Hyun Kyoung Ju

Characterization of increased phenolic compounds from fermented Bokbunja (Rubus coreanus Miq.) and related antioxidant activity.

This study examined the changes in the phenolic acid-content and antioxidant activity of Rubi Fructus (RF), the fruit of Rubus coreanus Miq., after fermentation with yeast (Saccharomyces cerevisiae). The phenolic acids were fractionated into three forms, free (Fr. A), ester (Fr. B), and insoluble-bound phenolic acids (Fr. C) and quantified by high performance liquid chromatography with a diode array detector (HPLC-DAD). This method was validated and allowed the successful identification of 11 phenolic acids in the RF extracts. HPLC-DAD analysis of the samples showed substantial increases in the levels of protocatechuic, vanillic and p-coumaric acid as the result of yeast fermentation. The total phenolic content (TPH) was also increased by fermentation. The total phenolics in Fr. A and Fr. B increased from 117 to 173 mg GAE/100 g and from 488 to 578 mg GAE/100 g, respectively. The total phenolics in Fr. C decreased from 264 to 175 mg GAE/100 g. The antioxidant activity of the fermented RF was measured as the 1,1-diphenoly-2-picrylhydrazyl (DPPH) radical scavenging capacity, which is expressed as the IC(50). The IC(50) for Fr. A and Fr. B decreased from 5.9 to 4.0 mg/ml (mg of dried RF equiv./ml) and from 1.2 to 0.8 mg/ml, respectively. In Fr. C, the IC(50) value increased from 2.1 to 2.8 mg/ml. In summary, the fermented RF had a higher total phenolic content and better DPPH radical-scavenging activity than the unfermented material.

Determination of bioactive compounds in fermented soybean products using GC/MS and further investigation of correlation of their bioactivities

The active ingredients and bioactivities (anti-oxidant, anti-tyrosinase, anti-proliferative and estrogenic activities) of soybean and soybean products (Cheonggukjang, Meju, Makjang, Doenjang and soy sauce) produced by different fermentation processes were compared. There were high correlations between active ingredients and bioactivities. Free phenolic acids extracted from soybean products were identified and quantified by gas chromatography/mass spectrometry (GC/MS). Overall, the components and activities in fermented soybean products were different than those in soybeans. Total phenolic content (TPC), protein content (PC) and anti-oxidant activity increased as fermentation time increased. TPC and PC showed strong negative correlations with anti-oxidant activity. Doenjang and soy sauce, two long-term fermented products, showed lower total flavonoid content (TFC) and estrogenic activities than short-term fermented soybean products. This might be explained by the decomposition and hydrolysis of flavonoids due to the long fermentation time and high temperature. Strong anti-proliferative activity against cancer cell lines, which was highly correlated with TFC, was found in Meju and Cheonggukjang. Soybean and all fermented products except Meju exhibited effective tyrosinase inhibitory activities. Fermented products showed stronger estrogenic activity than soybeans, which was highly correlated with syringic acid.

Determination of volatile biomarkers for apoptosis and necrosis by solid-phase microextraction–gas chromatography/mass spectrometry: A pharmacometabolomic approach to cisplatin's cytotoxicity to human lung cancer cell lines

In order to find potential new biomarkers of cisplatin-induced apoptosis and necrosis, volatile organic compounds (VOCs) from cisplatin-treated human lung cancer cell lines were investigated. The biological system employed was human non-small cell lung carcinoma A549 cell lines. The cell lines were treated with two different concentrations of cisplatin, 100 microM and 400 microM, and apoptosis and necrosis were determined by flow cytometric analysis. For each drug concentration, the VOCs from the treated cell lines were extracted by solid-phase microextraction (SPME), and subsequently analyzed by gas chromatography/mass spectrometry (GC/MS). The compounds that change during cisplatin-induced apoptosis and necrosis of lung cancer cell lines can serve as new biomarkers. The pharmacometabolomic approach presented in this study, significantly, implicates a non-destructive, sample-thrifty and time-saving tool for finding new biomarkers for the assessment of drug-induced cell death pathways.

Enhancement of Anti-Inflammatory Activity of Aloe vera Adventitious Root Extracts through the Alteration of Primary and Secondary Metabolites via Salicylic Acid Elicitation

Aloe vera (Asphodeloideae) is a medicinal plant in which useful secondary metabolites are plentiful. Among the representative secondary metabolites of Aloe vera are the anthraquinones including aloe emodin and chrysophanol, which are tricyclic aromatic quinones synthesized via a plant-specific type III polyketide biosynthesis pathway. However, it is not yet clear which cellular responses can induce the pathway, leading to production of tricyclic aromatic quinones. In this study, we examined the effect of endogenous elicitors on the type III polyketide biosynthesis pathway and identified the metabolic changes induced in elicitor-treated Aloe vera adventitious roots. Salicylic acid, methyl jasmonate, and ethephon were used to treat Aloe vera adventitious roots cultured on MS liquid media with 0.3 mg/L IBA for 35 days. Aloe emodin and chrysophanol were remarkably increased by the SA treatment, more than 10–11 and 5–13 fold as compared with untreated control, respectively. Ultra-performance liquid chromatography-electrospray ionization mass spectrometry analysis identified a total of 37 SA-induced compounds, including aloe emodin and chrysophanol, and 3 of the compounds were tentatively identified as tricyclic aromatic quinones. Transcript accumulation analysis of polyketide synthase genes and gas chromatography mass spectrometry showed that these secondary metabolic changes resulted from increased expression of octaketide synthase genes and decreases in malonyl-CoA, which is the precursor for the tricyclic aromatic quinone biosynthesis pathway. In addition, anti-inflammatory activity was enhanced in extracts of SA-treated adventitious roots. Our results suggest that SA has an important role in activation of the plant specific-type III polyketide biosynthetic pathway, and therefore that the efficacy of Aloe vera as medicinal agent can be improved through SA treatment.

