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Radix Polygalae, the dried roots of Polygala tenuifolia and P. sibirica, is one of the most well-known traditional Chinese medicinal plants. It is an important medicinal plant that has been used as a sedative and to improve memory for a number of years in most of Asia. However, the in vivo constituents of the multiple constituents from Radix Polygalae remain unknown. In the current study, ultra high performance liquid chromatography coupled to quadrupole time-of-flight mass spectrometry and the MarkerLynxTM software combined with multiple data processing approach were used to study the constituents in vitro and in vivo. A rapid and efficient method for the characterization of multiple constituents in the herbal medicine Radix Polygalae by ultra high performance liquid chromatography coupled to quadrupole time-of-flight mass spectrometry is described. In total, 35 compounds in the Radix Polygalae and 13 compounds absorbed into blood were characterized. Of the 35 compounds in vitro, ten were reported for first time. In the 13 compounds in vivo, six were prototype components and seven were metabolites were also elucidated for first time. This work narrowed the range of screening the potentially bioactive components and provided a basis for the quality control and mechanism of action.

Data-independent mass spectral acquisition is particularly powerful when combined with ultra-performance liquid chromatography (LC) that provides excellent separation of most components present in a given sample. Data-independent analysis (DIA) consists of alternating full MS scans and scans with fragmentation of all ions within a selected m/z range, providing precursor masses and structure information, respectively. Fragmentation spectra are acquired either by sequential isolation and fragmentation of sliding m/z ranges or fragmenting all ions entering the MS instrument with no ion isolation, termed broadband DIA. Previously, broadband DIA has only been possible using collision induced dissociation (CID). Here, we report the use of electron transfer dissociation (ETD) as the fragmentation technique in broadband DIA instead of traditional collision induced dissociation (CID) during MSE. In this approach, which we refer to as MSETD, we implement the inherent benefits provided by ETD, such as discrimination of leucine and isoleucine, in a DIA setup. The combination of DIA analysis and ETD fragmentation with supplemental CID energy provides a powerful platform to obtain information on all precursors and their sequence from a single experiment. Graphical Abstract ᅟ.

The fraudulent miss-description on food product labels regarding origin or composition is a widespread problem. In this work, a metabolomic approach based on the use of ultra-high performance liquid chromatography coupled to quadrupole time-of-flight mass spectrometry (UHPLC-QTOF-MS) has been applied to identify the differentiating chemical markers that allow geographic origin discrimination between different Spanish Extra Virgin Olive Oils (EVOOs). For this purpose, ninety EVOOs from 6 Spanish regions were analyzed. Data processing consisted on peak picking, retention time alignment and response normalization. Partial Least Square Discriminant Analysis (PLS-DA) and orthogonal PLS-DA (OPLS-DA) were applied to identify the most significant markers that allow groups separation. Twelve different compounds were found to correctly separate the EVOOs from their origin and 7 of them could be tentatively identified. The results of our work suggest that UHPLC-QTOF MS-based metabolomic analysis is a suitable approach for biomarker-detection in the food quality/safety field.

Ginseng is one of the most widely used natural medicines in the world. Recent studies have suggested Panax ginseng has a wide range of beneficial effects on aging, central nervous system disorders, and neurodegenerative diseases. However, knowledge about the specific bioactive components of ginseng is still limited. This work aimed to screen for the bioactive components in Panax ginseng that act against neurodegenerative diseases, using the target cell-based bioactivity screening method. Firstly, component analysis of Panax ginseng extracts was performed by UPLC-QTOF-MS, and a total of 54 compounds in white ginseng were characterized and identified according to the retention behaviors, accurate MW, MS characteristics, parent nucleus, aglycones, side chains, and literature data. Then target cell-based bioactivity screening method was developed to predict the candidate compounds in ginseng with SH-SY5Y cells. Four ginsenosides, Rg2, Rh1, Ro, and Rd, were observed to be active. The target cell-based bioactivity screening method coupled with UPLC-QTOF-MS technique has suitable sensitivity and it can be used as a screening tool for low content bioactive constituents in natural products.

The untargeted chemometric methodology for the impurity profiling of amisulpride pharmaceutical formulations was successfully applied and developed. For this purpose a fast, accurate and specific analytical method was elaborated with the use of ultra high pressure liquid chromatography coupled with high resolution hybrid electrospray ionization quadrupole time of flight mass spectrometer in a fast polarity switching mode. All the obtained chromatographic profiles were aligned and a multivariate chemometric analysis including principal component analysis and partial least square (PLS) was performed. The developed PLS-DA pattern recognition model allowed the identification of all the analyzed pharmaceutical formulations of amisulpride as well as their manufacturers. Additionally, six impurities of amisulpride were identified by the use of MS/MS fragmentation and one of them was found as the main impurity (Imp-1) and can be regarded as the primary impurity "marker" for the analyzed formulations. Furthermore, one new impurity of amisulpride was found and its chemical structure was proposed (4-amino-5-(ethylsulfinyl)-2-methoxy-N-[(1-ethylpyrrolidin-2-yl)methyl]benzamide).

RATIONALE As a herbal plant used in traditional Chinese medicine, Lamiophlomis rotata (Benth.) Kudo mainly displays its pharmacological effect by promoting blood circulation and hemostasis, dispelling wind, and acting as an analgesic. To identify the components contained in L. rotata, global detection and structural elucidation of both target and non-target components in the medicinal material was performed. METHODS L. rotata was ultrasonically extracted with methanol. Separation and analysis were achieved using liquid chromatography/quadrupole time-of-flight mass spectrometry (LC/QTOF-MS). A three-step protocol which included (1) potential components screening, (2) collection of qualitative information, and (3) database searching and structural elucidation was used for target and non-target identification. RESULTS A total of 42 components were tentatively identified, which included 12 iridoids (2 aglycones and 10 glucosides), 11 flavonoids (4 aglycones and 7 glucosides), and 13 phenylethanoid glycosides. Moreover, components of L. rotata extract belonging to the three main structural categories could be well separated in a 3D point plot according to their retention times, mass defects and degrees of unsaturation, facilitating the structural classification and identification in the subsequent studies. CONCLUSIONS The results provide a reasonable picture of the components contained in L. rotata extract and promote the further pharmacodynamic and/or pharmacokinetic characterization of this medical material, meanwhile demonstrating the utility of a universal methodology for the systematical study of herbal medicines. Copyright © 2016 John Wiley & Sons, Ltd.

