Abstract: INTRODUCTION Phytochemical analysis of phenolic acids and flavonols poses a challenge, necessitating the development of an efficient separation method. This facilitates the quantification of these compounds, yielding valuable insights into their benefits. OBJECTIVE To develop a highly effective separation of phenolic acids and flavonols by capillary electrophoresis and ultraviolet (UV) detection through the modification of the capillary surface using 3-aminopropyltriethoxysilane (APTES) at millimolar concentrations. METHODS The capillary surface is modified with 0.36 mM-APTES solution. The electrolyte is 20.0 mM borate buffer (pH 9.0). Separation performance (plate number N, resolution Rs ), stability, and reproducibility of the coating procedure are evaluated using the analysis of phenolic acids, rutin and quercetin. RESULTS The modified capillary provided efficient separation with plate numbers N ≥ 1.0 × 104 m-1 and resolution Rs ≥ 0.8 for all pairs of adjacent peaks of the separation of five selected phenolic acids, rutin, quercetin, caffeine and methylparaben (as internal standard). The precisions of the relative migration times for 17 consecutive analyses of samples over 3 h were 1% relative standard deviation (RSD) for rutin and 7% RSD for quercetin. The analysis of rutin and quercetin in 12 dietary supplement product samples only required a simple dilution step for sample preparation. CONCLUSION A straightforward modification technique utilising millimolar concentrations of APTES resulted in highly efficient separation of phenolic acids, rutin and quercetin, accompanied by high precision and surface stability. The modified capillary proved successful in analysing rutin and quercetin content in dietary supplements.

Abstract: INTRODUCTION In recent years, industrial production of Cannabis sativa has increased due to increased demand of medicinal products based on the plant. In these medicinal products, it is mainly the contents of cannabinoids like THCA and CBDA which are of interest, but also the flavonoids of C. sativa have pharmaceutical interest. OBJECTIVES The primary aim is to study the distribution of the different cannabinoids in leaves of C. sativa and specifically to which extent they are located on the trichomes found on the surface of C. sativa leaves. Desorption electrospray ionization (DESI) and matrix assisted laser desorption ionization (MALDI) mass spectrometry imaging (MSI) provide non-targeted imaging of numerous compounds in the same experiment. Therefore, the distribution of flavonoids is also mapped in the same experiments. MATERIAL AND METHODS Fan leaves from C. sativa were imaged in the lateral dimension using direct DESI-MSI as well as indirect DESI-MSI via a porous PTFE surface using pixel sizes of 150-200 μm. For cross sections of sugar leaves, MALDI-MSI was performed at 20 μm pixel size. RESULTS From indirect DESI-MSI experiments, a connection was made between the cannabinoid CBGA and capitate-stalked trichomes. Other cannabinoids like THCA/CBDA (isomers, which are not resolved in an MSI experiment) were also detected in the capitate-stalked trichomes, but in addition to this also in the small glandular trichomes. MALDI-MSI experiments on cross sections of sugar leaves confirmed that the cannabinoids were not an integral part of the leaf tissue itself, but originated from the trichomes on the surface of the leaf. CONCLUSION The study provides visual evidence that the cannabinoids are produced and accumulated in the trichomes of C. sativa leaves.

Abstract: INTRODUCTION Atractylodes chinensis is a Chinese herb that is used in traditional medicine; it contains volatile components that have enormous potential for pharmaceutical, food, and cosmetic applications. The destruction of wild resources demands significant improvement in the quality of artificial cultivation of Atractylodes chinensis. However, little is known about the compositional differences in the volatile substances derived from the wild and cultivated varieties of Atractylodes chinensis. OBJECTIVES We aimed to evaluate the specific components of Atractylodes chinensis and analyse the similarities and differences between the volatile components and metabolic pathways in the wild and cultivated varieties. MATERIAL AND METHODS Metabolomic analysis using gas chromatography-mass spectrometry (GC-MS) was employed following the extraction of volatile components from Atractylodes chinensis using headspace solid-phase microextraction (HS-SPME). RESULTS A total of 167 volatile metabolites were extracted, and 137 substances were matched with NIST and Wiley databases. Among them, 76 compounds exhibited significant differences between the two sources; these mainly included terpenes, aromatics, and esters. KEGG enrichment analysis indicated that the differential metabolites were primarily involved in the biosynthesis of secondary metabolites, terpene biosynthesis, and limonene and pinene degradation; all these pathways have geranyl diphosphate (GDP) as the common link. CONCLUSION The total content of volatile substances extracted from wild Atractylodes chinensis was 2.5 times higher than that from the cultured variety; however, each source had different dominant metabolites. This study underscores the necessity for protecting wild Atractylodes chinensis resources, while enhancing the quality of cultivated Atractylodes chinensis.

