Serum pharmacochemistry combined with multiple data processing approach to screen the bioactive components and their metabolites in Mutan Cortex by ultra‐performance liquid chromatography tandem mass spectrometry

Root cortex of Paeonia suffruticosa Andrews (Paeoniaceae), known as Moutan Cortex (MC), is known to have anti‐allergic and anti‐inflammatory properties. However, the constituents absorbed into blood after oral administration of MC remain unknown. A sensitive and rapid method by ultra‐high‐pressure liquid chromatography–electrospray ionization–quadrupole‐time‐of‐flight mass spectrometry (UPLC‐ESI‐Q‐TOF‐MS) technology and the MetaboLynx^TM software combined with multiple data processing approach (Mdpa) was established to investigate the absorbed constituents in rats after oral administration of MC, providing unique high‐throughput capabilities for drug metabolism study. A hyphenated electrospray ionization and quadrupole‐time‐of‐flight analyzer was used for the determination of accurate mass of the fragment ion in negative mode, with excellent MS mass accuracy and enhanced data acquisition. This rapid automated analysis method was successfully applied for screening and identification of the constituents absorbed and metabolized studies of MC after oral administration to rats. A total of 46 peaks were obtained from MC, 41 of which were tentatively characterized. In the VIP‐plot of orthogonal partial least‐squares discriminant analysis, 23 interesting ions in serum samples were extracted, and 16 parent components and seven metabolites were detected in vivo. The integrative serum pharmacochemistry technique, UPLC‐ESI‐Q‐TOF‐MS, and Mdpa method were successfully applied for rapid discovery of multiple components from MC. Copyright © 2013 John Wiley & Sons, Ltd.     

Ultra‐performance liquid chromatography coupled with electrospray ionization/quadrupole time‐of‐flight mass spectrometry for the rapid analysis of constituents in the traditional Chinese medical formula Danggui San

In this work, ultra‐performance LC with ESI quadrupole TOF‐MS (UPLC–ESI‐Q‐TOF‐MS) and automated MetaboLynx analysis was used to rapidly separate and identify the chemical constituents of Danggui San, a traditional Chinese medical formula. The analysis was performed on a Waters UPLC BEH C₁₈ column using a gradient elution system. A hyphenated ESI and Q‐TOF analyzer was used for the determination of the accurate mass of the protonated or deprotonated molecule and fragment ions in both positive and negative modes. Based on retention times, accurate mass, and the mass spectrometric fragmentation characteristics, a total of 47 compounds distributed over the chemical groups of phthalides, flavonoids, monoterpene glycosides, sesquiterpenoids, phenolics, and alkaloids, were simultaneously separated within 18 min and identified or tentatively elucidated in Danggui San for the first time. UPLC–ESI‐Q‐TOF‐MS analysis revealed the complexity of the chemical composition of this formula. The method developed is rapid, accurate, reliable, and highly sensitive to characterize the chemical constituents of Danggui San.     

Method for rapidly discovering active components in Yupingfeng granules by UPLC‐ESI‐Q‐TOF‐MS

Yupingfeng granules (YPFG) were isolated from a traditional Chinese medicine (TCM) formulation composed of three herbs (Astragali Radix, Atractylodis Macrocephalae Rhizoma, and Saposhnikoviae Radix). This formulation is used in TCM to tonify qi, and it can help strengthen exterior and reduce sweating. Nevertheless, the active components of YPFG remain unclear. In this study, the chemical constituents of YPFG were systematically characterized by ultra‐performance liquid chromatography coupled with electrospray ionization/ quadrupole time‐of‐flight mass spectrometry (UPLC‐ESI‐Q‐TOF‐MS). Fifty‐eight compounds, namely, 20 flavonoids, 19 saponins, nine organic acids, four volatile coumarins, three lactones, one alkaloid, and two other components, were identified. In addition, the constituents of YPFG with the potential for in vivo bioactivities following oral administration were investigated in Sprague–Dawley rats. Thirteen compounds, namely, 11 flavonoid‐related and 2 saponin‐related components, were detected in rat plasma. After enriching flavonoids and saponins in YPFG by extraction, the extracts and YPFG were administrated to immunosuppressed rats, respectively. Plasma samples were analyzed by UPLC‐ESI‐Q‐TOF‐MS, and principal component analysis (PCA) confirmed that the extracts had similar effects to YPFG. This method could discover active ingredients in YPFG quickly and provide a scientific basis for quality control and mechanism research.     

