Quantification and discovery of quality markers from Toddalia asiatica by UHPLC-MS/MS coupled with chemometrics.

INTRODUCTION: Toddalia asiatica (TA) is a classical traditional Chinese medicine used to treat rheumatoid arthritis and contusions. However, research regarding TA quality control is currently limited. OBJECTIVE: We aimed to establish a strategy for identifying quality markers that can be used for the evaluation of the quality of TA. METHOD: A rapid and efficient ultra-high-performance liquid chromatography coupled with triple quadrupole tandem mass spectrometry (UHPLC-MS/MS) method was developed for the quantitative determination of 19 compounds in TA from different regions. Then, the extraction process of TA was successively optimized by single-factor optimization and response surface methodology. Moreover, chemometrics was employed to confirm the correlation between quality and target compounds. RESULTS: Utilizing the UHPLC-MS/MS method, separation of the 19 bioactive compounds was achieved within 14 min. The method was validated in terms of linearity (r2 > 0.9982), precision (0.08%-3.70%), repeatability (0.50%-2.54%), stability (2.26%-5.46%), and recovery (95.8%-113%). The optimal extraction process (extraction solvent, 65% ethanol aqueous solution; solid-liquid ratio, 1:20; extraction time, 25 min) was determined with the total content of 19 bioactive compounds as indicator. Significant disparities were observed in the contents of target compounds across different batches of TA. Besides, all samples could be categorized into two distinct groups, and magnoflorine, (-)-lyoniresinol, nitidine chloride, norbraylin, skimmianine, and decarine were identified as quality markers. CONCLUSION: In the present study, we developed a strategy to improve the quality control of TA. In consideration of the pharmacodynamic activity and statistical differences, six compounds are proposed as quality markers for TA.

Limited co-benefits of protected areas in southwest China under current climate change and human modification.

An ambitious new Post-2020 Global Biodiversity Framework "Kunming-Montreal Global Biodiversity Framework" has been developed. However, the combined effects of climate change and human modification can undermine the potential benefits of the global post-2020 conservation efforts. The co-benefits of stabilizing the climate, conserving biodiversity, and maintaining intact wilderness areas may help to persuade the general public of the need to quickly expand existing protected areas (PAs). To maximize the co-benefits after 2020, the careful optimization of existing (PAs) network and scientific identification of conservation targets are both essential. Here, we mapped hotspots of biodiversity, climate vulnerability, and wilderness in Southwest China (SWC). By analyzing the representativeness and gaps of the existing PAs network in SWC, we devised post-2020 conservation targets and highlighted their implications for decision-makers. Our results showed that the incongruence between hotspots of different species exists, indicating that habitats suitable for one taxon may not fully harbor other taxa. According to our assessment, the five jurisdictions of SWC have warmed on average by 0.4°C-1.1 °C over the past 60 years alone. In particular, biodiversity hotspots in SWC are undergoing stark climatic changes. We uncovered prominent conservation gaps in SWC's network of PAs, especially in terms of climate vulnerability and biodiversity. Due to their insufficient number and unreasonable spatial distribution, the PAs network in SWC may be not capable of meeting its biodiversity, climate vulnerability, and wilderness conservation objectives. To rectify this, we proposed a 3-step mission: milestone 2025, milestone 2030, and goal 2050, which aims to protect 23%, 28%, and 60% of the terrestrial area in SWC, respectively. Taken together, our study derived conservation priority areas with relatively clear spatial boundaries and importance levels, thus providing detailed, timely information for decision-makers to expand the PAs network and implement conservation measures varying in strictness in post-2020 conservation practice.

Exploring the impact of urban form on urban land use efficiency under low-carbon emission constraints: A case study in China's Yellow River Basin.