Abstract: Abstract Introduction Natural products, the small organic molecules produced by plants, microbes and invertebrates, often present in the form of a mixture, this leads to the structural characterisation of natural extracts often requiring time‐consuming multistep purification procedures. Nuclear magnetic resonance (NMR) technology is traditionally utilised as a tool for the structural elucidation of pure compounds. Contemporarily, an up‐to‐date trend in the application of NMR in natural product research is shifting to the direct NMR analysis of crude mixtures, to obtain molecular structure and biological activity information without performing cumbersome separation. Objective To review works of literature on the evolution, principle and progress of NMR technologies for analysing mixtures, we focus on the successful application of NMR technologies in direct analysis of natural product extracts. Methodology Based on our research experience, academic tracking and extensive literature search, which involved, but not limited to, the use of various databases, like Web of Knowledge and PubMed. The keywords used, in various combinations, to retrieve recent literature on the successful application of NMR technologies to sheer natural product extracts, and excluded artificially natural product mixture and biofluid. Results NMR technologies for direct natural extracts analysis, including two‐dimensional J‐resolved spectroscopy (2D‐JRES), pure shift NMR, diffusion‐ordered NMR spectroscopy (DOSY), statistical correlation spectroscopy (STOCSY), concentration‐ordered NMR spectroscopy (CORDY), saturation transfer difference (STD) and water‐ligand observed via gradient spectroscopy (WaterLOGSY) were illustrated. Conclusions By these methods, molecular structure and biological activity information will be directly obtained from NMR analysis of natural products extract, aiming to save experimental time and expenses.

Abstract: INTRODUCTION Natural products and metabolomics are intrinsically linked through efforts to analyze complex mixtures for compound annotation. Although most studies that aim for compound identification in mixtures use MS as the main analysis technique, NMR has complementary advances that are worth exploring for enhanced structural confidence. OBJECTIVE This review aimed to showcase a portfolio of the main tools available for compound identification using NMR. MATERIALS AND METHODS COLMAR, SMART-NMR, MADByTE, and NMRfilter are presented using examples collected from real samples from the perspective of a natural product chemist. Data are also made available through Zenodo so that readers can test each case presented here. CONCLUSION The acquisition of 1 H NMR, HSQC, TOCSY, HSQC-TOCSY, and HMBC data for all samples and fractions from a natural products study is strongly suggested. The same is valid for MS analysis to create a bridged analysis between both techniques in a complementary manner. The use of NOAH supersequences has also been suggested and demonstrated to save NMR time.

Abstract: OBJECTIVES Oxidative stress is one of the carcinogenic mechanisms underlying the development of glioblastoma multiforme (GBM), a highly aggressive brain tumor type associated with poor prognosis. Curcumin is known to be an efficient antioxidant, anti-inflammatory, and anticancer compound. However, its poor solubility in water, inappropriate pharmacokinetics, and low bioavailability limit its use as an antitumor drug. We prepared PLGA-based curcumin nanoparticles changed with folic acid and chitosan (curcumin-PLGA-CS-FA) and evaluated its effects on GBM tumor cells' redox status. METHODS The nanoprecipitation method was used to synthesize CU nanoparticles (CU-NPs). The size, morphology, and stability were characterized by DLS, SEM, and zeta potential analysis, respectively. The CU-NPs' toxic properties were studied by MTT assay and measuring the intracellular reactive oxygen species (ROS) and malondialdehyde (MDA) concentrations. The study was completed by measuring the gene expression levels and activity of superoxide dismutase, catalase, glutaredoxin, and thioredoxin antioxidant enzymes. RESULTS The size, polydispersity index, and zeta potential of CU-NPs were 77.27 nm, 0.29, and -22.45 mV, respectively. The encapsulation efficiency was approximately 98%. Intracellular ROS and MDA levels decreased after CU-NP treatment. Meanwhile, the CU-NPs increased gene expression and activity of superoxide dismutase, catalase, glutaredoxin, and thioredoxin antioxidant enzymes. CONCLUSION CU-NPs might be effective in the prevention and treatment of glioblastoma cancer by modulating the antioxidant-oxidant balance.

Abstract: INTRODUCTION Chromatography and spectroscopy are nowadays well-validated techniques allowing to isolate and purify different class of natural products from the genus Codonopsis. Several categories of phytochemicals with drug like properties have been selectively extracted, isolated, characterised by this methodology. OBJECTIVES The present review aims to provide up-to-date and comprehensive information on the chromatography, phytochemistry and pharmacology of natural products of Codonopsis with an emphasis on the search for natural products having various biological activities and the semi-synthetic derivatives of bioactive ones and to highlight current gaps in knowledge. MATERIALS AND METHODS A literature search was performed in the SciFinder Scholar, PubMed, Medline, and Scopus databases. RESULTS During the period covered in this review, several classes of compounds have been reported from genus Codonopsis. Codonopsis pilosula and Codonopsis lanceolata are the most popular in the genus especially as per phytochemical and bioactive studies. Phytochemical investigation demonstrates that Codonopsis species contain mainly xanthones, flavonoids, alkaloids, polyacetylenes, phenylpropanoids, triterpenoids and polysaccharides, which contribute to numerous bioactivities. The major bioactive compounds isolated were used for semi-synthetic modification to increase the chance to discover lead compound. CONCLUSIONS It can be concluded that genus Codonopsis has been used as traditional medicines and food materials around the world over years due to chemical constituents with diverse structural types, exhibiting extensive pharmacological activities in immune system, blood system, cardiovascular system, central nervous system, digestive system, and so forth, with almost no obvious toxicity and side effect. Therefore, Codonopsis can be used as a promising ethnopharmacological plant source.