Xueshuantong Injection (XSTI), derived from Notoginseng total saponins, is a popular traditional Chinese medicine injection for the treatment of thrombus-resultant diseases. Current knowledge on its therapeutic basis is limited to five major saponins, whereas those minor ones are rarely investigated. We herein develop an offline two-dimensional liquid chromatography/quadrupole time-of-flight mass spectrometry-fast data directed analysis (offline 2D LC/QTOF-Fast DDA) approach to systematically characterize the saponins contained in XSTI. Key parameters affecting chromatographic separation in 2D LC (including stationary phase, mobile phase, column temperature, and gradient elution program) and the detection by QTOF MS (involving spray voltage, cone voltage, and ramp collision energy) were optimized in sequence. The configured offline 2D LC system showed an orthogonality of 0.84 and a theoretical peak capacity of 8976. Total saponins in XSTI were fractionated into eleven samples by the first-dimensional hydrophilic interaction chromatography, which were further analyzed by reversed-phase UHPLC/QTOF-Fast DDA in negative ion mode. The fragmentation features evidenced from 36 saponin reference standards, high-accuracy MS and Fast-DDA-MS(2) data, elemental composition (C

Both qualitative and quantitative analyses combined with activity evaluation were used for the integrated quality control study of Pien-Tze-Huang (PZH), a precious lozenge used for 460 years in China. In qualitative analysis, a high performance LC-quadrupole time-of-flight MS method was developed for the identification of chemical profiling in PZH. 27 compounds were unambiguously identified by comparing with reference substances, and 12 compounds were tentatively deduced based on their MS data and comparing with literatures. Then the in vitro anti-inflammatory activity of 27 compounds were evaluated, and 9 saponins, 11 bile acids, taurine and muscone exhibited significant inhibitory activities on TNF-α production with IC50 values ranging from 12.34 to 147.24μM. In quantitative analysis, 21 active compounds, selected as quality control markers, were simultaneously detected in 15 batches of PZH samples by ultra-performance LC-triple quadrupole MS. All the analytes were detected within 15min. Satisfactory linearity was achieved with wide linear range and fine determination coefficient (r(2)>0.9990). The respective relative standard deviations of intra- and inter-day precisions, repeatability, stability, and recovery were less than 4.32%, 4.94%, 4.78%, 4.87%, and 4.91%. This study established a high sensitive and efficient method for the integrated quality control of PZH, including qualitative and quantitative analyses combined with pharmacological activity evaluation.

Rationale There is a strong demand to develop a method capable of rapid screening for the adulteration of glucocorticoids (GCs) in cosmetics. An ambient ion source– direct analysis in real time (DART), coupled with quadrupole time of flight mass spectrometry (QTOF MS) was used for the rapid screening of GCs in essential oils. Methods Liquid–liquid extraction was employed prior to the DART-QTOF MS analysis. Calibration curves for eight GCs were obtained using methyltestosterone as an internal standard. MS/MS experiments and accurate mass measurements were carried out to provide reliable and powerful evidence for the adulteration of targeted GCs. Results Quantification results were obtained in terms of linearity (R2 for all GCs, 0.986–0.996), sensitivity (limit of detection, 2.0–50 ng/mL), and repeatability (RSD, 1.2–6.0%). The entire analytical process can be finished in 5 min, compared with the GC/MS or LC/MS methods for which typical analysis times range from tens of minutes to >1 h. In addition, comparison of the performance of DART and ESI ion sources showed that DART possessed a relatively low matrix effect when handling complex samples. Conclusions A new method for the rapid screening and quantification of GCs in essential oils was developed using DART-QTOF MS. Copyright © 2016 John Wiley & Sons, Ltd.

The dried fruit of Evodia rutaecarpa (Juss.) Benth. (ER) is a well-known traditional Chinese medicine and has been used in treatment of headache, abdominal pain, migraine, chill limbs, diarrhea, nausea, hyperbaropathy, dysmenorrheal and postpartum hemorrhage. In this work, a comprehensive characterization of chemical constituents in ER extract is carried out by high performance liquid chromatography coupled to electrospray ionization and quadrupole time-of-flight mass spectrometry. The increased power of higher mass resolution and exact mass measurements for precursor and fragment ions lead to detection of 3 acylgluconic acids, 3 flavonol glycosides, 6 limonoids and 24 alkaloids in ER, of which 6 quinolone alkaloids has not been reported previously to our best knowledge. This research enriches our knowledge of chemical constituents of ER and provides a reliable guide for further pharmacology and active mechanism study on this herb.

Pseudotargeted metabolomics is a novel strategy integrating the advantages of both untargeted and targeted methods. The conventional pseudotargeted metabolomics required two MS instruments, i.e., ultra-high performance liquid chromatography/quadrupole-time- of-flight mass spectrometry (UHPLC/Q-TOF MS) and UHPLC/triple quadrupole mass spectrometry (UHPLC/QQQ-MS), which makes method transformation inevitable. Furthermore, the picking of ion pairs from thousands of candidates and the swapping of the data between two instruments are the most labor-intensive steps, which greatly limit its application in metabolomic analysis. In the present study, we proposed an improved pseudotargeted metabolomics method that could be achieved on an UHPLC/Q-TOF/MS instrument operated in the multiple ion monitoring (MIM) mode with time-staggered ion lists (tsMIM). Full scan-based untargeted analysis was applied to extract the target ions. After peak alignment and ion fusion, a stepwise ion picking procedure was used to generate the ion lists for subsequent single MIM and tsMIM. The UHPLC/Q-TOF tsMIM MS-based pseudotargeted approach exhibited better repeatability and a wider linear range than the UHPLC/Q-TOF MS-based untargeted metabolomics method. Compared to the single MIM mode, the tsMIM significantly increased the coverage of the metabolites detected. The newly developed method was successfully applied to discover plasma biomarkers for alcohol-induced liver injury in mice, which indicated its practicability and great potential in future metabolomics studies.

Corticosteroids have been often found to be added to a dietary supplement for the purpose of illegally improving the effect of their products. Thus, it is imperative to develop or improve a method that enables one to rapidly and reliably analyze corticosteroids in health or dietary supplements, for the safety management purpose. In the present study, results from liquid chromatography-quadrupole time-of-flight mass spectrometry (LC-Q-TOF-MS) experiments for the selected 35 corticosteroid compounds are presented, which can be useful for the qualitative screening of corticosteroids in health or dietary supplements. Specifically, retention times, accurate mass data of the protonated steroids, m/z values of major fragment ions are given for the 35 corticosteroids. Further, fragmentation pathways for the selected steroids are also suggested. Based on the suggested fragmentation pathways, it was shown that an unknown steroid compound can be readily identified using the knowledge of a group of unique and specific common skeletal fragments. The high selectivity and sensitivity of the LC-Q-TOF-MS/MS results combined with the knowledge of the fragmentation pathways can offer a new opportunity for rapid and accurate screening of corticosteroids, thus preventing health-related incidents involving adulterated products and clamping down on illegally circulated health products.

