Determination of flurbiprofen in rat plasma using ultra-high performance liquid chromatography-tandem mass spectrometry and its application in a pharmacokinetic study.

A rapid and sensitive ultra-high performance liquid chromatography-tandem mass spectrometry method was developed to determine flurbiprofen in rat plasma. A triple quadrupole tandem mass spectrometer equipped with an electrospray ionization (ESI) source was used in negative ion mode. Acetonitrile precipitation was selected to prepare samples. Flurbiprofen and internal standard flurbiprofen-d5 were analyzed on an Acquity UPLC BEH C18 column with the mobile phase consisting of acetonitrile and water, and a gradient procedure was used for separation. The retention time of flurbiprofen was 0.67 min, and the whole running time was only 1.2 min. The detection was performed on a triple quadrupole tandem mass spectrometer using multiple reaction monitoring mode via an ESI source with optimized mass spectrometry parameters. The calibration curve was linear in the range of 25.0-1.00 × 104 ng/mL (r ≥ 0.99). The within-run and between-run relative standard deviations were not more than 13.9%. The within-run and between-run relative errors were from -9.0% to 3.4%. There was no significant matrix effect, and recovery was high. This method was fully validated, including whole blood stability in rat plasma, and successfully applied to the pharmacokinetic study in which 100% incurred sample reanalysis met the criteria.

Cretaceous diversity and disparity in a lacewing lineage of predators (Neuroptera: Mantispidae).

Mantidflies (Mantispidae) are an unusual and charismatic group of predatory lacewings (Neuroptera), whereby the adults represent a remarkable case of morphological and functional convergence with praying mantises (Mantodea). The evolutionary history of mantidflies remains largely unknown due to a scarcity of fossils. Here, we report the discovery of a highly diverse palaeofauna of mantidflies from the mid-Cretaceous (lowermost Cenomanian) of Myanmar. The raptorial forelegs of these mantidflies possess highly divergent morphological modifications, some of which are unknown among modern mantidflies, e.g. the presence of forked basal profemoral spines or even the complete loss of foreleg spine-like structures. A phylogenetic analysis of Mantispidae reveals a pattern of raptorial foreleg evolution across the family. The high species diversity and disparate foreleg characters might have been driven by diverse niches of predator-prey interplay in the complex tropical forest ecosystem of the mid-Cretaceous.

Metabonomic study of biochemical changes in urinary of type 2 diabetes mellitus patients after the treatment of sulfonylurea antidiabetic drugs based on ultra-performance liquid chromatography/mass spectrometry.

A metabonomic study on biochemical changes in the urine of type 2 diabetes mellitus (T2DM) patients after the treatment of sulfonylurea (SU) antidiabetic drugs was performed. An ultra-performance liquid chromatography/mass spectrometry (UPLC/MS) method was used to generate metabolic fingerprints for the metabonomic analysis of urinary samples obtained from 20 T2DM patients without any drug treatment and 20 T2DM patients treated with SU antidiabetic drugs and 20 normal glucose tolerance subjects. The resulting data were subjected to chemometric analysis (principal component analysis and partial least squares discriminant analysis) to investigate the effect of SU antidiabetic drugs on urinary metabolite profiles of T2DM patients. Biomarkers such as xanthine, phenylalanine, tryptophan, hippurate, phenylacetylglutamine, carnitine C8:1, carnitine C10:3, uric acid and citrate were found to be responsible for the separation of T2DM and SU-treated groups, which indicates a potential effect of SU on energy metabolism, Tricarboxylic acid (TCA) cycle, gut microflora metabolism and oxidative stress. The study may be helpful to the understanding of the action of mechanism of SU antidiabetic drugs.

Structure identification and elucidation of mosapride metabolites in human urine, feces and plasma by ultra performance liquid chromatography-tandem mass spectrometry method.

