Phytochemical Analysis最新文献

Introduction: Citri Reticulatae Pericarpium (CRP), also known as Chenpi in Chinese, is the dry mature peel of Citrus reticulata Blanco or its cultivated varieties. CRP as the health-care food and dietary supplement has been widely used in various diseases. The quality of CRP can be affected by various factors, which are closely related to the metabolite composition of CRP.

Objectives: The objective of this study is to conduct a comprehensive comparative analysis on the chemical profiling of 51 C. reticulata samples of eight medicinal varieties, grown in different areas, and provide a methodological reference for the study of pharmacodynamic material bases and quality control of C. reticulata.

Methodology: Initially, a comprehensive characterization was performed using quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS), and a heatmap visualization was employed for clarifying the distribution of the annotated active ingredients. Furthermore, obtained chemical profiles data were employed in multivariate statistical methods, comprising principal component analysis (PCA), and orthogonal partial least-squares-discrimination analysis (OPLS-DA).

Results: A total of 42 chemical components were annotated in positive ion mode. The relative contents were evident differences in the active ingredients of medicinal varieties of C. reticulata; mostly, polymethoxy flavones (PMFs) in C. reticulata "Dahongpao" were more abundant; among them, nobiletin and tangeretin are the main active ingredients in CRP. In addition, the relative contents of chemical constituents of C. reticulata "Dahongpao" and C. reticulata "Unshiu" from different areas were less variable. Compared with production origins, the varieties of C. reticulata had a greater impact on quality.

Conclusion: This work obtains a better understanding of the chemical profiles of medicinal varieties of C. reticulata, facilitated the reasonable applicability and quality control of medicinal varieties of C. reticulata.

Objectives: We used ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS), bioinformatics, and in vivo experiments to study the anti-colorectal cancer (CRC) effects of Wenzi Jiedu Decoction (WJD).

Methods: Detected the main components of WJD by UPLC-MS/MS. Obtained WJD targets and CRC targets through the open source database. Analyzed the WJD-CRC targets from a macro perspective by PPI, GO, and KEGG analyses. Validated bioinformatics findings by molecular docking and animal experiments.

Results: This study obtained 91 active compounds and 240 targets of WJD. Intersection with CRC genes (GSE32323 388 DEGs, GSE 215510 1253 DEGs), 36 WJD-CRC common targets were obtained. PPI and enrichment analyses indicated WJD exerted anti-CRC effects mainly through the chemokine signaling pathway and apoptosis. Quercetin, Luteolin, Kaempferol, Formononetin, Stigmasterol, and Hederagenin were the main compounds of WJD. CXCL8, BCL-2, BAX, BCL2L1, CASP3, AKT1, and TP53 were the core targets of WJD-CRC. Bulk molecular docking showed that core WJD compounds had good docking activity with WJD-CRC targets. Animal experiments had shown the tumor inhibition rate of the WJD group was 36.53%. WJD could regulate the ratio of CD4+, CD8+, and CD4+/CD8+, reduce the expression of CXCL8, and BCL-2, and increase the expression of BAX.

Conclusions: This study indicated that the potential mechanism of WJD in the prevention and treatment of CRC had the characteristics of the multi-target, multi-path, and multi-system mechanisms, which were mainly related to the regulation of chemokines and the promotion of apoptosis.

Introduction: The extraction of DNA is the basis of molecular biology research. The quality of the extracted DNA is one of the key factors for the success of molecular biology experiments.

Objective: To select a suitable DNA extraction method for Chinese medicinal herbs and seeds.

Methods: In this experiment, four commercial DNA extraction kits were used to extract the genomic DNA (gDNA) from the Pinellia ternata (Thunb.) Breit. powder; Arisaema amurense Maxim. powder as well as the seeds of Glycine max (L.) Merr. On the one hand, the concentration and purity of DNA extracted by these four kits were compared. On the other hand, nucleic acid amplification experiments were performed on three samples extracted by each of the four kits by Proofman-LMTIA methods, which is a novel nucleic acid isothermal amplification technique. The concentration and purity of DNA extracted by different kits were used to determine which methods were suitable for the dry powder of Chinese herbal medicines and seeds. The efficiency of the amplification curve to show whether the extracted DNA can be used in nucleic acid amplification experiments.