Ancient Chinese textile dyeing technology has been greatly developed since the Tang Dynasty. The types of plant that can be used in dyeing has also increased during that period of time. In this study, Ultra Performance Liquid Chromatography-Quadrupole-Time of Flight-Mass Spectrometry (UPLC-Q-TOF-MS) and Diode Array Detector methods were used for analyzing and identifying of the dyestuff used on the two pieces of textile from the Tang Dynasty. The results show that the dyestuffs of the red and brown samples are derived from madder; the dyestuffs of the blue sample is derived from indigo, while the dyestuffs of the purple is derived from Lithospermum rythrorhizon .

The Kang-Jing (KJ) formula is a compound preparation made from 12 kinds of herbs. So far, four different methods (M1-M4) have been documented for KJ preparation, but the influence of preparation methods on the holistic quality of KJ have remained unknown. In this study, a strategy was proposed to investigate the influence of different preparation methods on the holistic quality of KJ using ultra-high performance liquid chromatography coupled with quadrupole/time of flight mass spectrometry (UHPLC-QTOF-MS/MS) based chemical profiling. A total of 101 compounds mainly belonging to flavonoids, tanshinones, monoterpene glycosides, triterpenoid saponins, alkaloids, phenolic acids and volatile oils, were identified. Among these compounds, glaucine was detected only in M3/M4 samples, while two dehydrocorydaline isomers merely detected in M2/M3/M4 samples. Tetrahydrocolumbamine, ethylic lithospermic acid, salvianolic acid E and rosmarimic acid were only detected in M1/M3/M4 samples. In the subsequent quantitative analysis, 12 major compounds were determined by UHPLC-MS/MS. The proposed method was validated with respect to linearity, accuracy, precision and recovery. It was found that the contents of marker compounds varied significantly in samples prepared by different methods. These results demonstrated that preparation method does significantly affect the holistic quality of KJ. UHPLC-QTOF-MS/MS based chemical profiling approach is efficient and reliable for comprehensive quality evaluation of KJ. Collectively, this study provide the chemical evidence for revealing the material basis of KJ, and establish a simple and accurate chemical profiling method for its quality control.

Allii Macrostemonis Bulbus (named Xiebai in China) is a folk medicine with medicinal values for the treatment of thoracic obstruction and cardialgia, and a food additive as well. However, there is even no quantitative standard for Allii Macrostemonis Bulbus recorded in the current edition of the Chinese Pharmacopeia. Hence, simultaneous assay of multiple components is urgent. In this study, chemometric methods were firstly applied to discover the components with significant fluctuation among multiple Allii Macrostemonis Bulbus samples based on optimized fingerprints. Meanwhile, the major components and main absorbed components in rats were all selected as its representative components. Subsequently, a sensitive method was established for the simultaneous determination of 54 components (15 components for quantification and 39 components for semiquantification) by ultra high performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry. Moreover, the validated method was successfully applied to evaluate the quality of multiple samples on the market. It became known that multiple Allii Macrostemonis Bulbus samples varied significantly and showed poor consistency. This work illustrated that the proposed approach could improve the quality of Allii Macrostemonis Bulbus, and it also provided a feasible method for quality evaluation of other traditional Chinese medicines.

Fusarium oxysporum L11 is a non-pathogenic soil-borne fungal strain that yielded an extract that showed antifungal activity against phytopathogens. In this study, reversed-phase high-performance liquid chromatography (RP-HPLC) coupled to different atmospheric pressure ionization sources-quadrupole-time-of-flight mass spectrometry (API-QTOF-MS) was applied for the comprehensive profiling of the metabolites from the extract. The employed sources were electrospray (ESI), atmospheric pressure chemical ionization (APCI) and atmospheric pressure photoionization (APPI). Post-column addition of metal solutions of Ca, Cu and Zn(II) was also tested using ESI. A total of 137 compounds were identified or tentatively identified by matching their accurate mass signals, suggested molecular formulae and MS/MS analysis with previously reported data. Some compounds were isolated and identified by NMR. The extract was rich in cyclic peptides like cyclosporins, diketopiperazines and sansalvamides, most of which were new, and are reported here for the first time. The use of post-column addition of metals resulted in a useful strategy for the discrimination of compound classes since specific adducts were observed for the different compound families. This technique also allowed the screening for compounds with metal binding properties. Thus, the applied methodology is a useful choice for the metabolic profiling of extracts and also for the selection of metabolites with potential biological activities related to interactions with metal ions. Fil: Cirigliano, Adriana Monica. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Unidad de Microanálisis y Métodos Físicos en Química Orgánica. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Unidad de Microanálisis y Métodos Físicos en Química Orgánica; Argentina Fil: Rodriguez, Maria Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones en Biociencias Agrícolas y Ambientales. Universidad de Buenos Aires. Facultad de Agronomía. Instituto de Investigaciones en Biociencias Agrícolas y Ambientales; Argentina Fil: Gagliano, M. Laura. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Unidad de Microanálisis y Métodos Físicos en Química Orgánica. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Unidad de Microanálisis y Métodos Físicos en Química Orgánica; Argentina Fil: Bertinetti, Brenda Veronica. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Unidad de Microanálisis y Métodos Físicos en Química Orgánica. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Unidad de Microanálisis y Métodos Físicos en Química Orgánica; Argentina Fil: Godeas, Alicia Margarita. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones en Biociencias Agrícolas y Ambientales. Universidad de Buenos Aires. Facultad de Agronomía. Instituto de Investigaciones en Biociencias Agrícolas y Ambientales; Argentina Fil: Cabrera, Gabriela Myriam. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Unidad de Microanálisis y Métodos Físicos en Química Orgánica. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Unidad de Microanálisis y Métodos Físicos en Química Orgánica; Argentina

Liquid chromatography with quadrupole time-of-flight mass spectrometry coupled with automated data analysis by Peakview software was employed to systematically screen and characterize the astragalosides in Radix Astragali, a Chinese medical preparation. The separation was performed on a poroshell 120 SB-C18 column equipped in a conventional liquid chromatography system. After being separated using a general gradient elution, the analytes were detected by the triple quadrupole time-of-flight mass spectrometer in both positive- and negative-ion modes. The mass defect filtering function built in the Peakview software was utilized to rapidly screen the potential ions of interest, while some functions of Peakview such as Formula Finder, XIC manager, and IDA Explorer were employed to facilitate the assignment or characterization of the screened astragalosides. A total of 42 astragalosides were screened and tentatively characterized or assigned, and 20 of them were firstly detected in Radix Astragali. According to the screened astragalosides, acetylation, glycosidation, hydrogenation, oxidation, and hydration were considered to be the major secondary metabolic pathways involved in the formation of the astragalosides. The combination of liquid chromatography with quadrupole time-of-flight mass spectrometry and automated Peakview analysis is a feasible and efficient tool to screen and identify the constituents in complex matrices of herbal medicines.