1. Mosapride citrate (mosapride) is a potent gastroprokinetic agent. The only previous study on mosapride metabolism in human reported one phase I oxidative metabolite, des-p-fluorobenzyl mosapride, in human plasma and urine using HPLC method. Our aim was to identify mosapride phase I and phase II metabolites in human urine, feces and plasma using UPLC-ESI-MS/MS. 2. A total of 16 metabolites were detected. To the best of our knowledge, 15 metabolites have not been reported previously in human. 3. Two new metabolites, morpholine ring-opened mosapride (M15) and mosapride N-oxide (M16), alone with one known major metabolite, des-p-fluorobenzyl mosapride (M3), were identified by comparison with the reference standards prepared by our group. The chemical structures of seven phase I and six phase II metabolites of mosapride were elucidated based on UPLC-MS/MS analyses. 4. There were two major phase I reactions, dealkylation and morpholine ring cleavage. Phase II reactions included glucuronide, glucose and sulfate conjugation. The comprehensive metabolic pathway of mosapride in human was proposed for the first time. 5. The metabolites in humans were compared with those in rats reported previously. In addition to M10, the other 15 metabolites in humans were also found in rats. This result suggested that there was little qualitative species difference in the metabolism of mosapride between rats and humans. 6. In all, 16 mosapride metabolites including 15 new metabolites were reported. These results allow a better understanding of mosapride disposition in human.

[Study on in vitro and in vivo material base of Sini San by UPLC-PDA-MS/MS].

OBJECTIVE: To analyze chemical constituents of Sini San its migrating components in rat plasma and study its in vitro and in vivo material base using ultra-performance liquid chromatography coupled with photo-diode-array detector and tandem mass spetrometry (UPLC-PDA-MS/MS). METHOD: ACQUITY UPLC BEH C18 column (2.1 mm x 100 mm, 1.7 microm) was adopted, with gradient elution system of water containing 2 mmol x L(-1) ammonium acetate and acetonitrile at flow rate of 0.2 mL x min(-1). The column temperature was maintained at 35 degrees C. The mass spectra were obtained by electrospray ionization source operating in both positive and negative ion mode. Ions were scanned from the m/z 100 to 1 000, and the characteristic ions were schizolysised twice to obtain the secondary MS data. RESULT: Twenty chemical constituents were detected, including paeoniflorin, glycyrrhizic acid, saikosaponins a and naringin. In vivo, there were 8 ingredients directly absorbed into blood after the administration of Sini San decoction, such as paeoniflorin, naringin and hesperidin. Besides, 6 metabolites were also detected, involving glucuronides, sulfate and sulfoglucuronides. CONCLUSION: In vitro and in vivo chemical materials of Sini San decoction is analyzed by UPLC-PDA-MS/MS to reflect in vitro and in vivo material base of Sini San decoction in a comprehensive and rapid manner and provide basis for further study on efficacious material basis of Sini San decoction.

[Metabonomic study on protective effect of ethanol extracts of drynariae rhizoma on osteoporosis in rats urine by using UPLC-MS/MS].

This paper was designed to study metabonomic characters of the osteoporosis induced by high dose of hydrocortisone and the protective effects of Drynariae Rhizoma, which can replenish the kidney and strengthen the bones. A urinary metabonomics method based on ultra-performance liquid chromatography coupled with mass spectrometry (UPLC-MS/MS) was developed. Clear separation of healthy control group, model group and treatment group was achieved by using the principal components analysis (PCA) and 9 significantly changed metabolites were identified as potential biomarkers of osteoporosis. Compared with the health control group, the model group rats showed lower levels of creatinine, citric acid, azelaic acid, hippurate, tryptophan and indoxyl sulfate together with higher levels of phenylalanine, cresol sulfate and phenaceturic acid. These changes in urinary metabolites suggest that the disorders of amino acid metabolism, energy metabolism, gut microflora and anti-oxidative damage are related to osteoporosis induced by high dose of hydrocortisone and the potential effect of Drynariae Rhizoma on all the four metabolic pathways.

New Cretaceous Lacewings in a Transitional Lineage of Myrmeleontoidea and Their Phylogenetic Implications.