Results: The results showed that the Proofman-LMTIA methods were of high specificity and the optimal reaction temperatures were 63, 59, and 59°C for P. ternata (Thunb.) Makino; A. amurense Maxim. and G. max (L.) Merr., respectively. The concentration and purity of the gDNAs extracted with all kits were within the acceptable ranges; meanwhile, the amplification of the gDNA extracted by Kit II was of the highest efficiency.

Conclusion: In this experiment, the principle, concentration, purity, and time taken for extracting DNA with four kits were compared. The automated extraction kit based on the magnetic method is suitable for extracting DNA from Chinese medicinal herbs and seeds. The extracted DNA is suitable for nucleic acid amplification detection.

Introduction: The roots and rhizomes of Curcuma longa L. serve as distinct traditional Chinese medicines with varying therapeutic effects, likely attributed to differences in the accumulation and distribution of metabolites in these parts.

Objective: The study aims to investigate the differences and spatial distribution patterns of metabolites in C. longa L. roots and rhizomes.

Methods: Metabolite analysis of roots and rhizomes was conducted using ultra-high-performance liquid chromatography-quadruple orbitrap high-resolution mass spectrometry (UHPLC-Q-Orbitrap HRMS) combined with desorption electrospray ionization mass spectrometry imaging (DESI-MSI). Using principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA) to screen for differential metabolites. The relative contents of differential metabolites were visualized using heat maps. Additionally, the spatial distribution of differential metabolites was analyzed based on DESI-MSI.

Results: A total of 49 main chemical components were identified in roots and rhizomes using UHPLC-Q-Orbitrap HRMS. Through nontargeted metabolomics analysis combining UHPLC-Q-Orbitrap HRMS with PCA and OPLS-DA, 24 differential markers were identified; Additionally, using DESI-MSI alongside PCA and OPLS-DA, 18 differential markers were selected. Based on the DESI-MSI results, curcuminoids and sesquiterpenoids, including bisdemethoxycurcumin, demethoxycurcumin, furanodienone, furanogermenone, furanodiene, β-elemene, and curzerene, were more abundant in the rhizomes compared to the roots. And these differential compounds exhibited spatial distribution differences in the epidermis, phloem, and xylem between the roots and rhizomes.

Conclusion: The metabolomics analysis using UHPLC-Q-Orbitrap HRMS combined with DESI-MSI suggest differences in the accumulation and spatial distribution of metabolites in C. longa L. roots and rhizomes, possibly related to the biosynthesis of secondary metabolites.

Introduction: Phenolic compounds garner interest in developing medicines, nutraceuticals, and cosmeceuticals based on natural products. The quantity of phenolic compounds in a sample is commonly determined via spectrophotometry; however, this instrumented technique is relatively laborious and time consuming and requires a large amount of reagents.

Objective: This work aimed to develop a simple, point-of-need colorimetric sensor to rapidly determine total phenolic content (TPC) in tea extracts.

Methodology: We developed a radial paper-based analytical device (PAD) for TPC determination based on the established colorimetric reaction between the Folin-Ciocâlteu reagent and phenols. The PAD was designed to enable quantitative (with image capturing device and color processing software) and semiquantitative (using a color palette reference card) determinations. Analytical performance and stability of the PAD were evaluated based on the color responses.

Results: The PAD was successfully applied for the determination of phenolics in tea extracts obtained using several polar protic solvents, including water, methanol, and ethanol, with satisfactory accuracy (recovery of 95.5%-104%, 110%-116%, and 104%-110%, respectively) and precision (RSD < 9%). The obtained TPC values also agreed with those from visible spectrophotometry. Semiquantitative determination using the color reference card with three categories of TPC level (i.e., 0-100, 100-500, and 500-1000 mg gallic acid equivalent/L) provided > 95% accuracy. The devices were the most stable when stored at 4°C in a light-protected, vacuum-sealed container. The proposed PAD is promising for simple, rapid (~10-20 min), and accurate estimation of TPC in plant extracts.

