A non-invasive approach to explore the discriminatory potential of the urinary volatilome of invasive ductal carcinoma of the breast

Taunk, Khushman; Taware, Ravindra; More, Tushar H.; Porto-Figueira, Priscilla; Mohapatra, Rajkishore; Soneji, Dharmesh; Câmara, José S.; Nagarajaram, Hampapathalu A.; Rapole, Srikanth

doi:10.1039/c8ra02083c

Cited by 27 publications

(26 citation statements)

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“…Overall, the SPME approach is easy to use, sensitive (limits of detections in ppb range), reproducible and conserve the integrity of the biological sample [101]. Therefore, it's not surprising to find that SPME is widely used in volatomic analysis of various biospecimens [99,[101][102][103][104]. An overview about the typical experimental layout for the analysis of salivary VOCs using SPME-GC/MS is provided in Figure 2.…”

Section: Solid Phase Micro Extraction (Spme)mentioning

confidence: 99%

Unravelling the Potential of Salivary Volatile Metabolites in Oral Diseases. A Review

et al. 2020

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Fostered by the advances in the instrumental and analytical fields, in recent years the analysis of volatile organic compounds (VOCs) has emerged as a new frontier in medical diagnostics. VOCs analysis is a non-invasive, rapid and inexpensive strategy with promising potential in clinical diagnostic procedures. Since cellular metabolism is altered by diseases, the resulting metabolic effects on VOCs may serve as biomarkers for any given pathophysiologic condition. Human VOCs are released from biomatrices such as saliva, urine, skin emanations and exhaled breath and are derived from many metabolic pathways. In this review, the potential of VOCs present in saliva will be explored as a monitoring tool for several oral diseases, including gingivitis and periodontal disease, dental caries, and oral cancer. Moreover, the analytical state-of-the-art for salivary volatomics, e.g., the most common extraction techniques along with the current challenges and future perspectives will be addressed unequivocally.

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Section: Solid Phase Micro Extraction (Spme)mentioning

confidence: 99%

Unravelling the Potential of Salivary Volatile Metabolites in Oral Diseases. A Review

et al. 2020

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“…Urine is frequently analyzed using Liquid Chromatography-Mass Spectrometry (LC-MS), Gas Chromatography-Mass Spectrometry (GC-MS) and Nuclear Magnetic Resonance Spectroscopy (NMR) [5][6][7]. A specific class of metabolomics focused on the profile of volatile compounds (VCs) is termed volatolomics, the applications of which for diagnostic purposes is growing [8][9][10][11][12][13][14][15]. VCs are secreted by cells of the human body, as a result of their metabolism.…”

Section: Introductionmentioning

confidence: 99%

“…The changes in the profile of VCs in biological fluids, dependent on the metabolic changes, may reflect the presence of disease. Many studies have suggested that the profile of VCs in urine change in cancer [8][9][10], nephrological conditions [15], oxidative stress [11], gastrointestinal diseases [12][13][14] and other disease states.…”

Section: Introductionmentioning

confidence: 99%

“…An analysis of VCs with GC-MS typically requires a sample preparation step, particularly pre-concentration of analytes. These sample preparation methods for VC analysis in urine are comprised mostly of adsorption techniques, such as solid-phase microextraction (SPME) [8,10,[20][21][22] and thermal desorption with sorbent tubes [23]. These are selective techniques, which are also adversely affected by high water concentrations, limiting the number of compounds detected and reducing their usefulness for non-targeted profiling.…”

Section: Introductionmentioning

confidence: 99%

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An Optimization of Liquid–Liquid Extraction of Urinary Volatile and Semi-Volatile Compounds and Its Application for Gas Chromatography-Mass Spectrometry and Proton Nuclear Magnetic Resonance Spectroscopy

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Urinary volatile compounds (VCs) have been recently assessed for disease diagnoses. They belong to very diverse chemical classes, and they are characterized by different volatilities, polarities and concentrations, complicating their analysis via a single analytical procedure. There remains a need for better, lower-cost methods for VC biomarker discovery. Thus, there is a strong need for alternative methods, enabling the detection of a broader range of VCs. Therefore, the main aim of this study was to optimize a simple and reliable liquid–liquid extraction (LLE) procedure for the analysis of VCs in urine using gas chromatography-mass spectrometry (GC-MS), in order to obtain the maximum number of responses. Extraction parameters such as pH, type of solvent and ionic strength were optimized. Moreover, the same extracts were analyzed using Proton Nuclear Magnetic Resonance Spectroscopy (1H-NMR), to evaluate the applicability of a single urine extraction for multiplatform purposes. After the evaluation of experimental conditions, an LLE protocol using 2 mL of urine in the presence of 2 mL of 1 M sulfuric acid and sodium sulphate extracted with dichloromethane was found to be optimal. The optimized method was validated with the external standards and was found to be precise and linear, and allowed for detection of >400 peaks in a single run present in at least 50% of six samples—considerably more than the number of peaks detected by solid-phase microextracton fiber pre-concentration-GC-MS (328 ± 6 vs. 234 ± 4). 1H-NMR spectroscopy of the polar and non-polar extracts extended the range to >40 more (mainly low volatility compounds) metabolites (non-destructively), the majority of which were different from GC-MS. The more peaks detectable, the greater the opportunity of assessing a fingerprint of several compounds to aid biomarker discovery. In summary, we have successfully demonstrated the potential of LLE as a cheap and simple alternative for the analysis of VCs in urine, and for the first time the applicability of a single urine solvent extraction procedure for detecting a wide range of analytes using both GC-MS and 1H-NMR analysis to enhance putative biomarker detection. The proposed method will simplify the transport between laboratories and storage of samples, as compared to intact urine samples.

