Bulk derivatization and cation exchange restricted access media-based trap-and-elute liquid chromatography–mass spectrometry method for determination of trace estrogens in serum

Beinhauer, Jana; Bian, Linkan; Fan, Haibo; Šebela, Marek; Kukula, Maciej; Barrera, Jose; Schug, Kevin A.

doi:10.1016/j.aca.2014.11.032

Cited by 26 publications

(28 citation statements)

References 44 publications

(57 reference statements)

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“…After loading samples on an Oasis HLB (hydrophilic-lipophilic balance) cartridge (Waters, Milford, MA), unconjugated estrogens were first eluted with ethyl acetate, and conjugates were then eluted with methanol containing 0.1% ammonium hydroxide. A more recent study reported an automated on-line trap-and-elute sample treatment process by using a weak cation exchange (WCX) restricted access material (RAM) trap column (Waters) and on-line dilution [52]. This approach has potential application to older men, children, and postmenopausal women samples since the streamlined procedure requires only 100 μL of serum and an LLOQ of 3 pg/mL with excellent accuracy and precision.…”

Section: Lc-tandem Ms (Ms/ms)-based Assays For Estrogen Analysismentioning

confidence: 99%

Ultra-high sensitivity analysis of estrogens for special populations in serum and plasma by liquid chromatography–mass spectrometry: Assay considerations and suggested practices

Wang

Mesaros

Blair

2016

The Journal of Steroid Biochemistry and Molecular Biology

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Estrogen measurements play an important role in the clinical evaluation of many endocrine disorders as well as in research on the role of hormones in human biology and disease. It remains an analytical challenge to quantify estrogens and their metabolites in specimens from special populations including older men, children, postmenopausal women and women receiving aromatase inhibitors. Historically, immunoassays have been used for measuring estrogens and their metabolites in biological samples for risk assessment. However, the lack of specificity and accuracy of immunoassay-based methods has caused significant problems when interpreting data generated from epidemiological studies and across different laboratories. Stable isotope dilution (SID) methodology coupled with liquid chromatography-selected reaction monitoring-mass spectrometry (LC-SRM/MS) is now accepted as the ‘gold-standard’ to quantify estrogens and their metabolites in serum and plasma due to improved specificity, high accuracy, and the ability to monitor multiple estrogens when compared with immunoassays. Ultra-high sensitivity can be obtained with pre-ionized derivatives when using triple quadruple mass spectrometers in the selected reaction monitoring (SRM) mode coupled with nanoflow LC. In this review, we have examined the special issues related to utilizing ultra-high sensitivity SID LC-SRM/MS-based methodology to accurately quantify estrogens and their metabolites in the serum and plasma from populations with low estrogen levels. The major issues that are discussed include: sample preparation for both unconjugated and conjugated estrogens, derivatization, chromatographic separation, matrix effects, and assay validation.

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Section: Lc-tandem Ms (Ms/ms)-based Assays For Estrogen Analysismentioning

confidence: 99%

Ultra-high sensitivity analysis of estrogens for special populations in serum and plasma by liquid chromatography–mass spectrometry: Assay considerations and suggested practices

Wang

Mesaros

Blair

2016

The Journal of Steroid Biochemistry and Molecular Biology

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“…2‐Fluoro‐1‐methylpyridium p ‐toluenesulfonate (2‐FMP) is another reagent that has been used for ISD of estrogens in human and bovine serum . Dns‐Cl was excluded for consideration in this application due to its pH‐dependent charging; 2‐FMP is another reagent that will impart a permanent positive charge onto the derivatized analyte.…”

Section: Alcohols Phenols and Other Hydroxyl‐bearing Compoundsmentioning

confidence: 99%

Review of in situ derivatization techniques for enhanced bioanalysis using liquid chromatography with mass spectrometry