Identification of substances migrating from plastic baby bottles using a combination of low‐resolution and high‐resolution mass spectrometric analysers coupled to gas and liquid chromatography

This work presents a strategy for elucidation of unknown migrants from plastic food contact materials (baby bottles) using a combination of analytical techniques in an untargeted approach. First, gas chromatography (GC) coupled to mass spectrometry (MS) in electron ionisation mode was used to identify migrants through spectral library matching. When no acceptable match was obtained, a second analysis by GC‐(electron ionisation) high resolution mass spectrometry time of flight (TOF) was applied to obtain accurate mass fragmentation spectra and isotopic patterns. Databases were then searched to find a possible elemental composition for the unknown compounds. Finally, a GC hybrid quadrupole‐TOF‐MS with an atmospheric pressure chemical ionisation source was used to obtain the molecular ion or the protonated molecule. Accurate mass data also provided additional information on the fragmentation behaviour as two acquisition functions with different collision energies were available (MS^E approach). In the low‐energy function, limited fragmentation took place, whereas for the high‐energy function, fragmentation was enhanced. For less volatile unknowns, ultra‐high pressure liquid chromatography‐quadrupole‐TOF‐MS was additionally applied. Using a home‐made database containing common migrating compounds and plastic additives, tentative identification was made for several positive findings based on accurate mass of the (de)protonated molecule, product ion fragments and characteristic isotopic ions. Six illustrative examples are shown to demonstrate the modus operandi and the difficulties encountered during identification. The combination of these techniques was proven to be a powerful tool for the elucidation of unknown migrating compounds from plastic baby bottles. Copyright © 2015 John Wiley & Sons, Ltd.     

Profiling and analysis of multiple compounds in rhubarb decoction after processing by wine steaming using UHPLC–Q‐TOF‐MS coupled with multiple statistical strategies

Rhubarb is one of the most popular traditional Chinese medicines and has been used for thousands of years in many Asian countries. Prepared rhubarb is obtained by steaming raw rhubarb with glutinous rice wine until it turned black in appearance both inside and outside. After processing, the therapeutic effects of prepared rhubarb change a lot. To find out the exact compound changes of the chemical profile in a decoction of rhubarb after processing and to clarify the material basis of the changed therapeutic effects, an ultra‐high performance liquid chromatography with quadrupole time‐of‐flight mass spectrometry method coupled with automated data analysis software and statistical strategy was developed. As a result, 63 peaks in raw rhubarb and 54 peaks in prepared rhubarb were detected, and a total of 45 chemical compounds were identified. The analysis data were subjected to a principle component analysis and a t‐test. Based on the results, 16 peaks were found to be the main contributors to the significant difference (p < 0.05) between raw and prepared rhubarb. Compared with raw rhubarb, the content of 15 components in prepared rhubarb was lower, while only rhein (1,8‐dihydroxy‐3‐carboxy anthraquinone) showed a higher intensity.     

Simultaneous determination of six index constituents and comparative analysis of four ethnomedicines from genus Gentiana using a UPLC‐UV‐MS method

Many species from genus Gentiana (Gentianaceae) have a long history of applications as folk medicines in the world. A simple rapid UPLC‐UV‐MS/MS method has been developed and validated for the simultaneous determination of six index constituents (gentiopicroside, swertiamarin, loganic acid, sweroside, mangiferin and ferulic acid) from the four ethnomedicines (G. rigescens Franch. ex Hemsl., G. rhodantha Franch. ex Hemsl., G. scabra Bunge and G. farreri Balf. f.). The UPLC analysis was performed on Shim‐Pack XR‐ODS III (150 × 2.0 mm, 2.2 µm). The mobile phase was consisted of acetonitrile–0.1% formic acid water using gradient elution. The wavelength 242 nm was chose for the four iridoids as well as mangiferin and 320 nm was set for ferulic acid. Mass spectrometry was applied for identification and quantification for analytes with low concentration. All the regression equations revealed a good linear relationship (R² > 0.9993). Accuracy and precision were all within the required limits. The chromatogram fingerprints analysis combined with principal component analysis showed the similarity values of the four species were <0.788 while the similarity values of G. scabra Bunge and G. rigescens Franch. ex Hemsl. were >0.993, which provided reasonable foundation for utilization and clinical application of the four ethnomedicines. This developed method appears to be a useful tool for quality control of the four ethnomedicines. Copyright © 2014 John Wiley & Sons, Ltd.     