Abstract: INTRODUCTION Radix Angelicae sinensis (Danggui, DG) is known as one of the typical traditional Chinese medicines. DG material consists of a variety of volatile substances, polysaccharides, organic acids, ceramides, amino acids, vitamins, microelements, among others, and thus has been used for medicinal and edible purposes in a long history. The fragrance is of importance to assessing the DG material quality. OBJECTIVES This study was to determine volatile flavour compositions of DG materials and to reveal the odour complexity. MATERIAL AND METHODS Electronic nose (E-nose), integrated gas chromatography-mass spectrometry/olfactometry (GC-MS/O) and comprehensive two-dimensional gas chromatography-time-of-flight-mass spectrometry (GC × GC-TOF-MS), combined with solid-phase micro-extraction (SPME), were mainly used to address the flavour complexity of DG materials. RESULTS Using the E-nose sensor responses, a total of 105 batches of DG samples cultivated in six provinces of China were categorised according to their odour differentiations, and a principal component analysis (PCA) model was established for evaluating the sample quality through a combination of Hotelling's T2 and Q-residual values in a statistical quantitative sense. By the GC-MS/O and GC × GC-TOF-MS analyses, 196 volatile flavour compounds were identified, 51 odour-active areas discerned and 39 odourants determined. It was terpenes and aromatics of the flavour compounds that mainly contributed to the odour attributes of DG herb. CONCLUSION The SPME-GC × GC-TOF-MS method was the first time employed to analyse the volatile flavours of DG materials, and thus made a breakthrough in determining 196 flavour compounds, much more than those in any previous report. The work also made a significant step forward to link the flavour compositions and odour complexity of radix Angelicae sinensis by E-nose and GC-MS/O techniques. It not only provided a statistical PCA model that did not depend on any predetermined compositions or sensory properties for, but also a comprehensive insight into the quality evaluation of DG materials.

Abstract: INTRODUCTION Phytochemicals are used in many products, including foods, beverages, pharmaceuticals, and cosmetics. The extraction of phytochemicals is considered one of the best solutions to valorize these underestimated materials. Many methods have been developed to efficiently extract phytochemicals at high quality, high purity, and low costs without harming the environment. Recently, molecular hydrogen (H2 ) has shown its ability to improve the extraction of phytochemicals from plant materials. Due to its unique physicochemical and biological properties, H2 showed an efficient ability to extract phenolics and antioxidants at high yields with cost-effective potential. Without sophisticated equipment and high energy and solvent consumption, the hydrogen extraction method is a green and applicable alternative for the extraction of phytochemicals. OBJECTIVES This review aims to provide the latest knowledge and results concerning the studies on using hydrogen-rich solvents to extract phytochemicals from different agri-food wastes, by-products, and other plant materials. MATERIALS AND METHODS Recent literature relating to extracting phytochemicals by the hydrogen-rich solvent method and its potential mechanisms is summarized to provide a basic understanding of how hydrogen can improve the extraction of phytochemicals. RESULTS This review describes, for the first time, the practical procedure of how researchers and laboratories can apply the hydrogen extraction method under safe conditions at a low-budget scale. The review provides some examples of the hydrogen extraction method and the mechanisms and rationale behind its effectiveness. CONCLUSIONS It can be concluded that the hydrogen-rich solvent method is a green and cost-effective method for extracting phytochemicals from different plant materials.

Abstract: INTRODUCTION Flavonoids are active substances in many herbal medicines, and Areca catechu fruit (AF), an important component in traditional Chinese medicine (TCM), is rich in flavonoids. Different parts of AF, Pericarpium Arecae (PA) and Semen Arecae (SA), have different medicinal effects in prescription of TCM. OBJECTIVE To understand flavonoid biosynthesis and regulation in AF. METHODOLOGY The metabolomic based on liquid chromatography-tandem mass spectrometry (LC-MS/MS) and the transcriptome based on high-throughput sequencing technology were combined to comprehensively analyse PA and SA. RESULTS From the metabolite dataset, we found that 148 flavonoids showed significant differences between PA and SA. From the transcriptomic dataset, we identified 30 genes related to the flavonoid biosynthesis pathway which were differentially expressed genes in PA and SA. The genes encoding the key enzymes in the flavonoid biosynthesis pathway, chalcone synthase and chalcone isomerase (AcCHS4/6/7 and AcCHI1/2/3), were significantly higher expressed in SA than in PA, reflecting the high flavonoid concentration in SA. CONCLUSIONS Taken together, our research acquired the key genes, including AcCHS4/6/7 and AcCHI1/2/3, which regulated the accumulation of flavonol in AF. This new evidence may reveal different medicinal effects of PA and SA. This study lays a foundation for investigating the biosynthesis and regulation of flavonoid biosynthesis in areca and provides the reference for the production and consumption of betel nut.

Abstract: INTRODUCTION Ginger constitutes the rhizome part of the plant Zingiber officinale from the Zingiberaceae family. A large number of ginger varieties with high sensorial and functional quality are found in Northeast India. Hence, phytopharmacological screening of different ginger varieties is essential that will serve as a guideline in applied research to develop high-end products and improve economical margins. OBJECTIVE To determine the variation in total phenolics content (TPC), total flavonoids content (TFC), and antioxidant activities and correlate that with 6-gingerol contents of different ginger varieties collected from Northeast India using Pearson's correlation analysis. MATERIALS AND METHODS The TPC and TFC values were determined using standard methods. Antioxidant activities were measured using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) and hydroxyl radical scavenging assays, while reversed-phase high-performance liquid chromatography (RP-HPLC) analysis was utilised for quantitative determination of 6-gingerol content. RESULTS The result revealed that ginger variety 6 (GV6) contains the highest 6-gingerol content and TPC value showing maximum antioxidant activity, followed by GV5, GV4, GV9, GV3, GV2, GV8, GV1, and GV7. The findings also suggested that the antioxidant activity has much better correlations with TPC as compared with TFC values. Pearson's correlation analysis showed a significant correlation between 6-gingerol contents and TPC values. CONCLUSION This work underlines the importance of ginger varieties from Northeast India as a source of natural antioxidants with health benefits.

