Revealing microbial community assembly patterns and succession process in the blackening process of black-odor water.

Human-imported pollutants could induce water black, changing microbial community structure and function. Employed 16S rRNA high-throughput sequencing, field-scale investigations and laboratory-scale experiments were successively conducted to reveal mechanistic insights into microbial community assembly and succession of black-odor waters (BOWs). In the field-scale investigation, livestock breeding wastewater (56.7 ± 3.2%) was the most critical microbial source. Moreover, fermentation (27.1 ± 4.4%) was found to be the dominant function. Combined with laboratory experiments, the critical environmental factors, such as total organic carbon (30-100 mg/L), ammonia nitrogen (2.5-9 mg/L), initial dissolved oxygen (2-8 mg/L) and chlorophyll a (0-90 mg/L), impacted the intensity of blackening. The differentiation of ecological niches within the microbial community played a significant role in driving the blackening speed. In laboratory-scale experiments, the microbial ecological niche determined the blackening timing and dominations of the stochastic processes in the microbial assembly process (88 - 51%). The three stages, including the anaerobic degradation stage, blackening stage and slow recovery stage, were proposed to understand the assembly of the microbial communities. These findings enhance our understanding of microorganisms in BOWs and provide valuable insights for detecting and managing heavily organic polluted waters.

Sedentary behavior patterns and bone health among overweight/obesity older women: a cross-sectional study.

Background: Recent studies have indicated an association between sedentary behavior (SB), particularly patterns of SB, and bone health. However, it remains uncertain how different patterns of SB in overweight/obesity older women impact their bone health. This study aimed to investigate the association between objectively measured SB patterns and bone health in Chinese community-dwelling overweight/obesity older women. Methods: Cross-sectional data were obtained from a baseline survey of Physical Activity and Health in Older Women Study. Quantitative ultrasound was used to measure speed of sound (SOS), broadband ultrasound attenuation (BUA), bone quality index (BQI) and T value to evaluate bone health. SB patterns were measured using triaxial accelerometers, including sedentary time in SB bouts of ≥ 10, 30, and 60 min, number of SB bouts ≥ 10, 30, and 60 min. Multiple linear regression was used to examine the associations of different SB patterns with bone health. Results: After adjusting for confounders, sedentary time in SB bouts ≥ 60 min, number of SB bouts ≥ 60 min were significantly associated with bone health, with a lower SOS [ß = -2.75, 95% confidence interval (CI): -4.96 to -0.53, P = 0.015], BUA (ß = -1.20, 95% CI: -2.14 to -0.26, P = 0.013), BQI (ß = -1.56, 95% CI: -2.63 to -0.49, P = 0.004), T value (ß = -0.08, 95% CI: -0.14 to -0.03, P = 0.004) per 60 min increase of sedentary time in SB bouts ≥ 60 min, and a lower SOS (ß = -3.97, 95% CI: -7.54 to -0.40, P = 0.029), BUA (ß = -1.80, 95% CI: -3.44 to -0.16, P = 0.031), BQI (ß = -2.28, 95% CI: -4.08 to -0.47, P = 0.014) and T value (ß = -0.12, 95% CI: -0.22 to -0.03, P = 0.013) per bout increase of SB bouts ≥ 60 min, respectively. Conclusion: Limiting the duration of prolonged sedentary bouts and minimizing the occurrence of number of SB bouts ≥ 60 min could be essential in bone health management, especially for those older people who are overweight/obesity.

Appling fluorescence spectroscopy with absolute principal component coefficient to explore dynamic migration of DOM fractions from an urbanized river during torrential rainfall.

