Triple Quadruple Mass Spectrometry Research Articles

Simultaneous and on-line detection of organic and heavy metal components in water using a novel nebulization-assisted injection plasma ionization triple quadruple mass spectrometry instrument

BackgroundThe global release of organic and heavy metal components into natural water bodies is a major concern for the environment and human health. The assessment of water quality relies on analyzing organic and heavy metal components qualitatively and quantitatively. Real-time identification of organic and metal components in water systems requires different analytical techniques due to varying measurement requirements. Thus, on-line detecting both organic compounds and heavy metals in ambient water systems simultaneously using a single instrumentation setup presents a significant challenge. ResultsIn this study, an analytical technique of nebulization-assisted injection plasma ionization mass spectrometry (NI-PIMS) was developed. This novel method enables the simultaneous detection of heavy metals and organic compounds in water system with high sensitivity, which has been demonstrated by the limit of quantification (LOQ) values below 1.0 μg/L for the three sterols (Enrofloxacin, ciprofloxacin, and clenbuterol) and three heavy metals (Pb, Ba, and Cd). Moreover, the method was successfully applied to rapidly analyze real water samples from urban and rural areas in China. The analytical results are available in less than 0.5 min, and only a few microliters of sample are required for each analysis. Significance and noveltyAs far as we know, this is the first report of on-line simultaneous analysis of organic compounds and heavy metals in a water system using a single mass spectrometry instrument. Compared to traditional methods, NI-PIMS demonstrates higher efficiency, sensitivity, no or lower sample preparation, and less sample consumption. The advancement and widespread use of this technology are expected to enhance the effectiveness of mass spectrometers, broaden the applications, and play an important role in complex sample analysis in fields such as atmospheric science, environmental science, and earth science.

Loss of nephric augmenter of liver regeneration facilitates acute kidney injury via ACSL4-mediated ferroptosis.

Ferroptosis, characterized by lipid accumulation in intracellular compartments, is related to acute kidney injury (AKI), but the mechanism remains obscure. In our previous study, the protective effect of augmenter of liver regeneration (ALR) on AKI was not fully clarified. In this study, we established an AKI mouse model by knocking out proximal tubule-specific ALR and an AKI cell model by inducing hypoxia, as well as enrolled AKI patients, to investigate the effects of ALR on ferroptosis and the progression of AKI. We found that ALR knockout aggravated ferroptosis and increased ROS accumulation and mitochondrial damage, whereas ALR overexpression attenuated ferroptosis through clearance of ROS and maintenance of mitochondrial morphology. Mechanistically, we demonstrated that ALR could directly bind to long-chain-fatty-acid-CoA ligase 4 (ACSL4) and further inhibit the expression of ACSL4 by interacting with certain regions. By resolution liquid chromatography coupled with triple quadruple mass spectrometry, we found that ALR could reduce the contents of polyunsaturated fatty acids, especially arachidonic acid. In addition, we showed that ALR binds to ACSL4 and attenuates oxylipin accumulation, exerting a protective effect against ferroptosis in AKI. Therefore, targeting renal ALR can attenuate ferroptosis and can offer a promising strategy for the treatment of AKI.

N6-Methyladenosine (m6A) Methylation Is Associated with the Immune Microenvironments in Acute Intracerebral Hemorrhage (ICH).

Researchers have recently found that N6-methyladenosine (m6A) is a type of internal posttranscriptional modification that is essential in mammalian mRNA. However, the features of m6A RNA methylation in acute intracerebral hemorrhage (ICH) remain unknown. To explore differential methylations and to discover their functions in acute ICH patients, we recruited three acute ICH patients, three healthy controls, and an additional three patients and healthy controls for validation. The m6A methylation levels in blood samples from the two groups were determined by ultrahigh-performance liquid chromatography coupled with triple quadruple mass spectrometry (UPLC-QQQ-MS). Methylated RNA immunoprecipitation sequencing (MeRIP-seq) was employed to identify differences in m6A modification, and the differentially expressed m6A-modified genes were confirmed by MeRIP-qPCR. We found no significant differences in the total m6A levels between the two groups but observed differential methylation peaks. Compared with the control group, the coding genes showing increased methylation following acute ICH were mostly involved in processes connected with osteoclast differentiation, the neurotrophin signaling pathway, and the spliceosome, whereas genes with reduced m6A modification levels after acute ICH were found to be involved in the B-cell and T-cell receptor signaling pathways. These results reveal that differentially m6A-modified genes may influence the immune microenvironments in acute ICH.