Introduction: Liquid chromatography-mass spectrometry (LC-MS) has enhanced the rapid, accurate analysis of complex plant extracts, eliminating the need for extensive isolation. Tandem mass spectrometry (MS/MS) further enhances this process by providing detailed structural information. However, differentiating structural isomers remains a challenge due to their minor spectral and structural differences.

Objective: This study aimed to extend the applicability of LC-MS/MS for the structural identification of daphnane diterpenoids, with a particular focus on distinguishing functional isomers.

Methods: LC-MS analyses were performed using an UHPLC-Q-Exactive-Orbitrap MS. The MS conditions for distinguishing isomers were optimized using in-source CID and HCD modes with reference compounds. A qualitative analysis was then conducted on the extract of Daphne tangutica. The chemical structures of the detected daphnane diterpenoids were estimated by analyzing the fragmentation patterns in both the mass spectra and product ion spectra. These identifications were further validated by isolation and comparison with an in-house daphnane diterpenoid library.

Results: By optimizing MS conditions, especially in the negative ion mode, it was possible to accurately distinguish structural isomers such as yuanhuajine and gniditrin. Qualitative analysis of D. tangutica identified a total of 28 daphnanes, including seven previously unreported compounds. Furthermore, a novel geometric isomer of gniditrin was isolated by conducting isolation on the crude diterpenoid fraction.

Conclusion: This study demonstrated that LC-MS/MS analysis can effectively distinguish functional isomers of daphnane diterpenoids, thereby enhancing the identification of daphnanes in plant extracts and highlighting its potential as a powerful tool for phytochemical analysis.

Introduction: Fufang Shenqi Oral Liquid (FFSQOL) is an important Chinese medicine compound preparation with a wide range of clinical applications, which is mainly used to regulate immune function, improve cardiovascular function, and have anti-inflammatory and antibacterial effects. At present, it is of great importance to establish the quality evaluation method of FFSQOL and to investigate its quality markers (Q-markers).

Objectives: The aim of this study is to establish a quality evaluation method for FFSQOL and screen its Q-markers to provide a scientific basis for its quality control.

Methods: Fourteen batches of FFSQOL were subjected to high-performance liquid chromatography (HPLC) fingerprint and similarity analysis. The components of FFSQOL were identified, and their content was determined. This was combined with cluster analysis (CA) and principal component analysis (PCA) to determine the Q-markers of FFSQOL.

Results: In this study, an HPLC fingerprint was established for 14 batches of FFSQOL, identifying 12 common peaks and six major components. Four components were identified as stable and reproducible: gallic acid (504.94 ~ 1219.04 μg/mL), caffeic acid (452.15 ~ 783.01 μg/mL), 7-O-glucoside (1097.72 ~ 2440.41 μg/mL), and formononetin (176.2 ~ 177.51 μg/mL). Quality evaluation of the 14 batches was conducted using chemical pattern recognition analysis. CA results indicated two distinct groups, and PCA revealed that principal components 1 and 2 were the main factors influencing batch differences. A combination of HPLC fingerprint, content determination results, and chemical pattern recognition analysis was employed to identify Q-markers for FFSQOL. The markers identified were gallic acid, caffeic acid, calycosin 7-O-glucoside, and formononetin.

Conclusion: In this study, a quality evaluation method for FFSQOL was established through the implementation of fingerprint, content determination, and chemical pattern recognition analysis, resulting in the identification of four Q-Markers of FFSQOL, which laid the foundation for the formulation of FFSQOL quality standards.

Introduction: Huangma Tincture (HMT) is a Chinese patent medicine with a history of clinical use for more than 60 years, widely used for treating dermal chronic ulcer such as diabetic foot ulcer. However, the overall quality evaluation and control method of HMT has not yet been researched.

Objective: The aim of this study is to establish a comprehensive quality evaluation and control method for HMT based on high-performance liquid chromatography (HPLC) fingerprint, chemometrics, and multicomponent content determination.

