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• Metabolic profiles of BZYQT in rats were systematically characterized. • Flavonoids and triterpenoids of BZYQT showed the higher MS response in rat plasma. • Proposed metabolic pathways of atractylenolide III in rats were first explored. • BZYQT-related components underwent massive phase I and II metabolism in rats. • A total of 162 BZYQT-related xenobiotics were detected in rats biosamples. Bu-Zhong-Yi-Qi-Tang (BZYQT), a famous traditional Chinese medicine prescription (TCMP), has been extensively used for conditioning sub-health status and diseases caused by spleen-qi deficiency in China for over 700 years. BZYQT is prevalent not only in China, but also in Japan and South Korea for the clinical treatment of chronic diseases, such as fatigue, tuberculosis and loss of appetite after surgery. However, due to a lack of research on the holistic metabolism of BZYQT, the in vivo bioactive components of BZYQT remain unclear, hindering further study of its in vivo mechanism of action and quality control. In the present study, a four-step integrated strategy based on ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry (UHPLC-Q-TOF/MS) was established to systematically screen the in vivo xenobiotics of BZYQT. Ultimately, a total of 162 xenobiotics (59 prototypes and 103 metabolites) were identified or tentatively characterized, including 48 in plasma, 147 in urine and 58 in feces, while the in vivo metabolic profile of atractylenolide III (a major component of BZYQT) was elucidated for the first time. The xenobiotics of BZYQT mainly included flavonoids from Astragali Radix , Glycyrrhizae Radix et Rhizoma and Citrus reticulatae Pericarpium ; lactones from Angelicae Sinensis Radix and Atractylodis Macrocephalae Rhizoma ; and triterpenoid saponins from Cimicifugae Rhizoma. After oral administration, BZYQT-related components underwent diverse metabolic pathways. Among them, flavonoids mainly underwent glucuronidation, sulfation and demethylation, while lactones mainly underwent hydroxylation and acetylcysteine conjugation, and deglycosylation was the major metabolic reaction of saponins. Our investigation gives a comprehensive analysis of the metabolic characteristics of BZYQT and will provide an important basis for further studying the pharmacokinetics of BZYQT to explore its in vivo disposal features and discover its in vivo bioactive components. [ABSTRACT FROM AUTHOR]

In this study, we aimed to establish a method for determination of multiple mycotoxins such as aflatoxins, ochratoxin A, and zearalenone in red pepper powder samples based on the quick, easy, cheap, effective, rugged, and safe (QuEChERS) approach for extraction and cleanup, with detection and quantification by high-performance liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) in both positive- and negative-ion modes. The developed LC-MS/MS analytical method was compared with an enzyme-linked immunosorbent assay (ELISA) to improve the reliability of our developed method. The linearity, precision, and accuracy were validated for the LC-MS/MS methods. The results obtained with the LC-MS/MS were linear, with a correlation coefficient (R2) of >0.998. The limits of detection and quantification for mycotoxins were 0.07 to 0.71 μg/kg and 0.20 to 1.81 μg/kg, respectively. Intra- and interday precision tests (expressed as the relative standard deviation) for each analyte were 1.58 to 5.97% and 0.97 to 9.01%, respectively. Average recoveries were 85.70 to 94.99%. The validation results for the ELISA were linear (R2 > 0.995), and recoveries were 77.13 to 93.93%. Both analytical methods were applied to determine the presence of mycotoxins in commercial red pepper powder samples from South Korea. Four of the total 56 samples were contaminated with aflatoxins, and 6 samples were contaminated with ochratoxin A; these results were consistent for the two methods (P > 0.05). Therefore, our developed LC-MS/MS with QuEChERS approach for determination of multiple mycotoxins may be useful for controlling the quality and safety of red pepper powder. [ABSTRACT FROM AUTHOR]

The number of glyphosate intoxication cases has been increased after the regulation of paraquat. Unfortunately, there are no reports on the potential concentration of glyphosate for those acute intoxicated patients admitted to emergency rooms and the correlation between the concentration of glyphosate and clinical symptoms in Korea up to our knowledge. As a nonselective herbicide, analysis of glyphosate requires derivatization because of its amphoteric and strongly polar nature. In order to develop a method to determine the concentration of glyphosate and its metabolite, aminomethylphosphonic acid (AMPA) in blood samples without derivatization, liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) was utilized with a hydrophilic interaction chromatography (HILIC) column. The validation of this method showed that the limits of detection (LODs) and limits of quantitation (LOQs) for glyphosate and AMPA were 50 and 100ng/mL, respectively. In addition, matrix effect, recovery rate, and accuracy and precision in intra and inter-day were evaluated during the validation study of this method. Blood samples acquired from five glyphosate intoxicated patients were analyzed to investigate the correlation between the concentration of glyphosate and clinical symptoms. These patients were previously admitted to the emergency room at a University Hospital in Korea after glyphosate was self-administered in suicide attempts or by accident. As results of blood sample study, the concentration of glyphosate and AMPA were found in the range of 1.0-171.1 and 0.2-2.6μg/mL, respectively. The concentration ratio of glyphosate to AMPA was 55-71. According to the clinical reports for those patients, they were in the age between 47 and 82 years old and administered about 50-400mL. The blood samples were collected within 2-5h after administration of glyphosate. Among the intoxicated patients, the most common clinical symptom was metabolic acidosis, identified in four patients. The comparison between the concentration of glyphosate and administered dosage did not show the correlation, which suggests further investigation on the effects of surfactants in glyphosate from different vendors. [ABSTRACT FROM AUTHOR]