Traditional Chinese medicine (TCM) preparations have become effective treatments for many diseases. However, their active ingredients are still uncertain and difficult to identify. In this study, we propose a strategy that integrates ultra-performance liquid chromatography/quadrupole-time-of-flight mass spectrometry (UPLC/Q-TOF-MS) and bioactive (NF-κB inhibitor) luciferase reporter assay systems for the rapid determination of various anti-inflammatory compounds of TCM preparations. In this way, Bufei Granule (BFG), a TCM preparation used for the clinical therapy of asthma, was analyzed by the two ways of component identification and activity detection. Potential anti-inflammatory constituents were screened by NF-κB activity assay systems and simultaneously identified according to the mass spectrometry data. Three structural types of NF-κB inhibitors (caffeic acid derivatives, flavonoids and Pentacyclic triterpenes) were characterized. Further cytokine detection confirmed the anti-inflammatory effects of the potential NF-κB inhibitors. Compared with conventional chromatographic separation and inhibitory activity detection, integrating UPLC/Q-TOF-MS identification and virtual validation was more convenient and more reliable. This strategy clearly demonstrates that MS data-based fingerprinting is a meaningful tool not only in identifying constituents in complex matrix but also in directly screening for powerful trace ingredients in TCM preparations. Copyright © 2016 John Wiley & Sons, Ltd.

Graphical abstract, Pharmaceuticals, illicit drugs and personal care products are emerging contaminants widely distributed in water. Currently, a number of solid-phase extraction (SPE) procedures followed by liquid chromatography tandem mass spectrometry (LC–MS/MS) have been reported. However, target analysis of selected compounds is commonly used whereas other related contaminants present in the sample remain invisible. Carmona et al. [1] described a method for determining 21 emerging contaminants by LC–MS/MS with improved mobile phases. We tested this protocol in combination with high resolution mass spectrometry using a quadrupole time-of-flight (QqTOF) instrument to get a wide non-target screening approach in order to have a broader scope and more practical method for detecting licit and illicit drugs and personal care products than traditional target methods. The essential points in the method are: • The screening capabilities of QqTOF (ABSciex Triple TOF™) are used for detecting and identifying non-target pharmaceuticals and a large number of other emerging contaminants in water. • The quantitative features of the instrument, the Achilles heel of the QqTOF mass spectrometers, are established for few selected compounds. • The method may be applied to identify a large number of emerging contaminants in water. However, pre-validation will be needed to quantify them.

Pufferfish is worldwide known for its umami taste. Previous studies showed that umami peptide played a key role in taste perception of pufferfish. In order to more explain the umami taste of pufferfish in details, this study was aimed to identify more umami peptides from pufferfish. The extraction of Takifugu rubripes (T. rubripes) muscle after heating and ultrasound treatment was isolated by consecutive chromatography combined with sensory evaluation. The results showed that seven umami peptides from 300 to 3000 Da molecular weight range fractions were firstly identified by nano-liquid chromatography quadrupole time of flight mass spectrometry (Nano-LC/Q-TOF-MS), and peptide sequences are HLQLAIR, DPLRGGYY, AGLQFPVGR, LLLPGELAK, AGFAGDDAPR, GYSFTTTAER and DAGVIAGLNVLR. The taste characteristics of seven peptides were analyzed based on sensory evaluation, and their umami thresholds were in the ranges of 0.06–0.27 mM. Peptide sequences source analysis found that actin was one important taste-active precursor of the umami peptides in T. rubripes.

Grape dehydration is an oenological process used for production of high-quality reinforced and sweet wines. High-resolution mass spectrometry (ultrahigh-performance liquid chromatography/quadrupole time-of-flight [UHPLC/QTOF]) was used to deepen the characterization of some flavonoids previously proposed in Corvina and Raboso Piave withered grapes. By performing a targeted data analysis workflow and orthogonal identification approaches (tandem mass spectrometry [MS/MS]), in silico fragmentation, and calculation of putative retention time), elucidation on the structures of six compounds previously proposed was achieved (taxifolin-pentoside, two tetrahydroxyflavanone-hexoside derivatives, a tetrahydroxy-dimethoxyflavanone-hexoside derivative, a pentahydroxyflavone, and peonidin-O-pentoside); and the structures of four putative new grape flavonoids were characterized (dihydromyricetin-O-hexoside, taxifolin-di-O-hexoside, isorhamnetin, and a pinoquercetin isomer). Findings enlarge the panorama of flavonoids in grape and of their possible biosynthetic pathways.

In this research, a novel ultrasound-assisted extraction (USAE) method using a one-phase solvent (isopropanol, IPA) combined with liquid chromatography-electrospray ionization quadrupole time-of-flight mass spectrometry (LC-ESI-QToF-MS) was developed for exhaustive profiling of egg yolk lipids, which are considered to be essential nutrients that have important functions for human health. Compared with IPA-vortex extraction method, the optimized IPA-USAE method shows better performance in terms of extraction time, lipid losses, lipid recoveries and the number of lipids identified. With this established simple, high throughput and reliable analytical approach, untargeted comprehensive lipid profiling of four kinds of egg yolks was achieved, thus providing a powerful tool for nutritional research of egg lipids. Considering that the lipids contained in egg yolk are quite abundant in both quantity and species, the proposed IPA-USAE combined with LC-ESI-QToF-MS is also potentially applicable to comprehensive extraction and profiling of lipids in other biological samples.

In this paper, an ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF HRMS) method was developed and validated for screening, confirmation and quantitation of 31 anti-impotence compounds potentially illegally added to herbal-based dietary supplements. The analytes were well separated by the mobile phase consisted of 0.1% formic acid solution and acetonitrile with gradient elution at a flow rate of 0.3 mL/min. The MS analysis was operated in positive mode and the mass error of the 31 compounds were below 2.9 ppm. The method validation showed good linearity with coefficients of determination (r2) higher than 0.9973 for all analytes. LODs and LLOQs ranged from 0.005 to 0.50 μg/g or μg /mL and from 0.02 to 1.24 μg /g or μg/mL, respectively. The accuracy was in the range of 86.6% to 113.7%, while the intra-and inter-day precision were in the ranges of 0.9–7.6% and 0.9–11.4%, respectively. The absolute and relative matrix effect were in the range of 65.8–115.6% and 0.6–13.3%. The mean recoveries were in the range of 80.5–116.9%. The stability ranged from 0.4% to 8.5%. Among 200 batches of herbal-based dietary supplements, sildenafil and/or tadalafil were found to be added illegally in two samples, while not very high concentration of icariin was detected in one sample. The Q-TOF mass spectrometry has been proved to be a very powerful and efficient tool for rapid screening of 31 anti-impotence compounds potentially illegally added to herbal-based dietary supplements, ensuring food safety and public health.

