High-performance Liquid Chromatography-quadrupole Time-of-flight Mass Spectrometry Research Articles

MP22-06 A URINARY METABOLOMIC APPROACH CAN DIFFERENTIATE PATIENTS WITH BLADDER CANCER FROM NORMAL OR HEMATURIC CONTROLS, AND PREDICT CANCER-SPECIFIC SURVIVAL

You have accessJournal of UrologyBladder Cancer: Detection & Screening1 Apr 2014MP22-06 A URINARY METABOLOMIC APPROACH CAN DIFFERENTIATE PATIENTS WITH BLADDER CANCER FROM NORMAL OR HEMATURIC CONTROLS, AND PREDICT CANCER-SPECIFIC SURVIVAL Wun-Jae Kim, Phildu Jeong, Xing Jin, Young-Won Kim, In-Chang Cho, Won Tae Kim, Yong-June Kim, Sunghyouk Park, Sang-Cheol Lee, and Seok-Joong Yun Wun-Jae KimWun-Jae Kim More articles by this author , Phildu JeongPhildu Jeong More articles by this author , Xing JinXing Jin More articles by this author , Young-Won KimYoung-Won Kim More articles by this author , In-Chang ChoIn-Chang Cho More articles by this author , Won Tae KimWon Tae Kim More articles by this author , Yong-June KimYong-June Kim More articles by this author , Sunghyouk ParkSunghyouk Park More articles by this author , Sang-Cheol LeeSang-Cheol Lee More articles by this author , and Seok-Joong YunSeok-Joong Yun More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2014.02.854AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Metabolomics is the relatively new scientific study of the biochemical processes that involve metabolites. BC seems to be ideal for urinary metabolomics-based diagnosis, as urine can directly contact the cancer lesion in the bladder. Moreover, urine collection can be made conveniently and its metabolomics study is non-invasive. METHODS In the present study, high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (HPLC-QTOFMS) was used to profile urine metabolites of 138 patients with BC and 121 control subjects (69 healthy people and 52 patients with hematuria due to non-malignant disease), a patient population much larger than those assessed in other BC metabolomics studies. RESULTS Multivariate statistical analysis revealed that the cancer group could be clearly distinguished from the control groups on the basis of their metabolomic profiles, even when the hematuric control group was included. Patients with muscle-invasive BC could also be distinguished from patients with non-muscle-invasive BC on the basis of their metabolomic profiles. Successive analyses identified 12 differential metabolites that contributed to the distinction between the BC and control groups, and many of them turned out to be involved in glycolysis and beta-oxidation. The association of these metabolites with cancer was corroborated by microarray results showing that carnitine transferase and pyruvate dehydrogenase complex expressions is significantly altered in cancer groups. In terms of clinical applicability, the multivariate differentiation model diagnosed BC with a sensitivity and specificity of 91.3% and 92.5%, respectably, and comparable results were obtained by receiver operating characteristic analysis (AUC = 0.937). Multivariate regression also suggested that the metabolic profile correlates with cancer-specific survival time. CONCLUSIONS The excellent performance and simplicity of this metabolomics-based approach suggests that it has the potential to augment or even replace the current modalities for BC diagnosis © 2014FiguresReferencesRelatedDetails Volume 191Issue 4SApril 2014Page: e236 Advertisement Copyright & Permissions© 2014MetricsAuthor Information Wun-Jae Kim More articles by this author Phildu Jeong More articles by this author Xing Jin More articles by this author Young-Won Kim More articles by this author In-Chang Cho More articles by this author Won Tae Kim More articles by this author Yong-June Kim More articles by this author Sunghyouk Park More articles by this author Sang-Cheol Lee More articles by this author Seok-Joong Yun More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

Structural speculation and identification of alkaloids in Macleaya cordata fruits by high‐performance liquid chromatography/quadrupole‐time‐of‐flight mass spectrometry combined with a screening procedure

