Chenxiao Shan

Identification of liguzinediol metabolites in rats by ultra performance liquid chromatography/quadrupole-time-of-flight mass spectrometry

Ultra performance liquid chromatography/quadrupole-time-of-flight mass spectrometry (UPLC/QTOF MS) was employed to investigate the in vivo metabolism of liguzinediol. Urine, bile, feces and plasma samples were collected after intravenous administration of 10mg/kg liguzinediol to healthy rats. Altogether seven metabolites were detected and tentatively identified based on the characteristics of their protonated ions. The metabolites were mainly transformed by four main metabolic pathways including oxidation, sulfation, glycine conjugation and glucuronidation.

UFLC-Q-TOF/MS based screening and identification of the metabolites in plasma, bile, urine and feces of normal and blood stasis rats after oral administration of hydroxysafflor yellow A.

The dried flower of Carthamus tinctorius L. (honghua) is a widely used traditional Chinese medicine in clinics to treat coronary heart disease, hypertension, and cerebrovascular disease due to its functions of ameliorating circulation and removing blood stasis. Hydroxysafflor yellow A (HSYA) is an active marker component of honghua. In this paper, ultra-flow liquid chromatography coupled with quadrupole-time-of-flight mass-spectrometry (UFLC-Q-TOF/MS) was established and successfully applied to the detection and identification of the metabolites in bile, urine, plasma and feces samples of normal and model rats with orally administrated HSYA. A total of 8 metabolites were observed in normal rats, while 7 metabolites were detected in model rats. The distribution of metabolites in the plasma, bile, urine and feces of normal and model rats had obvious differences. The major in vivo metabolic pathways for HSYA included hydroxylation, hydroxylation+methylation, acetylation and glucuronidation, and there were also dehydration, hydrogenation, hydration, and hydroxylation+glucuronidation. All of these metabolites were reported for the first time, and these results are valuable and important for the understanding of the metabolic process and therapeutic mechanism of HSYA and some other pigments in honghua.

Metabolic profile of anhydrosafflor yellow B in rats by ultra-fast liquid chromatography/quadrupole time-of-flight mass spectrometry.

Anhydrosafflor yellow B (AHSYB) is one of the major active water-soluble pigments from Carthamus tinctorius, which has been found to inhibit ADP-induced platelet aggregation and possess significant antioxidant activity. However, the metabolic fate of AHSYB in vivo remains unknown. In order to explore whether AHSYB is extensively metabolized, the metabolites of AHSYB in plasma, urine, bile, and feces samples after intravenous administration to the rats were investigated by ultra-fast liquid chromatography/quadrupole time-of-flight mass spectrometry (UFLC/Qq-TOF-MS/MS) combined with Metabolitepilot™ software. In total, AHSYB and 22 metabolites including both phase I and phase II metabolism processes were found and tentatively identified from the bio-samples. The metabolic pathways were involved in oxidation, reduction, hydroxylation, methylation, dimethylation, O-acetylation, hydrolyzation, sulfation, glucuronidaton, glutathionation and combination with glucose. The results showed that the renal and biliary routes play an important role in the clearance and excretion of AHSYB as well as hepatocyte metabolism. All of these results were reported for the first time and would contribute to a further understanding of the in vivo intermediated processes and metabolic mechanism of AHSYB and its analogs.

Metabolite identification of liguzinediol in dogs by ultra-flow liquid chromatography/tandem mass spectrometry

Ultra-flow liquid chromatography/quadrupole-time-of-flight mass spectrometry (UFLC/Q-TOF MS) method combined with metabolitepilot(MT) software was used for analysis of the metabolites of liguzinediol in dogs. Urine, bile, feces and plasma samples were collected after intravenous administration of 8 mg/kg liguzinediol to healthy dogs. Besides liguzinediol, seven metabolites were detected and identified by UFLC/Q-TOF MS method. The results showed that liguzinediol had some main metabolic pathways in dogs including oxidation, sulfation, cysteine conjugation, N-acetylcysteine conjugation and glucuronidation.