Identifying hepatoprotective mechanism and effective components of Yinchenzhufu decoction in chronic cholestatic liver injury using a comprehensive strategy based on metabolomics, molecular biology, pharmacokinetics, and cytology

Ethnopharmacological relevanceIn Traditional Chinese Medicine (TCM), cholestasis liver disease belongs to jaundice. Yinchenzhufu decoction (YCZFD) is a classic formula used for treating jaundice. Aim of the studyThis study was aimed to investigate the potential mechanism and effective components of YCZFD in chronic cholestatic liver injury (CCLI). Materials and methodsA chronic cholestatic mouse model induced by 3, 5-diethoxycarbonyl-1, 4-dihydroxychollidine was used to investigate the effect of YCZFD. Then, metabolomics was used to investigate the metabolites influenced by YCZFD. Serum and liver bile acid (BA) levels were measured using liquid chromatography coupled with triple quadruple mass spectrometry (LC-MS/MS), and the gene and protein expressions of BA transporters and metabolic enzymes were detected. Additionally, the pharmacokinetics of multiple components of YCZFD was explored to clarify the potential effective components. The effects of absorbed components of YCZFD on BA metabolism and transporter function, inflammation, and farnesoid X receptor (FXR) and pregnane X receptor (PXR) activation were analyzed using sandwich cultured rat hepatocytes, AML12 cells, and dual-luciferase receptor systems, respectively. ResultsYCZFD decreased the liver damage in chronic cholestatic mice. Serum metabolomics results indicated that the main pathways influenced by YCZFD involved primary BA biosynthesis and arachidonic acid metabolism. YCZFD upregulated the expression of FXR, PXR, and BA efflux transporters and the metabolic enzymes of liver tissues, promoting BA excretion and metabolism in cholestatic mice. Additionally, YCZFD downregulated the expression of genes and proteins of the toll-like receptor 4 (TLR4)/nuclear factor kappa-B (NF-κB) pathway and decreased liver inflammation. The pharmacokinetic study indicated that multiple components showed different pharmacokinetic properties. Among the absorbed components of YCZFD, multiple components activated the transcription of FXR and PXR, regulated BA transporters and metabolic enzyme function, and reduced the gene expression of TLR4 and NF-κB1. ConclusionYCZFD can ameliorate CCLI by promoting the excretion and metabolism of BAs and inhibiting inflammation via the TLR4/NF-κB signaling pathway. The multiple components of YCZFD could act on BA homeostasis regulation and anti-inflammation, exhibiting a combined effect against CCLI.

Two-Stage Supramolecular Self-Assembly-Directed Collagen-Peptide-Decorated Liposomal Complexes of Curcumin Microspheres with Enhanced Solubility and Bioavailability.