Alkaloids with significant therapeutic effects are the main active constituents of Macleaya cordata, which is a perennial herb plant in the Papaveraceae family. A systematic and novel method for speculating and identifying the structures of alkaloids in M. cordata fruits by high-performance liquid chromatography/quadrupole-time-of-flight mass spectrometry (HPLC/Q-TOF-MS) with a screening procedure was reported. Investigation of mass spectral fragmentation of alkaloids was carried out based on the tandem mass spectrometry (MS/MS) data analyses of eight reference substances. The skeletons of alkaloids were determined by their ultraviolet spectra (UV) and MS/MS data. The substituent groups of the alkaloids were acquired through a screening procedure developed in our laboratory and MS/MS data. The substituent linkage sites were deduced by MS/MS fragmentation behavior, as well as biosynthetic pathways of related alkaloids. The structures of 21 alkaloids were speculated in this study, 10 of which were reported for the first time in M. cordata. Furthermore, benzyltetrahydroisoquinoline and N-methyltetrahydroprotoberberine-type alkaloids were discovered, which indirectly proved that the biosynthetic pathways of benzophenanthridine alkaloids reported in Eschscholtzia california existed in M. cordata as well. HPLC/Q-TOF-MS combined with a screening procedure was a systematic and reliable method for speculating and elucidating the structures of alkaloids. This study might be useful for the identification of other compounds in herbal medicines.

Determination of three exogenous plant hormone residues in bean sprout by high performance liquid chromatography-quadrupole-time of flight mass spectrometry

This study was aimed to the establishment of an analytical method for the determination of three exogenous plant hormone residues in bean sprout by high performance liquid chromatography-quadrupole-time of flight mass spectrometry (HPLC-Q-TOF-MS). The target compounds were gibberellins, 6-benzylaminopurine and parachlorophenoxyacetic acid. The QuEChERS (quick, easy, cheap, effective, rugged, and safe) method was used for sample preparation. The analytes were extracted with a solution containing 1% (v/v, if not specified) acetic, 50% ethanol, 49% acetonitrile, and cleaned-up by dispersive solid-phase extraction with diatomite dispersant, then degreased by hexane. The three target compounds were separated on an Eclipse Plus C18 column (100 mm x 3.0 mm, 1.8 microm) with mobile phases A (water containing 0.1% formic acid) and B (methanol) by gradient elution within 15 min, and detected under negative electrospray ionization (ESI) mode. The quantitative analysis was carried out by extracting the peak area with accurate mass. The confirmatory analysis of the target compounds was performed with the qualitative fragments. The results showed that the limits of quantification (LOQs, S/N = 10) for the three target compounds were from 5.0 microg/kg to 10 microg/kg. The respective mean recoveries were found to be in the range of 79.1%-96.1%, and the RSDs were 5.7%-10.4%. It was applicable to the analysis of the three exogenous plant hormones in bean sprout samples. This method is simple, fast and efficient.

Study on intervention effect of Jieduquyuziyin prescription systemic lupus erythematosus by HPLC-Q-TOF/MS

To establish a metabonomic method based on high performance liquid chromatography-quadrupole-time of flight mass spectrometry (HPLC-Q-TOF/MS), in order to study the changes in serum metabolites of systemic lupus erythematosus (SLE) mice after treatment of Jieduquyuziyin prescription, the pathogenesis of SLE and mechanism of drug action. The orthogonal partial least squares (OPLS) was applied for the pattern recognition of experimental data, finding a significant difference in the control group, the SLE model group, the Jieduquyuziyin prescription-treated group and the prednisone acetate-treated group. According to the OPLS load diagram, 12 differential metabolites, including traumatic acid, PAF, 12 (S)-HEPE, 15(S)-HETrE and Hepoxilin B3 were identified by using accurate mass combined with MS/MS data After treatment with Jieduquyuziyin prescription, the relative contents of PAF, 12 (S)-HETE were close to the level of the control group. According to the analysis on metabolic pathway, SLE could cause significant changes in unsaturated fatty acid and amino acid metabolism pathway, while Jieduquyuziyin prescription has a effect in regulating disorder of unsaturated fatty acid metabolism pathway.

