Biomarker Discovery in Low-Grade Breast Cancer Using Isobaric Stable Isotope Tags and Two-Dimensional Liquid Chromatography-Tandem Mass Spectrometry (iTRAQ-2DLC-MS/MS) Based Quantitative Proteomic Analysis

2009 ◽  
Vol 8 (1) ◽  
pp. 362-373 ◽  
Author(s):  
Pavel Bouchal ◽  
Theodoros Roumeliotis ◽  
Roman Hrstka ◽  
Rudolf Nenutil ◽  
Borivoj Vojtesek ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Mass Spectrometry ◽  
Liquid Chromatography ◽  
Stable Isotope ◽  
Biomarker Discovery ◽  
Low Grade ◽  
Tandem Mass ◽  
Two Dimensional ◽  
Chromatography Tandem Mass Spectrometry ◽  
Liquid Chromatography Tandem Mass

Proteomic profiling of breast cancer serum via two-dimensional liquid-chromatography—tandem mass spectrometry

2006 ◽  
Vol 24 (18_suppl) ◽  
pp. 542-542
Author(s):  
Q. C. Ru ◽  
L. A. Zhu ◽  
J. Silberman ◽  
R. Mural ◽  
C. D. Shriver
Keyword(s):  
Breast Cancer ◽  
Mass Spectrometry ◽  
Liquid Chromatography ◽  
Growth Factor ◽  
Protein Kinase ◽  
Atypical Hyperplasia ◽  
Tandem Mass ◽  
Two Dimensional ◽  
Chromatography Tandem Mass Spectrometry ◽  
Liquid Chromatography Tandem Mass

542 Background: Discovering biomarkers for early cancer detection is a major goal of translational research. We have applied modern proteomic technology, two-dimensional liquid-chromatography-tandem mass spectrometry (2-D LC-MS/MS), to large-scale breast cancer serum profiling. Methods: 189 human sera from breast disease or cancer patients (14 atypical hyperplasia, 63 benign, 29 DCIS, and 83 invasive CA) were analyzed via 2-D LC-MS/MS method. Patient ages ranged from 19 to 97 years, with a mean of 55. Ethnicities included African American, Asian, Hispanic, Native American, and Caucasian. This methodology utilizes only 10 μL of human serum per specimen. Every sample was run three times to ensure reproducibility and to maximize identification. The protein identification was conducted using the Bioworks Browser 3.1 and the IPI_human protein database (version 6.23.04) and filtered via HUPO criteria to ensure confidence. Results: 230 high-confidence proteins were identified from analyses of 189 sera. 26 common serum proteins were detected in all samples and 17 were found in two- or three- diagnostic stages. 83 proteins such as insulin receptor substrate 4, serine/threonine-protein kinase WNK4, DNA mismatch repair protein MSH6, and signal recognition particle 72 kDa protein were detected only in invasive sera. 30 proteins including breast carcinoma amplified sequence 3, uncharacterized bone marrow protein BM032, fibroblast growth factor receptor 2, and tumor necrosis factor receptor superfamily member 4 were detected only in DCIS sera. The 15 proteins detected only in atypical hyperplasia sera included cholecystokinin type A receptor and cell division cycle 2-related protein kinase 7. 59 proteins including serine/threonine protein phosphatase 6, probable transcription factor CST, insulin-like growth factor IA, and connective tissue growth factor were found in benign samples. Conclusions: Identification of signaling pathway/transcriptions proteins and cell cycle related proteins in breast cancer sera, and identification of breast cancer amplified sequence 3 proteins in the DCIS sera, suggest that high-throughput proteomic technologies like 2-D LC - MS/MS may facilitate the discovery of diagnostically useful breast cancer biomarkers. No significant financial relationships to disclose.

A sensitive method to determine melatonin in saliva by automated online in-tube solid-phase microextraction coupled with stable isotope-dilution liquid chromatography-tandem mass spectrometry

2017 ◽  
Vol 9 (21) ◽  
pp. 3134-3140 ◽  
Author(s):  
Atsushi Ishizaki ◽  
Akiko Uemura ◽  
Hiroyuki Kataoka
Keyword(s):  
Mass Spectrometry ◽  
Liquid Chromatography ◽  
Stable Isotope ◽  
Solid Phase Microextraction ◽  
Solid Phase ◽  
Tandem Mass ◽  
Stable Isotope Dilution ◽  
Chromatography Tandem Mass Spectrometry ◽  
Liquid Chromatography Tandem Mass ◽  
Sensitive Method

Melatonin (MLT) plays important roles in regulating the sleep-wake cycle, and has many beneficial effects on health. A simple, rapid, and sensitive method was developed for the determination of MLT in human saliva by automated online in-tube solid-phase microextraction coupled with stable isotope-dilution liquid chromatography-tandem mass spectrometry.

scholarly journals Measurement of Urinary d- and l-2-Hydroxyglutarate Enantiomers by Stable-Isotope-Dilution Liquid Chromatography–Tandem Mass Spectrometry after Derivatization with Diacetyl-l-Tartaric Anhydride

2004 ◽  
Vol 50 (8) ◽  
pp. 1391-1395 ◽  
Author(s):  
Eduard A Struys ◽  
Erwin E W Jansen ◽  
Nanda M Verhoeven ◽  
Cornelis Jakobs
Keyword(s):  
Mass Spectrometry ◽  
Differential Diagnosis ◽  
Liquid Chromatography ◽  
Stable Isotope ◽  
Tandem Mass Spectrometry ◽  
Tandem Mass ◽  
Internal Standards ◽  
Chromatography Tandem Mass Spectrometry ◽  
Liquid Chromatography Tandem Mass

Abstract Background: The differential diagnosis of d-2-hydroxyglutaric aciduria (d-2-HGA), l-2-hydroxyglutaric aciduria (l-2-HGA), and the combined d/l-2-hydroxyglutaric aciduria (d/l-2-HGA) can be accomplished only by the measurement of the corresponding 2-hydroxyglutarate (2-HG). Available methods for the determination of d- and l-2-HG in urine are either time-consuming and expensive or have not been extensively validated. We aimed to develop a method for their rapid and sensitive measurement. Methods: We used liquid chromatography–tandem mass spectrometry (LC-MS/MS) for the determination of d- and l-2-HG with stable-isotope-labeled internal standards. Urine samples of 20 μL were mixed with 250 μL of methanol containing the internal standards and subsequently dried under nitrogen. The analytes were derivatized by use of diacetyl-l-tartaric anhydride (DATAN) to obtain diastereomers, which were separated on an achiral C18 HPLC column and detected by MS/MS in multiple-reaction-monitoring mode. Results: The use of DATAN as chiral derivatization reagent provided very well separated peaks of the formed diastereomers of d- and l-2-HG, with a total runtime of 5 min. The inter- and intraassay CVs for d- and l-2-HG ranged from 3.4% to 6.2%. Mean recoveries of d- and l-2-HG, evaluated on two concentrations, were 94%. Detection limit of the presented method was 20 pmol for a sample volume of 20 μL. Method comparison of the LC-MS/MS method with a gas chromatography–mass spectrometry method, in which d- and l-2-HG were derivatized with R-(−)-butanol, showed good agreement between the two methods. Conclusions: Urinary d- and l-2-HG can be analyzed by MS/MS after derivatization with DATAN. The presented method may be suitable for the differential diagnosis of 2-HGA.

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