Mass spectrometry based proteomic analysis of human stem cells: A brief review

Stem cells can give rise to various cell types and are capable of regenerating themselves over multiple cell divisions. Pluripotency and self-renewal potential of stem cells have drawn vast interest from different disciplines, with studies on the molecular properties of stem cells being one example. Current investigations on the molecular basis of stem cells pluripotency and self-renewal entail traditional techniques from chemistry and molecular biology. In this mini review, we discuss progress in stem cell research that employs proteomics approaches. Specifically, we focus on studies on human stem cells from proteomics perspective. To our best knowledge, only the following types of human stem cells have been examined via proteomics analysis: human neuronal stem cells, human mesenchymal stem cells, and human embryonic stem cells. Protein expression serves as biomarkers of stem cells and identification and expression level of such biomarkers are usually determined using two-dimensional electrophoresis coupled mass spectrometry or non-gel based mass spectrometry.

Metabolomic Investigation of the Anti-Platelet Aggregation Activity of Ginsenoside Rk1 Reveals Attenuated 12-HETE Production

Comprehensive metabolomics analysis is an effective method of measuring metabolite levels in the body following administration of a pharmaceutical compound and can allow for monitoring of the effects of the compound or assessment of appropriate treatment options for individual patients. In the present metabolomics study, samples pretreated with antiplatelet compounds were extracted and subjected to ultraperformance liquid chromatography/quadrupole time-of-flight mass spectrometry. The acquired data were processed using peak clustering and evaluated by partial least-squares (PLS) and orthogonal projections to latent structures discriminant analyses (OPLS-DA). As a result, meaningful endogenous metabolites, namely eicosanoids and thromboxane B(2) (TXB(2)), were identified. TXB(2), a key element in platelet aggregation, was decreased upon ginsenoside Rk(1) treatment via inhibition of cyclooxygenase (COX) activity. One of the arachidonic acid (AA) metabolites, 12-hydroxy-5,8,10,14-eicosatetraenoic acid (12-HETE), was decreased significantly in the ginsenoside Rk(1)-treated platelets compared to the AA-induced group. In the mechanism study of ginsenoside Rk(1), a strong linkage to intracellular calcium levels, which induce platelet activation, was found. Additionally, the translocation of 12-LOX from cytosol to membrane, which is related with the intracellular calcium levels, was determined. Therefore, a decreased 12-HETE level induced by ginsenoside Rk(1) on antiplatelet aggregation is related to 12-LOX translocation resulting from decreased Ca(2+) levels. This study shows that global metabolomic analysis has potential for use in understanding the biological behavior of antiplatelet drugs.

Evaluation of headspace‐gas chromatography/mass spectrometry for the analysis of benzene in vitamin C drinks; pitfalls of headspace in benzene detection

Recently, there have been reports regarding the presence of benzene in vitamin C drinks. This is caused by sodium benzoate and ascorbic acid (vitamin C), which can react together to induce benzene formation. While the headspace gas chromatography method is well known for the detection of benzene, there could be pitfalls in the process of benzene extraction. This study was performed to check if benzene could be generated under high-temperature incubation conditions. As a result, the amount of benzene detected by headspace-gas chromatography/mass spectrometry (HSGC/MS) was affected by temperature changes. As the temperature of the sample vial was increased, newly generated benzene from the headspace also increased, causing false-positive determination of benzene. Although 80 degrees C is generally accepted for the temperature of headspace sample vials, lower temperatures, such as 40 degrees C, minimize the false-positive identification of benzene. Considering that this minimization allows benzene to be quantified at around 5 ppb, this lower temperature should definitely be considered since benzene, which is formed in sodium benzoate, can appear in vitamin C drinks under certain circumstances. The proposed analysis method of benzene in vitamin C drinks by HSGC/MS at 40 degrees C is an accurate and universal method for the monitoring of benzene without false-positive identification.

Investigation of metabolite alteration in dimethylnitrosamine-induced liver fibrosis by GC–MS

BACKGROUND A metabolomic study of biomarkers associated with dimethylnitrosamine (DMN)-induced hepatic fibrosis in Sprague-Dawley rats was performed using GC-MS. The clinical chemistry of the collected blood and the histopathology of excised liver samples were examined, and urine samples were prepared by solvent extraction. RESULTS Through pattern analysis, the DMN-treated group was divided into two subgroups based on the aspartate aminotransferase (AST) levels compared with the control, a moderately higher group (DMN subgroup A) and a significantly higher group (DMN subgroup B). Uric acid, orotic acid, N-phenylacetylglycine and glutaric acid were biomarkers for DMN subgroup A, aminomalonic acid was a biomarker for DMN subgroup B, and arabitol level distinguished control versus DMN treatment regardless of AST level. CONCLUSION This study suggests that the identification and profiling of AST level-related metabolites may be useful as a diagnostic tool and for the study of the mechanism of liver fibrosis induced by DMN.

Characterization of Carbon Monoxide Fatalities by Metabolomics

ABSTRACT Metabolite profiling of postmortem blood in carbon monoxide poisoning cases was performed with gas chromatography-mass spectrometry based on metabolomics. All data were processed with an automated mass spectral deconvolution and identification system and the metabolites were characterized using the National Institute of Standards and Technology library. Discriminant analysis was performed and the carbon monoxide-induced biomarkers were determined with a variable importance parameter in the projection plot obtained from partial least square discriminant analysis. This metabolomics approach, including global metabolites profiling, is useful in improving the interpretation of metabolism in carbon monoxide poisoning deaths.