The extinct neuropteran families Cratosmylidae and Babinskaiidae hitherto only known from the Cretaceous represent the transitional lineage between Nymphidae and advanced myrmeleontoids (e.g., Nemopteridae and Myrmeleontidae) in the superfamily Myrmeleontoidea. Here, we describe two new species, which respectively belong to Cratosmylidae and Babinskaiidae, namely, Araripenymphes burmanus sp. nov. and Paradoxoleon chenruii gen. et sp. nov., from the mid-Cretaceous Kachin amber of Myanmar. Cratosmylidae, which was previously only recorded from the Lower Cretaceous of Brazil (Crato Formation), is first reported from the mid-Cretaceous Kachin amber of Myanmar, and the co-occurrence of Araripenymphes Menon, Martins-Neto and Martill, 2005 across South America and Asia further documents the Gondwanan origin of the northern Myanmar amber lacewing paleofauna. The first finding of a deeply bifurcated forewing MP with two free branches in Babinskaiidae (viz., Paradoxoleon chenruii gen. et sp. nov.) highlights the morphological diversity of this extinct family. The phylogenetic positions of Araripenymphes burmanus sp. nov. and Paradoxoleon chenruii gen. et sp. nov. were recovered on the basis of a morphology-based phylogenetic analysis, and the monophyly of Cratosmylidae + Babinskaiidae was corroborated. Given the paraphyly of Cratosmylidae, its familial status is discussed.

The first mitochondrial genome of spongillafly from Asia (Neuroptera: Sisyridae: Sisyra aurorae Navás, 1933) and phylogenetic implications of Osmyloidea.

The spongillafly species Sisyra aurorae Navás, 1933 (Neuroptera: Sisyridae) is an endemic species in China and is first recorded from Shanghai. The mitogenome of this species is sequenced, representing the first mitogenome of Sisyridae from Asia. The nearly complete mitogenome is 15,634 bp, which contains 13 protein-coding genes (PCGs), 22 transfer RNA genes (tRNAs), two ribosomal RNA genes (rRNAs), and a control region. The gene order and arrangement are similar to other lacewing mitogenomes. Both Bayesian and maximum likelihood analyses based on 13 PCGs recovered the interfamilial phylogeny within Osmyloidea as Sisyridae + (Nevrorthidae + Osmylidae).

New Cretaceous antlion-like lacewings promote a phylogenetic reappraisal of the extinct myrmeleontoid family Babinskaiidae.

Babinskaiidae is an extinct family of the lacewing superfamily Myrmeleontoidea, currently only recorded from the Cretaceous. The phylogenetic position of this family is elusive, with inconsistent inferences in previous studies. Here we report on three new genera and species of Babinskaiidae from the mid-Cretaceous Kachin amber of Myanmar, namely Calobabinskaia xiai gen. et sp. nov., Stenobabinskaia punctata gen. et sp. nov., and Xiaobabinskaia lepidotricha gen. et sp. nov. These new babinskaiids are featured by having specialized characters, such as the rich number of presectoral crossveins and the presence of scaly setae on forewing costal vein, which have not yet been found in this family. The exquisite preservation of the Kachin amber babinskaiids facilitate a reappraisal of the phylogenetic placement of this family based on adult morphological characters. Our result from the phylogenetic inference combining the data from fossil and extant myrmeleontoids recovered a monophyletic clade composed of Babinskaiidae and another extinct family Cratosmylidae, and further assigned this clade to be sister group to a clade including Nemopteridae, Palaeoleontidae, and Myrmeleontidae. Babinskaiidae appears to be a transitional lineage between Nymphidae and advanced myrmeleontoids, with ancient morphological diversification.

Liquid chromatography-mass spectrometry for analysis of a novel beta(2)-adrenoceptor agonist trantinterol and its metabolites in beagle dog urine.

A liquid chromatography-tandem mass spectrometry method was developed for the identification of metabolites of trantinterol, a novel beta(2)-adrenoceptor agonist, in beagle dog urine. The separation of metabolites was performed on a reversed-phase C(8) column using 0.1% formic acid in water and methanol (70 : 30, v/v) as the mobile phase. The structural information and elemental information of metabolites were acquired by an electrospray ionization tandem mass spectrometer and a quadrupole time-of-flight mass spectrometer, respectively. A total of 13 metabolites were detected and characterized on the basis of their tandem MS/MS fragmentation patterns. The accurate masses of nine metabolites were determined and two metabolites were further confirmed by comparing with reference standards. The metabolic pathways of trantinterol in beagle dog are proposed.

A rapid and reliable UPLC-MS/MS method for the identification and quantification of fourteen synthetic anti-diabetic drugs in adulterated Chinese proprietary medicines and dietary supplements.