This research delves into the dynamic interplay between urbanization and the characteristics of Dissolved Organic Matter (DOM) in the Anyang River, particularly under the stress of torrential rain. The motivation stems from a critical need to decipher how urban landscapes influence water quality, focusing on the intricate transformations and movements of DOM. Employing advanced fluorescence spectroscopy techniques like Excitation-Emission Matrices (EEM) and Parallel Factor Analysis (PARAFAC), the study meticulously differentiates DOM compositions in urban and agricultural settings. It unveils a pronounced distinction, with urban streams showing elevated proteinaceous DOM from wastewater, contrasting with the humic substances prevalent in agricultural runoff. The analysis also captures how intense rainfall events catalyze significant shifts in DOM profiles, thereby emphasizing the need for tailored water quality management strategies in urbanized catchments. This comprehensive approach not only bridges gaps in understanding the urban impact on riverine ecosystems but also sets a foundation for future research and policy development in the face of escalating environmental changes.

Mitigated N2O emissions from submerged-plant-covered aquatic ecosystems on the Changjiang River Delta.

Submerged plants affect nitrogen cycling in aquatic ecosystems. However, whether and how submerged plants change nitrous oxide (N2O) production mechanism and emissions flux remains controversial. Current research primarily focuses on the feedback from N2O release to variation of substrate level and microbial communities. It is deficient in connecting the relative contribution of individual N2O production processes (i.e., the N2O partition). Here, we attempted to offer a comprehensive understanding of the N2O mitigation mechanism in aquatic ecosystems on the Changjiang River Delta according to stable isotopic techniques, metagenome-assembly genome analysis, and statistical analysis. We found that the submerged plant reduced 45 % of N2O emissions by slowing down the dissolved inorganic nitrogen conversion velocity to N2O in sediment (Vf-[DIN]sed). It was attributed to changing the N2O partition and suppressing the potential capacity of net N2O production (i.e., nor/nosZ). The dominated production processes showed a shift with increasing excess N2O. Meanwhile, distinct shift thresholds of planted and unplanted habitats reflected different mechanisms of stimulated N2O production. The hotspot zone of N2O production corresponded to high nor/nosZ and unsaturated oxygen (O2) in unplanted habitat. In contrast, planted habitat hotspot has lower nor/nosZ and supersaturated O2. O2 from photosynthesis critically impacted the activities of N2O producers and consumers. In summary, the presence of submerged plants is beneficial to mitigate N2O emissions from aquatic ecosystems.

Maintaining the working state of firefighters by utilizing self-concept clarity as a resource.

The working state of firefighters is important for their own safety as well as that of the general public. The purpose of this study is to investigate the correlations between self-concept clarity, resilience, work engagement, and job burnout among firefighters, as well as the impacts of self-concept clarity and resilience as resources that can maintain their working state. Based on data from 2,156 firefighters, analysis showed that self-concept clarity was negatively associated with job burnout and positively associated with work engagement. The results also showed that self-concept clarity had a direct effect on job burnout and work engagement, and an indirect effect by improving the firefighters' resilience. Maintaining and improving their self-concept clarity and resilience promises to be an effective strategy for guaranteeing the working state of firefighters.

The ecological role of microbiome at community-, taxonomic - and genome-levels in black-odorous waters.

Black-odorous waters (BOWs) are heavily polluted waters where microbial information remains elusive mechanistically. Based on gene amplicon and metagenomics sequencing, a comprehensive study was conducted to investigate the microbial communities in urban and rural BOWs. The results revealed that microbial communities' assembly in urban and rural BOWs was predominantly governed by stochastic factors at the community level. At the taxonomic level, there were 62 core species (58.48%) in water and 207 core species (44.56%) in sediment across urban and rural areas. Notably, significant differences were observed in the functional genetic composition of BOWs between urban and rural areas. Specifically, rural areas exhibited an enhanced abundance of genes involved in nitrogen fixation, Fe2+ transport, and sulfate reduction. Conversely, urban areas showed higher abundances of some genes associated with carbon fixation, nitrification and denitrification. A sulfur-centered ecological model of microbial communities was constructed by integrating data from the three levels of analysis, and 14 near-complete draft genomes were generated, representing a substantial portion of the microbial community (35.04% in rural BOWs and 29.97% in urban BOWs). This research provides significant insights into the sustainable management and preservation of aquatic ecosystems affected by BOWs.

