Renowned for their nutritional benefits, citrus fruits are harvested at various stages in China for functional food production. This study introduces an innovative analytical method, DART-MS, enabling direct qualitative analysis of citrus samples without the need for preprocessing. Simultaneously, the combination of chemometrics can be applied to distinguish between three different citrus samples: Citri Reticulatae Pericarpium, Citri Reticulatae Pericarpium Viride, and Citri Reticulatae "Chachi". Notably, given the international regulatory concerns surrounding synephrine, a precise quantitative analysis method for synephrine was developed. The limit of detection (LOD) and the limit of quantification (LOQ) were 39 ng mL-1 and 156 ng mL-1, respectively. The recovery rates obtained varied from 98.46% to 100.71%. Furthermore, the intra-day and inter-day precision demonstrated robust consistency, with values spanning 5.0-6.1% and 5.03-6.08%, respectively, offering quicker results compared to those from HPLC-MS, promising a safer assessment of herbal and food products.

Agarwood holds significant importance as a valuable resource for aromatic purposes, however, key components responsible for aroma and the differences between regions and grades remain to be deeply elucidated. Thus, the odors of agarwood sourced from typical zones, as well as the renowned Kynam agarwood, were analyzed by HS-SPME Arrow GC-MS in SCAN and MRM modes. The integrated strategy proposed herein exploits the respective advantages of non-targeted and targeted analysis. In addition to a total of 55 volatile components identified from the NIST database, 114 odor components were matched according to the Smart Aroma Database, and a series of differential compounds was also unearthed and quantified.

BackgroundAcute lung injury (ALI) and acute respiratory distress syndrome (ARDS) are the leading causes of acute respiratory failure in many critical diseases and are among the main respiratory diseases with high clinical mortality. The global outbreak of coronavirus disease 2019 (COVID-19) can cause severe ARDS, resulting in a steep rise in the number of patient deaths. Therefore, it is important to explore the pathogenesis of ALI and find effective therapeutic agents. In recent years, thanks to modern biomedical tools, some progress has been made in the application of traditional Chinese medicine (TCM) treatment principles based on syndromic differentiation and holistic concepts in clinical and experimental studies of ALI. More and more TCM effective components and formulae have been verified to have significant curative effects, which have a certain guiding significance for clinical practice.PurposeIt is hoped to provide reference for the clinical research of ALI/ARDS and provide theoretical basis and technical support for the scientific application of TCM in respiratory related diseases.MethodsWe performed a literature survey using traditional books of Chinese medicine and online scientific databases including PubMed, Web of Science, Google Scholar, ScienceDirect, China National Knowledge Infrastructure (CNKI), and others up to January 2023.ResultsIn recent years, thanks to modern biomedical tools, some progress has been made in the application of TCM treatment principles based on syndromic differentiation and holistic concepts in clinical and experimental studies of ALI. This paper mainly reviews the research progress of ALI/ARDS mechanism, the understanding of its etiology and pathogenesis by TCM, and the therapeutic effects of TCM formulae and active ingredients of Chinese medicine. A large number of studies have shown that the effective components and formulae of TCM can prevent or treat ALI/ARDS in vivo and in vitro experiments.ConclusionTCM effective components and formulae play an important role in the prevention and treatment of ALI/ARDS through multiple approaches and multiple targets, and provide necessary theoretical support for the further development and utilization of TCM resources.

Pyrrolizidine alkaloids (PAs) in food and natural preparations have received widespread attention due to their hepatotoxicity, genotoxicity, and embryotoxicity. Mass spectrometry (MS), as a high resolution, high sensitive, and high throughput detection tool, has been the most commonly used technique for the determination of PAs. The continuous advancement of new technologies, methods, and strategies in the field of MS has contributed to the improvement of the analytical efficiency and methodological enhancement of PAs. This paper provides an overview of the structure, toxicity properties and commonly employed analytical methods, focusing on the concepts, advances, and novel techniques and applications of MS-based methods for the analysis of PAs. Additionally, the remaining challenges, future perspectives, and trends for PA detection are discussed. This review provides a reference for toxicological studies of PAs, content monitoring, and the establishment of quality control and safety standards for herbal and food products.