An ultraperformance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was developed for simultaneous qualitative and quantitative analysis of 14 synthetic anti-diabetic drugs in adulterated Chinese proprietary medicines (CPMs) and dietary supplements. The samples were prepared by ultrasonic extraction with methanol and separated on a C18 column with mobile phase consisting of acetonitrile and water (both containing 0.10% formic acid). Gradient elution was applied with a flow rate of 0.20 mL/min. Two transitions from protonated molecules were monitored for each synthetic anti-diabetic drug in positive mode of electrospray ionization (ESI). The two transitions, the peak area ratio of the two transitions and the retention time were used for identification. The more intensive transition was used for quantification. The analysis time was 6 min per sample. Satisfactory linear relationships were estimated between the peak area and the concentration with correlation coefficients higher than 0.995. The limit of detection ranged from 0.03 to 5.45 ng/mL. The relative standard deviation of intra-day precision was below 7.6%, the RSD of inter-day precision was below 15% and the relative error of accuracy was between -10 and 7.8%. The proposed method is rapid, selective, reliable and was successfully applied to the analysis of 30 real samples of 22 CPMs and eight dietary supplements from the local market in China.

A simple and low-energy method to prepare loratadine nanosuspensions for oral bioavailability improvement: preparation, characterization, and in vivo evaluation.

The effervescent method, as a simple and effective technology to prepare nanosuspensions, has gained great attention. In this present research, loratadine (LTD) nanosuspensions were successfully prepared by the effervescent method using Soluplus as stabilizer to improve the bioavailability of LTD in vivo. The mean particle size was about 100 nm. And the LTD nanosuspensions were lyophilized for further study. The freeze-dried powders could be dissolved quickly, and the mean particle size remained almost unchanged after powders were re-dissolved. By transmission electron microscope (TEM), differential scanning calorimetry (DSC), scanning electron microscopy (SEM), Fourier transform infrared (FT-IR) spectroscopy, and X-ray diffraction (X-RD), the characterizations of LTD nanosuspensions and freeze-dried powders were studied. Commercial tablets were used as the reference to investigate the dissolution behaviors in different release media and of bioavailability in vivo of LTD freeze-dried powders. The cumulative dissolution of the LTD freeze-dried powders was superior in different release media compared with commercial tables. In addition, for the evaluation of the bioavailability of LTD nanosuspensions, the LTD concentration in rat plasma was determined using LC-MS/MS method. The results showed that the AUC0-24 and Cmax of LTD freeze-dried powders were about 2.14- and 2.01-fold higher than those of commercial tablets. In short, the effervescent method has been successfully applied to the preparation of LTD nanosuspensions to improve the bioavailability of LTD in vivo with the advantage of low energy consumption. This simple technology also provides an idea for the preparation of the other nanosuspensions.

Simultaneous quantification of enalapril and enalaprilat in human plasma by high-performance liquid chromatography-tandem mass spectrometry and its application in a pharmacokinetic study.

A rapid, selective and sensitive high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method was developed to simultaneously determine enalapril and enalaprilat in human plasma. With benazepril as internal standard, sample pretreatment involved in a one-step protein precipitation (PPT) with methanol of 0.2 ml plasma. Analysis was performed on an Ultimate XB-C(18) column (50 mm x 2.1 mm, i.d., 3 microm) with mobile phase consisting of methanol-water-formic acid (62:38:0.2, v/v/v). The detection was performed on a triple quadrupole tandem mass spectrometer by multiple reaction-monitoring (MRM) mode via electrospray ionization (ESI) source. Each plasma sample was chromatographed within 2.5 min. The linear calibration curves for enalapril and enalaprilat were both obtained in the concentration range of 0.638-255 ng/ml (r(2) > or = 0.99) with the lower limit of quantification (LLOQ) of 0.638 ng/ml. The intra-day precision (R.S.D.) was below 7.2% and inter-day R.S.D. was less than 14%, while accuracy (relative error R.E.) was within +/-8.7 and +/-5.5%, determined from QC samples for enalapril and enalaprilat which corresponded to requirement of the guidance of FDA. The HPLC-MS/MS method herein described was fully validated and successfully applied to the pharmacokinetic study of enalapril maleate capsules in 20 healthy male volunteers after oral administration.