In the Urban Anthropocene, how to meet the demands of growing urban populations on limited urban land is a key global challenge. Unreasonable urban planning and land use has brought about undesirable consequences including huge carbon emissions. However, research on the spatial impact of urban form on urban land use efficiency (ULUE) under low-carbon emission constraints is limited. This study analyzes 91 cities located in China's Yellow River Basin (YRB). First, we define a new comprehensive indicator system to measure ULUE under low-carbon constraints using the SBM-UN model. We then select nine landscape indicators to quantify the sprawl, complexity, and aggregation of urban form. Finally, we use Spatial Durbin Model to reveal the relationship between urban form and ULUE. We find that carbon emissions in the YRB increased steadily during the study period. The average value of ULUE increased from 0.469 in 1994 to 0.772 in 2018. Efficiency improved most in the provinces of Shaanxi, Henan, Ningxia, and Shandong, with growth rates of 234.15%, 102.40%, 93.09%, and 66.24%, respectively. Positive global Moran's I indices suggest that the spatial distribution of ULUE is positively correlated at basin level. Moreover, urban form metrics in the YRB demonstrated significant regional differences from 1994 to 2018. The regression results showed irregular urban form can negatively impact ULUE while compact and aggregated urban forms can improve ULUE under low carbon constrains. In addition, there are both positive and negative correlations between urban sprawl and ULUE in different regions. Today's choices on urban form can restrict the development pattern of cities and lock in pathways of carbon emissions in the future. Based on the findings in this study, the government should pursue optimal city sizes, avoid scattered patterns and aim for compact urban form.

Continuous EEG detection of DCI and seizures following aSAH: a systematic review.

Objective: Delayed cerebral ischemia (DCI) and seizures are dreaded neurological complications following aneurysmal subarachnoid hemorrhage (SAH). They may lead to severe mortality or mobility. Continuous electroencephalography (cEEG) is quite useful for secondary brain injury monitoring. This study aims to assess the effectiveness and sensitivity of cEEG in detecting DCI and seizures.Methods: Reports published up to July 31, 2018 were retrieved from PubMed, Medline, Cochrane and EMABSE. A systematic review was carried out on eligible studies.Results: 1. DCI was diagnosed in 20%-62% of aSAH patients, which was detected by a decrease on alpha/delta ratio; DCI can be predictive from 7 h to 1.9 days prior to standard diagnosis by CT, TCD and MRI. 2. Nonconvulsive status epilepticus (NCSE) occurred in 2.9%-30.8% of aSAH patients, and nonconvulsive seizures (NCSZ) in 6-23%; poor outcomes (high disability and mortality) were associated with NCSE.Conclusions: cEEG is an effective monitoring tool for early detection of DCI and seizures, which may help to improve the diagnostic accuracy and provide early treatment for patients with aSAH.

Anti-inflammatory diterpenes from the fruits of Vitex trifolia L. var. simplicifolia Cham.

Two new labdane-type diterpenes, named viterotulin C (1) and vitexilactone D (2), together with five known diterpenes (3-7), were isolated from the fruits of Vitex trifolia L. var. simplicifolia Cham. Their structures were elucidated by detailed analysis of spectroscopic data. All the compounds were evaluated for their inhibitory effects on nuclear factor-kappa B (NF-κB) pathway in HEK 293 cell line. These compounds presented inhibition on TNF-α-induced NF-κB activation, with inhibition rates ranging from 42.52 ± 10.69% to 68.86 ± 10.76% at the concentration of 50 µM.

A Beta/ZSM-22 Zeolites-Based-Mixed Matrix Solid-Phase Dispersion Method for the Simultaneous Extraction and Determination of Eight Compounds with Different Polarities in Viticis Fructus by High-Performance Liquid Chromatography.

