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Featured researches published by Mary Bedner.


Analytical Chemistry | 2010

Development of a Candidate Reference Measurement Procedure for the Determination of 25-hydroxyvitamin D3 and 25-hydroxyvitamin D2 in Human Serum Using Isotope-Dilution Liquid Chromatography/Tandem Mass Spectrometry

Susan S.-C. Tai; Mary Bedner; Karen W. Phinney

Vitamin D exists in two major forms, vitamin D(3) and vitamin D(2). Vitamin D helps the body absorb calcium and promote optimal bone health. Both forms of vitamin D are metabolized to 25-hydroxyvitamin D in the body, and the levels of 25-hydroxyvitamin D(3) [25(OH)D(3)] and 25-hydroxyvitamin D(2) [25(OH)D(2)] in serum are considered the best indicators of vitamin D status. A candidate reference measurement procedure for serum 25(OH)D(3) and 25(OH)D(2) has been developed and critically evaluated. The deuterated compounds 25(OH)D(3)-d(3) and 25(OH)D(2)-d(3) are used as internal standards for 25(OH)D(3) and 25(OH)D(2), respectively. The 25(OH)D(3) and 25(OH)D(2) and their respective labeled internal standards are simultaneously extracted from serum using liquid-liquid extraction prior to reversed-phase liquid chromatography-tandem mass spectrometry (LC-MS/MS). Chromatographic separation was performed using a cyano (CN) column for both 25(OH)D(3) and 25(OH)D(2). Atmospheric pressure chemical ionization (APCI) in the positive ion mode and multiple reaction monitoring (MRM) were used for LC-MS/MS. The accuracy of the method was evaluated by recovery studies of measuring 25-hydroxyvitamin D [25(OH)D] in spiked samples with known 25(OH)D levels. The recoveries of the added 25(OH)D(3) and 25(OH)D(2) ranged from 99.0% to 101.0%. The absolute recoveries with this method were 97% and 92% for 25(OH)D(3) and 25(OH)D(2), respectively. Excellent precision was obtained with between-set coefficients of variation (CVs) of 0.2-0.6% for 25(OH)D levels >1 ng/g and within 2% for the level of <1 ng/g. Chromatographic separation of 25(OH)D(3) and 25(OH)D(2) from their respective isomers 3-epi-25(OH)D(3) and 3-epi-25(OH)D(2) was achieved. The limit of detection at a signal-to-noise ratio of approximately 3 was 40 pg of 25(OH)D on column (or approximately 0.15 ng/g as expressed as a concentration). This candidate reference measurement procedure for serum 25(OH)D(3) and 25(OH)D(2) demonstrates good accuracy and precision and low susceptibility to interferences. It can be used to provide an accuracy base to which clinical methods for 25(OH)D(3) and 25(OH)D(2) can be compared and that will serve as a standard of higher order for measurement traceability.


Analytical Chemistry | 2012

Development and Certification of a Standard Reference Material for Vitamin D Metabolites in Human Serum

Karen W. Phinney; Mary Bedner; Susan S.-C. Tai; Veronica Vamathevan; Lane C. Sander; Katherine E. Sharpless; Stephen A. Wise; James H. Yen; Rosemary L. Schleicher; Madhulika Chaudhary-Webb; Christine M. Pfeiffer; Joseph M. Betz; Paul M. Coates; Mary Frances Picciano

The National Institute of Standards and Technology (NIST), in collaboration with the National Institutes of Healths Office of Dietary Supplements (NIH-ODS), has developed a Standard Reference Material (SRM) for the determination of 25-hydroxyvitamin D [25(OH)D] in serum. SRM 972 Vitamin D in Human Serum consists of four serum pools with different levels of vitamin D metabolites and has certified and reference values for 25(OH)D(2), 25(OH)D(3), and 3-epi-25(OH)D(3). Value assignment of this SRM was accomplished using a combination of three isotope-dilution mass spectrometry approaches, with measurements performed at NIST and at the Centers for Disease Control and Prevention (CDC). Chromatographic resolution of the 3-epimer of 25(OH)D(3) proved to be essential for accurate determination of the metabolites.


