33%, with 3-caffeoylquinic acid being the majority. Caffeine was not influenced by either maturation or roasting, with values between 2.16 and 2.41%. The volatile compounds furfural, 5-methylfurfural, and 2-ethyl-5-methylpyrazine were positively correlated with the evaluated sensory attributes and 5-methylfurfural was the only one significantly correlated with all attributes. Ethylpyrazine, furfuryl acetate, 1-furfurylpyrrole, 4-ethyl-2-methoxyphenol, and difurfuryl ether were negatively correlated. The stripping did not affect the quality and composition of this new cultivar, however, the roasting caused changes in both the chemical and sensorial profiles, appropriately indicated by the principal component analysis.

Densitometric method for assessment of six specialized metabolites in four Sida sp. and its congener Abutilon indicum: Targeted metabolomics, greenness assessment, and chemometrics analysis.

Sida is one of the most diverse genera, with about 200 species distributed in tropical and subtropical regions of the world. Among 18 species distributed in India, Sida acuta, Sida cordifolia, Sida rhombifolia, and Sida cordata are used in traditional medicines along with its possible adulterant Abutilon indicum for several therapeutic uses. The non-availability of marker-based validated methods for the identification and classification of these species leads to adulteration. Indoloquinoline and quinazoline are the major bioactive alkaloids distributed in Sida spp. First time, a simple, economical and high throughput method was developed and validated for the simultaneous determination of 20-hydroxyecdysone (1), vasicine (2), vasicinone (3), cryptolepine (4), quindolinone (5), and cryptolepinone (6) using HPTLC-UV densitometry. The method was validated to meet globally accepted ICH guidelines. The method was sensitive with LOD and LOQ ranging from 0.38-0.63 and 1.57-2.12 µg/band. The samples were spiked at 3 different concentrations, the recovery values were 93.49-98.88%. In addition, the greenness index of the HPTLC method was estimated using four different greenness assessment techniques. Targeted HPTLC analysis indicated the distribution of specialized metabolites in Sida spp. and A. indicum. However, the occurrence of cryptolepine in A. indicum was not reported in the literature, so this was further confirmed by liquid chromatographic studies of the samples from different locations. The chromatographic data was statistically evaluated by principal component analysis (PCA) and hierarchical clustering (HCA). HPTLC-based targeted metabolite quantitation explains the adulteration/substitution in Sida raw material and derived herbal preparations.

Identification of Adulteration of Flaxseed Oil From QINGHAI Area Using GC-MS Profiling of Phytosterol Composition and Chemometrics.

Flaxseed oil is an important source of vegetable oil with a polyunsaturated fatty acid. It is significant to establish a method to quickly identify adulterated flaxseed oil. In the present study, the qualitative and quantitative analysis of phytosterol of flaxseed oil from different varieties and different production areas in the Qinghai area was first performed by gas chromatography-mass spectrometry (GC-MS) and the phytosterol standard profile of flaxseed oil was established. Then, a combination of similarity evaluation and cluster analysis was used to distinguish pure flaxseed oil from flaxseed oil adulterated with concentrations of 10-50% rapeseed oil, peanut oil, sunflower oil, and sesame oil, and discriminant analysis was used to identify the types of adulterated flaxseed oil. The results showed that similarity evaluation combined with cluster analysis can distinguish pure and adulterated flaxseed oil when the concentration of the adulterant was greater than 10%. Discriminant analysis models accurately identified the types of adulterating oil in flaxseed oil when the concentration of rapeseed, peanut, or sunflower oil was greater than 20%, and that of sesame oil was greater than 30%. This study shows that the determination of the phytosterol composition and chemometrics is a valuable tool to evaluate the purity of flaxseed oil.

Investigation of Phytochemical, Antioxidant and Antidiabetic Potentials of Scabiosa L. (Caprifoliaceae) Species with Chemometric Methods.

In this research, the total phenolic and flavonoid amounts, phenolic compositions, in vitro antioxidant, antibacterial and antidiabetic properties of the methanol extracts obtained from Scabiosa L. (Caprifoliaceae) species distributed in the flora of Türkiye were investigated using chemometric methods. For this purpose, principal component (PCA) and agglomerative hierarchical clustering analysis were performed as chemometric methods. Chlorogenic acid, quinic acid and cyranoside were determined in the extracts. According to chemometric analysis, S. columbaria subsp. ochroleuca var. ochroleuca and S. triniifolia species were found to be valuable in terms of methanol extract yields, total phenolic and flavonoid contents, antioxidant and antidiabetic activities while S. columbaria subsp. ochroleuca var. webbiana species were found to be valuable in terms of phenolic composition. The methanol extracts of Scabiosa species showed high antioxidant activity, with high phenolic and flavonoid contents. Among the tested 13 bacteria, Scabiosa extracts showed only low activity against Klebsiella pneumoniae, Streptococcus pneumoniae and Pseudomonas aeruginosa. The extracts showed high α-amylase and α-glucosidase inhibitory activity. The results show that Scabiosa methanol extracts may be a source of alternative antioxidants that may be beneficial in slowing or preventing the progression of various oxidative stress-related diseases.

