Joon Hyuk Suh

Identification and Quantification of Potential Anti-inflammatory Hydroxycinnamic Acid Amides from Wolfberry

Wolfberry or Goji berry, the fruit of Lycium barbarum, exhibits health-promoting properties that leads to an extensive study of their active components. We synthesized a set of hydroxycinnamic acid amide (HCCA) compounds, including trans-caffeic acid, trans-ferulic acid, and 3,4-dihydroxyhydrocinnamic acid, with extended phenolic amine components as standards to identify and quantify the corresponding compounds from wolfberry and to investigate anti-inflammatory properties of these compounds using in vitro model. With optimized LC-MS/MS and NMR analysis, nine amide compounds were identified from the fruits. Seven of these compounds were identified in this plant for the first time. The amide compounds with a tyramine moiety were the most abundant. In vitro studies indicated that five HCCA compounds showed inhibitory effect on NO production inuded by lipopolysaccharides with IC50 less than 15.08 μM (trans-N-feruloyl dopamine). These findings suggested that wolfberries demonstrated anti-inflammatory properties.

Lipidomic analysis for carbonyl species derived from fish oil using liquid chromatography–tandem mass spectrometry

Lipid peroxidation gives rise to carbonyl species, some of which are reactive and play a role in the pathogenesis of numerous human diseases. Oils are ubiquitous sources that can be easily oxidized to generate these compounds under oxidative stress. In this present work, we developed a targeted lipidomic method for the simultaneous determination of thirty-five aldehydes and ketones derived from fish oil, the omega-3 fatty acid-rich source, by using liquid chromatography-tandem mass spectrometry (LC-MS/MS). The analytes include highly toxic reactive carbonyl species (RCS) such as acrolein, crotonaldehyde, trans-4-hydroxy-2-hexenal (HHE), trans-4-hydroxy-2-nonenal (HNE), trans-4-oxo-2-nonenal (ONE), glyoxal and methylglyoxal, all of which are promising biomarkers of lipid peroxidation. They were formed using in vitro Fe(II)-mediated oxidation, and derivatized using 2,4-dinitrophenylhydrazine (DNPH) for the feasibility of quantitative assay. Before analysis, solid phase extraction (SPE) was used to clean samples further. Uniquely different patterns of carbonyl compound generation between omega-3 and 6 fatty acids were observed using this lipidomic approach. The method developed was both validated, and successfully applied to monitor formation of carbonyl species by lipid peroxidation using ten different fish oil products. Hypotheses of correlations between the monitored dataset of analytes and their parent fatty acids were also tested using the Pearsons correlation test. Results indicate our method is a useful analytical tool for lipid peroxidation studies.

Online turbulent flow extraction coupled with liquid chromatography–tandem mass spectrometry for high throughput screening of anabolic steroids in horse urine

Graphical abstract Figure. No Caption available. HighlightsOnline turbulent flow extraction for equine urine was developed for the first time.Target compounds were androgenic anabolic steroids and their metabolites.Detection of analytes was performed using tandem mass spectrometry.This method simplifies the tedious steps of conventional extractions.The analytes were successfully determined in horse urine after drug administration. Abstract A high throughput method for simultaneous screening of anabolic steroids and their metabolites (4‐esterendione, trenbolone, boldenone, oxandrolone, nandrolone, methandrostenolone, testosterone, 1‐androstendione, ethisterone, normethandrolone, methyltestosterone, 16&bgr;‐Hydroxystanozolol, epitestosterone, bolasterone, norethandrolone, danazol, stanozolol and androstadienone) in equine urine by online turbulent flow extraction coupled with liquid chromatography‐tandem mass spectrometry was developed. The use of turbulent flow chromatography could simplify pretreatment of horse urine, which has complex matrices as well as high viscosity. The urine was extracted by mixed‐mode cation exchange solid phase extraction, and hydrolyzed using &bgr;‐glucuronidase/arylsulfatase. Then, the sample was automatically loaded on the TurboFlow Cyclone extraction column for removal of further matrix, followed by separation on a fused core C18 column before MS/MS detection. Optimization and validation of the method were discussed in detail. All analytes were rapidly detected within 10 min with high sensitivity (picogram to nanogram per milliliter level), and no interference was observed. The linearity range was from 0.1–10 ng/mL for nine steroids and 1.0–50 ng/mL for the others, with correlation of coefficient values over 0.995. Precision and accuracy ranged from 0.1 to 14.5% and 1.7 to 12.4%, respectively. The developed method was successfully applied to the analysis of anabolic steroids in horse urine after administration of a model drug.