Viticis Fructus (VF) was named Manjingzi as a commonly used traditional Chinese medicine (TCM) targeting various pains and inflammation for more than 2000 years. To guarantee the quality of Viticis Fructus, a simple, quick and eco-friendly Beta/ZSM-22 zeolites-based-mixed matrix solid-phase dispersion method (B/Z-MMSPD) was established for simultaneous extraction and determination of eight compounds (two phenolic acids, two iridoid glycosides, vanillin and three flavonoids) with different polarities from Viticis Fructus by high performance liquid chromatography coupled with a diode array detector (HPLC-DAD). Beta and ZSM-22 were mixed as the sorbent. Water, tetrahydrofuran and methanol were blended with certain ratio as the eluent. Several parameters including types of sorbents, mass ratio of Beta to ZSM-22, mass ratio of matrix to sorbent, grinding time, types, concentration and volume of eluent were optimized. The recoveries of eight analytes were within the range of 95.0%-105% (RSDs ≤ 4.13%). The limits of detection and limits of quantitation ranged from 0.5 to 5.5 µg/g and from 1.5 to 16 µg/g, respectively. Compared to the traditional extract methods, it was a simple, rapid, efficient and green method. The results demonstrated that a simple, rapid, efficient and green B/Z-MMSPD was developed for the simultaneous extraction and determination of eight target analytes with different polarities for quality control of Viticis Fructus.

Discovery of Alternative Producers of the Enediyne Antitumor Antibiotic C-1027 with High Titers.

The potent cytotoxicity and unique mode of action make the enediyne antitumor antibiotic C-1027 an exquisite drug candidate for anticancer chemotherapy. However, clinical development of C-1027 has been hampered by its low titer from the original producer Streptomyces globisporus C-1027. Here we report three new C-1027 alternative producers, Streptomyces sp. CB00657, CB02329, and CB03608, from The Scripps Research Institute actinomycetes strain collection. Together with the previously disclosed Streptomyces sp. CB02366 strain, four C-1027 alternative producers with C-1027 titers of up to 11-fold higher than the original producer have been discovered. The five C-1027 producers, isolated from distant geographic locations, are distinct Streptomyces strains based on morphology and taxonomy. Pulsed-field gel electrophoresis and Southern analysis of the five C-1027 producers reveal that their C-1027 biosynthetic gene clusters (BGCs) are all located on giant plasmids of varying sizes. The high nucleotide sequence similarity among the five C-1027 BGCs implies that they most likely have evolved from a common ancestor.

Determination of ecliptasaponin A in rat plasma and tissues by liquid chromatography-tandem mass spectrometry.

A sensitive, rapid and specific high-performance liquid chromatography tandem mass spectrometry method (HPLC-MS/MS) was developed to determine ecliptasaponin A in rat plasma and tissues after oral administration. Ginsenoside Rg1 was used as the internal standard (IS). The plasma and tissues samples were prepared by liquid-liquid extraction with ethyl acetate and separated on an Eclipse Plus C18 column (2.1 mm × 150 mm, 5 µm) at a flow rate of 0.4 mL/min using acetonitrile and water (containing 0.05% acetic acid) as the mobile phase. The tandem mass detection was carried out with eletrospray ionization in negative mode. Quantification was performed by using multiple reaction monitoring (MRM), which monitored the fragmentation of m/z 633.4→587.2 for ecliptasaponin A and m/z 859.4→637.4 for the IS. The calibration curves obtained were linear in different matrices, and the lower limit of quantification (LLOQ) achieved was 0.5 ng/mL both for rat plasma and tissues. The intra- and inter-day precisions were below 15%. This method was successfully applied to pharmacokinetic study of ecliptasaponin A in rat plasma and tissues after oral administration. Copyright © 2015 John Wiley & Sons, Ltd.

A Review on the Terpenes from Genus Vitex.

The genus Vitex, which belongs to the Verbenaceae family, includes approximately 250 species. Some species of the genus Vitex have traditionally been used for the treatment of headaches, ophthalmodynia, coughs, asthma, premenopausal syndrome, etc. Chemical investigations indicate that the characteristic constituents of the genus Vitex are terpenes, and 210 of these compounds, including monoterpenoids, sesquiterpenoids, diterpenoids and triterpenoids, have been obtained from 12 species. Pharmacological studies had shown that these terpenes possess anti-inflammatory, antitumor, antibacterial, antioxidant activities, and so on. In this paper, the identity of these terpenes and their pharmacological effects are reviewed, which can provide references for further research regarding the chemistry and utilization of the Vitex species.

Thermosensitive in situ nanogel as ophthalmic delivery system of curcumin: development, characterization, in vitro permeation and in vivo pharmacokinetic studies.