Association of daily step counts and step intensity with fear of falling in Chinese community-dwelling older women.

This study aimed to investigate the relationship between objectively measured step counts, step intensity and FOF in community-dwelling older Chinese women. Cross-sectional analyses were conducted on pooled data (n = 1101) from the baseline survey of the Physical Activity and Health Study (PAHIOWS). Step counts and step intensity were measured using wGT3X-BT accelerometers, and FOF was categorized into two levels (low and high) for logistic regression analysis. Higher step counts were significantly and negatively associated with FOF; however, after adjusting for step intensity, the association was no longer statistically significant. On the contrary, higher step intensity was negatively associated with FOF independent of step counts. This finding may provide new insights into the physical activity management of FOF in older women.

Characterizing dissolved organic matter and bacterial community interactions in a river network under anthropogenic landcover.

Anthropogenic landcover could rise nutrient concentrations and impact the characteristics and bioavailability of dissolved organic matter (DOM) in a river network. Exploring the interactions between DOM and microbials might be conducive to revealing biogeochemistry behaviors of organic matter. In this study, synchronous fluorescence spectra (SFS) with Gaussian band fitting and two-dimensional correlation spectroscopy (2D-COS) were employed to identify DOM fractions and reveal their interactions with bacterial communities. DOM was extracted from a river network under eco-agricultural rural (RUR), eco-residential urban (URB), eco-economical town (TOW), and eco-industrial park (IND) regions in Jiashan Plain of eastern China. The overlapping peaks observed in the SFS were successfully separated into four fractions using Gaussian band fitting, i.e., tyrosine-like fluorescence (TYLF), tryptophan-like fluorescence (TRLF), microbial humic-like fluorescence (MHLF), and fulvic-like fluorescence (FLF) materials. Across all four regions, TRLF (44.79% ± 7.74%) and TYLF (48.09% ± 8.85%) were the dominant components. Based on 2D-COS, variations of TYLF and TRLF were extremely larger than those of FLF in RUR-TOW. However, in URB-IND, the former exhibited lower variations compared to the latter. These suggested that FLF be likely derived continuously from lignin and other residue of terrestrial plant origin along the river network, and TYLF and TRLF be originated discontinuously from domestic wastewater in RUR-TOW. By high-throughput sequenced OTUs, the number of organisms in RUR-TOW could be higher than those in URB-IND, while genes associated with carbohydrate metabolism were lower in former than those in the latter. According to co-occurrence networks, microbes could promote the production of TYLF and TRLF in RUR-TOW. In contrast, microbial communities in URB-IND might contribute to decompose FLF. The obtained results could not only reveal interactions between DOM fractions and bacterial communities in the river network, but this methodology may be applied to other water bodies from different landscapes.

Mechanistic insights into dissolved organic matter-driven protistan and bacterial community dynamics influenced by vegetation restoration.

Vegetation restoration projects can not only improve water quality by absorbing and transferring pollutants and nutrients from non-vegetation sources, but also protect biodiversity by providing habitat for biological growth. However, the mechanism of the protistan and bacterial assembly processes in the vegetation restoration project were rarely explored. To address this, based on 18 S rRNA and 16 S rRNA high-throughput sequencing, we investigated the mechanism of protistan and bacterial community assembly processes, environmental conditions, and microbial interactions in the rivers with (out) vegetation restoration. The results indicated that the deterministic process dominated the protistan and bacterial community assembly (94.29% and 92.38%), influenced by biotic and abiotic factors. For biotic factors, microbial network connectivity was higher in the vegetation zone (average degree = 20.34) than in the bare zone (average degree = 11.00). For abiotic factors, the concentration of dissolved organic carbon ([DOC]) was the most important environmental factor affecting the microbial community composition. [DOC] was lower significantly in vegetation zone (18.65 ± 6.34 mg/L) than in the bare zone (28.22 ± 4.82 mg/L). In overlying water, vegetation restoration upregulated the protein-like fluorescence components (C1 and C2) by 1.26 and 1.01-folds and downregulated the terrestrial humic-like fluorescence components (C3 and C4) by 0.54 and 0.55-folds, respectively. The different DOM components guided bacteria and protists to select different interactive relationships. The protein-like DOM components led to bacterial competition, whereas the humus-like DOM components resulted in protistan competition. Finally, the structural equation model was established to explain that DOM components can affect protistan and bacterial diversity by providing substrates, facilitating microbial interactions, and promoting nutrient input. In general, our study provides insights into the responses of vegetation restored ecosystems to the dynamics and interactives in the anthropogenically influenced river and evaluates the ecological restoration performance of vegetation restoration from a molecular biology perspective.