Abstract: INTRODUCTION Alismatis rhizoma (AR), a distinguished diuretic traditional Chinese herbal medicine, is widely used for the treatment of diarrhea, edema, nephropathy, hyperlipidemia, and tumors in clinical settings. Most beneficial effects of AR are attributed to the major triterpenoids, whose contents are relatively high in AR. To date, only 25 triterpenoids in AR have been characterized by LC-MS because the low-mass diagnostic ions are hardly triggered in MS, impeding structural identification. Herein, we developed an advanced data post-processing method with abundant characteristic fragments (CFs) and neutral losses (NLs) for rapid identification and classification of the major triterpenoids in AR by UPLC-Q-TOF-MSE . OBJECTIVE We aimed to establish a systematic method for rapid identification and classification of the major triterpenoids of AR. METHODS UPLC-Q-TOF-MSE coupled with an advanced data post-processing method was established to characterize the major triterpenoids of AR. The abundant CFs and NLs of different types of triterpenoids were discovered and systematically summarized. The rapid identification and classification of the major triterpenoids of AR were realized by processing the data and comparing with information described in the literature. RESULTS In this study, a total of 44 triterpenoids were identified from AR, including three potentially new compounds and 41 known ones, which were classified into six types. CONCLUSION The newly established approach is suitable for the chemical profiling of the major triterpenoids in AR, which could provide useful information about chemical constituents and a basis for further exploration of its active ingredients in vivo.

Abstract: INTRODUCTION Hesperidin (hesperetin 7-rutinoside, HP), a flavonoid glycoside found in Citrus unshiu Marcowicz or Citrus reticulata Blanco (Rutaceae), has been reported to exert a variety of pharmacological effects. As the efficacies and qualities of their dried peel, Chinpi and its derived Kampo medicines can be evaluated by their HP contents, a method for HP detection must be developed. OBJECTIVES To produce a specific monoclonal antibody against HP (mAb 5D12) to detect the HP contents in Japanese traditional medicines via indirect competitive enzyme-linked immunosorbent assay (icELISA). METHOD BALB/c mice were immunised with many haptens of HP-bovine serum albumin (BSA) conjugates that were prepared using sodium periodate (NaIO4 ) to cause an immune response. In addition, conventional hybridoma techniques were utilised to generate mAb 5D12. RESULTS The detection range of HP by the mAb 5D12-based icELISA was 1.56-25.0 ng/mL, with a detection limit of 1.12 ng/mL. The maximum coefficient of variation, as evaluated from the intra- and inter-assays, was <10.0%, and the percentages of recovery, as determined by the spike-recovery tests, were 105%-115%. Moreover, the HP content, which was obtained from the developed icELISA, correlated well with that obtained via high-performance liquid chromatography-ultraviolet (HPLC-UV). CONCLUSION These validation analyses revealed that the established icELISA technique exhibited high precision and accuracy. Notably, this is the first report on the development of icELISA for the HP content-based quality control of Chinpi and its derived Kampo medicines.

Abstract: INTRODUCTION Origanum onites has antibacterial, antifungal, and antioxidant properties due to its essential oil. The fact that these oils are sensitive to environmental conditions causes the need for advanced processing technologies. OBJECTIVE This study was aimed to encapsulate essential oil of O. onites by spray drying to extend its shelf life. METHODS Spray drying was used to encapsulate the essential oil of O. onites, which was then mixed with maltodextrin, arabic gum, and β-cyclodextrin using response surface methodology. After 60 days of storage, aroma analysis was performed on the essential oil of O. onites and the essential oil powder that was produced under ideal conditions. RESULTS O. onites essential oil powder was subjected to GC-MS analysis to identify 72 aroma components. Carvacrol and linalool concentrations varied from 22.66 to 30.11 mg/g and from 32.25 to 53.57 mg/g, respectively. The spray-dried microcapsules' D4.3 values ranged from 40.75 to 48.95 μm. For every trial, the drying yield and solubility values were higher than 60% and 90%, respectively. The results of the surface optimization showed that the ideal inlet air temperature was 190°C and the ideal concentration of β-cyclodextrin in the wall material was 1%. CONCLUSION The findings indicate that O. onites essential oil was successfully encapsulated via spray drying. The preservation of critical aroma compounds in the resulting capsules is high.

Abstract: INTRODUCTION Broccoli sprouts have great health and commercial value because they are rich in sulforaphane, a special bioactive compound that helps to prevent chronic diseases, such as cancer and cardiovascular disease. OBJECTIVE The aim of this study was to increase the levels of active substances in broccoli sprouts and understand their metabolic mechanisms. METHODOLOGY Metabolomics based on liquid chromatography-tandem mass spectrometry and transcriptome analysis were combined to analyse the enrichment of metabolites in broccoli sprouts treated with cold plasma. RESULTS After 2 min of cold plasma treatment, the contents of sulforaphane, glucosinolates, total phenols, and flavonoids, as well as myrosinase activity, were greatly improved. Transcriptomics revealed 7460 differentially expressed genes in the untreated and treated sprouts. Metabolomics detected 6739 differential metabolites, including most amino acids, their derivatives, and organic acids. Enrichment analyses of metabolomics and transcriptomics identified the 20 most significantly differentially expressed metabolic pathways. CONCLUSIONS Overall, cold plasma treatment can induce changes in the expression and regulation of certain metabolites and genes encoding active substances in broccoli sprouts.