The severe respiratory dysfunctions associated with acute lung injury (ALI) and its sequelae have a high morbidity and mortality rate, are multifactorial, and lack a viable treatment. Considering the critical function that amino acids and derivatives play in the genesis of illnesses and the regulation of metabolic processes, monitoring the levels of metabolites associated with amino acids in biological matrices is necessary and interesting to study their pathological mechanisms. Exploring the dynamics of amino acids and derivatives level and searching for biomarkers provides improved clinical ideas for the diagnosis and treatment of ALI. Therefore, we developed an ultra-high-performance liquid chromatography-electrospray tandem mass spectrometry (UHPLC-MS/MS) method that can simultaneously determine the amino acid and derivatives metabolic levels to study amino acid profiles in different biological samples to facilitate clinical research of ALI. In this study, 48 amino acids and derivatives, including neurotransmitters, polyamines, purines, and other types, were quantified simultaneously in a fast, high-throughput, sensitive, and reliable manner within a 15-minute run time without derivatization. No relevant studies have been reported to quantify these 48 amino acid metabolites in three biological samples simultaneously. Satisfactory linearity (R > 0.995), inter-day and intra-day accuracy (85.17-112.67 % and 85.29-111.60 %, respectively), inter-day and intra-day precision (RSD < 13.80 % and RSD < 12.01 %, respectively), matrix effects (81.00 %-118.00 %), recovery (85.09 %-114.65 %) and stability (RSD < 14.72 %) were all demonstrated by the optimized method's successful validation for all analytes. In addition, the suggested method was effectively implemented in plasma, urine, and lung tissue from normal mice and mice with ALI, with the aim of finding potential biomarkers associated with ALI. Potential biomarkers were screened through multivariate statistical analysis and volcanic map analysis, and the changes of markers in ALI were again identified through heat map analysis and correlation analysis with biochemical indicators, which provided ideas and references for subsequent mechanism studies. Here, the technique created in this work offers a quick and dependable way to perform an integrated analysis of amino acids in a variety of biological materials, which can provide research ideas for understanding the physiopathological state of various diseases.

目的 明确金丝桃苷对吡咯里西啶生物碱(PAs)诱导小鼠急性肝损伤的作用及机制.方法 40只C57BL/6雄性小鼠随机分成5组,即空白对照组、模型组以及金丝桃苷低、中、高剂量组,每组8只.除空白对照组外,其余小鼠采用PAs建立急性肝损伤模型,造模6 h、30 h后金丝桃苷低、中、高剂量组小鼠分别灌胃给予20、40、80 mg/kg金丝桃苷各1次,造模48 h后,收集血清、肝脏、回肠、粪便.检测小鼠血清生化指标丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)和总胆汁酸(TBA)水平;苏木精-伊红(HE)染色法观察小鼠肝脏组织病理学形态变化;测定小鼠血清、肝脏、回肠、粪便中胆汁酸含量;实时荧光定量逆转录聚合酶链式反应(RT-qPCR)法及Western blot法检测小鼠肝脏组织中法尼酯X受体(Fxr)、小异源二聚体伴侣(Shp)、胆盐输出泵(Bsep)、胆固醇-7α-羟化酶(Cyp7a1)、甾醇-12α-羟化酶(Cyp8b1)mRNA与蛋白表达水平.结果 与空白对照组比较,模型组血清生化指标ALT、AST、TBA水平显著升高(P<0.05),肝细胞大面积坏死,同时伴有肝窦淤血;与模型组比较,金丝桃苷各剂量组血清生化指标ALT、AST、TBA水平显著降低(P<0.05),金丝桃苷中、高剂量组未见明显肝细胞坏死及肝窦淤血.与空白对照组比较,模型组胆汁酸代谢异常,小鼠肝脏Fxr、Shp、Bsep、Cyp7a1、Cyp8b1 mRNA及蛋白表达水平明显降低(P<0.05);与模型组比较,金丝桃苷中剂量组胆汁酸稳态失衡有所改善,小鼠肝脏Fxr、Shp、Bsep、Cyp7a1、Cyp8b1 mRNA及蛋白表达水平明显升高(P<0.05).结论 金丝桃苷可改善PAs诱导的急性肝损伤,其机制与调控胆汁酸相关基因、维护胆汁酸稳态平衡有关.

Pyrrolizidine alkaloids (PAs) are potent hepatotoxins that can cause liver damage. Hyperoside (Hyp), a natural flavonoid, can be extracted from medicinal plants. Hyp displays hepatoprotective activity in various liver diseases. However, the potential effect and mechanism of action of Hyp in ameliorating PA-induced liver injury remain obscure. This study aimed to explore the protective effect of Hyp against PA-induced hepatotoxicity and its underlying mechanism. We established an in vitro model of PAs in mouse primary hepatocytes and developed a mouse model of acute PA toxicity to investigate the protective effect of Hyp. We found that Hyp notably attenuated PA-induced hepatotoxicity. RNA-sequencing showed that the beneficial effect of Hyp against PA-induced hepatotoxicity was associated with the transcription factor EB (TFEB)-peroxisome proliferator-activated receptor-γ coactivator-1-α (PGC1α) pathway. Our results confirmed that both the autophagy-lysosomal pathway and mitochondrial biogenesis were induced by Hyp through TFEB nuclear translocation in PA-induced liver injury. Furthermore, we demonstrated that activation of the mechanistic target of rapamycin complex 1 (mTORC1) by MHY 1485 decreased TFEB nuclear translocation and abrogated the protective effect of Hyp against PA-induced liver injury in mice. In contrast, inhibition of mTORC1 activity increased the level of TFEB and reduced hepatotoxicity induced by PAs in mouse livers. Likewise, Hyp-induced TFEB activation was validated in vitro. In conclusion, Hyp can activate the TFEB-mediated autophagy-lysosomal pathway and mitochondrial biogenesis through inhibition of mTORC1 activity, alleviating the liver injury induced by PAs, thus suggesting the potential value of Hyp in the treatment of PA-induced hepatotoxicity.