Natural Dietary Products and Their Effects on Appetite Control

Natural dietary products have been thoroughly studied for their effects of antiadipogenesis to prevent and treat obesity for decades. Nevertheless, in the past few years appetite control for the treatment of obesity has attracted much attention as a new target. Homeostatic control of energy intake involves a complex system that conveys peripheral signals to the central nervous system where multiple signals are integrated and then provide feedback to regulate satiation. This perspective aims at elucidating the neuronal mechanisms of food intake and energy balance as well as providing an alternative pathway of controlling weight using natural dietary products.

Boswellia serrata resin extract alleviates azoxymethane (AOM)/dextran sodium sulfate (DSS)‐induced colon tumorigenesis

SCOPE Boswellia serrata (BS) resin is a popular dietary supplement for joint nourishment. In this study, we investigated the chemopreventive effects of dietary BS extract and its impact of gut microbiota on azoxymethane/dextran sulfate sodium (AOM/DSS)-induced colitis-associated colon cancer in mice. METHODS AND RESULTS Male ICR mice were injected with AOM and 2% DSS via drinking water. The mice were fed with 0.25 or 0.5% BS extract, and colonic tissue were collected at 15 weeks. The main effective components of BS supercritical CO2 extraction were analyzed by LC-MS/MS are boswellic acids. We found that treatment with BS extract significantly reduce the colonic tumor formation. Western blot and histological analysis revealed that dietary BS extract could markedly reduce the inflammation associated protein levels expression. Furthermore, BS extract reduced cell proliferation via inhibiting phosphorylation level of protein kinase B (Akt), glycogen synthase kinase 3β (GSK3β), and downregulation of cyclin D1. In addition, BS extract also altered the composition of gut microbiota by enhancing the proportion of Clostridiales and reducing the percentage of Bacteroidales. CONCLUSION In summary, BS extract decreased the protein levels of inflammative enzymes such as inducible nitric oxide synthase and cyclooxygenase-2 in colonic mucosa. It also mediated Akt/GSK3β/cyclin D1 signaling pathway and altered the composition of gut microbiota to alleviate tumor growth. Taken together, this study suggests that BS extract has great potential to suppress colon tumorigenesis.

Development of aqueous mobile phase using chaotrope for the chromatographic determination of melamine in infant formula

Direct analysis of melamine using reverse phase chromatography is a challenge because this compounds small size and strong polar nature leads to abnormal peak symmetry as well as poor retention. Here, we introduce a simple and reliable reverse phase liquid chromatographic method using sodium hexafluorophosphate to modify an acidic aqueous eluent, resulting in improved chromatographic behaviors of melamine in complex food matrices. Variables affecting the retention mechanism, including chaotrope type, concentration and stationary phase, were investigated. Under optimum conditions, melamine retention, separation efficiency, peak shape and reproducibility were significantly improved as compared to other methods that use ionic liquids or a micellar mobile phase. No interference affected melamine detection when infant formula was applied as the food matrix. Analytical reliability was demonstrated through estimation of validation parameters such as specificity, linearity, precision, accuracy and recovery. This method is suitable for routine analysis of melamine in infant formula. More noteworthy, this is the first time that an organic solvent-free mobile phase using chaotropic salt, meeting the concept of green chemistry, has been proposed.