Methods: Establishing chemical fingerprint of HMT and carrying out similarity analysis comprehensively reflect the consistency of the formulation in terms of chemical composition. Chemometrics analyses, including hierarchical cluster analysis (HCA), principal component analysis (PCA), and orthogonal partial least squares-discriminant analysis (OPLS-DA), were performed to identify components crucial to quality differences. The content of five potential bioactive ingredients (brucine, strychnine, jatrorrhizine, coptisine, and berberine hydrochloride) was determined as a supplementary quality control measure.

Results: HPLC fingerprint of HMT with similarity index > 0.990 was established, in which five common compounds (brucine, strychnine, jatrorrhizine, coptisine, and berberine hydrochloride) were identified. HCA, PCA, and OPLS-DA results, validated through 200 permutation tests, were basically consistent. The contents of brucine, strychnine, jatrorrhizine, coptisine, and berberine hydrochloride in 20 batches of HMT samples were 0.2088-0.5556, 0.2599-0.9868, 0.1358-0.2092, 0.2634-0.6843, and 1.8301-2.7826 mg/mL, respectively.

Conclusion: HPLC fingerprint combined with chemometrics and multicomponent content determination considering both effect and toxicity provides a robust method for the comprehensive quality evaluation and control of HMT.

The increasing demand for global food resources and over-dependence on terrestrial agroecosystems pose a significant challenge to the sustainable production of food commodities. Macroalgae are an essential source of food production in the marine environment, and their cultivation is a promising approach to alleviate the impending global food insecurity due to key factors, such as independence from terrestrial agriculture, rapid growth rate, unique biochemical composition, and carbon capture potential. Moreover, in many countries, seaweed has been used as food for decades because of its health and nutritional benefits. Seaweed contains bioactive components that are beneficial against various pathological conditions, including cancer, type 2 diabetes, and neurological disorders. Furthermore, the natural products derived from macroalgae have also been found to have immunostimulatory and antimicrobial properties. Macroalgae are also a significant source of rare sugars such as L-fucose, L-rhamnose, and glucuronic acid. Besides sugars, other bioactive components have been widely reported for their potential in cosmeceuticals. We have outlined the nutrient composition and functional properties of different species of macroalgae, with an emphasis on their potential as value-added products to the functional food market. Beyond being nutritional powerhouses, the variety of biological activities in human health and biomedicine makes them excellent candidates for developing novel drugs. Therefore, this review summarizes the pharmaceutical applications of macroalgae and suggests potential strategies for incorporating macroalgae-derived bioactive compounds into therapeutic products.

Introduction: Cyanobacterium Arthrospira platensis (AP) (Nordstedt) Gomont contains high content of phycobiliproteins (PBP), which are an important source for food industry. Methods effectively extracting proteins contained in AP cells are demanded to provide a supply of the material. Water-based extraction methods are advisable due to the high solubility of the PBP.

Objectives: Extraction techniques such as ultrasound assisted extraction (UAE) and pressurized water extraction (PWE) are popular due to their environmental friendliness, better extraction efficiency, and faster extraction process. In this paper, efficiency of the two methods is compared.

Materials and methods: PWE along with UAE is utilized for release of PBP from the AP cells. The extraction parameters including time, temperature, pressure, and ultrasound intensity are tested to obtain the most efficient setup. The methods were evaluated using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), and the replicates of PWE extracts were further analyzed by capillary isoelectric focusing with laser-induced fluorescence (cIEF-LIF).

Results: The developed PWE method using higher pressure treatment at lower temperature was significantly faster than UAE methods, and the SDS-PAGE results showed a high content of phycobiliproteins in the extracts. cIEF-LIF analysis showed that the sequential PWE of individual samples was repeatable, and the mild extraction provided a fluorescent profile similar to the commercially available C-phycocyanin standard.

Conclusion: Pressurized water extraction was shown to be an efficient, rapid, and well-automated extraction method for AP proteins in general, including bioactive phycobiliproteins. Obtained results encourage the use of PWE in small-scale analytical applications for primary extraction of proteins.