Background Previous studies have demonstrated that high-density lipoprotein cholesterol (HDL-C) plays an anti-atherosclerosis role through reverse cholesterol transport. Several studies have validated the efficacy and safety of natural products in treating atherosclerosis (AS). However, the study of raising HDL-C levels through natural products to treat AS still needs to be explored. Methods The gene sets associated with AS were collected and identified by differential gene analysis and database query. By constructing a protein–protein interaction (PPI) network, the core submodules in the network are screened out. At the same time, by calculating node importance (Nim) in the PPI network of AS disease and combining it with Kyoto Encyclopedia of genes and genomes (KEGG) pathways enrichment analysis, the key target proteins of AS were obtained. Molecular docking is used to screen out small natural drug molecules with potential therapeutic effects. By constructing an in vitro foam cell model, the effects of small molecules on lipid metabolism and key target expression of foam cells were investigated. Results By differential gene analysis, 451 differential genes were obtained, and a total of 313 disease genes were obtained from 6 kind of databases, then 758 AS-related genes were obtained. The enrichment analysis of the KEGG pathway showed that the enhancement of HDL-C level against AS was related to Lipid and atherosclerosis, Cholesterol metabolism, Fluid shear stress and atherosclerosis, PPAR signaling pathway, and other pathways. Then we intersected 31 genes in the core module of the PPI network, the top 30 genes in Nims, and 32 genes in the cholesterol metabolism pathway, and finally found 3 genes. After the above analysis and literature collection, we focused on the following three related gene targets: APOA1 , LIPC , and CETP . Molecular docking showed that Genistein has a good binding affinity for APOA1 , CETP , and LIPC . In vitro, experiments showed that Genistein can up-regulated APOA1 , LIPC , and CETP levels. Conclusions Based on our research, Genistein may have the effects of regulating HDL-C and anti-atherosclerosis. Its mechanism of action may be related to the regulation of LIPC , CETP , and APOA1 to improve lipid metabolism.

The characterization and quality control of natural herbal medicines, such as traditional Chinese medicines (TCMs), is of great significance to ensure their potential efficacy and avoid severe side ef-fects. Thin-layer chromatography (TLC) is a simple and classic approach for examining quality marker of natural products. Nevertheless, it is more difficult to further characterize the compounds adsorbed on the TLC plate. Herein, we reported a simple setup of laser ablation-assisted direct analysis in real-time mass spectrometry (LA-DART-MS), in which the coupling of mass spectrometry information to provide a pre-dominant dimension in the identification of unknown chemical compositions separated on standard TLC plates, and it was applied for rapid characterization of various kinds of natural herbal medicines. The re-sults showed that the introduction of low-cost small laser pointer had significantly improved the desorp-tion process. The system was successfully applied in the analysis of alkaloids, flavonoids, anthraquinones, volatile oils, glycosides, organic acids, and eight different TCMs including Sophorae Flavescentis Radix, Angelicae Sinensis Radix, Acori Tatarinowii Rhizoma, Phellodendri Chinensis Cortex, Picrasmae Ramulus et Folium, Gynura Japonica, Rhei Radix et Rhizoma and Dendrobii Caulis. The obtained limits of detection (LODs) of this method for various types of reference substances were in the range of 4.6-162.2 ng/band on TLC plates. Furthermore, the quality control and identification of different species of Dendrobii Caulis herb was achieved. This study combines the advantages of TLC and ambient mass spectrometry to pro-vide a good choice for the screening and identification of active ingredients and the quality evaluation of botanical samples. (c) 2021 Elsevier B.V. All rights reserved.