Abstract: INTRODUCTION Scrophulariae Radix (SR) has been extensively used in traditional Chinese medicine (TCM) for thousands of years. However, the processing methods and production areas of Scrophularia ningpoensis have undergone notable historic changes. Thus, their effects on the bioactive constituents of SR still need to be studied further. OBJECTIVES This study aimed to establish an objective and comprehensive method to identify the correlation of bioactive constituents of SR with variety, place of origin and processing method for evaluating their qualities. METHODOLOGY An accurate and rapid high-performance liquid chromatography-diode array detector (HPLC-DAD) method for the simultaneous determination of 11 marker components (aucubin, harpagide, 6-O-methyl-catalpol, harpagoside, verbascoside, isoverbascoside, angoroside C, cinnamic acid, l-tyrosine, l-phenylalanine, and l-tryptophan) was established to evaluate the quality of SR for the first time. In addition, the effects of different production areas and processed methods on the target compounds were studied by analysing 66 batches of SR samples with chemometrics methods, including similarity evaluation of chromatographic fingerprints of TCM, principal component analysis (PCA), and partial least squares-discriminant analysis (PLS-DA). RESULTS Compared with "sweating", short-term "steaming" and "slice-drying" could largely preserve the bioactive constituents of SR. When using the model established through PLS-DA, five components were identified as the most significant variables for discrimination. Furthermore, the score plots of PCA and the similarity evaluation revealed that variety had a more notable influence on the quality of SR than the place of origin. CONCLUSION An objective approach of HPLC fingerprint coupled with chemometrics analysis and quantitative assessment could be applied to discriminate different processed SR and evaluate the qualities of SR rapidly.

Abstract: INTRODUCTION Qingjin Yiqi granule (QYG) is a prescription medicine of traditional Chinese medicine which is widely used clinically for the recovery of coronavirus patients. However, there is currently limited research on the quality control of QYG. OBJECTIVE To evaluate the quality of QYG qualitatively and quantitatively by making full use of advanced chromatography-mass spectrometry techniques. METHODS Firstly, a multicomponent characterisation of QYG was performed by ultrahigh-performance liquid chromatography coupled with a Q Exactive™ hybrid quadrupole-Orbitrap mass spectrometry (UHPLC-Q-Orbitrap-MS) system using a rapid negative/positive switching mode. Secondly, the co-condition fingerprint analysis of constituted herbal medicines of QYG was performed to unveil active ingredients as the quality markers of QYG. Thirdly, the marker compounds in 10 batches of QYG were quantified by ultrahigh-performance liquid chromatography coupled with a Waters Xevo TQ-S triple quadrupole mass spectrometry (UPLC-QQQ-MS) system. RESULTS A comprehensive method that combined the inclusion list and data-dependent acquisition (DDA) to achieve a systematic characterisation of QYG was established by UHPLC-Q-Orbitrap-MS. After analysis based on Compound Discoverer software and Global Natural Products Social (GNPS) platform, a total of 332 compounds were detected. Eleven Q-markers were determined for the quality evaluation of QYG by comparison with the fingerprint of nine constituted herbal medicines. An adjusted multiple reaction monitoring (MRM) quantification method was further established to simultaneously determine the 11 Q-markers for holistic quality evaluation of QYG. CONCLUSION This is the first study to report comprehensive multicomponent characterisation, identification, and quality assessment of QYG, which could be used for effective guarantee of the quality of QYG.

Abstract: INTRODUCTION Solanum nigrum L. is a traditional medicinal herb and edible plant. Many studies provide evidence that S. nigrum L. is a nutritious vegetable. Polyphenols and steroidal glycoalkaloids are the main components. OBJECTIVES This study aimed to systemically evaluate the phytochemical profile, quantification, and bioactivities of polyphenolics and glycoalkaloids in different parts of S. nigrum L. RESULTS Total polyphenols (TPC) and total glycoalkaloids (TGK) were determined using the Folin-Ciocalteu and acid dye colorimetric methods, respectively. A total of 55 polyphenolic constituents (including 22 phenolic acids and 33 flavonoids) and 24 steroidal glycoalkaloids were identified from different parts using ultrahigh-performance liquid chromatography Q-exactive high-resolution mass spectrometry (UHPLC-QE-HRMS), of which 40 polyphenols (including 15 phenolic acids and 25 flavonoids) and one steroidal glycoalkaloid were characterised for the first time in S. nigrum L. Moreover, typical polyphenols and glycoalkaloids were determined using HPLC-UV and HPLC-evaporative light-scattering detector (ELSD), respectively. In addition, the TPC and TGK and their typical constituents were compared in different anatomical parts. Finally, the antioxidant capacities of polyphenolic extracts from different parts of S. nigrum L. were evaluated by ·OH, 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging and ferric-reducing antioxidant power (FRAP) assay in vitro. In addition, the antitumour effects of TGK from different parts of S. nigrum L. on the proliferation of PC-3 cells were investigated using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Polyphenolic and glycoalkaloid extracts from different parts of S. nigrum L. showed different antioxidant and cytotoxic capacities in vitro. CONCLUSION This is the first study to systematically differentiate between polyphenolic and glycoalkaloid profiles from different parts of S. nigrum L.