Influence of sulfur fumigation on the chemical profiles of Atractylodes macrocephala Koidz. evaluated by UFLC–QTOF–MS combined with multivariate statistical analysis

&NA; In the present study, the chemical compositions of Atractylodes macrocephala Koidz. (AMK) were analyzed systematically and influence of sulfur fumigation on the chemical profiles was evaluated by ultrafast flow liquid chromatography coupled with quadrupole‐time‐of‐flight mass spectrometry (UFLC–QTOF–MS) combined with multivariate statistical analysis. 52 components were detected from non‐fumigated AMK (NF‐AMK) and 28 components were newly produced after sulfur fumigation, out of which 59 major peaks were identified. The concentrations of 20 compounds significantly decreased and 37 compounds obviously increased. The potential structural transformation mechanism of terpenoids was explored to illustrate the correlation of the components contents before and after sulfur fumigation. Eight sulfur‐containing/dehydrated‐integrated atractylenolides that evolved from the NF‐AMK were screened out as potential characteristic chemical markers to examine the post‐harvest handling procedures of commercial AMK with excessive sulfur fumigation and maintain consistent quality. HighlightsThe chemical compositions of Atractylodes macrocephala Koidz. (AMK) were analyzed systematically and influence of sulfur fumigation on the chemical profiles was evaluated by UFLC–QTOF–MS combined with multivariate statistical analysis.52 components were examined from non‐fumigated AMK (NF‐AMK) and 28 compounds were newly produced after sulfur fumigation, out of which 59 major peaks were identified.Eight sulfur‐containing/dehydrated‐integrated atractylenolides that evolved from the NF‐AMK were screened out as potential characteristic chemical markers to examine the post‐harvest handling procedures of commercial AMK with excessive sulfur fumigation.

Contents variation analysis of free amino acids, nucleosides and nucleobases in semen sojae praeparatum fermentation using UFLC-QTRAP MS

UFLC-QTRAP MS was used to develop a sensitive and rapid method of evaluating content variation during Semen sojae praeparatum (SSP) fermentation. It did this through the simultaneous quantification of 22 free amino acids and 16 nucleosides and nucleobases in the raw materials and processed products of SSP. The method was shown to be reproducible and accurate. The limits of detection and quantity values were 0.09-168.75 and 0.31-562.50 ng/mL for the 38 analytes, respectively. The data were examined through principal components analysis to compare the content variations. The quantitative results showed that the ingredients were properly determined in most of the samples and were converted regularly throughout the SSP fermentation process. These results correspond to the morphologic changes and principal components analysis results.

Occurrence and Seasonal Variations of Lipophilic Marine Toxins in Commercial Clam Species along the Coast of Jiangsu, China

Recent studies have examined lipophilic marine toxins (LMTs) in shellfish and toxic algae worldwide, but the occurrence and seasonal variations of LMTs in commercial clams (including Mactra veneriformis, Ruditapes philippinarum, Meretrix meretrix, and Cyclina sinensis) at their major culturing area in Jiangsu, China, remain largely unexplored. In this study, a new solid phase extraction (SPE) in combination with an ultra-fast liquid chromatography and triple-quadrupole linear ion trap mass spectrometry (UFLC-TQ-MS) method was developed to determine the presence of 10 typical LMTs (okadaic acid (OA), yessotoxins (YTXs), azaspiracids (AZA1-3), pectenotoxins (PTX2), gymnodimine (GYM), dinophysistoxins (DTX1&2), and spirolides (SPX1)) in the aforementioned four clam matrices. After confirmation of its sensitivity and precision, this method was used to evaluate the amounts of LMTs in clam samples harvested in five aquaculture zones of the Jiangsu coastal area. Monthly variations of GYM, PTX2, OA, and DTX1&2 in 400 clam samples from the sample areas were determined from January 2014 through August 2015. Peak values were observed during May and August. This is the first systematic report of LMTs detected in clam samples harvested in Jiangsu. Follow-up research and the implementation of protective measures are needed to ensure the safety of clams harvested in this area.