[Simultaneous determination of 22 antibiotics in environmental water samples by solid phase extraction-high performance liquid chromatography-tandem mass spectrometry].

The widespread and frequent use of antibiotics to treat diseases or encourage animal growth has resulted in their persistence and accumulation in water, soil, and sediments. As a typical emerging pollutant in the environment, antibiotics have become an important research focus in recent years. Antibiotics are commonly found at trace levels in water environments. Unfortunately, the determination of various types of antibiotics, all of which exhibit different physicochemical properties, remains a challenging endeavor. Thus, developing pretreatment and analytical techniques to achieve the rapid, sensitive, and accurate analysis of these emerging contaminants in various water samples is an essential undertaking.In this paper, a solid phase extraction-high performance liquid chromatography-tandem mass spectrometry (SPE-HPLC-MS/MS) method for the simultaneous determination of 22 antibiotics including 4 penicillins, 12 quinolones and 6 macrolides in environmental water samples was developed. Based on the characteristics of the screened antibiotics and sample matrix, the pretreatment method was optimized, focusing on the SPE column, pH of the water sample, and amount of ethylene diamine tetra-acetic acid disodium (Na2EDTA) added to the water sample. Prior to extraction, a 200 mL water sample was added with 0.5 g of Na2EDTA and pH-adjusted to 3 using sulfuric acid or sodium hydroxide solution. Water sample enrichment and purification were achieved using an HLB column. HPLC separation was carried out on a C18 column (100 mm×2.1 mm, 3.5 µm) via gradient elution with a mobile phase composed of acetonitrile and 0.15% (v/v) formic acid aqueous solution. Qualitative and quantitative analyses were performed on a triple quadrupole mass spectrometer in multiple reaction monitoring mode using an electrospray ionization source. The results showed correlation coefficients greater than 0.995, indicating good linear relationships. The method detection limits (MDLs) and limits of quantification (LOQs) were in the ranges of 2.3-10.7 ng/L and 9.2-42.8 ng/L, respectively. The recoveries of target compounds in surface water at three spiked levels ranged from 61.2% to 157%, with relative standard deviations (RSDs) of 1.0%-21.9%. The recoveries of target compounds in wastewater at three spiked levels were 50.1%-129%, with RSDs of 1.2%-16.9%. The method was successfully applied to the simultaneous determination of antibiotics in reservoir water, surface water, sewage treatment plant outfall, and livestock wastewater. Most of the antibiotics were detected in watershed and livestock wastewater. Lincomycin was detected in 10 surface water samples, with a detection frequency of 90%, and ofloxacin showed the highest contents (127 ng/L) in livestock wastewater. Therefore, the present method exhibits excellent performance in terms of MDLs and recoveries compared with previously reported methods. The developed method presents the advantages of small water sample volumes, wide applicability, and fast analysis times; thus, it can be considered a rapid, efficient, and sensitive analytical method with excellent potential for monitoring emergency environmental pollution. The method could also provide a reliable reference for formulating antibiotic residue standards. The results provide strong support for and an improved understanding of the environmental occurrence, treatment, and control of emerging pollutants.

Applying synchronous fluorescence spectra with Gaussian band fitting and two-dimensional correlation to characterize structural composition of DOM from soils in an aquatic-terrestrial ecotone.