Evaluation of the influence of mirabilite on the absorption and pharmacokinetics of the ingredients in Dahuang‐mudan decoction by a validated UPLC/QTOF–MS/MS method

Dahuang‐mudan decoction (DMD) has been widely used for disease treatment in China for 1700 years. The formula consists of Rhubarb, moutan bark, Prunus persica, wax gourd kernel and mirabilite, which have been well studied by multidisciplinary approaches. However, the role of the mineral mirabilite in DMD is unclear. The objective of this study was to investigate the effects of mirabilite on the absorption and pharmacokinetics of the ingredients in DMD. The constituents were identified in DMD extract and the plasma of mirabilite–DMD (MDMD, 50 g kg^?1) treated rats and nonmirabilite–DMD (NMDMD, 50 g kg^?1) treated rats. The plasma was also used to investigate the effects of mirabilite on the pharmacokinetics of active ingredients in DMD using a new validated UPLC–MS/MS method. The results showed that 63 compounds were identified in the extract of DMD, 27 and 22 of which were found in the plasmas of MDMD‐ and NMDMD‐treated rats, respectively. Furthermore, the results of a pharmacokinetic study suggested that mirabilite influenced the absorption of the five constituents by decreasing the absorption of emodin and rhein while increasing the absorption of aloe‐emodin, paeoniflorin and amygdalin; the pharmacokinetic parameters, including the T_max, C_max, AUC_0–t, MRT_0–t, CLz and t_1/2 of five constituents, significantly changed in MDMD‐treated rats compared with the NMDMD. The method validation for selectivity, precision, accuracy, matrix effect, recovery and stability met the acceptance criteria. These findings uncover the roles of mirabilite in DMD and demonstrate the application of scientific principles to the study of DMD in human health care.     

Development and validation of a solid‐phase extraction method coupled with LC–MS/MS for the simultaneous determination of 16 antibiotic residues in duck meat

The present study was carried out to determine 16 antibiotics belonging to seven different groups (tetracyclines, sulfonamides, penicillins, fluoroquinolones, macrolides, lincosamides and trimethoprims) in duck meat. A solid‐phase extraction method based on Oasis HLB cartridges coupled with liquid chromatography–electrospray ionization tandem mass spectrometry was developed. Solutions of 0.1 m ethylenediaminetetraacetic acid disodium salt and 2% trifluoroacetic acid were used for the preliminary extraction of the target antibiotics from duck meat and n‐hexane was used for purification prior to solid‐phase extraction. Mobile phases composed of 0.1% trifluoroacetic acid in distilled water (solvent A) and 0.1% trifluoroacetic acid in methanol (solvent B), combined with a reversed‐phase C₁₈ analytical column, provided the optimal separation and signal intensity. The linearity of the method was assessed using six concentrations (5, 10, 20, 30, 40, and 50 μg/kg), and the recoveries, which were calculated at three spiking concentrations (5, 10 and 20 μg/kg), were in the range 69.8–103.3% with relative standard deviations (RSDs) ≤ 6.9% for the 16 tested antibiotics. Matrix effects ranging from ?47.2 to ?13.5% were observed for all the analytes, and the limits of quantitation (LOQ), which ranged from 4.93 to 26.21 μg/kg, were much lower than the maximum residue limits (MRLs) set by various regulatory authorities. Ten samples from a market were tested, and none of the target analytes were detected. Thus, a simple and versatile protocol has been developed to detect and quantify 16 antibiotics in duck meat samples.     

Comprehensive characterization and quantification of multiple components in Dan‐Huang‐Qu‐Yu capsule using a multivariate data processing approach based on microwave‐assisted extraction with UHPLC and Q Exactive quadrupole‐orbitrap high‐resolution mass spectrometry

Dan‐Huang‐Qu‐Yu capsule, a Chinese herbal medicine compound preparation, is widely used for chronic pelvic inflammatory disease. In this study, a rapid, selective, and sensitive microwave‐assisted extraction ultra‐high‐performance liquid chromatography‐Q Exactive quadrupole‐orbitrap high‐resolution mass spectrometry method was developed for analyzing its chemical compositions. A total of 85 compounds, including 22 flavonoids, 8 terpenoids, 5 quinones, 5 phthaleolactone, 23 organic acids, and 22 other compounds were identified from Dan‐Huang‐Qu‐Yu capsule. Among them, 35 major compounds were unambiguously detected by comparing them with reference standards and selected as quality control markers, which were simultaneously determined in Dan‐Huang‐Qu‐Yu capsule. The established method was successfully validated and applied for simultaneous determination of 35 bioactive compounds in Dan‐Huang‐Qu‐Yu capsule from ten sample batches. The quantitative data of the analytes were analyzed by principal component analysis for quality assessment of Dan‐Huang‐Qu‐Yu capsule. Six compounds (e.g., astragaloside IV, salvianolic acid B, ellagic acid, chlorogenic acid, N‐butylidenephthalide, and luteolin) were screened out and regarded as chemical markers for quality control of Dan‐Huang‐Qu‐Yu capsule. The established method has been proved to be a novel and useful tool for rapid research of Dan‐Huang‐Qu‐Yu capsule. This research will provide reference for the scientific research of traditional Chinese medicines.