Green formulations of phytonutrients with enhanced solubility and bioavailability are of great significance in nutrition therapy. In the present contribution, we hypothesized that the collagen peptides could be a safe, natural, food-grade, and cost-effective functional agent for the surface decoration and stabilization of liposomes in powder form and hence a "green" solution for the oral delivery of phytonutrients. The present study reports a two-stage supramolecular self-assembly-directed process for the preparation of collagen peptide-decorated liposomal complexes of curcumin (CCL) [10% (w/w)] as microspheres (125 ± 25 μm) with improved solubility (1.46 × 105-fold) and sustained-release properties under gastrointestinal pH conditions. The molecular self-assembly of collagen peptides around the lipid bilayers and the various noncovalent interactions and conformational changes leading to the supramolecular assembly to act as a matrix for the encapsulation of lipid vesicles of curcumin were clear from the spectroscopic studies (UV-vis, fluorescence, FTIR, and circular dichroism). Further investigation of pharmacokinetics following a randomized double-blinded controlled trial on healthy volunteers (n = 15) demonstrated that the oral administration of 2.5 g of CCL sachet (250 mg of curcumin) enhanced the plasma concentration (Cmax: 118 vs. 4.3 ng/mL), the elimination half-life (4.2 vs. 0.7 h), and bioavailability as per the area under the curve over 12 h [AUC0-12h (CCL) = 506·8 vs. AUC0-12h (C95) = 9.47 (53-fold)], when the plasma concentration of curcumin was estimated with triple quadruple tandem mass spectrometry (UPLC-ESI-MS/MS).

Determination of Ibuprofen in Pharmaceutical Preparations by UPLC-MS/MS Method

Ibuprofen (IBU) is a non-steroidal anti-inflammatory drug that is extensively prescribed. For the determination of IBU in pharmaceutical formulations, a sensitive, simple, accurate, and rapid ultra-performance liquid chromatography method in combination with tandem triple quadruple mass spectrometry (UPLC-MS/MS) has been used and validated. The chromatographic separation was accomplished using a C18 UPLC column, 95 Å, 2.1 x 50 mm, 1.8 µm, and 0.1 percent formic acid in conjunction with acetonitrile (25:75 v/v. The flow rate was 0.15 ml min-1, with a run duration of 2.0 minutes. The injection volume was 5 μL and the column temperature was held constant at 40 °C.The mass transitions of İbuprofen and IS were m/z 205.1 → 159.0 and 249.9 → 229.1, respectivelty. According to ICH guidelines, the approach was thoroughly verified. The linear range 1-5000 ng mL-1 calibration curve has a strong correlation coefficient (0.9921). Within and between days precision were expressed as relative standard deviation and were lower than 6.24%. This method has been used to determine IBU in both pure form and pharmaceutical formulations with great success.

Sensitive and rapid simultaneous quantitation of leucovorin and its major active metabolite 5-methyl-tetrahydrofolate in human plasma using a liquid chromatography coupled with triple quadruple mass spectrometry.

Bioanalysis of an endogenous compound such as leucovorin is never an easy task on a liquid chromatography tandem mass spectrometer (LC-MSMS). Unless it is necessary, regulatory guidance discourages working with surrogate matrices for calibration curve standard preparation. Herein, a selective and sensitive liquid chromatography-tandem mass spectrometry method for simultaneous determination of leucovorin and 5-methyl tetrahydrofolic acid in human plasma was developed and validated. Stable labeled internal standards, i.e. leucovorin D4 and 5- methyl tetrahydrofolic acid 13 C5 , were used as internal standards to track and compensate the parent compounds during processing and extraction from plasma. The method involves a rapid solid-phase extraction from plasma followed by reverse-phase gradient chromatography and mass spectrometry detection with a total run time of 5min. The method was developed and validated from 5 to 2,202 ng/ml for leucovorin and from 5 to 1,300 ng/ml for 5-methyl tetrahydrofolic acid. The mean recoveries for leucovorin and 5-methyl tetrahydrofolic acid were 100.4 and 100.9% respectively. The validated method enabled the simultaneous analysis of leucovorin and 5-methyl tetrahydrofolic acid in samples from clinical pharmacokinetic studies of leucovorin. The peak concentrations of leucovorin and 5-methyl tetrahydrofolic acid were 651-883 and 518-635ng/ml, respectively, in fasted and fed conditions. The terminal half-life values for leucovorin and 5-methyl tetrahydrofolic acid were 9.3-10.5 and 9.2-17.6h, respectively.

Forced Degradation Study of Zanubrutinib: An LC-PDA and LC-MS Approach.