The method of ultra-high-performance liquid chromatography/quadrupole time-of-flight mass spectrometry (UHPLC-Q/TOF-MS) was established and combined with principal component analysis (PCA) to identify natural Calculus Bovis, in vitro cultured Calculus Bovis and artificial Calculus Bovis. PCA, which was particularly powerful in dealing with multicollinearity and variables that outnumber the samples, was used to analyze the UHPLC-MS data of the processed samples, and potential markers were analyzed and described based on orthogonal partial least-squares discriminant analysis. According to the results in this study, the approach of combining UHPLC-QTOF-MS with PCA was proven to be credible and could be used to identify Calculus Bovis from in vitro cultured Calculus Bovis and artificial Calculus Bovis and to determine if there is Calculus Bovis in patented Chinese medicines that should contained Calculus Bovis medicinal materials.

Cicadae Periostracum, which is derived from the slough of Cicadidae insects, is a commonly used crude drug in traditional Chinese medicine (TCM). As specified in Chinese Pharmacopoeia, Cryptotympana atrata (CA) is the only official species of this crude drug. However, the slough of other three species, i.e., Auritibicen flammatus (AF), Cryptotympana mandrina (CM) and Platypleura kaempferi (PK), have been also used as the origins of Cicadae Periostracum in Chinese herbal market, although whether the quality of these four origins is consistent or not is still unknown. In present study, ultra-high performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS/MS) was employed to qualitatively and quantitatively compare the chemical profiles of the four origins. Totally, 34 N-acetyldopamine polymers were identified from the four origins, including 4 N-acetyldopamine dimers, 11 N-acetyldopamine trimers, 10 N-acetyldopamine tetramers, and 9 N-acetyldopamine pentamers. AF, CM and PK had similar chemical profiles with that of CA. The contents and compositional ratio of the four types of polymers in CA, AF and CM were consistent with each other, but significantly lower or different in PK. All these results suggested that AF and CM might be considered as the potential resources of Cicadae Periostracum concerning their consistent holistic quality, whereas whether PK could be used as potential origin of Cicadae Periostracum or not need further evaluation for their different compositional ratios and contents of the four types of N-acetyldopamine polymers. This is the first study on chemical profiling and comparison of N-acetyldopamine polymers in four origins of Cicadae Periostracum, which is beneficial for potential resources utilization and quality standard improvement of Cicadae Periostracum.

The aim of this study was to simultaneously determine the presence of unauthorized drug substances in health foods and herbal products used in the treatment of conditions such as gout and anti-osteoporosis. Therefore, we developed and optimised a rapid and accurate method to simultaneously measure 20 anti-gout and anti-osteoporosis drug substances using an ultra-high-performance liquid chromatography (UPLC) system equipped with a photodiode array (PDA) detector. The method was validated to fully meet internationally accepted standards. LODs and LOQs spiked in solid and liquid negative samples were ranged from 0.12 to 1.50 μg/mL, and ranged from 0.36 to 4.50 μg/mL. Linearities (R2> 0.999), stabilities (RSD ≤ 2.92%), accuracies (84.25∼106.62%, intra-day; 84.56∼105.85%, inter-day), precisions (RSD ≤ 3.71% on the intra-day; RSD ≤ 3.47% on the inter-day), recoveries spiked in various type of blank samples such as powder, liquid, tablet, and capsule were determined within 81.20–116.20 %, respectively. From a confirmation of matrix effects (88.06∼110.50% in solid blank; 89.16∼110.52% in liquid blank), it was confirmed that this method was not significantly affected by a sample matrix. The validated method was used to analyse 116 samples containing health foods, herbal products, and seized forensic samples advertised to be effective anti-gout and anti-osteoporosis agents. Of the 20 drug substances screened, dexamethasone was detected and confirmed by comparing the tandem mass spectrometry (MS/MS) fragment ion patterns of a reference standard and the sample using LC-quadrupole-time-of-flight (Q-TOF)/MS. The concentrations of adulterants in seized forensic samples ranged from 0.013 to 0.022 %.

Using ultra high performance liquid chromatography quadrupole/time-of-flight mass spectrometry (UPLC-QTOFMS) based metabolomics, we focused on developing a method for the comprehensive distinction between Citri Reticulatae Blanco Pericarpium(CRBP) and Citri Reticulatae Chachi Pericarpium (CRCP), as well as the CRCP within different storage years in this study. Through this, we hope to enhance Citri Reticulatae Pericarpium (CRP) Quality Control system. Using UNIFI software and an online database identified chemical components in the 3-30 years CRCP(40 batches) and CRBP (10 batches)samples, and multivariate statistical analysis methods and heat-map were applied to distinguish between CRCP and CRBP and CRCP in different storage years. The results showed that a total of 92 compounds were identified from CRCP and CRBP samples, most of which were flavonoids. Principal component analysis (PCA) and orthogonal partial least squares discrimination analysis (OPLS-DA) indicated that it can effectively distinguish between CRBP and CRCP and various storage years CRCP, and 19 metabolites were identified as potential markers for distinguishing between CRBP and CRCP, and 15 potential markers showed a higher level of CRCP than CRBP. At the same time, 31 metabolites were identified to distinguish CRCP in different storage years, metabolite levels increased in 3-10 years and decreased after 15-30 years. Therefore, this approach can effectively distinguish between CRCP and CRBP and CRCP with different storage years, and may also provide a feasible strategy for the certification of Chinese herbal medicines from different species and storage years.

Scopolia tangutica is a traditional Chinese medicine used for antispasmodic, anesthesia, analgesia, and sedation. Its medicinal activity is associated to alkaloid constituents, including tropane and cinnamamide types. Low content of alkaloids in plant makes them difficult to be isolated and identified. The present work developed an effective method to quickly characterize alkaloids from Scopolia tangutica by high-performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry. Thirteen reference compounds were studied for their fragmentation pathways, including five tropane alkaloids and eight cinnamamide ones. Alkaloid constituent was analyzed by an optimized high-performance liquid chromatography method and mass spectrometry analysis to achieve systematic characterization of alkaloids from Scopolia tangutica. As a result, 53 compounds were identified, including 21 tropane alkaloids (eight new ones), 18 caffeoyl ones (ten new ones) and 14 dicaffeoyl ones (seven new ones). It was important to provide rich information in phytochemical study and structure-guided isolation of important compounds from this plant.