Analytical Chemistry | 2013

Development of a Standard Reference Material for Metabolomics Research

Karen W. Phinney; Guillaume Ballihaut; Mary Bedner; Brandi S. Benford; Johanna E. Camara; Steven J. Christopher; W. Clay Davis; Nathan G. Dodder; Gauthier Eppe; Brian E. Lang; Stephen E. Long; Mark S. Lowenthal; Elizabeth A. McGaw; Karen E. Murphy; Bryant C. Nelson; Jocelyn L. Prendergast; Jessica L. Reiner; Catherine A. Rimmer; Lane C. Sander; Michele M. Schantz; Katherine E. Sharpless; Lorna T. Sniegoski; Susan S.-C. Tai; Jeanice M. Brown Thomas; Thomas W. Vetter; Michael J. Welch; Stephen A. Wise; Laura J. Wood; William F. Guthrie; Charles Hagwood

The National Institute of Standards and Technology (NIST), in collaboration with the National Institutes of Health (NIH), has developed a Standard Reference Material (SRM) to support technology development in metabolomics research. SRM 1950 Metabolites in Human Plasma is intended to have metabolite concentrations that are representative of those found in adult human plasma. The plasma used in the preparation of SRM 1950 was collected from both male and female donors, and donor ethnicity targets were selected based upon the ethnic makeup of the U.S. population. Metabolomics research is diverse in terms of both instrumentation and scientific goals. This SRM was designed to apply broadly to the field, not toward specific applications. Therefore, concentrations of approximately 100 analytes, including amino acids, fatty acids, trace elements, vitamins, hormones, selenoproteins, clinical markers, and perfluorinated compounds (PFCs), were determined. Value assignment measurements were performed by NIST and the Centers for Disease Control and Prevention (CDC). SRM 1950 is the first reference material developed specifically for metabolomics research.


Analytical and Bioanalytical Chemistry | 2008

Certification of standard reference materials containing bitter orange

Lane C. Sander; Karsten Putzbach; Bryant C. Nelson; Catherine A. Rimmer; Mary Bedner; J. Brown Thomas; Barbara J. Porter; Laura J. Wood; Michele M. Schantz; Karen E. Murphy; Katherine E. Sharpless; Stephen A. Wise; James H. Yen; P. H. Siitonen; R. L. Evans; A. Nguyen Pho; Mark Roman; Joseph M. Betz

A suite of three dietary supplement standard reference materials (SRMs) containing bitter orange has been developed, and the levels of five alkaloids and caffeine have been measured by multiple analytical methods. Synephrine, octopamine, tyramine, N-methyltyramine, hordenine, total alkaloids, and caffeine were determined by as many as six analytical methods, with measurements performed at the National Institute of Standards and Technology and at two collaborating laboratories. The methods offer substantial independence, with two types of extractions, two separation methods, and four detection methods. Excellent agreement was obtained among the measurements, with data reproducibility for most methods and analytes better than 5% relative standard deviation. The bitter-orange-containing dietary supplement SRMs are intended primarily for use as measurement controls and for use in the development and validation of analytical methods.


Clinica Chimica Acta | 2013

An assessment of 25-hydroxyvitamin D measurements in comparability studies conducted by the Vitamin D Metabolites Quality Assurance Program.