CONTEXT: The nanogel combining cationic nanostructured lipid carriers (CNLC) and thermosensitive gelling agent could enhance preocular retention and ocular permeation capacity of curcumin (CUR). OBJECTIVE: The purpose of the study was to develop and characterize a thermosensitive ophthalmic in situ nanogel of CUR-CNLC (CUR-CNLC-GEL) and evaluate in vitro and in vivo properties of the formulations. MATERIALS AND METHODS: The physicochemical properties, in vitro release and corneal permeation, were evaluated. Ocular irritation and preocular retention capacity were also conducted. Finally, pharmacokinetic study in the aqueous humor was investigated by microdialysis technique. RESULTS: The solution-gel transition temperature of the optimized formulation diluted by simulated tear fluid was 34 ± 1.0 °C. The CUR-CNLC-GEL displayed zero-order release kinetics. The apparent permeability coefficient (Papp) and the area under the curve (AUC0→∞) of CUR-CNLC-GEL were 1.56-fold and 9.24-fold, respectively, than those of curcumin solution (CUR-SOL, p < 0.01). The maximal concentration (Cmax) was significantly improved (p < 0.01). The prolonged mean residence time (p < 0.01) indicated that CUR-CNLC-GEL is a controlled release formulation. DISCUSSION AND CONCLUSION: Those results demonstrated that CUR-CNLC-GEL could become a potential formulation for increasing the bioavailability of CUR in the aqueous humor by enhancing corneal permeation and retention capacity.

[Chemical Constituents from Caragana changduensis].

Objective: To study the chemical constituents of the red heartwood of the stems and roots of Caragana changduensis. Methods: The chemical constituents were isolated and purified by means of several column chromatographic techniques,and their structures were determined by spectroscopic methods. Results: Ten compounds were isolated and identified as kushenin( 1),( 6aR,11aR)-3-hydroxy-4,9-dimethoxy-pterocarpan( 2),(-)-4-methoxymaackiain( 3),(-)-homopterocarpin( 4),2,4-dimethoxybenzoic acid( 5),2-methoxy-4-ethoxybenzoic acid( 6),3-acetyl-oleanolic acid( 7),7-hydroxy-2,3-dimethylchromone( 8),liquiritigenin( 9),and ß-sitosterol( 10). Conclusion: Compounds 1,3,5,7,and 8 are obtained from this genus for the first time. All the compounds are obtained from this plant for the first time.

MicroRNA-340 Inhibits Esophageal Cancer Cell Growth and Invasion by Targeting Phosphoserine Aminotransferase 1.

BACKGROUND/AIMS: Emerging evidence indicates that microRNA (miR)-340 is downregulated in various human cancers, suggesting that it acts as a tumor suppressor. The aim of the present study was to evaluate the expression and role of miR-340 in human esophageal squamous cell carcinoma (ESCC). METHODS: The expression of miR-340 was examined in 64 paired ESCC and adjacent non-tumor tissues by quantitative real time PCR. The effects of miR-340 on ESCC cell proliferation and metastasis were examined by MTT and Matrigel invasion assays. Tumor growth was assessed by subcutaneous inoculation of cells into BALB/c nude mice. Targets of miR-340 were identified by bioinformatics and verified by luciferase reporter assays, quantitative real-time PCR, and western blotting. RESULTS: MiR-340 was significantly downregulated in ESCC tumor tissues compared to adjacent non-tumor tissues and in ESCC cell lines compared to esophageal endothelial cells. Overexpression of miR-340 inhibited ESCC cell growth, colony formation, and invasion, and tumor growth in a xenograft mouse model. PSAT1 was identified as a direct target of miR-340 and its ectopic expression partially reversed the miR-340 mediated inhibition of viability, invasion and EMT in ESCC cells. The expression of miR-340 was negatively correlated with that of PSAT1 in human ESCC samples. CONCLUSION: MiR-340 functions as a tumor suppressor by modulating the expression of PSAT1 and may contribute to the progression and invasiveness of ESCC.