Baghdady

Schug

2015

J of Separation Science

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Accurate and specific analysis of target molecules in complex biological matrices remains a significant challenge, especially when ultra-trace detection limits are required. Liquid chromatography with mass spectrometry is often the method of choice for bioanalysis. Conventional sample preparation and clean-up methods prior to the analysis of biological fluids such as liquid-liquid extraction, solid-phase extraction, or protein precipitation are time-consuming, tedious, and can negatively affect target recovery and detection sensitivity. An alternative or complementary strategy is the use of an off-line or on-line in situ derivatization technique. In situ derivatization can be incorporated to directly derivatize target analytes in their native biological matrices, without any prior sample clean-up methods, to substitute or even enhance the extraction and preconcentration efficiency of these traditional sample preparation methods. Designed appropriately, it can reduce the number of sample preparation steps necessary prior to analysis. Moreover, in situ derivatization can be used to enhance the performance of the developed liquid chromatography with mass spectrometry-based bioanalysis methods regarding stability, chromatographic separation, selectivity, and ionization efficiency. This review presents an overview of the commonly used in situ derivatization techniques coupled to liquid chromatography with mass spectrometry-based bioanalysis to guide and to stimulate future research.

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“…It is therefore crucial to clean the samples prior to LC-MS. This is often accomplished with relatively laborious pretreatments, such as liquid-liquid extraction (LLE) and solid phase extraction (SPE) [41, 42]. These off-line techniques can also suffer from poor recovery and accurancy.…”

Section: Introductionmentioning

confidence: 99%

“…As an example, a reversed phase-based RAM LC-MS/MS system has been used for quantitative determination of dansylated bisphenol in human saliva [65]. A similar approach with RAM trap-and-elute system has been used for trace quantification of estrogens in cerebrospinal fluid and human serum samples [41, 46, 47]. Rao et al .…”

Section: Introductionmentioning

confidence: 99%

Targeted quantification of lipid mediators in skeletal muscles using restricted access media-based trap-and-elute liquid chromatography-mass spectrometry

Wang

Bian

et al. 2017

Analytica Chimica Acta

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Lipid mediators (LMs) are a class of bioactive metabolites of the essential polyunsaturated fatty acids (PUFA), which are involved in many physiological processes. Their quantification in biological samples is critical for understanding their functions in lifestyle and chronic diseases, such as diabetes, as well allergies, cancers, and in aging processes. We developed a rapid, and sensitive LC-MS/MS method to quantify the concentrations of 14 lipid mediators of interest in mouse skeletal muscle tissue without time-consuming liquid-liquid or solid-phase extractions. A restricted-access media (RAM) based trap was used prior to LC-MS as cleanup process to prevent the analytical column from clogging and deterioration. The system enabled automatic removal of residual proteins and other biological interferences presented in the tissue extracts; the target analytes were retained in the trap and then eluted to an analytical column for separation. Matrix evaluation tests demonstrated that the use of the combined RAM trap and chromatographic separation efficiently eliminated the biological or chemical matrix interferences typically encountered in bioanalytical analysis. Using 14 LM standards and 12 corresponding deuterated compounds as internal standards, the five-point calibration curves, established over the concentration range of 0.031 to 320 ng mL−1, demonstrated good linearity of r2 > 0.9903 (0.9903 to 0.9983). The lower detection limits obtained were 0.016, 0.031, 0.062, and 0.31 ng mL−1 (0.5, 1, 2, and 10 pg on column), respectively, depending on the specific compounds. Good accuracy (87.1–114.5%) and precision (<13.4%) of the method were observed for low, medium, and high concentration quality control samples. The method was applied to measure the amount of 14 target LMs in mouse skeletal muscle tissues. All 14 analytes in this study were successfully detected and quantified in the gastrocnemius muscle samples, which provided crucial information for both age and gender-related aspects of LMs signaling in skeletal muscles previously unknown. This method could be applied to advance the understanding of skeletal muscle pathophysiology to study the role of LMs in health and disease. Furthermore, we will expand the application of this methodology to humans and other tissues/matrices in the near future.

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Bulk derivatization and cation exchange restricted access media-based trap-and-elute liquid chromatography–mass spectrometry method for determination of trace estrogens in serum

Cited by 26 publications

References 44 publications

Ultra-high sensitivity analysis of estrogens for special populations in serum and plasma by liquid chromatography–mass spectrometry: Assay considerations and suggested practices

Ultra-high sensitivity analysis of estrogens for special populations in serum and plasma by liquid chromatography–mass spectrometry: Assay considerations and suggested practices

Review of in situ derivatization techniques for enhanced bioanalysis using liquid chromatography with mass spectrometry

Targeted quantification of lipid mediators in skeletal muscles using restricted access media-based trap-and-elute liquid chromatography-mass spectrometry

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