Zanubrutinib is an unfamiliar second generation selective Brutson's Tyrosine Kinase inhibitor used to treat mantle cell lymphoma. In the present analysis, a new, stability indicating reverse-phase, high performance liquid chromatography method was developed and validated for the determination of Zanubrutinib succeeding degradation studies as pert the International Conference on Harmonization guidelines. The chromatographic separation of Zanubrutinib was achieved in a C18 column (250 × 4.6mm, 5-μm particle size) using a mobile phase of Acetonitrile: 0.1% Tri Ethyl Amine (65:35v/v) monitored at 219nm. The forced degradation studies were conducted by exposing the analyte to acidic, alkaline and neutral hydrolysis, oxidative, reductive, photolytic, and thermal stress conditions and the degradation behavior was studied. The analyte showed degradation under acidic, alkaline, oxidative and reductive stress conditions with additional peaks but, it was stable under neutral, photolytic and thermal stress conditions. The developed method was extended to triple quadruple mass spectrometry to characterize degradation products and to study the fragmentation pattern. Total four degradants were characterized including DP1 in acid &base hydrolysis, DP2 in oxidative and DP3, DP4 in reductive stress condition. As no substantial method was available for quantification of Zanubrutinib and to characterize zanubrutinib degradants, this method can be used for regular analysis in quality control labs.

Comprehensive Analysis of N6-Methyladenosine (m6A) Methylation in Neuromyelitis Optica Spectrum Disorders.

Background: N6-Methyladenosine (m6A) methylation is the most prevalent internal posttranscriptional modification on mammalian mRNA. But its role in neuromyelitis optica spectrum disorders (NMOSD) is not known. Aims: To explore the mechanism of m6A in NMOSD patients. Methods: This study assessed the m6A methylation levels in blood from two groups: NMOSD patients and healthy controls. Methylated RNA immunoprecipitation Sequencing (MeRIP-seq) and RNA-seq were performed to assess differences in m6A methylation between NMOSD patients and healthy controls. Ultra-high performance liquid chromatography coupled with triple quadruple mass spectrometry (UPLC-QQQ-MS) method was performed to check m6A level. Differential m6A methylation genes were validated by MeRIP-qPCR. Results: Compared with that in the control group, the total m6A level was decreased in the NMOSD group. Genes with upregulated methylation were primarily enriched in processes associated with RNA splicing, mRNA processing, and innate immune response, while genes with downregulated methylation were enriched in processes associated with the regulation of transcription, DNA-templating, and the positive regulation of I-kappa B kinase/NF-kappa B signalling. Conclusion: These findings demonstrate that differential m6A methylation may act on functional genes to regulate immune homeostasis in NMOSD.

Validation of a Liquid Chromatography/Tandem Mass Spectrometry Assay for the Quantification of Plasma Dihydroartemisinin

Background: Insufficient plasma level of dihydroartemisinin (DHA) can select resistance and will further hinder malaria elimination program. We investigated clinical applicability of a validated liquid chromatography/tandem mass spectrometry (LC-MS/MS) assay to quantify plasma concentration of DHA in healthy subjects from a single oral administration of fixed dose combination of Dihydroartemisinin-Piperaquine.Materials and Methods: Micro-elution solid-phase extraction in a 96-well plate format was used to prepare the samples. DHA separation happened in Acquity UPLCTM BEH C18 column (50 × 2.1 mm, 1.7 µm). Mobile phase was a mixture of acetonitrile-ammonium acetate 10 mM pH 3.5 (50:50, v/v) at 0.3 mL/minute flow rate. Waters Acquity UPLC™ H-Class system coupled with triple quadruple mass spectrometry in positive electrospray ionization mode was used for detection. The internal standard was a stable isotope labelled DHA.Results: Calibration curve was linear with a correlation coefficient >0.995 over a concentration range of 1–1,000 ng/mL. Bias and variation for accuracy and precision were in the range of 15% (20% at the lower limit of quantification). Using 5 µL sample, lower limit of quantification was 1 ng. Matrix effect was less than 15%. The method was successfully applied to investigate the pharmacokinetics of DHA from five healthy subjects, although carry over and the role of anticoagulants were not tested.Conclusion: The LC-MS/MS assay for the quantification of plasma DHA was validated for selectivity, linearity, lower limit of quantitation, accuracy, precision, matrix effect and stability. Although clinical applicability was demonstrated, this method was to be improved to address the not-tested validation parameters.Keywords: dihydroartemisinin, liquid chromatography/tandem mass spectrometry assay (LC-MS/MS), malaria, Indonesia