Qualitative and quantitative analyses of bioactive secolignans from folk medicinal plant Peperomia dindygulensis using UHPLC-UV/Q-TOF-MS

Peperomia dindygulensis, with secolignans (SLs) as major bioactive constituents, is a commonly used traditional folk medicine in mainland China for treatment of stomach, liver, mammary, and esophageal cancers. However, to date, there is no method available for the qualitative and quantitative analyses of SLs in this medicinal plant. The purpose of this study was to establish a sensitive, selective, and reproducible method for rapidly profiling, identifying, and determining SLs in the whole plant of P. dindygulensis. Ultra high-performance liquid chromatography (UHPLC) coupled with ultraviolet detector (UV) and quadrupole tandem time-of-flight mass spectrometry (Q-TOF-MS) were used for this analyses. The fragmentation behaviors of different types of SLs were described. A total of thirteen SLs, including two new derivatives, were identified or tentatively characterized in P. dindygulensis samples. In addition, seven major SLs in herbal samples from different regions in China were successfully determined. The method developed in this study is suitable for the qualitative and quantitative analyses of SLs in P. dindygulensis, and may be applicable for determining or identifying SLs from other Pepermia genus plants.

Development and validation of a UFLC–MS/MS method for the determination of anhydrosafflor yellow B in rat plasma and its application to pharmacokinetic study

A sensitive ultrafast liquid chromatography coupled with triple quadrupole mass spectrometric (UFLC-MS/MS) method for the quantification of anhydrosafflor yellow B (AHSYB), a major active water-soluble pigment from Carthamus tinctorius, in rat plasma has been developed and validated. Sample preparation was achieved by protein precipitation of plasma with four volumes of methanol. Rutin was used as the internal standard (IS). The analytes were separated using a C18 column with an 8min gradient elution, followed by mass spectrometric detection using negative electrospray ionization (ESI(-)) in multiple reaction monitoring (MRM) mode. The method was linear in the concentration range of 25-10,000ng/mL for AHSYB. Intra-day and inter-day precision variation was less than 6.5%. The relative error of accuracy was within ±9.4%. The mean recovery of AHSYB was higher than 70.9%. The established method was successfully applied to the pharmacokinetic study after intravenous (2.5mg/kg) and oral (30mg/kg) dosing of AHSYB in normal rats. And the pharmacokinetic properties of AHSYB in rats with acute blood stasis and the differences between normal and acute blood stasis syndrome rats were also investigated. The results showed that the compound was poorly absorbed (∼0.3%) and the AUC0-t, AUC0-∞ and F were all significantly lower (P<0.05) in acute blood stasis syndrome rats, suggesting that disease condition may alter the body metabolism by enhancing metabolite enzyme activity.

A comprehensive in vitro and in vivo metabolism study of hydroxysafflor yellow A

As the most important marker component in Carthamus tinctorius L., hydroxysafflor yellow A (HSYA) was widely used in the prevention and treatment of cardiovascular diseases, due to its effect of improving blood supply, suppressing oxidative stress, and protecting against ischemia/reperfusion. In this paper, both an in vitro microsomal incubation and an in vivo animal experiment were conducted, along with an LC-Q-TOF/MS instrument and a 3-step protocol, to further explore the metabolism of HSYA. As a result, a total of 10 metabolites were searched and tentatively identified in plasma, urine, and feces after intravenous administration of HSYA to male rats, although no obvious biotransformation was found in the simulated rat liver microsomal system. The metabolites detected involving both phase I and phase II metabolism including dehydration, deglycosylation, methylation, and glucuronic acid conjugation. A few of the metabolites underwent more than one-step metabolic reactions, and some have not been reported before. The study would contribute to a further understanding of the metabolism of HSYA and provide scientific evidence for its pharmacodynamic mechanism research and clinical use.