Mary Bedner; Katrice A. Lippa; Susan S.-C. Tai

BACKGROUND The National Institute of Standards and Technology (NIST), in collaboration with the National Institutes of Health Office of Dietary Supplements, established the first accuracy-based program for improving the comparability of vitamin D metabolite measurements, the Vitamin D Metabolites Quality Assurance Program. METHODS The study samples were human serum or plasma Standard Reference Materials (SRMs) with 25-hydroxyvitamin D values that were determined at NIST. Participants evaluated the materials using immunoassay (IA), liquid chromatography (LC) with mass spectrometric detection, and LC with ultraviolet absorbance detection. NIST evaluated the results for concordance within the participant community as well as trueness relative to the NIST value. RESULTS For the study materials that contain mostly 25-hydroxyvitamin D3 (25(OH)D3),the coefficient of variation (CV) for the participant results was consistently in the range from 7% to 19%, and the median values were biased high relative to the NIST values. However, for materials that contain significant concentrations of both 25-hydroxyvitamin D2 (25(OH)D2) and 25(OH)D3, the median IA results were biased lower than both the LC and the NIST values, and the CV was as high as 28%. The first interlaboratory comparison results for SRM 972a Vitamin D Metabolites in Human Serum are also reported. CONCLUSIONS Relatively large within-lab and between-lab variability hinders conclusive assessments of bias and accuracy.


Journal of Chromatography A | 2008

Development of liquid chromatographic methods for the determination of phytosterols in Standard Reference Materials containing saw palmetto.

Mary Bedner; Michele M. Schantz; Lane C. Sander; Katherine E. Sharpless

Liquid chromatographic (LC) methods using atmospheric pressure chemical ionization/mass spectrometric (APCI-MS) detection were developed for the separation and analysis of the phytosterols campesterol, cycloartenol, lupenone, lupeol, beta-sitosterol, and stigmasterol. Brassicasterol and cholesterol were also included for investigation as internal standards. The methods were used to identify and quantify the phytosterols in each of two Serenoa repens (saw palmetto) Standard Reference Materials (SRMs) developed by the National Institute of Standards and Technology (NIST). Values obtained by LC-MS were compared to those obtained using the more traditional approach of gas chromatography with flame ionization detection. This is the first reported use of LC-MS to determine phytosterols in saw palmetto dietary supplement materials.


Journal of Chromatography A | 2012

Development and Comparison of Three Liquid Chromatography-Atmospheric Pressure Chemical Ionization/Mass Spectrometry Methods for Determining Vitamin D Metabolites in Human Serum

Mary Bedner; Karen W. Phinney

Liquid chromatographic methods with atmospheric pressure chemical ionization mass spectrometry were developed for the determination of the vitamin D metabolites 25-hydroxyvitamin D₂ (25(OH)D₂), 25-hydroxyvitamin D₃ (25(OH)D₃), and 3-epi-25-hydroxyvitamin-D₃ (3-epi-25(OH)D₃) in the four Levels of SRM 972, Vitamin D in Human Serum. One method utilized a C18 column, which separates 25(OH)D₂ and 25(OH)D₃, and one method utilized a CN column that also resolves the diastereomers 25(OH)D₃ and 3-epi-25(OH)D₃. Both methods utilized stable isotope labeled internal standards for quantitation of 25(OH)D₂ and 25(OH)D₃. These methods were subsequently used to evaluate SRM 909c Human Serum, and 25(OH)D₃ was the only vitamin D metabolite detected in this material. However, SRM 909c samples contained matrix peaks that interfered with the determination of the [²H₆]-25(OH)D₃ peak area. The chromatographic conditions for the C18 column were modified to remove this interference, but conditions that separated the matrix peaks from [²H₆]-25(OH)D₃ on the CN column could not be identified. The alternate internal standard [²H₃]-25(OH)D₃ did not suffer from matrix interferences and was used for quantitation of 25(OH)D₃ in SRM 909c. During the evaluation of SRM 909c samples, a third method was developed using a pentafluorophenylpropyl column that also separates the diastereomers 25(OH)D₃ and 3-epi-25(OH)D₃. The 25(OH)D₃ was measured in SRM 909c using all three methods, and the results were compared.


