Identification and quality evaluation of Chinese rice wine using UPLC-PDA-QTOF/MS with dual-column separation.

BACKGROUND: Chinese rice wine (CRW) is a well-known drink and functional food that is used in traditional Chinese medicine. However, there is still a lack of quality control and evaluation methods for CRWs. PURPOSE: The study aimed to establish a new method that can serve both as quality control and evaluation method and, as well as an identification method for CRWs. METHOD: Compound identification in different CRW samples and determination of uracil, xanthine, uridine, adenine, guanosine, 5-hydroxymethylfurfural, and adenosine contents from 29 CRW samples from 14 brands were performed using UPLC-PDA/TOF-MS. The dual-column chromatographic separation of CRW was performed using CORTECS T3 coupled to HSS T3. The optimal mobile phase consisted of water with 0.1% formic acid, 40 mM ammonium acetate, and methanol: acetonitrile (2:1). Furthermore, to compare the UPLC fingerprints between CRWs of different brands, a similarity analysis was performed to classify the CRW samples. Finally, network pharmacology and in vitro efficacy and toxicity tests were used to investigate the biological function of the seven components and CRWs. RESULTS: A total of 55 compounds were unambiguously or tentatively identified. Among them, nucleoside, pyrimidines and purines were reported in CRW for the first time. The seven components were successfully determined, and their contents showed large variations among different brands of CRW, which was consistent with the results of the chromatographic fingerprint similarities. The results of in vitro efficacy and toxicity tests indicated that CRWs and seven components had obvious protective effect on H9c2 cell injury induced by the H2O2 model. Network pharmacology analysis showed that these seven compounds might be the main active components of CRW that promote blood circulation and ventilation. CONCLUSION: This study revealed that dual-column chromatographic separation is an effective method for quantitative and chromatographic fingerprint analyzes of complex samples, and seven compounds can be used for the quality evaluation and control of CRWs.

An integrated chemical analysis and network pharmacology approach to identify quality markers of Actinidia eriantha Benth radix on gastric cancer.

INTRODUCTION: Actinidia eriantha Benth radix (AEBR) is one of the most commonly used medicines by the She people in China, used primarily for the treatment of tumours of the digestive tract. There is currently limited to no data on the quality control of AEBR. OBJECTIVES: The aim of this study was to identify quality markers of AEBR. MATERIAL AND METHODS: An ultra-performance lquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS) method was used to identify and analyse the components of AEBR from water extracts. An ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was also established for the simultaneous determination of 13 active components in the water extracts. The network pharmacology method was used to screen for quality markers of AEBR in gastric cancer. RESULTS: This study tentatively identified 199 chemical constituents and isomers, including 67 pentacyclic triterpenoids, 20 flavonoids, 39 phenolic acids, 18 coumarins, and other compounds. The 13 active components in the water extracts were successfully determined using a validated UPLC-MS/MS method. Based on the network pharmacology method, four compounds were selected as quality markers of AEBR. CONCLUSION: This study provides an important reference for the quality control of AEBR. Chemical analysis combined with network pharmacology provides an effective strategy for the discovery of quality markers in traditional Chinese/herb medicine.

Toxicokinetics, in vivo metabolic profiling, and in vitro metabolism of gelsenicine in rats.

Gelsenicine, mainly isolated from Gelsemium elegans Benth., is one of the most toxic alkaloids. The lack of information on gelsenicine leads to inaccurate risk and poisoning evaluation. In this study, the metabolic profiling and toxicokinetics of gelsenicine was studied by ultra-high performance liquid chromatography (UPLC) with quadrupole time-of-flight (Q-ToF) and tandem mass spectrometry in rats after intraperitoneal (i.p., 40 µg/kg) and intragastric (i.g., 60 µg/kg) administration. After i.p. administration, the area under the curve (AUC), the apparent volume of distribution (V), and the total body clearance (CL/F) of gelsenicine in plasma were 3.79 µg/L h, 38.47 L/kg, and 11.87 mL/h kg, respectively. After i.g. administration, the corresponding values were slightly increased (5.49 µg/L h; 53.10 mL/kg, and 12.66 mL/h kg). The toxicokinetic results indicated that the hepatic first-pass effect was predominant after i.p. administration. The UPLC-Q-ToF-MS data revealed nine metabolites in plasma, urine, and bile which were largely obtained by demethylation, hydroxylation, acetylation and glycine conjugation. Metabolites were mainly excreted through urine and bile, most of which in urine was basically eliminated in 24 h. Molecular docking and liver microsome experiments further showed that gelsenicine was metabolized by cytochrome P450 3A4 and 3A5. Summarizing, the present study provides metabolic and toxicokinetic information on gelsenicine which in turn may help in future risk assessment and forensic identification after poisonings.

An UPLC-MS/MS method for quantification of D-pinitol in rat plasma and its application to a pharmacokinetic and bioavailability study.