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“…Though, metabolic investigation of cancer is a routinely followed tool to get meaningful insights of differential metabolomics response but unfortunately, volatile component of the metabolome is often ignored. Volatile organic metabolites (VOMs) comprise significant part of the metabolome which could complement and shed light on crucial aspects of metabolic dysregulations that occur during various pathophysiologies (Cavaco et al 2018;Silva et al 2011;Taunk et al 2018;Taware et al 2017Taware et al , 2018.Therefore, in this study, we have carried out characterization of the VOMs produced by the MCF-7 (non-aggressive/non-metastatic) and MDA-MB-231( aggressive/metastatic)BC cell lines grown under normoxic and hypoxic conditions to identify the hypoxia driven specific volatilomic signature. Moreover, we also attempted to establish the hypoxia mediated volatilomic alterations as well as related metabolic pathways specific to the aggressive phenotype of breast cancer cells.…”

Section: Introductionmentioning

confidence: 99%

Extracellular volatilomic alterations induced by hypoxia in breast cancer cells

et al. 2020

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IntroductionThe metabolic shift induced by hypoxia in cancer cells has not been explored at volatilomic level so far. The volatile organic metabolites (VOMs) constitute an important part of the metabolome and their investigation could provide us crucial aspects of hypoxia driven metabolic reconfiguration in cancer cells. Objective To identify the altered volatilomic response induced by hypoxia in metastatic/aggressive breast cancer (BC) cells. Methods BC cells were cultured under normoxic and hypoxic conditions and VOMs were extracted using HS-SPME approach and profiled by standard GC-MS system. Univariate and multivariate statistical approaches (p < 0.05, Log 2 FC ≥ 0.58/≤ − 0.58, PC1 > 0.13/< − 0.13) were applied to select the VOMs differentially altered after hypoxic treatment. Metabolic pathway analysis was also carried out in order to identify altered metabolic pathways induced by the hypoxia in the selected BC cells. Results Overall, 20 VOMs were found to be significantly altered (p < 0.05, PC1 > 0.13/< − 0.13) upon hypoxic exposure to BC cells. Further, cell line specific volatilomic alterations were extracted by comparative metabolic analysis of aggressive (MDA-MB-231) vs. non-aggressive (MCF-7) cells incubated under hypoxia and normoxia. In this case, 15 and 12 VOMs each were found to be significantly altered in aggressive cells when exposed to hypoxic and normoxic condition respectively. Out of these, 9 VOMs were found to be uniquely associated with hypoxia, 6 were specific to normoxia and 6 were found common to both the conditions. Formic acid was identified as the most prominent molecule with higher abundance levels in aggressive as compared to non-aggressive cells in both conditions. Furthermore, metabolic pathway analyses revealed that fatty acid biosynthesis and nicotinate and nicotinamide metabolism were significantly altered in aggressive as compared to non-aggressive cells in normoxia and hypoxia respectively. Conclusions Higher formate overflow was observed in aggressive cells compared to non-aggressive cells incubated under both the conditions, reinforcing its correlation with aggressive and invasive cancer type. Moreover, under hypoxia, aggressive cells preferred to be bioenergetically more efficient whereas, under normoxia, fatty acid biosynthesis was favoured when compared to non-aggressive cells.

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A non-invasive approach to explore the discriminatory potential of the urinary volatilome of invasive ductal carcinoma of the breast

Abstract: A non-invasive urinary volatilomics approach for exploring the IDC type breast cancer.

Cited by 27 publications

References 70 publications

Unravelling the Potential of Salivary Volatile Metabolites in Oral Diseases. A Review

Unravelling the Potential of Salivary Volatile Metabolites in Oral Diseases. A Review

An Optimization of Liquid–Liquid Extraction of Urinary Volatile and Semi-Volatile Compounds and Its Application for Gas Chromatography-Mass Spectrometry and Proton Nuclear Magnetic Resonance Spectroscopy

Extracellular volatilomic alterations induced by hypoxia in breast cancer cells

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