Multi-pharmaceuticals analysis in fish by continuous series solid-phase extraction coupled with UPLC-MS/MS

Abstract Pharmaceuticals which are widely used in aquatic can easily migrate into the environment and aquatic animals, and can increase the risk of drug resistance and allergic symptoms if consumed by humans. In order to achieve high-throughput analysis of pharmaceuticals with different physical and chemical properties from complex matrices, we developed a new method for various types pharmaceuticals in fish and shrimp tissue. Series solid-phase extraction (s-SPE) with different adsorbents was selected for extracting and purifying analytes with different paddings. s-SPE were combined with ultra performance liquid chromatography triple quadruple tandem mass spectrometry (UPLC-MS/MS) for the detection of 30 pharmaceuticals antibiotics in fish samples. This method was stabilized and reliable to determinate the pharmaceuticals in fish and shrimp samples. As the method combined multiple Chinese national standards method, it could be easily treat the multi-pharmaceuticals from the fish and shrimp samples once time. It provided for both quantitative and qualitative methods and they could be applied to single- or multi-residue methods.

Detect 15 pesticides fungicide in edible fungi by multiplug filtration clean-up pretreatment technology with ultra-high performance liquid chromatography-tandem mass spectrometry

To establish a method for determination of 15 pesticides residue in edible fungi by multiplug filtration clean-up(m-PFC) pretreatment technique with ultra-high performance liquid chromatography-tandem triple quadruple mass spectrometry(UPLC-MS/MS). The interferences of edible fungus samples were removed by extracting with acetonitrile and filtration type extraction column, which were fat, carbohydrates, water-soluble vitamins. Samples were separated with column of Waters ACQUITY UPLC®HSS T3(2.1 mm×100 mm, 1.8 μm), and were scanned by multiple reaction monitoring mode(MRM). Samples were quantified with matrix matching standard curve external standard method. The recoveries of 15 target compounds at the spiked levels of 10, 20, 50 μg/kg were 82.5%-118.5%, and the relative standard deviations were between 6.1% and 23.1%(n=6). The detection limit of 15 target compounds was 1-3 μg/kg, and the limit of quantification was 3-10 μg/kg. This method improves the efficiency of pretreatment, has good stability and high sensitivity, and could be used for the detection of 15 pesticides in edible fungi.

Pharmacokinetics-based comprehensive strategy to identify multiple effective components in Huangqi decoction against liver fibrosis

BackgroundHuangqi decoction (HQD) has been used to treat chronic liver diseases since the 11th century, but the effective components in HQD against liver fibrosis have not been definitively clarified. PurposeTo investigate and identify multiple effective components in HQD against liver fibrosis using a pharmacokinetics-based comprehensive strategy. MethodsThe absorbed representative components in HQD and their metabolites were detected in human plasma and urine using high-resolution mass spectrometry combined with a database-directed method, and then pharmacokinetics in multiple HQD components in human plasma was analyzed by ultra-performance liquid chromatography coupled with triple-quadruple mass spectrometry. Furthermore, the anti-fibrotic effect of potential effective HQD components was studied in LX-2 cells and that of a multi-component combination of HQD (MCHD) was verified in a mouse CCl4-induced hepatic fibrosis model. ResultsTwenty-four prototype components in HQD and 17 metabolites were identified in humans, and the pharmacokinetic characteristics of 14 components were elucidated. Among these components, astragaloside IV, cycloastragenol, glycyrrhizic acid, glycyrrhetinic acid, liquiritigenin, and isoliquiritigenin downregulated the mRNA expression of α-SMA; cycloastragenol, calycosin-7-O-β-D-glucoside, formononetin, glycyrrhetinic acid, liquiritin, and isoliquiritin downregulated the mRNA expression of Col I; and calycosin, liquiritigenin, isoliquiritigenin, cycloastragenol, and glycyrrhetinic accelerated the apoptosis of LX-2 cells. MCHD reduced serum aminotransferase activity and hepatic collagen fibril deposition in mice with CCl4-induced hepatic fibrosis. ConclusionUsing the pharmacokinetics-based comprehensive strategy, we revealed that multiple effective HQD components act together against liver fibrosis.