Homovanillic acid (HVA) and vanillylmandelic acid (VMA) are end-stage metabolites of catecholamine and are clinical biomarkers for the diagnosis of neuroblastoma. For the first time in Korea, we implemented and validated a liquid chromatography tandem mass spectrometry (LC–MS/MS) assay to measure urinary concentrations of HVA and VMA according to Clinical and Laboratory Standards Institute guidelines. Our LC–MS/MS assay with minimal sample preparation was validated for linearity, lower limit of detection (LOD), lower limit of quantification (LLOQ), precision, accuracy, extraction recovery, carryover, matrix effect, and method comparison. A total of 1209 measurements was performed to measure HVA and VMA in spot urine between October 2019 and September 2020. The relationship between the two urinary markers, HVA and VMA, was analyzed and exhibited high agreement (89.1% agreement, kappa's k = 0.6) and a strong correlation (Pearson's r = 0.73). To our knowledge, this is the first study to utilize LC–MS/MS for simultaneous quantitation of spot urinary HVA and VMA and analyze the clinical application of both markers on a large scale for neuroblastoma patients. [ABSTRACT FROM AUTHOR]

Insamyangpye decoction (ISYPD) is an oriental herbal prescription used in Korea to treat lung-related diseases such as chronic obstructive pulmonary disease. ISYPD is a complex prescription consisting of 13 herbal medicines, and ISYPD sample was obtained by adding 50 L of distilled water to a mixture (5 kg) of 13 herbal medicines, extracting at 100 °C for 2 h using an electric extractor, and freeze-drying. In this study, an accurate and sensitive liquid chromatography tandem mass spectrometry (LC–MS/MS) method based on multiple reaction monitoring (MRM) was developed and verified for quality assessment of ISYPD using 10 marker components: mulberroside A (1), amygdalin (2), liquiritin apioside (3), naringin (4), poncirin (5), platycodin D (6), ginsenoside Rb1 (7), glycyrrhizin (8), saikosaponin A (9), and schizandrin (10). These marker compounds were separated using an Acquity UPLC BEH C18 column (2.1 mm × 50 mm, 1.7 μm) maintained at 30 °C with a mobile phase elution gradient of acetonitrile in distilled water, both containing 0.1% (v/v) trifluoroacetic acid. Marker components were quantified using the LC–MS/MS MRM method developed and validated, and found at 0.09–7.47 mg/g. [ABSTRACT FROM AUTHOR]

The Sogunjung decoction (SGJD) is a traditional herbal formula that has been used to treat constipation and improve the constitution of infirm children in Korea. In this study, simultaneous quantification of gallic acid (1), magnoflorine (2), albiflorin (3), paeoniflorin (4), liquiritin apioside (5), liquiritin (6), liquiritigenin (7), coumarin (8), cinnamaldehyde (9), benzoylpaeoniflorin (10), and glycyrrhizin (11) was conducted using fast and sensitive liquid chromatography–tandem mass spectrometry (LC–MS) multiple-reaction monitoring to develop a quality-control protocol for the SGJD. A Waters Acquity UPLC BEH C18 column (2.1 × 100 mm, 1.7 μm) was used for the chromatographic separation of the 11 marker compounds in the SGJD using two mobile phases (5 mM ammonium acetate in distilled water containing 0.1% (v/v) formic acid, and acetonitrile). The MS parameters for a simultaneous analysis were capillary voltage (3.0 kV), source temperature (150 °C), desolvation temperature (500 °C), desolvation gas flow (700 L/h), and cone gas flow (50 L/h). The developed LC–MS method was validated by the evaluation of linearity, limits of detection, limits of quantification, recovery and precision. By using the developed and validated assay, the 11 marker components in the SGJD were detected in amounts of 0.01–51.83 mg/g. [ABSTRACT FROM AUTHOR]