Background: Snake bile originates from nearly 20 species from three families (Elapidae, Colubridae, and Viperidae). However, the components of various snake bile were not compared with one another. Objecive: The aim of the study was to develop a TLC-MS method for differentiation of various snake bile derived from a different genus. Method: As a type of traditional identification method for Chinese herbs, TLC has considerable advantages as a multicomponent separation system. Results: In this paper, TLC was used to separate the components in snake bile. It was found that the snake bile from Colubridae family and Cobra species is different from the others using TLC. The molecular formulas of the bands were elucidated with TLC coupled with quadrupole–time-of-flight–MS by the TLC-MS interface. Two bands on the plate were identified with the reference substance, and the other three bands were analyzed by the TLC-MS method without the reference substance. The corresponding molecular formulas of these bands were given according to accurate molecular weights. Conclusions: The results from this study indicate that the proposed method is reliable, and it has been successfully applied to the identification of snake bile samples.

The flowers of Juglans regia L. have demonstrated their medicinal value as a part of edible plants. In this study, an analytical methodology for identification of flavonoid chemical constituents in flowers of Juglans regia L. was established using ultra-high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF-MS). Thirty-six flavonoid compounds were identified based on highly accurate mass measurements, and the mass spectrometric fragmentation pathways of eleven representative compounds were proposed, which was helpful for the identification of different types of flavonoids. The study has laid the foundation for the research and development of effective drugs from flowers of Juglans regia L.

Drug screening using high-resolution mass spectrometers, including quadrupole time-of-flight mass analyzers (QqTOFs), is becoming increasingly popular due to the additional flexibility that these instruments offer laboratories. Liquid chromatography (LC) coupled to TOF, as in an LC-QqTOF, offers comparable sensitivity and a shortened method development time relative to triple quadrupole-based mass spectrometry. In addition, LC-QqTOF data that are collected in untargeted mode can be analyzed retrospectively to detect additional compounds that were not predefined targets. Much of the power of LC-QqTOF lies in data processing, and the data analysis workflow that a lab uses must be adequately validated.

High-throughput simultaneous quantitative and qualitative (Quan/Qual) analysis is attractive to combine targeted with non-targeted analysis, e.g. in pharmacometabolomics and drug metabolism studies. This study aimed to investigate the possibilities and limitations of high-throughput Quan/Qual analysis by ultra-high performance liquid chromatography (UHPLC) coupled with high-resolution mass spectrometry (HRMS), to develop a widely applicable Quan/Qual UHPLC-HRMS method and to provide recommendations for Quan/Qual method development. A widely applicable 4.25-min UHPLC method for small-molecules was used to investigate and optimize mass spectrometric parameters of a Synapt G2S for Quan/Qual analysis. The method was applied on a rat metabolomics study investigating the effect of the fasting state and administration of a dosing vehicle on the rat plasma metabolic profile. scan mode with a short scan time of 30 ms. This 4.25-min Quan/Qual analysis method enabled quantification with accuracy and precision values ≤ 20% at the lowest quality control (QC) level and ≤15% at higher QC levels for 16 out of 19 tested analytes. It provided both parent m/z values and fragmentation spectra for compound identification with limited loss of chromatographic resolution and it revealed biologically relevant metabolites in its application to the metabolomics study. Quan/Qual method development requires balancing between the amount of qualitative data, the quality of the quantitative data and the analysis time. Recommendations are provided for MS resolution, scan mode, scan rate, smoothing and peak integration in Quan/Qual method development and analysis. BACKGROUND METHODS RESULTS CONCLUSION

In the food industry and herbal markets, it is critical to control the quality of processed Panax ginseng products. In this study, ultra-performance liquid chromatography coupled to quadrupole time of flight mass spectrometry (UPLC-QTOF/MS)-based metabolomics was applied for the quality evaluation of white ginseng (WG), tae-geuk ginseng (TG), red ginseng (RG), and black ginseng (BG). Diverse metabolites including ginsenosides were profiled by UPLC-QTOF/MS, and the datasets of WG, TG, RG, and BG were then subjected to multivariate analyses. In principal component analysis (PCA), four processed ginseng products were well-differentiated, and several ginsenosides were identified as major components of each product. S-plot also characterized the metabolic changes between two processed ginseng products, and the major ginsenosides of each product were found as follows: WG (M-Rb1, M-Rb2, M-Rc, Re, Rg1), TG (Rb2, Rc, Rd, Re, Rg1), RG (Rb1, Rb2, Rc, Rd, Re, Rg1), and BG (Rd, Rk1, Rg5, Rg3). Furthermore, the quantitative contents of ginsenosides were evaluated from the four processed ginseng products. Finally, it was indicated that the proposed metabolomics approach was useful for the quality evaluation and control of processed ginseng products.

Necrostatins have been shown to retard necroptosis, a programmed necrotic-like cell death, which has been shown to underlie pathophysiology of various diseases. Nec-1s, a novel highly effective necrostatin, overcomes some drawbacks of former necrostatin analogues. The determination of Nec-1s in biological system, however, has not been carried out so far. Therefore, this study was undertaken to optimize and validate the HPLC-DAD-Q-TOF method for the assessment of Nec-1s levels in the plasma what is the necessity for designing its proper dosing regimen for in vivo studies. Benefits of the proposed analytical protocol include: (i) simple sample preparation (precipitation of plasma proteins, evaporation of acetonitrile, reconstitution in mobile phase), (ii) fast, selective and sensitive analysis due to a highly orthogonal LC-MS system providing less than 8 min analysis time, (iii) detection of Nec-1s without any matrix interferences, and quantitation of very low concentration levels of Nec-1s (LLOQ ~ 20 ng/mL), (iv) high reliability of Nec-1s determination with precision and accuracy values meeting the FDA criteria for biomedical analysis. The proposed analytical protocol is suitable for routine use in relevant biological studies, and, in this work, it was successfully applied for monitoring of Nec-1s plasma levels in rats providing reproducible and consistent results. Based on pharmacokinetic features, which can also be assessed due to the results of this study, there will be efforts to perform both acute and chronic in vivo studies and potential clinical safety studies first.