Xanthoceraside Could Ameliorate Alzheimer's Disease Symptoms of Rats by Affecting the Gut Microbiota Composition and Modulating the Endogenous Metabolite Levels.

Xanthoceraside (XAN) is a natural-derived compound with anti-Alzheimer activity from the husks of Xanthoceras sorbifolia. Although its therapeutic effect had been confirmed in previous studies, the mechanism was still unclear due to its poor solubility and low permeability. In this study, the pharmacological effect of XAN on Alzheimer's disease (AD) was confirmed by behavior experiments and H&E staining observation. Fecal microbiota transplantation (FMT) experiment also replicated the therapeutic effects, which indicates the potential targets of XAN on gut microbiota. The sequencing of 16S rRNA genes in fecal samples demonstrated that XAN reversed gut microbiota dysbiosis in AD animals. XAN could change the relative abundances of several phyla and genus of bacterial, particularly the ratio of Firmicutes/Bacteroidetes. Among them, Clostridium IV, Desulfovibrio, Corynebacterium, and Enterorhabdus had been reported to be involved in the pathologic developments of AD and other central nervous system disease. In metabolomics study, a series of host endogenous metabolites were detected, including amino acids, lysophosphatidylcholine, dihydrosphingosine, phytosphingosine, inosine, and hypoxanthine, which were all closely associated with the development of AD. Combined with the Spearman's correlation analysis, it was confirmed that the increases of five bacterial strains and decreases of six bacterial strains were closely correlated with the increases of nine host metabolites and the decreases of another five host metabolites. Therefore, XAN can modulate the structure of gut microbiota in AD rats; the changes of gut microbiota were significantly correlated with endogenous metabolites, and symptom of AD was ultimately alleviated. Our findings suggest that XAN may be a potential therapeutic drug for AD, and the gut microbiota may be potential targeting territory of XAN via microbiome-gut-brain pathway.

In vivo evaluation of risperidone-SAIB in situ system as a sustained release delivery system in rats.

The objective of this study was to evaluate a sustained release sucrose acetate isobutyrate (SAIB) in situ system formulation of risperidone (RSP) in vivo. The formulation contained SAIB, ethanol, and polylactic acid (PLA) as a release regulator. In vivo pharmacokinetics (PK) studies have shown that PLA is effective in reducing the burst effect. After a 12.5mg/kg IM injection of a 25mg/g RSP-SAIB in situ system, the C(max) was markedly reduced from 944.1+/-80.2 to 330.4+/-33.6ng/ml by increasing PLA from 1% to 10% (w/w), the T(max) were prolonged from 2 to 4.3+/-2.0h, and the area under the curve from day 0 to 2 (AUC(0-2day)) was reduced significantly from 16294.8+/-3946.4 to 7025.3+/-1979.2ngh/ml. For the RSP-SAIB in situ system including 10% PLA, the high release rates over a short period allowed therapeutic plasma concentrations to be achieved in the initial stages after activation, and sustained release of the drug led to a stable plasma concentration (by day 25, the plasma concentration was 8% of the C(max)). These initial in vivo studies suggest that RSP-SAIB in situ system is effective as a sustained delivery system.

Simultaneous enantioseparation of four beta2-agonists by capillary electrophoresis with cyclodextrin additives. Study of the enantioselective mechanism.

The simultaneous capillary electrophoretic enantioseparation of adrenergic beta(2)-agonists enantiomers (trantinterol, mabuterol, clenbuterol, bambuterol) was studied with beta-cyclodextrin, ethyl-beta-CD, methyl-beta-CD, hydroxypropyl-beta-CD, and hydroxyethyl-beta-CD as chiral selector. The type and concentration of the chiral selector and buffer pH played a very important role in the enantioseparation of the analyzed compounds. Hydroxypropyl-beta-CD was found to be the most effective complexing agent and allowed excellent chiral/achiral resolutions compared to the other CDs. The simultaneous enantioseparation of four beta(2)-agonists was achieved using 100 mM citric acid-10 mM Na(2)HPO(4) buffer at pH 2.5 containing 120 mM hydroxypropyl-beta-CD with an applied voltage of 20 kV. Method validation in terms of repeatability, linearity, and limits of detection and quantification was performed. The effect of structural features of analytes on R(s) and t(m) was studied. Complexation binding constants for the interactions between the four compounds and three different CDs were evaluated for elucidating the enantioseparation mechanism. It was found that very small differences in the chemical structure of the analytes resulted in significant changes in stereoselective recognition.