Analysis of the Constituents in "Zhu She Yong Xue Shuan Tong" by Ultra High Performance Liquid Chromatography with Quadrupole Time-of-Flight Mass Spectrometry Combined with Preparative High Performance Liquid Chromatography.

"Zhu She Yong Xue Shuan Tong" lyophilized powder (ZSYXST), consists of a series of saponins extracted from Panax notoginseng, which has been widely used in China for the treatment of strokes. In this study, an ultra-high performance liquid chromatography with quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF/MS) combined with preparative high performance liquid chromatography (PHPLC) method was developed to rapidly identify both major and minor saponins in ZSYXST. Some high content components were removed through PHPLC in order to increase the sensitivity of the trace saponins. Then, specific characteristic fragment ions in both positive and negative mode were utilized to determine the types of aglycone, saccharide, as well as the saccharide chain linkages. As a result, 94 saponins, including 20 pairs of isomers and ten new compounds, which could represent higher than 98% components in ZSYXST, were identified or tentatively identified in commercial ZSYXST samples.

[Isoflavonoids from Caragana changduensis and their nitric oxideinhibitory activities].

Ten isoflavonoids were isolated from the heartwoods of Caragana changduensis Lion f. by means of various column chromatographic techniques. Based on the detailed spectral data analysis (MS and NMR), as well as comparison with the literatures, their chemical structures were determined as 7,2'-dihydroxy-8,4'-dimethoxyisoflavone (1), 4'-hydroxy-7,3'-dimethoxyisoflavone (2), 5, 7, 4'-trihydroxy-2',5'-dimethoxyisoflavone (3), prunetin (4), afrormosin (5), odoratin (6), genistein (7), texasin (8), pratensein (9), and 6,7,3'-trihydroxy-4'-methoxyisoflavone (10). Among them, compounds 1-3 and 9-10 were isolated from the Caragana genus for the first time. All the compounds were obtained from this species for the first time. In the preliminary assays, compounds 1, 2, 6, and 7 possessed significant inhibitory effects on NO production, with IC50 values of 48.12, 25.32, 62.71, 43.59 µmol x L(-1), respectively.

A practical strategy to discover new antitumor compounds by activating silent metabolite production in fungi by diethyl sulphate mutagenesis.

Many fungal biosynthetic pathways are silent in standard culture conditions, and activation of the silent pathways may enable access to new metabolites with antitumor activities. The aim of the present study was to develop a practical strategy for microbial chemists to access silent metabolites in fungi. We demonstrated this strategy using a marine-derived fungus Penicillium purpurogenum G59 and a modified diethyl sulphate mutagenesis procedure. Using this strategy, we discovered four new antitumor compounds named penicimutanolone (1), penicimutanin A (2), penicimutanin B (3), and penicimutatin (4). Structures of the new compounds were elucidated by spectroscopic methods, especially extensive 2D NMR analysis. Antitumor activities were assayed by the MTT method using human cancer cell lines. Bioassays and HPLC-photodiode array detector (PDAD)-UV and HPLC-electron spray ionization (ESI)-MS analyses were used to estimate the activated secondary metabolite production. Compounds 2 and 3 had novel structures, and 1 was a new compound belonging to a class of very rare natural products from which only four members are so far known. Compounds 1-3 inhibited several human cancer cell lines with IC50 values lower than 20 µM, and 4 inhibited the cell lines to some extent. These results demonstrated the effectiveness of this strategy to discover new compounds by activating silent fungal metabolic pathways. These discoveries provide rationale for the increased use of chemical mutagenesis strategies in silent fungal metabolite studies.

A review of recent research progress on the astragalus genus.

Astragalus L., is one of the largest genuses of flowering plants in the Leguminosae family. Roots of A. membranaceus Bge. var. mongholicus (Bge.) Hsiao, A. membranaceus (Fisch.) Bge. and its processed products are listed in the China Pharmacopeia for "qi deficiency" syndrome treatment. However, more and more researches on other species of Astragalus have been conducted recently. We summarize the recent researches of Astragalus species in phytochemistry and pharmacology. More than 200 constituents, including saponins and flavonoids, obtained from 46 species of Astragalus genus were collected for this article. In pharmacological studies, crude extracts of Astragalus, as well as isolated constituents showed anti-inflammatory, immunostimulant, antioxidative, anti-cancer, antidiabetic, cardioprotective, hepatoprotective, and antiviral activities. The goal of this article is to provide an overview of chemical and pharmacological studies on the Astragalus species over the last 10 years, which could be of value to new drug or food supplement research and development.