A method for simultaneous analysis of egg and milk allergens using LC-QTOF-MS was developed. The proteins measured were α-casein, β-lactoglobulin, and ovalbumin, which are the main protein allergens in milk and eggs. The proteins were digested using trypsin, and the digests were analyzed by LC-QTOF-MS. Sixteen peaks were detected that confirmed the amino acid sequences of the digests, and a MRM method with high resolution (MRM-HR) using product ions of these peaks was applied for quantification. Next, validation studies were performed using beverage products to which milk and egg standard protein solutions had been added. Good linearity was achieved over the concentration range of 1.25 to 20 μg/g of milk and egg protein, and acceptable reproducibility and accuracy were obtained at 10 μg/g. Moreover, good agreement was also observed between LC-QTOF-MS and ELISA. These findings suggest that this LC-QTOF-MS method may be useful for determining the milk and egg protein contents of beverages.

Fabry disease is a rare lysosomal storage disorder resulting from the lack of &alpha, Gal A gene activity. Globotriaosylceramide (GB3, ceramide trihexoside) is a novel endogenous biomarker which predicts the incidence of Fabry disease. At the early stage efficacy/biomarker study, a rapid method to determine this biomarker in plasma and in all relevant tissues related to this disease simultaneously is required. However, the limited sample volume, as well as the various levels of GB3 in different matrices makes the GB3 quantitation very challenging. Hereby we developed a rapid method to identify GB3 in mouse plasma and various tissues. Preliminary stability tests were also performed in three different conditions: short-term, freeze-thaw, long-term. The calibration curve was well fitted over the concentration range of 0.042&ndash, 10 &mu, g/mL for GB3 in plasma and 0.082&ndash, 20 &mu, g/g for GB3 in various tissues. This method was successfully applied for the comparison of GB3 levels in Fabry model mice (B6, 129-Glatm1Kul/J), which has not been performed previously to the best of our knowledge.

Here, time-decoupled comprehensive two-dimensional ultra-high liquid chromatography (UHPLC) coupled with an ion mobility (IM)-high resolution mass spectrometer (HRMS) was established and used to analyze ginsenosides from the main roots of white ginseng (WG) and red ginseng (RG), which enabled the separation of complex samples in four dimensions (2D-LC, ion mobility, and mass spectrometry). The incompatibility of mobile phases, dilution effect, and long analysis time, which are the main shortcomings of traditional comprehensive 2D-LC methods, were largely avoided in this newly established 2D-UHPLC method. The orthogonality of this system was 55%, and the peak capacity was 4392. Under the optimized 2D-UHPLC-IM-MS method, 201 ginsenosides were detected from white and red ginseng samples. Among them, 10 pairs of co-eluting isobaric ginseng saponins that were not resolved by 2D-UHPLC-HRMS were further resolved using 2D-UHPLC-IM-MS. In addition, 24 ginsenoside references were analyzed by UHPLC-IM-MS to obtain their collision cross section (CCS) values and ion mobility characteristics. Finally, the established new method combined with multivariate statistical analysis was successfully applied to differentiate WG and RG, and 9 ginsenosides were found to be the potential biomarkers by S-Plot and the values of max fold change, which could be used for classifying WG and RG samples. Overall, the obtained results demonstrate the applicability and potential of the established time-decoupled online comprehensive 2D-UHPLC-IM-MS system, and it will be extended to the analysis of other targeted or untargeted compounds, especially co-eluting isomers in more herbal extracts.

Metabonomic techniques have been used to discover subtle differences in the small molecule profiles of chicken\ud eggs, which could help to combat fraud within the egg industry. High Performance Liquid Chromatography Quadrupole Time-of-Flight Mass Spectrometry (HPLC-Q-ToF-MS) was used to obtain profiles of the small molecules present in the yolks of chicken eggs stored for different lengths of time. Statistical analysis, including the use of XCMS Online and further exploratory statistics, was able to uncover differences in the abundances of several of the small molecules found in these egg yolks. One of these small molecules was identified, through the use of METLIN and MS/MS analysis, as choline. A targeted study was then carried out over a longer storage period, using the same instrumentation and analytical techniques, in order to observe how the concentration of choline in egg yolk changes over a longer period of time.

Hongjingtian injection is made from Rhodiola wallichiana and used in the treatment of stable angina pectoris associated with coronary heart disease. In this study, the chemical constituents in Hongjingtian injection were comprehensively studied using liquid chromatography quadrupole time-of-flight mass spectrometry. A total of 49 compounds were identified or assumed, including 10 organic acids, nine phenylethanoids, 10 phenylpropanoids, two flavonoid glycosides, seven monoterpene glycosides, seven octylglycosides and four other types of compounds. The structures of seven compounds were confirmed by comparing their retention times, MS and UV spectra with the corresponding authentic standards. Amongst the 49 compounds, 35 were firstly found in R. wallichiana, while they have been reported in other species of the genus Rhodiola, including Rhodiola crenulata, Rhodiola sacra, Rhodiola rosea and Rhodiola kirilowii. The possible fragmentation pathways in the mass spectrometry of the major types of compounds are proposed and summarized. Our study demonstrates a rapid method for characterizing the chemical constituents present in the Hongjingtian injection, which could also be applied to the identification of chemical constituents in other TCM formulae containing R. wallichiana.

The implementation of whole-exome sequencing in clinical diagnostics has generated a need for functional evaluation of genetic variants. In the field of inborn errors of metabolism (IEM), a diverse spectrum of targeted biochemical assays is employed to analyze a limited amount of metabolites. We now present a single-platform, high-resolution liquid chromatography quadrupole time of flight (LC-QTOF) method that can be applied for holistic metabolic profiling in plasma of individual IEM-suspected patients. This method, which we termed “next-generation metabolic screening” (NGMS), can detect >10,000 features in each sample. In the NGMS workflow, features identified in patient and control samples are aligned using the “various forms of chromatography mass spectrometry (XCMS)” software package. Subsequently, all features are annotated using the Human Metabolome Database, and statistical testing is performed to identify significantly perturbed metabolite concentrations in a patient sample compared with controls. We propose three main modalities to analyze complex, untargeted metabolomics data. First, a targeted evaluation can be done based on identified genetic variants of uncertain significance in metabolic pathways. Second, we developed a panel of IEM-related metabolites to filter untargeted metabolomics data. Based on this IEM-panel approach, we provided the correct diagnosis for 42 of 46 IEMs. As a last modality, metabolomics data can be analyzed in an untargeted setting, which we term “open the metabolome” analysis. This approach identifies potential novel biomarkers in known IEMs and leads to identification of biomarkers for as yet unknown IEMs. We are convinced that NGMS is the way forward in laboratory diagnostics of IEMs. Electronic supplementary material The online version of this article (10.1007/s10545-017-0131-6) contains supplementary material, which is available to authorized users.

