Clinical applications of gas chromatography and gas chromatography–mass spectrometry of steroids

Wolthers, B.G.; Kraan, G.P.B.

doi:10.1016/s0021-9673(99)00153-3

Cited by 117 publications

(77 citation statements)

References 112 publications

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“…Due to efficient utilization of the micellar sweeping effect, the LODs and RSD% values for peak areas for all five steroids were also lower than the previous values (73 mg/L and 5.7% for testosterone) obtained by the sequential PF-MEKC [23]. Although the method is not as sensitive as GC-MS [6,12,13] or LC-MS [45], the obtained LOQ (35 mg/L) enables determination of testosterone in small volumes of male urine after microscale IA-SPE. A minimum of 0.8-1.0 mL of male urine is required per analysis.…”

Section: Discussioncontrasting

confidence: 55%

“…Traditionally, testosterone concentration in urine has been determined by GC-MS [12,13]. Enzymatic hydrolysis of the testosterone conjugates followed by SPE or liquid-liquid extraction (LLE) is required prior to the GC-MS analysis [12].…”

Section: Introductionmentioning

confidence: 99%

“…Enzymatic hydrolysis of the testosterone conjugates followed by SPE or liquid-liquid extraction (LLE) is required prior to the GC-MS analysis [12]. Moreover, the presence of polar hydroxyl and carbonyl groups in the testosterone structure makes derivatization compulsory [12,13].…”

Section: Introductionmentioning

confidence: 99%

“…Laborious and time-consuming sample pretreatment is a notable disadvantage of the GC-MS technique. Moreover, nonspecific LLE and SPE performed, for example, with neutral polystyrene resin or stronger RP bound silica particles [12] are not designed for the separation of testosterone from related steroids during the sample pretreatment.…”

Section: Introductionmentioning

confidence: 99%

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Microscale immunoaffinity SPE and MEKC in fast determination of testosterone in male urine

et al. 2007

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Conventional methods for the determination of testosterone in body fluids typically suffer from poor recovery, lack of specificity, complex sample pretreatment, or the need for derivatization. Here, a simple, specific, and fast analysis method for testosterone was developed, with a methodology based on testosterone-specific immunoaffinity SPE (IA-SPE) and subsequent analysis by partial filling MEKC (PF-MEKC). An immunosorbent consisting of a recombinant antitestosterone Fab fragment covalently attached to activated Sepharose was prepared. IA-SPE and PF-MEKC were set up in hyphenated and off-line constructions, and the applicability of the two constructions in analysis of testosterone in male urine was investigated. The results obtained with the hyphenated construction proved to be only indicative of the presence of testosterone. The off-line IA-SPE and PF-MEKC construction, however, was successfully used in the determination of free testosterone in male urine samples after enzymatic hydrolysis of the glucuronide conjugates. Except for the hydrolysis reaction, no sample pretreatment was required. After hydrolysis, the overall analysis time per sample was only 14 min. The off-line IA-SPE and PF-MEKC method proved to be robust, sensitive (LOQ 35 mg/L), and specific, enabling separation of testosterone from four related steroids. Thus, it provides attractive features when compared to traditional methods for determination of testosterone in male urine.

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Section: Discussioncontrasting

confidence: 55%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Microscale immunoaffinity SPE and MEKC in fast determination of testosterone in male urine

et al. 2007

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“…Metabolite quantitation in traditional chromatography-MS-based targeted analyses has relied on the use of stable-isotope dilution to effectively minimize the uncertainty in the measurement (reviewed in [19,[135][136][137]). In this approach, stable-isotope-labeled analogues (typically labeled with 2 H, 13 C, or 15 N) of the target analyte(s) are spiked into the sample prior to treatment, in order to account for both systematic errors due to the sample processing approach and systematic errors encountered during sample analysis by GC-or LC-MS.…”

Section: Stable-isotope Labeling Techniquesmentioning

confidence: 99%

Future of Liquid Chromatography–Mass Spectrometry in Metabolic Profiling and Metabolomic Studies for Biomarker Discovery

et al. 2007

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SUMMARYThe future utility of liquid chromatography-mass spectrometry (LC-MS) in metabolic profiling and metabolomic studies for biomarker discover will be discussed, beginning with a brief description of the evolution of metabolomics and the utilization of the three most popular analytical platforms in such studies: NMR, GC-MS, and LC-MS. Emphasis is placed on recent developments in highefficiency LC separations, sensitive electrospray ionization approaches, and the benefits to incorporating both in LC-MS-based approaches. The advantages and disadvantages of various quantitative approaches are reviewed, followed by the current LC-MS-based tools available for candidate biomarker characterization and identification. Finally, a brief prediction on the future path of LC-MS-based methods in metabolic profiling and metabolomic studies is given.

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Internal standard strategies for relative and absolute quantitation of peptides in biological matrices by liquid chromatography tandem mass spectrometry

Pailleux

Beaudry

2012

Biomedical Chromatography

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The development of LC-MS/MS instruments and related applications improved the large-scale analyses of proteins and peptides in complex biological mixtures. The historical factor limiting these types of studies was the lack of sensitivity and reproducibility. However, the capacity of these analyses to detect proteins and peptides was significantly enhanced to a point where they are routinely performed in specialized laboratories in support to drug development programs as well as prognostic and diagnostic investigations. The analytical strategy used in peptidomic analyses needs to minimize the fluctuation in data measurements that might mask or reduce the precision of the determinations and consequently reduce the sensitivity of the assay. Inherently, it outlines the importance of careful standardization to reduce technical and instrumental variation. Therefore, this review will focus on the strengths and the limitations of the different experimental approaches used for the integration of internal standards in peptidomic studies. This review will examine a wide variety of methods, reagents, instrumentations and data analysis tools available to design peptidomic experiments. Moreover, this review will focus on the importance of precision and accuracy in order to adequately establish analysis threshold to detect peptide expression differences.

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Clinical applications of gas chromatography and gas chromatography–mass spectrometry of steroids

Cited by 117 publications

References 112 publications

Microscale immunoaffinity SPE and MEKC in fast determination of testosterone in male urine

Microscale immunoaffinity SPE and MEKC in fast determination of testosterone in male urine

Future of Liquid Chromatography–Mass Spectrometry in Metabolic Profiling and Metabolomic Studies for Biomarker Discovery

Internal standard strategies for relative and absolute quantitation of peptides in biological matrices by liquid chromatography tandem mass spectrometry

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