A strategy of accuracy quantification by extending the concentration monitoring coverage based on online double collision energy of ultra-high performance liquid chromatography tandem mass spectrometry: The pharmacokinetics of Toddalia asiatica as a case study.

To facilitate the safety, efficacy and rationality of clinical application of traditional Chinese medicines (TCMs), pharmacokinetic research played an indispensable role. The key challenge during pharmacokinetic investigation lied at the substantial fluctuation of compound concentrations in the plasma over the course of absorption. Taking the pharmacokinetics of six compounds after administration of Toddalia asiatica (TA) as an example, an efficient strategy was established by introducing the online double collision energy (ODCE) into the quantification process applying ultra-high performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS). During the analytical program, double collision energy (DCE) was optimized to establish the dual calibration curve (DCC) with large concentration monitoring coverage (CMC) for meeting the wide content range of certain target compounds. Method validation test was performed in terms of linearity, precision, sensitivity, matrix effect, recovery, etc. The results displayed that the CMC of todarolactone with high exposure in plasma was extended from 1.25-2,500 ng/mL to 1.25-125,000 ng/mL. Furthermore, a rapid UHPLC-MS/MS method integrated with ODCE was successfully applied to the determination of six compounds in rat plasma, revealing an extremely high plasma concentration of todarolactone (16,662 ng/mL). This strategy could expand the range of quantification while retaining extraordinary sensitivity. Consequently, it could be a fit-for-purpose strategy to quantify compounds over a wide concentration range for in vivo process monitoring.

A review of the ethnopharmacology, phytochemistry, pharmacology, pharmacokinetics and quality control of Paeonia lactiflora Pall.

ETHNOPHARMACOLOGICAL RELEVANCE: Paeonia lactiflora Pall. (called Shaoyao in China) is a common herb cultivated all over the world. In some Asian and European countries, such as China, Japan, South Korea and Britain, P. lactiflora has a long history of ethnomedical uses, which is widely used to relieve pain, treat gynecological diseases, anti-infection and so on. It is attributed to the extensive pharmacological activities of total glucosides of P. lactiflora. Up to now, it is still commonly used in clinical medicine. THE AIM OF THE REVIEW: The paper aims to make a comprehensive review on the botanical characterization and distribution, ethnopharmacology, phytochemistry, biosynthesis pathway, pharmacology, pharmacokinetics and quality control of P. lactiflora, so as to provide new insights and scientific evidence for the subsequent research. MATERIALS AND METHODS: The information of P. lactiflora was obtained from books related to traditional Chinese medicine and electronic databases, including Scifinder, PubMed, Web of Science, CNKI and Google Scholar. RESULTS: P. lactiflora is a kind of herb with a long history and it is used for medicine, food and ornamental, and shows high utilization value. There are 200 compounds have been identified from it, including terpenoids, flavonoids, polyphenols, organic acids and others, among those paeoniflorin, a monoterpenoid glycoside, has multiple activities and is currently the focus of pharmacological research. A great deal of pharmacological experiments supported the anti-inflammatory, anti-oxidant, hepatoprotective, neuroprotective, antibacterial, antitumor, dermatosis treating and other effects of P. lactiflora. In addition, evaluating the quality of P. lactiflora is essential to safe use of drug in humans. CONCLUSIONS: The chemical components of P. lactiflora are diverse and have a wide range of activities. Modern pharmacological studies have provided reliable evidence for the traditional efficacy, such as suppressing liver yang, regulating menstruation and relieving pain. However, there are still some problems to be solved, such as part of the pharmacological mechanism has not been clarified and the biosynthetic pathway of cage-like monoterpenoids remains poorly defined. In addition, further studies on compounds other than paeoniflorin are clearly warranted. It is hoped that P. lactiflora will serve the clinic better in the future.