Dissolved organic matter (DOM), the primary participant of carbon and nitrogen cycle, has a great impact on the behavior and fate of organic pollutants and heavy metals in eutrophic lakes. The dynamic spectral properties of DOM fractions were revealed in an aquatic-terrestrial ecotone under the different types of land use. Composite soil samples of different depths (0-20, 20-40 and 40-60 cm) were collected from four different land uses along a disturbed-impact gradient in Taihu Lake, China, i.e., grassland (GRL), forest land (FOL), paddy field (PAF), and vegetable field (VEF). DOM mainly consisted of tyrosine-like material (TYLF), tryptophan-like material (TRLF), microbial humic-like material (MHLF), fulvic-like material (FLF) and humic-like material (HLF) within all soil profiles, where TRLF was the dominant component (61.30 %) using synchronous fluorescence spectroscopy (SFS) combined with principal component analysis and Gaussian band fitting. Based on two-dimensional correlation spectroscopy with SFS and Fourier transform infrared, the variation order of DOM fractions was FLF → MHLF → HLF → TRLF → TYLF within the GRL soil profile, and MHLF exhibited an oppositive change with aliphatic OH and amide I in protein. The order of DOM fractions was MHLF → FLF → HLF → TYLF → TRLF within the FOL soil profile, and the change trend of MHLF remained oppositive with aliphatic OH and CO in ester. The order of DOM within the PAF soil profile fractions was TRLF → MHLF → HLF → TYLF → FLF, and changing trends of TYLF were oppositive to aliphatic OH, CH bending vibration, CH bending vibration and CO in ester. The order of DOM fractions was HLF → TYLF → TRLF → FLF → MHLF within the VEF soil profile, where the changing trend TYLF remained oppositive to aliphatic OH, CH deformations in lignin and aliphatic group and amide I in protein. This study may provide important support for alleviating lake water eutrophication or pollution.

Turnover of dissolved organic carbon fuels nocturnal CO2 emissions from a headwater catchment reservoir, Southeastern China: Effects of ecosystem metabolism on source partitioning of CO2 emissions.

Dam reservoirs in headwater catchments, as critical zones for their proximity to terrestrial sources, play important roles in dissolved organic carbon (DOC) cycling. However, the effects of ecosystem metabolism (EM) on DOC cycling are not well known. Here, in-situ diurnal and monthly observations were conducted to measure EM (including gross primary production (GPP), ecosystem respiration (ER) and heterotrophic respiration (HR)), DOC turnover and CO2 emissions in a headwater catchment reservoir in Southeastern China in 2020. Our study showed the nocturnal CO2 emission rate was about twice as high as in daytime, and was strongly driven by EM. The values for DOC turnover velocity ranged from 0.10 to 1.59 m/day, and the average DOC turnover rate was 0.13 day-1, with the average removal efficiency of 12%. The contribution of respired DOC to daily CO2 emissions ranged from 17% to 61%. The accumulated efficiencies were estimated to be 13% for the selected 15 reservoirs throughout the Changjiang River network, corresponding to about 0.34 Tg C/year of the respired DOC. The modified CO2 flux was 0.75 Tg C/year, and respired DOC accounted for about 45% of total emitted CO2 from the 15 larger reservoirs. Our research emphasizes the necessity of incorporating the effects of EM into studies of reservoir DOC removal and CO2 emissions.

Enhanced chemodiversity, distinctive molecular signature and diurnal dynamics of dissolved organic matter in streams of two headwater catchments, Southeastern China.