The first green lacewing (Insecta: Neuroptera: Chrysopidae) from the mid-Cretaceous amber of Myanmar.

The lacewing family Chrysopidae is here reported from the mid-Cretaceous amber of Myanmar for the first time. A new genus and species, Parabaisochrysa xingkei gen. et sp. nov., is described. The new genus belongs to the extinct subfamily Limaiinae and is characterized by the presence of three gradate series of crossveins in both fore- and hind wing, and by the MP1 coalescent with RP+MA in the hind wing.

Taxonomic notes on Babinskaiidae from the Cretaceous Burmese amber, with the description of a new species (Insecta, Neuroptera).

Babinskaiidae is an extinct lacewing family of the superfamily Myrmeleontoidea. Hitherto, nine species of seven genera are described from the Lower and mid-Cretaceous. Here a new species of Babinskaiidae is described from Cretaceous Burmese amber, namely Parababinskaia makarkinisp. n. The new species possesses an A2 vein in the hind wing, suggesting that the loss of this vein might not be an autapomorphy of Babinskaiidae. The female of Electrobabinskaia burmana Lu, Zhang & Liu, 2017 is also described for the first time based on two specimens with their abdomens perfectly preserved, exhibiting a specialised sternum VI with paired elongate projections. A brief discussion of female genital characters is provided, which may increase our understanding of the morphology and phylogenetic position of Babinskaiidae.

Taxonomic notes on Cretarophalis patrickmuelleri Wichard, 2017 (Insecta: Neuroptera: Nevrorthidae) from the mid-Cretaceous of Myanmar, and its phylogenetic significance.

Nevrorthidae is a family of Neuroptera with low species diversity, disjunct geographic distributions, and a controversial phylogenetic status. Previous fossil records of the family are from the Eocene except for the recently described species Cretarophalis patrickmuelleri Wichard, 2017 from mid-Cretaceous Burmese amber, following earlier records of an undescribed species. However, such a significant finding of this family from Mesozoic was originally presented only with a preliminary description. Here we re-describe the species based on exquisitely preserved materials from mid-Cretaceous Burmese amber that provides for a much more detailed description, in particular of the hindwing, female genitalia, and larval head. Furthermore, distinctive morphological characters of C. patrickmuelleri with significant phylogenetic relevance are discussed. The general morphology, particularly that of the larva, has changed little between the Mesozoic and Cenozoic, and it reflects an evolutionary stasis that might correspond to their aquatic larval life-style. The present finding also supports the relictual nature of the modern nevrorthids.

Liverwort Mimesis in a Cretaceous Lacewing Larva.

Camouflage and mimicry are staples among predator-prey interactions, and evolutionary novelties in behavior, anatomy, and physiology that permit such mimesis are rife throughout the biological world [1, 2]. These specializations allow for prey to better evade capture or permit predators to more easily approach their prey, or in some cases, the mimesis can serve both purposes. Despite the importance of mimesis and camouflage in predator-avoidance or hunting strategies, the long-term history of these traits is often obscured by an insufficient fossil record. Here, we report the discovery of Upper Cretaceous (approximately 100 million years old) green lacewing larvae (Chrysopoidea), preserved in amber from northern Myanmar, anatomically modified to mimic coeval liverworts. Chrysopidae are a diverse lineage of lacewings whose larvae usually camouflage themselves with a uniquely constructed packet of exogenous debris, conveying greater stealth upon them as they hunt prey such as aphids as well as evade their own predators [3, 4]. However, no lacewing larvae today mimic their surroundings. While the anatomy of Phyllochrysa huangi gen. et sp. nov. allowed it to avoid detection, the lack of setae or other anatomical elements for entangling debris as camouflage means its sole defense was its mimicry, and it could have been a stealthy hunter like living and other fossil Chrysopoidea or been an ambush predator aided by its disguise. The present fossils demonstrate a hitherto unknown life-history strategy among these "wolf in sheep's clothing" predators, one that apparently evolved from a camouflaging ancestor but did not persist within the lineage.