Abstract: INTRODUCTION Although the Tibetan medicine Triphala (THL) is widely used in many countries, insufficient progress has been made in quality control. OBJECTIVES The present study aimed to propose a methodology for quality control of THL based on HPLC fingerprinting combined with an orthogonal array design. METHODS Seven identified peaks were used as indicators to examine the effects of temperature, extraction time, and solid-liquid ratio on the dissolution of active ingredients in THL. Fingerprint analysis was performed on 20 batches of THL from four geographical areas (China, Laos, Thailand, and Vietnam). For further chemometric assessment, analysis techniques including similarity analysis, hierarchical clustering analysis, principal component analysis, and orthogonal partial least squares discrimination analysis (OPLS-DA) were used to classify the 20 batches of samples. RESULTS Fingerprints were established and 19 common peaks were identified. The similarity of 20 batches of THL was more than 0.9 and the batches were divided into two clusters. Four differential components of THL were identified based on OPLS-DA, including chebulinic acid, chebulagic acid, and corilagin. The optimal extraction conditions were an extraction time of 30 min, a temperature of 90°C, and a solid-liquid ratio of 30 mL/g. CONCLUSION HPLC fingerprinting combined with an orthogonal array design could be used for comprehensive evaluation and quality assessment of THL, providing a theoretical basis for further development and utilization of THL.

Abstract: INTRODUCTION Shenlingbaizhu granule, a Traditional Chinese Medicine prescription comprising Renshen, Gancao, and Shanyao, is widely consumed in China nowadays. OBJECTIVE The study tries to propose pharmacopoeia quality markers (Q-markers) to prevent counterfeiting involving Renshen, Gancao, and Shanyao. METHODOLOGY A novel strategy, that is, library-based ultra-high-performance liquid chromatography-quadrupole-orbitrap mass spectrometry, was used to analyse the lyophilised aqueous powder of Shenlingbaizhu granule. Subsequently, quantum chemistry calculation and UV-vis spectra scanning were also performed through theoretical or experimental approaches. RESULT Thirty-two isomers have been strictly distinguished, especially positional isomeric isochlorogenic acid B versus isochlorogenic acid C, positional isomeric schaftoside versus isoschaftoside, positional isomeric ginsenoside Rg2 versus 20S-ginsenoside Rg3, and stereoisomeric 20S-ginsenoside Rg3 versus 20R-ginsenoside Rg3. Seventeen compounds were unexpectedly observed, particularly scoparone and pectolinarigenin, while a total of 76 bioactive compounds have been putatively identified in the study. The quantum chemistry calculation and UV-vis spectra scanning results revealed that glycyrrhizic acid, ginsenoside Re, ginsenoside Rb1, and diosgenin displayed different dipole moment values and maximum absorption wavelengths from each other. CONCLUSION The study recommends glycyrrhizic acid, ginsenoside Re, ginsenoside Rb1, and diosgenin as four anti-counterfeiting Q-markers for the pharmacopoeia. The anti-counterfeiting Q-markers can be detected using conventional HPLC. The observation of 17 unexpected compounds updates our knowledge regarding the bioactives of Shenlingbaizhu granule.

Abstract: Agarwood, a fragrant resinous wood mainly formed by Aquilaria spp., is used worldwide as a natural fragrance and traditional medicine. A large amount of Aquilaria sinensis (Lour.) Gilg leaves are underutilised during the process of the agarwood industry, and the development of A. sinensis leaves as tea has recently attracted more and more attention. However, the small molecule profile of A. sinensis leaves and their bioactivities has been rarely reported.To conduct a rapid untargeted liquid chromatography-mass spectrometry (LC-MS) analysis of A. sinensis leaves with a molecular networking (MN) strategy and evaluate its antioxidant and antidiabetic value.A MN-assisted tandem mass spectrometry (MS/MS) analysis strategy was used to investigate the small molecule profile of A. sinensis leaves. Additionally, the integration of antioxidant and α-glucosidase inhibitory assays with MN analysis was executed to expeditiously characterise the bioactive compounds for potential prospective application.Five main chemical groups including phenol C-glycosides, organic acids, 2-(2-phenylethyl) chromones, benzophenone O-glycosides and flavonoids were rapidly revealed from the A. sinensis leaves. Eighty-one compounds were provisionally identified by comparing their MS/MS fragments with canonical pathways. The featured xanthone C-glycosides and benzophenone C-glycosides were recognised as the primary components of A. sinensis leaves. Several dimers and a trimer of mangiferin were reported firstly in A. sinensis leaves. Furthermore, 17 and 14 potential bioactive molecules were rapidly annotated from antioxidant and α-glucosidase inhibitory fraction, respectively.Our findings will help expand the utilisation of A. sinensis leaves and thus promote the high-quality development of agarwood industry.