Dissolved organic matter (DOM) is a complicated assembly of organic molecules, including thousands of molecules with various structures and properties. However, how the stream DOM sources respond to carbon compositions and the transformation processes remains unclear. In this study, the chemical characteristics and spectral and mass spectrometry (FT-ICR MS) of DOM were analyzed. Six sampling points of headwater stream (HWSs) were sampled, and an effluent polluted stream (WSR) and a main stream of the Changjiang River (DT) were also sampled for comparison. In situ degradation experiments and FT-ICR MS analysis were also performed to observe the dynamic processes of DOM in HWS. The results showed that the anthropogenic markers of sewage (i.e. sulfur (S) compounds and marker from antibiotics and estrogen) in HWS were higher than those in DT. The molecular weight decreased while the degradation products (S-containing compounds and unsaturated compounds (HU)) increased after in situ degradation due to the influence of both the photodegradation and biodegradation process. In addition, the KMD plots showed that the DOM homologue intensities in range 400-600 Da changed significantly after demethylation by biodegradation. The components of highly refractory substances and the degradation degree of DOM in DT was higher than that in HWS. We extracted the refractory DOM pool in HWS, which was mainly small molecular with molecular weights < 600 Da. These molecular will be difficult to remove in traditional drinking water treatment processes and easily produced disinfection byproducts (DBPs). This study emphasized the necessity of identifying the sources and transformation processes of DOM in HWS and clarified the types and characteristics of DOM that should be considered in future drinking water treatment.

Dominance of nitrous oxide production by nitrification and denitrification in the shallow Chaohu Lake, Eastern China: Insight from isotopic characteristics of dissolved nitrous oxide.

In recent decades, most lakes in Eastern China have suffered unprecedented nitrogen pollution, making them potential "hotspots" for N2O production and emission. Understanding the mechanisms of N2O production and quantifying emissions in these lakes is essential for assessing regional and global N2O budgets and for mitigating N2O emissions. Here, we measure isotopic compositions (δ15N-N2O and δ18O-N2O) and site preference (SP) of dissolved N2O in an attempt to differentiate the relative contribution of N2O production processes in the shallow, eutrophic Chaohu Lake, Eastern China. Our results show that the bulk isotope ratios for δ15N-N2O, δ18O-N2O, and SP were 5.8 ±â€¯3.9‰, 29.3 ±â€¯13.4‰, and 18.6 ±â€¯3.2‰, respectively. More than 76.8% of the dissolved N2O was produced via microbial processes. Findings suggest that dissolved N2O is primarily produced via nitrification (between 27.3% and 48.0%) and denitrification (between 31.9% and 49.5%). In addition, isotopic data exhibit significant N2O consumption during denitrification. We estimate the average N2O emission rate (27.5 ±â€¯26.0 µg N m-2 h-1), which is higher than that from rivers in the Changjiang River network (CRN). We scaled-up the regional N2O emission (from 1.98 Gg N yr-1 to 4.58 Gg N yr-1) using a N2O emission factor (0.51 ±â€¯0.63%) for shallow lakes in the middle and lower region of the CRN. We suggest that beneficial circumstances for promoting complete denitrification may be helpful for reducing N2O production and emissions in fresh surface waters.

Enhanced nitrogen imbalances in agroecosystems driven by changing cropping systems in a coastal area of eastern China: from field to watershed scale.

Agricultural activities exacerbate nitrogen (N) imbalances in the agroecosystem by disturbing the N inputs and outputs, yet the influence of changes in cropping systems on the N balance of agroecosystems remains unclear. In this study, at the field scale, we calculated the N balance of four cropping systems, (1) traditional crops with traditional crop rotation (G-G), (2) vegetables with traditional crop rotation (V-G), (3) vegetables with vegetable rotation (V-V), and (4) greenhouse vegetables (GHV); then analyzed the influence of changes in cropping systems from 1995 to 2015 on the N balances in the agroecosystems in sub-watersheds of the Dagu River. The results indicate that N balances were higher in GHV, V-V, and V-G than G-G, due to significantly higher inputs of N fertilizers and lower N use efficiency (NUE) in vegetable cultivation compared to traditional crops. Driven by economic benefits between 1995 and 2015, V-G, V-V, and GHV replaced G-G in a considerable number of cultivation areas in the sub-watersheds. These changes resulted in an increase of 109.9-170.1% in the N balance in the agroecosystem in the sub-watersheds between 1995 and 2015. In the entire watershed, the total N surplus contribution by V-V, V-G, and GHV increased from 39.3% to 79.1% between 1995 and 2015. These findings suggest that increased vegetable cultivation contributed to the increased risk of N pollution in agricultural production. Thus, there should be a focus on the management of cropping systems to control N loss from agricultural lands.