Deng-Zhan-Xi-Xin injection is a well-known traditional Chinese medicine prescription for the treatment of cardiovascular and cerebral vessel diseases. However, there have been few reports on its chemical constituents and metabolic pathway, which has blocked its further quality control and studies on its pharmacology and mechanism of action. In this study, an integrative method was established to rapidly explore the chemical constituents and metabolites of Deng-Zhan-Xi-Xin injection using ultra high performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry and the UNIFI™ software combined with multiple data processing approaches. As a result, a total of 40 compounds, including 9 flavonoids and 31 phenolic acids were identified or tentatively characterized, and five compounds were first reported in Deng-Zhan-Xi-Xin injection. Under the same analysis conditions, 70 compounds have been detected in rats, including 25 prototypes and 45 metabolites. This was the first systematic research study on the metabolic profiling of Deng-Zhan-Xi-Xin injection. This study provides valuable chemical information for the quality control and research on pharmacology and mechanism of action of Deng-Zhan-Xi-Xin injection. Moreover, it provides a valuable strategy for analyzing the chemical components and metabolites of other traditional Chinese medicine prescriptions.

Rationale: The World Anti-Doping Agency (WADA) encourages drug-testing laboratories to develop screening methods that can detect as many doping substances as possible in urine. The use of full-scan high-resolution acquisition (FS/HR) with gas chromatography/mass spectrometry (GC/MS) for the detection of known and unknown trimethylsilyl (TMS) derivatives of anabolic-androgenic steroids (AAS) provides anti-doping testing bodies with a new analytical tool. Methods: The AAS were extracted from urine samples by generic liquid–liquid extraction, after enzymatic hydrolysis, and TMS derivatization. The extracted urine was analyzed by GC/Q-TOF and GC/Q-Orbitrap to compare the performance of the two instrument types for the detection of 46 AAS in human urine. The quantitation of endogenous anabolic steroids and the ability of the two analytical platforms to comply with the requirements for testing as part of the WADA Athlete Biological Passport (ABP) were also assessed. Results: The data presented show that the analytical performance for both instruments complies with the WADA specifications. The limits of detection (LODs) for both instruments are well below the WADA 50% Minimum Required Performance Levels. The mass errors in the current study for the GC/Q-Orbitrap platform are lower than those obtained for the GC/Q-TOF instrument. Conclusions: The data presented herein proved that both molecular profiling platforms can be used for antidoping screening. The mass accuracies are excellent in both instruments; however, the GC/Q-Orbitrap performs better as it provides higher resolution than the GC/Q-TOF platform.

This paper uses the LC-quadrupole-time-of-flight MS technique to evaluate the behavioral characteristics of MS of 485 pesticides under different conditions and has developed an accurate mass database and spectra library. A high-throughput screening and confirmation method has been developed for the 485 pesticides in fruits and vegetables. Through the optimization of parameters such as accurate mass number, time of retention window, ionization forms, etc., the method has improved the accuracy of pesticide screening, thus avoiding the occurrence of false-positive and false-negative results. The method features a full scan of fragments, with 80% of pesticide qualitative points over 10, which helps increase pesticide qualitative accuracy. The abundant differences of fragment categories help realize the effective separation and qualitative identification of isomer pesticides. Four different fruits and vegetables-apples, grapes, celery, and tomatoes-were chosen to evaluate the efficiency of the method at three fortification levels of 5, 10, and 20 μg/kg, and satisfactory results were obtained. With this method, a national survey of pesticide residues was conducted between 2012 and 2015 for 12 551 samples of 146 different fruits and vegetables collected from 638 sampling points in 284 counties across 31 provincial capitals/cities directly under the central government, which provided scientific data backup for ensuring pesticide residue safety of the fruits and vegetables consumed daily by the public. Meanwhile, the big data statistical analysis of the new technique also further proves it to be of high speed, high throughput, high accuracy, high reliability, and high informatization.

An ultra-performance convergence chromatography (UPC2) method coupled with quadrupole time-of-flight mass spectrometry (Q-TOF-MS), was developed for the determination of complicated triacylglycerols (TAGs) in human milk fat. The use of supercritical carbon dioxide as the mobile phase improved the chromatographic separation of the TAGs significantly. By optimizing the scan modes of Q-TOF-MS and selecting parent ions for MS/MS ionization, the fragment ions of each TAG including TAG isomers with overlapping retention time were adequately resolved for identification and quantification. A total of 95 different TAGs were identified in the human milk samples from Chinese mother volunteers, with O-P-L representing the main TAG, followed by O-P-O and O-L-L. In addition, the compositions and contents of TAGs in different fats and oils were successfully measured. The developed method can serve as an advanced and reliable analytical tool for the determination of complicated TAGs in various biological samples.

According to traditional Chinese medical theory, Ziziphi Spinosae Semen needs to be stir-fried before clinical application for its sedative-hypnotic effect enhancement. A rapid and comprehensive analysis strategy of ultra high performance liquid chromatography with quadrupole time-of-flight mass spectrometry and multiple data analysis platforms was developed for the efficient and sensitive identification of components in crude and parched Ziziphi Spinosae Semen to explore the composition changes that happen during the stir-frying process. Both positive and negative ion modes were applied for mass spectrometry detection, and 40 components were identified from crude and parched Ziziphi Spinosae Semen, respectively. Principal component analysis and t-test were applied to find differences between crude and parched Ziziphi Spinosae Semen. As a result, Ziziphi Spinosae Semen samples could be clearly divided into two groups according to their processing methods, and 19 key markers that contributed to the classification significantly (P < 0.05) were found. This kind of change in contents of components might be responsible for the recommended clinical application of parched Ziziphi Spinosae Semen.

Efficient targeted identification of chemical constituents from traditional Chinese medicine is still a major challenge. In this study, we used a characteristic ion filtering strategy to characterize compounds of Eucommia ulmoides Oliv. by ultra-high performance liquid chromatography quadrupole time of flight tandem mass spectrometry (UHPLC-ESI-Q-TOF-MS/MS). By using the ion filtering approach, target constituents of Eucommia ulmoides Oliv. were easily tentatively identified from the enormous LC/MS data set. The strategy consisted of the following three steps: 1) To establishing a characteristic ion database by diagnostic product ions or neutral loss fragments; 2) To evaluate the structural information of the compounds by high-resolution diagnostic characteristic ion filtering; 3) To confirm the different classes by chemical profiling according to their MS/MS spectra. In this study, characteristic ions are summarized as five major groups of compounds in Eucommia ulmoides Oliv. In total, 113 compounds were tentatively identified, including 23 potentially novel compounds. The results form a foundation for the quality control and chemical basis of Eucommia ulmoides Oliv.