Chemometrics-based Chemical Analysis of Myrrh and Its Vinegar-processed Products by UPLC-MS/MS.

Myrrh is widely used in clinical practice but accompanied by obvious toxicity. According to traditional Chinese medicines theory, processing with vinegar can effectively reduce its toxicity. However, the detoxification processing technology of Myrrh and the corresponding mechanism have been unclear. The objective of this study is to systematically analyze the variation in chemical composition of raw Myrrh and its processed products using UPLC-Q-TOF-MS/MS coupled with chemometrics. A total of 75 compounds including 56 sesquiterpenoids, 2 diterpenoids, 15 triterpenoids and 2 other types were identified. Raw Myrrh and its processed products were divided into two major groups, and 14 chemical markers were selected out by principal component analysis and partial least square discriminant analysis. Additionally, the exact content of 5 representative chemical markers was determined to be significantly reduced after vinegar-processing by UPLC-QQQ-MS/MS. Moreover, multivariate statistical analysis and the quantitative results comprehensively indicated that the optimized processing method was processing at a ratio of 200 : 5 (Myrrh:vinegar). This research provides not only a reliable foundation for the study of Myrrh, but also a scientific reference for clinical use of this herb.

Comprehensive metabolome characterization and comparison between two sources of Dragon's blood by integrating liquid chromatography/mass spectrometry and chemometrics.

Dragon's Blood (DB) serves as a precious Chinese medicine facilitating blood circulation and stasis dispersion. Daemonorops draco (D. draco; Qi-Lin-Jie) and Dracaena cochinchinensis (D. cochinchinenesis; Long-Xue-Jie) are two reputable plant sources for preparing DB. This work was designed to comprehensively characterize and compare the metabolome differences between D. draco and D. cochinchinenesis, by integrating liquid chromatography/mass spectrometry and untargeted metabolomics analysis. Offline two-dimensional liquid chromatography/ion mobility-quadrupole time-of-flight mass spectrometry (2D-LC/IM-QTOF-MS), by utilizing a powerful hybrid scan approach, was elaborated for multicomponent characterization. Configuration of an XBridge Amide column and an HSS T3 column in offline mode exhibited high orthogonality (A0 0.80) in separating the complex components in DB. Particularly, the hybrid high-definition MSE-high definition data-dependent acquisition (HDMSE-HDDDA) in both positive and negative ion modes was applied for data acquisition. Streamlined intelligent data processing facilitated by the UNIFI™ (Waters) bioinformatics platform and searching against an in-house chemical library (recording 223 known compounds) enabled efficient structural elucidation. We could characterize 285 components, including 143 from D. draco and 174 from D. cochinchinensis. Holistic comparison of the metabolomes among 21 batches of DB samples by the untargeted metabolomics workflows unveiled 43 significantly differential components. Separately, four and three components were considered as the marker compounds for identifying D. draco and D. cochinchinenesis, respectively. Conclusively, the chemical composition and metabolomic differences of two DB resources were investigated by a dimension-enhanced analytical approach, with the results being beneficial to quality control and the differentiated clinical application of DB.

Development of chemometric-assisted supercritical fluid extraction of effective and natural tyrosinase inhibitor from Syzygium aqueum leaves.

Tyrosinase is a key enzyme in enzymatic browning, causing quality losses in food through the oxidation process. Thus, the discovery of an effective and natural tyrosinase inhibitor via green technology is of great interest to the global food market due to food security and climate change issues. In this study, Syzygium aqueum (S. aqueum) leaves, which are known to be rich in phenolic compounds (PC), were chosen as a natural source of tyrosinase inhibitor, and the effect of the sustainable, supercritical fluid extraction (SFE) process was evaluated. Response surface methodology-assisted supercritical fluid extraction (RSM-assisted SFE) was utilized to optimize the PCs extracted from S. aqueum. The highest amount of PC was obtained at the optimum conditions (55 °C, 3350 psi, and 70 min). The IC50 (661.815 µg/mL) of the optimized extract was evaluated, and its antioxidant activity (96.8 %) was determined. Gas chromatography-mass spectrometry (GC-MS) results reveal that 2',6'-dihydroxy-4'-methoxychalcone (2,6-D4MC) (82.65 %) was the major PC in S. aqueum. Chemometric analysis indicated that 2,6-D4MC has similar chemical properties to the tyrosinase inhibitor control (kaempferol). The toxicity and physiochemical properties of the novel 2,6-D4MC from S. aqueum revealed that the 2,6-D4MC is safer than kaempferol as predicted via absorption, distribution, metabolism, and excretion (ADME) evaluation. Enzyme kinetic analysis shows that the type of inhibition of the optimized extract is non-competitive inhibition with Km = 1.55 mM and Vmax = 0.017 µM/s. High-performance liquid chromatography (HPLC) analysis shows the effectiveness of S. aqueum as a tyrosinase inhibitor. The mechanistic insight of the tyrosinase inhibition using 2,6-D4MC was successfully calculated using density functional theory (DFT) and molecular docking approaches. The findings could have a significant impact on food security development by devising a sustainable and effective tyrosinase inhibitor from waste by-products that is aligned with the United Nation's SDG 2, zero hunger.