Identification of plasma lipid species as promising diagnostic markers for prostate cancer

BackgroundProstate cancer is a very common and highly fatal in men. Current non-invasive detection methods like serum biomarker are unsatisfactory. Biomarkers with high accuracy for diagnostic of prostate cancer are urgently needed. Many lipid species have been found related to various cancers. The purpose of our study is to explore the diagnostic value of lipids for prostate cancer.ResultsUsing triple quadruple liquid chromatography electrospray ionization tandem mass spectrometry, we performed lipidomics profiling of 367 lipids on a total 114 plasma samples from 30 patients with prostate cancer, 38 patients with benign prostatic hyperplasia (BPH), and 46 male healthy controls to evaluate the lipids as potential biomarkers in the diagnosis of prostate cancer. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway database was used to construct the potential mechanism pathway. After statistical analysis, five lipids were identified as a panel of potential biomarkers for the detection of prostate cancer between prostate cancer group and the BPH group; the sensitivity, specificity, and area under curve (AUC) of the combination of these five lipids were 73.3, 81.6%, and 0.800, respectively. We also identified another panel of five lipids in distinguishing between prostate cancer group and the control group with predictive values of sensitivity at 76.7%, specificity at 80.4%, and AUC at 0.836, respectively. The glycerophospholipid metabolism pathway of the selected lipids was considered as the target pathway.ConclusionsOur study indicated that the identified plasma lipid biomarkers have potential in the diagnosis of prostate cancer.

Rapid screening of glycoalkaloids in Solanum scabrum and S. nigrum berries using ultra-high-performance liquid chromatography with pathway-specified in-source fragmentation tandem mass spectrometry.

The safe consumption of Solanum scabrum and S. nigrum berries (SNBs) depends on a reliable and rapid chemical screen for the testing of the fruit and/or final food and industrial products for the presence and level of toxic glycoalkaloids. Such a rapid and sensitive screen could also be used by those involved in food safety and forensics, industry, research labs and those in agriculture production, breeding and food processing. Significant variation in the content and composition of glycoalkaloids across SNBs has been reported. To facilitate high-throughput targeted analysis, this work overcame the slow scan speed of a traditional triple quadruple mass spectrometry (QqQ) method by development of a pseudo-MS3 method. In-source fragmentation functioned as a pseudo-MS or pseudo-hydrolysis to trim down the structurally diverse and complex glycosides into five types of aglycone ions, which were then analyzed using multiple reaction monitoring (MRM). Characteristic product ions were selected based on the aglycone skeleton and substitution pattern and associated fragmentation pathway. A compact method with only 15 MRM transitions were developed for high-throughput screening of very diverse glycoalkaloids. Glycosides of the same aglycone type were readily identified in the same transition window without the need for mass spectra interpretation. Validated using solamargine, the sole available standard, the accuracy was 99.7-101.3%, the intra- and inter-day precision were, respectively, 2.5-5.0% and 8.0-9.2%, and the lower limit of detection and quantification were, respectively, 3.1 and 10.2 ng/mL (with 1 μL injection volume). The peudo-MS3 method allowed for high-throughput targeted analysis with compact MRM transitions to address a large number of glycoalkaloids with diverse structures. This method could serve to meet the most heavy-duty demand for rapid inspection of glycoalkaloids in SNBs. This method can be adopted and used by those involved in food safety and forensics, in developing food and industrial products and in genetics and breeding.