Nitrogen pollution and sources in an aquatic system at an agricultural coastal area of Eastern China based on a dual-isotope approach.

Nitrogen (N) pollution of water courses is a major concern in most coastal watersheds in eastern China with intensive agricultural production. We use hydrogeological and dual-isotopic approaches to analyze the N concentrations, pollution, transformations, and sources of surface water and groundwater in an agricultural watershed of the Jiaozhou Bay (JZB) area. Results showed that dissolved total N (DTN) concentrations in sub-rivers (SRs) ranged from 6.0 to 25.3 mg N L-1 in the dry season and 9.1-26.7 mg N L-1 in the wet season, which indicated a positive relationship with the percentages of agricultural land. Meanwhile, the dominant dissolved N species in SRs changed from nitrate (NO3-, 64-100%) to dissolved organic N (DON, 52-77%) from the dry season to the wet season and the increased DON concentrations showed a positive relationship with the planted proportions of vegetable production systems. The NO3- concentrations of groundwaters ranged from 10.6 to 121.4 mg N L-1, which were over the limit for drinking water by the World Health Organization. Isotopic analysis indicated that most NO3- originated from the microbiological conversion via nitrification, whereas the deletion of denitrification was insignificant in this area. The results of the stable isotope analysis in R mixing model showed the contributions of potential NO3- sources which were in order of manure fertilizers (20.6-69.0%) > soil organic matter (19.5-53.2%) > chemical fertilizers (5.5-34.3%) > atmospheric deposition (1.3-18.8%). This study suggests that the management of crop productions and reasonable manure fertilizer application should be implemented to protect the quality of aquatic systems in the JZB area.

A UHPLC-MS/MS method for simultaneous determination of twelve constituents from Erigeron breviscapus extract in rat plasma: Application to a pharmacokinetic study.

A rapid, sensitive and specific ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method was developed and validated to simultaneously determine the twelve major bioactive ingredients (neochlorogenic acid, chlorogenic acid, caffeic acid, cynarin, scopoletin, scutellarin, isochlorogenic acid A, apigenin-7-o-glucuronide, isochlorogenic acid C, scutellarein, luteolin, and apigenin) in rat plasma. Gallic acid and wogonoside were used as internal standards (IS1 and IS2). The plasma samples were pretreated and extracted by liquid-liquid extraction and protein precipitation with ethyl acetate-acetonitrile (95:5, v/v). Chromatographic separation was accomplished on Agilent ZORBAX RRHD Eclipse Plus C18 column (2.1mm×50mm, 1.8µm) utilizing 0.1% formic acid aqueous solution and acetonitrile as mobile phase under gradient conditions at a flow rate of 0.3mL·min-1. Mass spectrometric detection was performed in multiple reaction monitoring (MRM) mode using electrospray ionization (ESI) in positive and negative mode. The whole intra- and inter-day precision (as relative standard deviation) of all analytes were less than 11.03%, and the accuracy (as relative error) were in the range from -10.43% to 9.76% and from -10.14% to 10.33%. The lower limits of quantification (LLOQ) were 20, 3.0, 100, 7.0, 0.30, 2.0, 70, 1.0, 20, 30, 10, and 2.0ngmL-1 for neochlorogenic acid, chlorogenic acid, caffeic acid, cynarin, scopoletin, scutellarin, isochlorogenic acid A, apigenin-7-o-glucuronide, isochlorogenic acid C, scutellarein, luteolin, and apigenin, respectively. Extraction recovery, matrix effect and stability were found to be the required limits. This method was selective and sensitive for the investigation of the pharmacokinetics of twelve constituents following oral administration to research study about in Erigeron breviscapus of clinical practices for separately analytes on rats.