Sample preparation is a crucial bottleneck in the whole analytical scheme, especially for the determination of trace analytes in complex matrices. To achieve high extraction and clean-up performance with a short analysis time, various functionalized materials are developed as solid-phase sorbents. In recent years, metal-organic frameworks (MOFs) have emerged as promising stationary phases for pharmaceutical separation and analysis. In this review, we summarize recent advances in applications of MOFs and their composites for analyzing different pharmaceuticals including antibiotics, anti-inflammatory agents, steroid hormones, abused substances, and other kinds of drugs in sample preparation. Both representative MOFs and the preparation methods are introduced. Corresponding analytical approaches have been established through different extraction modes or devices, mainly focusing on solid-phase extraction, solid-phase microextraction, and other MOFs-based microextraction techniques. These pretreatment techniques are successfully applied in handling diverse complex samples such as biological fluids, plant extracts, edible matrix and environmental samples. Moreover, the remaining challenges of MOFs as advanced sorbents, the future perspectives and trends for pharmaceutical analysis are discussed. (C) 2020 Elsevier B.V. All rights reserved.

Progesterone is the representative progestogens in five major classes of steroid hormones and plays important roles in mammalian pregnancy and animal growth and development. Conventional available analytical methods for progesterone involve immunoassay, gas chromatography-mass spectrometry (GC-MS), and high-performance liquid chromatography-mass spectrometry (HPLC-MS), which lack specificity or usually require sophisticated operations and relatively long time. Herein, we developed a novel strategy for rapid analysis of progesterone via direct analysis in real time mass spectrometry (DART-MS) combined with solid-phase extraction (SPE) using an amino functionalized metal-organic frameworks (MOFs). Under optimized conditions, a wide linear range of 0.5–500 ng mL−1 was achieved, with a satisfactory correlation coefficient (R2 = 0.9992). The relative standard deviations (RSDs) were in the range from 2.4 to 8.4%, demonstrating good precision. The applicability was then confirmed by analyzing spiked lake water and synthetic urine samples, and recoveries are between 92.0 and 117.8% in all three spiked levels (5, 25, and 100 ng mL−1). The sensitivity was notably improved compared with solely DART-MS and obtained detection limit decreased by about 50 times. This research provided a rapid, simple, highly sensitive, and efficient approach for analysis of hormones through combination of advantages of ambient mass spectrometry and porous nanomaterials.

Pyrrolizidine alkaloids (PAs) are naturally occurring plant toxins associated with severe liver damage if excessive ingestion. Herein, a novel analytical strategy on utilizing direct analysis in real-time mass spectrometry (DART-MS) was developed, and applied in analysis of six representative PAs. The calibration curves in the range of 10-1000 ng.mL(-1) were established, and relative standard deviations (RSDs) were less than 10%. The limits of detection (LODs) and limits of quantization (LOQs) were 0.55-0.85 ng.mL(-1) and 1.83-2.82 ng.mL(-1), respectively. The feasibility of method was indicated by analysing real samples including Gynura japonica, drug tablets, granules, and fresh cow's milk. Moreover, the results of DART-MS were in good agreement with those observed by high performance liquid chromatography mass spectrometry (HPLC-MS), but consumed less time without chromatographic separation. This research provides a facile fashion for safety assessment of herbal and food products containing PAs and presents promising applications in food, pharmaceutical and clinical analysis.

INTRODUCTION:Agarwood is a highly valuable fragrant resinous wood which is widely used as traditional Chinese medicines, perfumes, incense and decorations. Due to its high economic value and excessive demand, this leads to a rising price and proliferation of fake commodities. Thus, strict authenticity identification and quality evaluation of agarwood are of great significance.OBJECTIVE:To establish a simple, rapid and non-destructive technique for identifying the authenticity of agarwood.METHODS:Liquid extraction surface analysis mass spectrometry (LESA-MS) was firstly proposed to identify the authenticity of 62 agarwood samples without sample preparation. In addition, multivariate statistical models and thin-layer chromatography (TLC) method were used to analyse and verify the results of LESA-MS.RESULTS:Representative compounds of agarwood were detected by LESA-MS. A characteristic 2-(2-phenylethyl)chromone compound (m/z 319.1) was treated as a key chemical marker to identify agarwood and its counterfeits rapidly. Several other chromones ions were identified and used as additional evidence for authentic samples. A total of 62 samples were visually discriminated as two groups by principal component analysis (PCA) and orthogonal projection to latent structures discriminant analysis (OPLS-DA), and the specific characteristic marker was highlighted. Moreover, the qualitative results of the conventional TLC method were in agreement with the LESA-MS approach.CONCLUSION:The proposed LESA-MS method was successfully applied in the direct qualitative analysis of agarwood from different sources. This study indicated great feasibility and practicality of LESA-MS in the rapid identification of agarwood, and provided a non-destructive and meaningful preliminary screening tool for the agarwood industry.