D-pinitol could be a potential therapeutic agent for the treatment of diabetes mellitus (DM) type II. In this work, a sensitive and rapid ultra performance liquid chromatography coupled with tandem mass spectrometry method was firstly developed and validated for the determination and pharmacokinetic study of D-pinitol in rat plasma. D-pinitol and 5,7-dihydroxychromone (Internal Standard, IS) were completely separated on a BEH C18 column. The plasma samples were deproteinated with acetonitrile: ethanol (1:1). The MRM transitions for D-pinitol was m/z 179.125 â†’ 105.049, and for IS was m/z 195.085 â†’ 109.031. The method linearity ranges was 5-200 ng/mL. The precision, accuracy, recovery, matrix effect, stability under different conditions, were all within the required criteria. After intragastric (50 mg/kg) administration of D-pinitol to the rats, the maximum plasma concentration (Cmax) was 77.8 ± 19.5 ng/mL. The time to reach the maximum plasma concentration (Tmax) was 2.2 ± 0.98 h. Apparent distribution volume (Vz) was 1557.5 ± 1329.1 L/kg and the plasma centration time curve (AUC0-t) was 1265.5 ± 479.3 µg/L*h. After intravenous (5.0 mg/kg) administration, Vz was 325.2 ± 107.8 L/kg and AUC(0-t) was 693.0 ± 89.9 µg/L*h. Our study indicated D-pinitol had a slow elimination phase and might be the high affinity binding to blood protein in vivo, which are helpful for its further drug development and clinical application.

Pharmacokinetic and metabolic profiling studies of sennoside B by UPLC-MS/MS and UPLC-Q-TOF-MS.

Sennoside B is a specific dianthrone compound extracted from senna, which is widely used as a stimulant laxative but has potential side effects. This study aimed to obtain the metabolic and pharmacokinetic data of sennoside B. The metabolic profiles of sennoside B were obtained from rat plasma, urine, bile and feces by an ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS). As a result, 14 metabolites were structurally identified and the proposed metabolic pathways of sennoside B included hydrolysis to aglycones, release of rhein-type anthrone, and extensive conjugation. As the only compound detected in the plasma samples after intravenous and intragastric administrations, the prototype was selected as the plasma marker in the pharmacokinetic study. A simple and sensitive ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was developed for the quantitation of sennoside B in rat plasma. The linear range of sennoside B was 5-1000 ng/mL (R2 ≥ 0.991) and the lowest limit of quantification (LLOQ) was 5 ng/mL. The intra- and inter- precisions of the assay were less than 10%, whereas accuracy ranged from 85.80% to 103.80%. The extraction recovery, matrix effect and stability of sennoside B were within acceptable limits. The established method was well validated and successfully applied to the pharmacokinetic study of sennoside B. The oral absolute bioavailability of sennoside B was calculated as 3.60% and the value apparent volume of distribution of intravenous and intragastric administrations were 32.47 ±â€¯10.49 L/kg and 7646 ±â€¯1784 L/kg, respectively. The maximum plasma concentrations were 212.6 ±â€¯50.9 µg/L and 14.06 ±â€¯2.73 µg/L for intravenous and intragastric dosing groups, respectively. According to the current results of pharmacokinetic and metabolic profiling studies, metabolites with high abundance in tissues would be the next object in the pharmacokinetic study of sennoside B.

Metabolic Profiling of Alpinetin in Rat Plasma, Urine, Bile and Feces after Intragastric Administration.

Alpinetin, a bioactive flavonoid, has been known to have a diverse therapeutic effect, with namely anti-inflammatory, anticancer and antioxidant effects with low systemic toxicity. This study aimed to obtain metabolic profiles of alpinetin in orally administrated rats. The metabolites of alpinetin were systematically analyzed and identified by ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS). The chromatographic separation was achieved on a High Strength Silica (HSS) T3 (1.8 µm, 2.1 × 100 mm) column with the mobile phase consisting of water containing 0.1% formic acid and acetonitrile with 0.1% formic acid via gradient elution. An extracted ion chromatogram strategy based on multiple prototype/metabolite intermediate templates and 71 typical metabolic reactions was proposed to comprehensively profile the metabolites of alpinetin. With the metabolite profiling strategy, altogether 15 compounds were recognized from urine, plasma, bile and feces of rats after intragastric administration of alpinetin for the first time. The prototype, glucuronide conjugates and phenolic acids metabolites were the probable predominant form of alpinetin in rats. This work showed a comprehensive study of the probable metabolic pathways of alpinetin in vivo, which could provide meaningful information for future pharmacological studies.

A Rapid UPLC-MS Method for Quantification of Gomisin D in Rat Plasma and Its Application to a Pharmacokinetic and Bioavailability Study.

Gomisin D, a lignan compound isolated from Fructus Schisandra, is a potential antidiabetic and anti-Alzheimer's agent. Recently, gomisin D was used as a quality marker of some traditional Chinese medicine (TCM) formulas. In this study, a rapid ultra-performance liquid chromatography/tandem mass spectrometry method (UPLC-MS/MS) was developed and validated to quantify gomisin D in rat plasma for a pharmacokinetic and bioavailability study. Acetonitrile was used to precipitate plasma proteins. Separations were performed on a BEH C18 column with a gradient mobile phase comprising of acetonitrile and water (0.1% formic acid). An electrospray ionization source was applied and operated in the positive ion mode. The multiple reaction monitoring mode (MRM) was utilized to quantify gomisin D and nomilin (internal standard, IS) using the transitions of m/z 531.2 → 383.1 and m/z 515.3 → 161.0, respectively. The calibration curve was linear over the working range from 1 to 4000 ng/mL (R² = 0.993). The intra- and interday precision ranged from 1.9% to 12.9%. The extraction recovery of gomisin D was in the range of 79.2-86.3%. The validated UPLC-MS/MS method was then used to obtain the pharmacokinetic characteristics of gomisin D after intravenous (5 mg/kg) and intragastric (50 mg/kg) administration to rats. The bioavailability of gomisin D was 107.6%, indicating that this compound may become a promising intragastrical medication. Our results provided useful information for further preclinical studies on gomisin D.