Comprehensive Metabolomics Analysis of Mandarins (Citrus reticulata) as a Tool for Variety, Rootstock, and Grove Discrimination

The metabolite profile responsible for the quality of mandarin fruit is influenced by preharvest factors including genotype, rootstock, grove location, etc. In this paper, mandarin varieties were discriminated using metabolomics. Additionally, effects on metabolic profiles due to grove location and rootstock differences were also investigated. Results revealed that mandarin varieties could be differentiated using the metabolite profile, while the compositions of flavonoids have the potential for variety differentiation. With regard to fruits of the same variety, grove location might determine the overall profile of metabolites, whereas rootstock possibly affected composition of secondary metabolites. Pathway enrichment analysis demonstrated that biosynthesis pathways of terpenoids and steroids involving limonene and linalool were highly influenced by variety diversity. Moreover, the flavonoid biosynthesis pathway, involving hesperetin, naringenin, eriodictyol, and taxifolin, was indicated to have a close relationship with rootstock differentiation. This study provides useful and important information with depth for breeding and optimizing preharvest practices.

Differentiation between Flavors of Sweet Orange (Citrus sinensis) and Mandarin (Citrus reticulata)

Pioneering investigations referring to citrus flavor have been intensively conducted. However, the characteristic flavor difference between sweet orange and mandarin has not been defined. In this study, sensory analysis illustrated the crucial role of aroma in the differentiation between orange flavor and mandarin flavor. To study aroma, Valencia orange and LB8-9 mandarin were used. Their most aroma-active compounds were preliminarily identified by aroma extract dilution analysis (AEDA). Quantitation of key volatiles followed by calculation of odor activity values (OAVs) further detected potent components (OAV ≥ 1) impacting the overall aromatic profile of orange/mandarin. Follow-up aroma profile analysis revealed that ethyl butanoate, ethyl 2-methylbutanoate, octanal, decanal, and acetaldehyde were essential for orange-like aroma, whereas linalool, octanal, α-pinene, limonene, and (E,E)-2,4-decadienal were considered key components for mandarin-like aroma. Furthermore, an unreleased mandarin hybrid producing fruit with orange-like flavor was used to validate the identification of characteristic volatiles in orange-like aroma.

Metabolic analysis reveals altered long-chain fatty acid metabolism in the host by Huanglongbing disease

Candidatus Liberibacter asiaticus (CLas) is the presumed causal agent of Huanglongbing, one of the most destructive diseases in citrus. However, the lipid metabolism component of host response to this pathogen has not been investigated well. Here, metabolic profiling of a variety of long-chain fatty acids and their oxidation products was first performed to elucidate altered host metabolic responses of disease. Fatty acid signals were found to decrease obviously in response to disease regardless of cultivar. Several lipid oxidation products strongly correlated with those fatty acids were also consistently reduced in the diseased group. Using a series of statistical methods and metabolic pathway mapping, we found significant markers contributing to the pathological symptoms and identified their internal relationships and metabolic network. Our findings suggest that the infection of CLas may cause the altered metabolism of long-chain fatty acids, possibly leading to manipulation of the hosts defense derived from fatty acids.

Development of an improved sample preparation platform for acidic endogenous hormones in plant tissues using electromembrane extraction

Despite their importance in pivotal signaling pathways due to trace quantities and complex matrices, the analysis of plant hormones is a challenge. Here, to improve this issue, we present an electromembrane extraction technology combined with liquid chromatography-tandem mass spectrometry for determination of acidic plant hormones including jasmonic acid, abscisic acid, salicylic acid, benzoic acid, gibberellic acid and gibberellin A4 in plant tissues. Factors influencing extraction efficiency, such as voltage, extraction time and stirring rate were optimized using a design of experiments. Analytical performance was evaluated in terms of specificity, linearity, limit of quantification, precision, accuracy, recovery and repeatability. The results showed good linearity (r2 > 0.995), precision and acceptable accuracy. The limit of quantification ranged from 0.1 to 10 ng mL-1, and the recoveries were 34.6-50.3%. The developed method was applied in citrus leaf samples, showing better clean-up efficiency, as well as higher sensitivity compared to a previous method using liquid-liquid extraction. Organic solvent consumption was minimized during the process, making it an appealing method. More noteworthy, electromembrane extraction has been scarcely applied to plant tissues, and this is the first time that major plant hormones were extracted using this technology, with high sensitivity and selectivity. Taken together, this work gives not only a novel sample preparation platform using an electric field for plant hormones, but also a good example of extracting complex plant tissues in a simple and effective way.