Abstract: INTRODUCTION Cannabis sativa L. inflorescences are rich in secondary metabolites, particularly cannabinoids. The most common techniques for elucidating cannabinoid composition are expensive technologies, such as high-pressure liquid chromatography (HPLC). OBJECTIVES We aimed to develop and evaluate the performance of a novel fluorescence spectroscopy-based method coupled with N-way partial least squares regression (N-PLS-R) and partial least squares discriminant analysis (PLS-DA) models to replace the expensive chromatographic methods for preharvest cannabinoid quantification. METHODOLOGY Fresh medicinal cannabis inflorescences were collected and ethanol extracts were prepared. Their excitation-emission spectra were measured using fluorescence spectroscopy and their cannabinoid contents were determined by HPLC-PDA. Subsequently, N-PLS-R and PLS-DA models were applied to the excitation-emission matrices (EEMs) for cannabinoid concentration prediction and cultivar classification, respectively. RESULTS The N-PLS-R model was based on a set of EEMs (n = 82) and provided good to excellent quantification of (-)-Δ9-trans-tetrahydrocannabinolic acid, cannabidiolic acid, cannabigerolic acid, cannabichromenic acid, and (-)-Δ9-trans-tetrahydrocannabinol (R2 CV and R2 pred > 0.75; RPD > 2.3 and RPIQ > 3.5; RMSECV/RMSEC ratio < 1.4). The PLS-DA model enabled a clear distinction between the four major classes studied (sensitivity, specificity, and accuracy of the prediction sets were all ≥0.9). CONCLUSIONS The fluorescence spectral region (excitation 220-400 nm, emission 280-550 nm) harbors sufficient information for accurate prediction of cannabinoid contents and accurate classification using a relatively small data set.

Abstract: INTRODUCTION Propolis is a resinous natural substance collected by honeybees from buds and exudates of various trees and plants; it is widely accepted that the composition of propolis depends on the phytogeographic characteristics of the site of collection. OBJECTIVES The aim of this study was to determine the phytochemical composition of ethanolic extracts from eight propolis batches collected in different regions of Benin (north, center, and south) and Congo, Africa. MATERIAL AND METHODS Characterization of propolis samples was performed by using different hyphenated chromatographic methods combined with carbon-13 nuclear magnetic resonance (13 C NMR) dereplication with MixONat software. Their antioxidant or anti-advanced glycation end-product (anti-AGE) activity was then evaluated by using diphenylpicrylhydrazyl and bovine serum albumin assays, respectively. RESULTS Chromatographic analyses combined with 13 C NMR dereplication showed that two samples from the center of Benin exhibited, in addition to a huge amount of pentacyclic triterpenes, methoxylated stilbenoids or phenanthrenoids, responsible for the antioxidant activity of the extract for the first one. Among them, combretastatins might be cytotoxic. For the second one, the prenylated flavanones known in Macaranga-type propolis were responsible for its significant anti-AGE activity. The sample from Congo was composed of many triterpene derivatives belonging to Mangifera indica species. CONCLUSION Therefore, propolis from the center of Benin seems to be of particular interest, due to its antioxidant and anti-AGE properties. Nevertheless, as standardization of propolis is difficult in tropical zones due to its great chemodiversity, a systematic phytochemical analysis is required before promoting the use of propolis in food and health products in Africa.

Abstract: INTRODUCTION Miroestrol (Mi) and deoxymiroestrol (Dmi) are trace, yet potent, phytooestrogens found in white Kwao Krua [Pueraria candollei var. mirifica (Airy Shaw & Suvat.) Niyomdham, PM]. However, the analysis of these substances is difficult because of complex matrix effects and their various analogues. In addition, alteration in the cross-reactivity of a gold nanoparticle (AuNP)-based immunochromatographic assay (ICA) resulting from the electrostatic adsorption between antibodies and AuNPs has not yet been evaluated. OBJECTIVES This study aims to develop, characterise, and validate ICA with a monoclonal antibody exhibiting similar reactivity against Mi and Dmi (MD-mAb). MATERIALS AND METHODS The ICA performance was validated for cross-reactivity and performance in comparison with those of indirect competitive enzyme-linked immunosorbent assays (icELISAs) with MD-mAb and mAb exhibiting specificity against Mi (Mi-mAb). RESULTS The ICA showed a limit of detection (LOD) at 1 and 16 μg/mL for Mi and Dmi, respectively. The cross-reactivity of the ICA with Dmi was lower (6.25%) than that observed with the icELISA (120%). Cross-reactivity of ICA against other compounds of the PM was also correlated with those of icELISA; no false-positive/negative results were observed. The repeatability and reproducibility of the ICA were confirmed. The results obtained using ICA in samples of PM are correlated with the concentrations determined through icELISAs. CONCLUSION An ICA with MD-mAb was constructed and validated. However, direct conjugation via the electrostatic adsorption of mAb-AuNPs was expected to alter the cross-reactivity of ICA, especially that of the analyte analogue Dmi.