Metabonomics study of lung cancer cells based on liquid chromatography-mass spectrometry

The metabolic profiles of the polar metabolites and the non-polar metabolites in lung tumor cell lines H358, A549, HCC827, H1299, Calu-3, Calu-l, PC-9 and normal cell line MRC-5 were analyzed separately using high performance liquid chromatography-quadrupole time-of flight mass spectrometry (HPLC-Q-TOF/MS). Partial least square discriminant analysis ( PLS-DA) was used to process the metabolic data. The results showed that the metabolites of the lung cancer cell lines and normal cell line have significant differences. Further, 10 polar metabolites and 21 non-polar metabolites which had a significant contribution to classification were selected and preliminarily identified due to the accurate mass. Comparing with the normal cell line, the lung tumor cell lines present an abnormal metabolism in protein, fatty acid, and phospholipids. These results may provide important information for the early diagnosis of lung cancer.

Identification of ozonation by-products of 4- and 5-methyl-1H-benzotriazole during the treatment of surface water to drinking water

During the treatment of surface water to drinking water, ozonation is often used for disinfection and to remove organic trace substances, whereby oxidation by-products can be formed. Here we use the example of tolyltriazole to describe an approach for identifying relevant oxidation by-products in the laboratory and subsequently detecting them in an industrial-scale process. The identification process involves ozonation experiments with pure substances at laboratory level (concentration range mg L −1). The reaction solutions from different ozone contact times were analyzed by high performance liquid chromatography - quadrupole time-of-flight mass spectrometry (HPLC-QTOF-MS) in full scan mode. Various approaches were used to detect the oxidation by-products: (i) target searches of postulated oxidation by-products, (ii) comparisons of chromatograms (e.g., UV/VIS) of the different samples, and (iii) color-coded abundance time courses (kinetic) of all detected compounds were illustrated in a kind of a heat map. MS/MS, H/D exchange, and derivatization experiments were used for structure elucidation for the detected by-product. Due to the low contaminant concentrations (ng L −1-range) of contaminants in the untreated water, the conversion of results from laboratory experiments to an industrial-scale required the use of HPLC-MS/MS with sample enrichment (e.g., solid phase extraction.) In cases where reference substances were not available or oxidation by-products without clear structures were detected, reaction solutions from laboratory experiments were used to optimize the analytical method to detect ng L −1 in the samples of the industrial processes. We exemplarily demonstrated the effectiveness of the methodology with the industrial chemicals 4- and 5-methyl-1H-benzotriazole (4- and 5-MBT) as an example. Moreover, not only did we identify several oxidation by-products in the laboratory experiments tentatively, but also detected three of the eleven reaction products in the outlet of the full-scale ozonation unit.

Serial mixed-mode cation- and anion-exchange solid-phase extraction for separation of basic, neutral and acidic pharmaceuticals in wastewater and analysis by high-performance liquid chromatography–quadrupole time-of-flight mass spectrometry

A novel solid-phase extraction (SPE) method is presented whereby 15 basic, neutral and acidic pharmaceuticals in wastewater were simultaneously extracted and subsequently separated into different fractions. This was achieved using mixed-mode cation- and anion-exchange SPE (Oasis MCX and MAX) in series. Analysis was performed by high-performance liquid chromatography–quadrupole time-of-flight mass spectrometry (HPLC/QTOF-MS). A fast separation was achieved, with all compounds eluting within 6 min, narrow chromatographic peaks, with a peak base width of 6 s on average, and a high mass accuracy of quantified wastewater sample ions, with average mass errors in absolute value of 0.7 mDa or 2.7 ppm. The recovery of the SPE method in the analysis of sewage treatment plant (STP) influent and effluent wastewater was on average 80% and the ion suppression 30%. For less demanding samples Oasis MCX used alone may be an alternative method, although for STP influent waters containing high loads of organic compounds the clean-up achieved using only Oasis MCX was insufficient, leading to unreliable quantitation. Furthermore, serial SPE separation according to molecular charge added an additional degree of analyte confirmation. For quantitation, an approach combining external standard calibration curves, isotopically labelled surrogate standards and single-point standard addition was used. The applicability of the method was demonstrated in the analysis of influent and effluent wastewater from an STP, using small sample volumes (25–50 mL). The effluent wastewater had been subjected to three different treatments; activated sludge, activated sludge followed by ozonation, and a membrane bioreactor (MBR). Ozone treatment proved superior in removal of the analysed pharmaceuticals, while the MBR provided higher removal efficiencies than the activated sludge process.