Unveiling hepatotoxicity distinction of coumarin-related compounds from glycosides to aglycones in Fructus Psoraleae by integrating UPLC-Q-TOF-MS and high content analysis.

ETHNOPHARMACOLOGICAL RELEVANCE: Fructus Psoraleae (FP), the dried and ripe fruit of Cullen corylifolium (L.) Medik., is widely used due to its various clinical pharmacological effects, but its hepatotoxicity restricts its clinical application. So far, its hepatotoxic components and their underlying mechanism have not been systematically elucidated. AIM OF THE STUDY: This study was undertaken to reveal the hepatotoxicity distinction of coumarin-related compounds from glycosides to aglycones in FP and elucidate their potential mechanism. METHODS: Rats were administrated with the aqueous extract of Fructus Psoraleae (AEFP), in which eight coumarin-related compounds were focused. Subsequently, compounds exposed in rats' livers were detected by UPLC-Q-TOF-MS, and the identified hepatotoxic compounds were evaluated to elaborate their possible mechanism by the aid of high content analysis (HCA). RESULTS: Eight coumarin-related compounds were identified, among which psoralenoside (PO), isopsoralenoside (IPO), psoralen (P), and isopsoralen (IP) were the principally exposed compounds in rats' livers. Furocoumarinic acid glucoside (FAG), (E)-3-(4-(((2S, 3R, 4S, 5S, 6R)-3,4,5-trihydroxy-6-(hydroxymethyl) tetrahydro-2H-pyran-2-yl) oxy) benzofuran-5-yl) acrylic acid (isofurocoumarinic acid glucoside, IFAG), furocoumarinic acid (FA), and (E)-3-(4-hydroxybenzofuran-5-yl) acrylic acid (isofurocoumarinic acid, IFA) were also detected in low abundance. P, IP, FA, and IFA were identified as the hepatotoxic compounds, while their glycosides were almost non-hepatotoxic. The HCA's results showed that hepatotoxic compounds disrupted the balance in reactive oxygen species (ROS), nuclear area, and mitochondrial membrane potential of HepG2 cells, leading to the occurrence of hepatotoxicity. CONCLUSIONS: P, IP, FA, and IFA were identified as hepatotoxic compounds, from which P and IP were proposed as the important risk components for hepatotoxicity. The conversion from glycosides to aglycones played an essential role in FP-induced hepatotoxicity.

Simultaneous determination of multiple components in rat plasma by UHPLC-sMRM for pharmacokinetic studies after oral administration of Qingjin Yiqi Granules.

As a Traditional Chinese Medicine prescription, Qingjin Yiqi Granules (QJYQ) provides an effective treatment for patients recovering from COVID-19. However, the pharmacokinetics characteristics of the main components of QJYQ in vivo are still unknown. An efficacious ultra-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) was developed and validated for the simultaneous determination of 33 components in rat plasma after oral administration of QJYQ. The plasma samples were precipitated with 400 µL methanol/acetonitrile (1/1, v/v) and analyzed in scheduled multiple reaction monitoring mode. The linear relationship of the 33 components was good (r > 0.9928). The lower limit of quantification for 33 components ranged from 0.4-60.5 ng/mL. The average recoveries and matrix effects of the analytes ranged from 72.9% to 115.0% with RSD of 1.4%-15.0%. All inter-day and intra-day RSDs were within 15.0%. After oral administration (3.15 g/kg), the validated approach was effectively applied to the pharmacokinetics of main components of QJYQ. Finally, fifteen main constituents of QJYQ with large plasma exposure were obtained, including baicalin, wogonoside, wogonin, apigenin-7-O-glucuronide, verbenalin, isoferulic acid, hesperidin, liquiritin, harpagide, protocatechuic acid, p-Coumaric acid, ferulic acid, sinapic acid, liquiritin apioside and glycyrrhizic acid. The present research lays a foundation for clarifying the therapeutic material basis of QJYQ and provides a reference for further scientific research and clinical application of QJYQ.