Abstract: Abstract Introduction Dahlia pinnata Cav. is a flower native to Mexico that has many applications; in particular, its petals have been used for ornamental, food, and medicinal purposes, for example to treat skin rashes and skin cracks. It has been reported that the medicinal properties of plants are generally related to the phytochemical constituents they possess. However, there are few studies on black D. pinnata . Objectives The present study was aimed at qualitatively and quantitatively determining the phytochemical profile of petals from black D. pinnata . Methodology Phytochemicals from Dahlia petals were extracted by consecutive maceration (hexane, dichloromethane, and methanol); then, the extracts were analyzed through colorimetric assays and UV–Vis spectroscopy for qualitative identification and quantification of phytochemical compounds, respectively. The methanolic extract was analyzed by flow injection analysis–electrospray ionization–Fourier transform ion cyclotron resonance mass spectrometry (FIA‐ESI‐FTICR‐MS) in negative and positive mode. Results Quantitative phytochemical profiling of the methanolic extract by UV–Vis spectroscopy indicated high contents of phenolic compounds (34.35 ± 3.59 mg EQ/g plant) and sugars (23.91 ± 1.99 mg EQ/g plant), while the qualitative profiling by FIA‐ESI‐FTICR‐MS allowed the tentative identification of several flavonoids and phenolic acids. Kaempferol‐3‐rutinoside, pelargonidin‐3‐(6″‐malonylglucoside)‐5‐glucoside, rutin, kaempferol‐3‐(2″,3″‐diacetyl‐4″‐ p ‐coumaroylrhamnoside), and myricetin‐3‐(2‴‐galloylrhamnoside) were the main compounds detected. Conclusion The results expand our knowledge of the phytochemical constituents of petals from black D. pinnata .

Abstract: INTRODUCTION Olive leaves, abundant by-products of the olive oil industry, are a rich source of oleuropein, an important polyphenol in olive leaves. So far, no published methods have been validated using matrix standards for oleuropein quantification in olive leaves. OBJECTIVES The study aimed to develop an HPLC method for oleuropein determination in olive leaves using spiked matrix standards prepared from a blank olive leaf matrix, to validate the method with respect to aqueous standards, and cross-validate the HPLC method with UPLC-MS and UPLC-UV techniques. METHODOLOGY Oleuropein was extracted into methanol and analysed by HPLC with fluorescence detection (FLD; excitation and emission wavelengths 281 and 316 nm, respectively) and by UPLC-MS-UV. For validation, calibration curves of spiked matrix standards (0.4 to 4.8 mg/g) were analysed by the three methods over several days. Oleuropein was then analysed in French olive varieties. RESULTS For the HPLC-FLD method, repeatability and intermediate precision were less than 5% RSD and linearity was demonstrated by the Fischer test. Differences in results of the spiked placebos by the three methods were non-significant, as confirmed by ANOVA. Extraction recovery was >90%, and there was a strong linear relationship between authentic and spiked matrix standards. The determination of oleuropein in French olive varieties is reported, including analysis in "Olivière" cultivar for the first time, leaves of which contained twice the amount of oleuropein compared with "Picholine". CONCLUSION Accurate quantification of oleuropein is possible using aqueous standards. Cross-validation indicates that selective analysis can equally be carried out by HPLC or by UPLC-MS techniques.

Abstract: INTRODUCTION The spores of the medicinal fungus Ganoderma lucidum possess hepatoprotective properties. The main components, triterpenes, are particularly beneficial, making the screening and preparation of active triterpenes from Ganoderma lucidum significant. OBJECTIVES We aimed to screen and verify cyclooxygenase-2 inhibitors from G. lucidum spores, establish a rapid online hyphenated technique for the preparation of active ingredients, and analyze the structures of the active ingredients. METHODS Ultrafiltration LC combined with an enzyme inhibition assay and molecular docking was employed to screen and evaluate cyclooxygenase-2 ligands, which were prepared by pressurized liquid extraction coupled online with countercurrent chromatography and semi-preparative LC. The structures of the compounds were identified by nuclear magnetic resonance spectroscopy. RESULTS Six cyclooxygenase-2 inhibitors, namely, ganoderic acids I, C2 , G, B, and A and ganoderenic acid A, were screened and evaluated. They were prepared using the online hyphenated technique, following which their structures were identified. CONCLUSION This study provides opportunities for the discovery and development of new therapeutic drugs from other natural resources, as the present instrumental setup achieved efficient and systematic extraction and isolation of natural products compared with reference separation methods, thus exhibiting significant potential for industrial applications.

Abstract: INTRODUCTION Azadirachta indica A. Juss. is a well-known medicinal plant that has been used traditionally to cure various ailments in every corner of the globe. There are many in vitro and in vivo experimental evidences in connection with the bioactivity of the extracts of this plant. Lung cancer is the deadliest form of cancer and contributes to the most cancer related deaths. The mode of action of anticancer components of this plant is still to be established explicitly. OBJECTIVE The objective of this study is to identify druggable targets of active constituents of A. indica A. Juss. for non-small cell lung cancer (NSCLC) using network pharmacology and validation of activity through molecular docking analysis. METHODOLOGY Targets of all the active phytochemicals from A. indica were predicted and genes related to NSCLC were retrieved. A protein-protein interaction (PPI) network of the overlapping genes were prepared. Various databases and servers were employed to analyse the disease pathway enrichment analysis of the clustered genes. Validation of the gene/protein activity was achieved by performing molecular docking, and ADMET profiling of selected phytocompounds was performed. RESULT Gene networking revealed three key target genes as EGFR, BRAF and PIK3CA against NSCLC by the active components of A. indica. Molecular docking and ADMET analysis further validated that desacetylnimbin, nimbandiol, nimbin, nimbinene, nimbolide, salannin and vepinin are the best suited anti- NSCLC among all the phytocompounds present in this plant. CONCLUSION The present study has provided a better understanding of the pharmacological effects of active components from A. indica and its potential therapeutic effect on NSCLC.