Extraction Optimization of Six Alkaloids and Four Lignans in Zanthoxylum armatum by Orthogonal Design and Ultra-fast Liquid Chromatography–tandem Quadrupole Mass Spectrometry

Alkaloids and lignans in Zanthoxylum armatum display important biological activities, but the quanitification method of alkaloids has not been reported. In this study, an effective extraction method was developed through an orthogonal design, and ten compounds in roots, stems, branches and leaves of Zanthoxylum armatum were simultaneously quantified by ultra-fast liquid chromatography–tandem quadrupole mass spectrometry (UFLC–MS/MS). The roots, stems, branches and leaves of Z. armatum were ultrasonically extracted with methanol (solvent–to–sample ratio 100 : 1, v/w) for 10 min. An UFLC–MS/MS method was developed with a gradient UFLC mobile phase and triple quadruple tandem mass spectrometry with electrospray ionization in the positive ion mode. The method was validated for linearity, precision, repeatability and accuracy. The limits of detection and quantification were within 0.01–7.5 and 0.04–30 ng/mL, respectively. The root samples collected from Tian’e County were abundant in N-methylanhydrotetrahydroberberrubine A, escholidine perchlorate and pinoresinol monomethyl ether, eudesminthe.

Simultaneous Determination of 17 Constituents of Chinese Wild Radix Salvia miltiorrhiza from Different Geographical Areas by Ultra-High Performance Liquid Chromatography Coupled to Triple Quadrupole Mass Spectrometry

Background and Objective: Radix Salvia miltiorrhiza (RSM) has been used clinically for the prevention and treatment of cardiovascular diseases; therefore, it is important to strengthen its quality management. Considering multiple constituents when assessing RSM quality is essential. We established a simple, rapid method to identify and quantify the major bioactive constituents in RSM using ultra high performance liquid chromatography (UPLC) coupled to a triple quadruple mass spectrometry (QqQ-MS). Methods: We analyzed 17 markers from 50 batches of wild S. miltiorrhiza samples that were collected from different locations in China. The ultrasonic extracts of all samples were determined using the UPLC-QqQ-MS method and were assessed by hierarchical cluster analysis (HCA). Results: We used this method to analyze 50 sample batches of the 17 compounds and obtained results with excellent linearity (R2, 0.9915-0.9997), precision (relative standard deviation, RSD, 0.15-1.94%), repeatability (RSD, 1.28-4.71%), stability (RSD, 0.97-5.60%) and recovery (RSD, 0.305-6.40%). The hierarchical cluster analysis was used to classify the 50 samples based on the characteristics of the 17 compound markers. Conclusion: We demonstrated that the developed method was simple, reproducible and sensitive, and it is capable of systematic and scientific evaluation for quality control of RSM. The HCA clearly demonstated that the RSM samples from different locations were significantly different and the quality of wild Radix S. miltiorrhiza could generally be judged according to its geographical origin.

A new method of ultra-performance liquid chromatography/tandem mass spectrometry for determination of chlorogenic acid in human plasma and urine.

Chlorogenic acid (CA) is well known for its various biological activities. Here, a clinical study was performed in patients with advanced malignant cancer to explore its therapeutic effects. We aimed to develop a method to quantify CA in human plasma and urine to assist the clinical pharmacokinetic study. Ultra-performance liquid chromatography (UPLC) coupled with a triple quadruple mass spectrometry was used to separate and detect CA, with puerarin serving as the internal standard. The method presents an excellent linearity ranging from 5 to 2000 ng/mL for plasma analysis and 50 to 20 000 ng/mL for urine analysis. Intra- and inter-day precision and accuracy were both less than 15% for plasma and urine. These results showed that the novel UPLC method was robust and sensitive, and fulfilled the requirements for a clinical pharmacokinetic study of CA in patients with advanced cancer.

Treatment Effect and Ecological Risk Assessment of Typical Antibiotics in Wastewater from Large-scale Dairy Farms in Tianjin

To obtain the content level and treatment efficiency of typical antibiotics in wastewater treatment in large-scale dairy farms in Tianjin, the SPE-UPLC-MS/MS (ultra-high performance liquid chromatography tandem triple quadruple mass spectrometry combined with solid-phase extraction for pretreatment) technology was utilized to investigate and monitor seven typical antibiotics in wastewater from 12 large-scale dairy farms in Tianjin. Antibiotic residues were detected in 12 large-scale dairy farms before and after wastewater treatment. In the wastewater before treatment, the detection rates of tilmicosin (TIL), oxytetracycline hydrochloride (OTC), sparfloxacin (SPA), sulfathiazloe (STZ), ofloxacin (OFL), and sarafloxacin hydrochloride (SAR) were all 100%, whereas the detection rate of sulfadiazine (SDZ) was 83.33%. Among them, TIL and OTC were the main antibiotic components in untreated wastewater, the concentrations were 25.21 μg·L-1 and 9.87 μg·L-1, respectively. The detection rates of SDZ and OFL in the treated wastewater dropped to 25.00% and 41.66%, respectively. The main components were TIL and OTC and the concentrations were 11.30 μg·L-1 and 3.71 μg·L-1, respectively. There were significant decreases in the concentrations. The treatment effect on antibiotics from different farms ranged from 24.95% to 81.05%. The comprehensive treatment effect of the anaerobic-anoxic-oxic (AAO) treatment process was better than that of the anoxic-oxic (AO) treatment process. OFL, SAR, and OTC were the main high-risk pollutants in treated wastewater. Each large-scale dairy farm contained one or more antibiotic with RQs>1, and their emissions pose an ecological risk to the environment.

Pharmacokinetic comparison of five xanthones in rat plasma after oral administration of crude and processed Garcinia hanburyi extracts

Gamboge, a dried resin secreted by Garcinia hanburyi Hook. f. (Guttiferae), possesses remarkable anticancer activity. However, due to toxicity, it must be processed before use in clinics. Xanthones are the main bioactive ingredients in gamboge. In order to elucidate the influence of processing technology on pharmacological properties of gamboge, an efficient, sensitive, and selective ultra-performance liquid chromatography coupled with triple quadruple mass spectrometry (UPLC-MS/MS) method of five critical xanthones, including β-morellic acid (β-MA), isogambogenic acid (IGNA), gambogenic acid (GNA), R-gambogic acid (GA), and S-GA in rat plasma was established for a comparative pharmacokinetics study of these xanthones after oral administration of crude and processed G. hanburyi extracts. The chromatographic separation of these five xanthones along with an internal standard (I.S.) was carried out on a Waters Acquity UPLC BEH C8 column with a gradient elution method using acetonitrile/0.1% formic acid-water as mobile phases. The eluate was detected by multiple-reaction monitoring (MRM) scanning with an electrospray ionization source operating in the positive ionization mode. Sample preparation involved a liquid-liquid extraction of the five analytes with ethyl acetate. Deoxyschizandrin was employed as an internal standard. This assay method was validated for selectivity, linearity, intra-day and inter-day precision, accuracy, recovery, matrix effect, and stability. The results revealed that the calibration curves displayed good linear regression (r > 0.995), and the lower limit of quantification (LLOQ) was <5.52 ng/mL for each analyte. The intra-day and inter-day precision (RSD) of the five xanthones at low, medium, and high levels was <10.58%, and the bias of the accuracy ranged from −8.54 to 10.2%. All other parameters fulfilled the FDA criteria for bioanalytical validation. In addition, the assay was successfully applied to the determination and pharmacokinetic study of these five xanthones after oral administration of crude and processed gamboge. Furthermore, Cmax of GNA and AUC0−t of IGNA were increased significantly (P < 0.05) after processing, while AUC0−t of β-MA, R-GA, and S-GA decreased remarkably (P < 0.05), which suggested that processing exerted different effects on the absorption of xanthones. The results might be valuable for the clinical reasonable application and understanding the processing mechanism of gamboge.