Analytical and Bioanalytical Chemistry | 2008

Development of saw palmetto (Serenoa repens) fruit and extract standard reference materials

Michele M. Schantz; Mary Bedner; Stephen E. Long; John L. Molloy; Karen E. Murphy; Barbara J. Porter; Karsten Putzbach; Catherine A. Rimmer; Lane C. Sander; Katherine E. Sharpless; Jeanice M. Brown Thomas; Stephen A. Wise; Laura J. Wood; James H. Yen; Takashi Yarita; Agnes Nguyenpho; Wendy R. Sorenson; Joseph M. Betz

As part of a collaboration with the National Institutes of Health’s Office of Dietary Supplements and the Food and Drug Administration’s Center for Drug Evaluation and Research, the National Institute of Standards and Technology has developed two standard reference materials (SRMs) representing different forms of saw palmetto (Serenoa repens), SRM 3250 Serenoa repens fruit and SRM 3251 Serenoa repens extract. Both of these SRMs have been characterized for their fatty acid and phytosterol content. The fatty acid concentration values are based on results from gas chromatography with flame ionization detection (GC-FID) and mass spectrometry (GC/MS) analysis while the sterol concentration values are based on results from GC-FID and liquid chromatography with mass spectrometry analysis. In addition, SRM 3250 has been characterized for lead content, and SRM 3251 has been characterized for the content of β-carotene and tocopherols. SRM 3250 (fruit) has certified concentration values for three phytosterols, 14 fatty acids as triglycerides, and lead along with reference concentration values for four fatty acids as triglycerides and 16 free fatty acids. SRM 3251 (extract) has certified concentration values for three phytosterols, 17 fatty acids as triglycerides, β-carotene, and γ-tocopherol along with reference concentration values for three fatty acids as triglycerides, 17 fatty acids as free fatty acids, β-carotene isomers, and δ-tocopherol and information values for two phytosterols. These SRMs will complement other reference materials currently available with concentrations for similar analytes and are part of a series of SRMs being developed for dietary supplements.


Analytical and Bioanalytical Chemistry | 2012

Development and certification of green tea-containing standard reference materials.

Lane C. Sander; Mary Bedner; M. C. Tims; James H. Yen; David L. Duewer; Barbara J. Porter; Steven J. Christopher; Russell D. Day; Stephen E. Long; John L. Molloy; Karen E. Murphy; Brian E. Lang; R. Lieberman; Laura J. Wood; M. J. Payne; Mark Roman; Joseph M. Betz; A. NguyenPho; Katherine E. Sharpless; Stephen A. Wise

AbstractA suite of three green tea-containing Standard Reference Materials (SRMs) has been issued by the National Institute of Standards and Technology (NIST): SRM 3254 Camellia sinensis (Green Tea) Leaves, SRM 3255 Camellia sinensis (Green Tea) Extract, and SRM 3256 Green Tea-Containing Solid Oral Dosage Form. The materials are characterized for catechins, xanthine alkaloids, theanine, and toxic elements. As many as five methods were used in assigning certified and reference values to the constituents, with measurements carried out at NIST and at collaborating laboratories. The materials are intended for use in the development and validation of new analytical methods, and for use as control materials as a component in the support of claims of metrological traceability. FigureGreen Tea - Camellia sinensis


Analytical Chemistry | 2011

Dynamic calibration approach for determining catechins and gallic acid in green tea using LC-ESI/MS.

Mary Bedner; David L. Duewer

Catechins and gallic acid are antioxidant constituents of Camellia sinensis, or green tea. Liquid chromatography with both ultraviolet (UV) absorbance and electrospray ionization mass spectrometric (ESI/MS) detection was used to determine catechins and gallic acid in three green tea matrix materials that are commonly used as dietary supplements. The results from both detection modes were evaluated with 14 quantitation models, all of which were based on the analyte response relative to an internal standard. Half of the models were static, where quantitation was achieved with calibration factors that were constant over an analysis set. The other half were dynamic, with calibration factors calculated from interpolated response factor data at each time a sample was injected to correct for potential variations in analyte response over time. For all analytes, the relatively nonselective UV responses were found to be very stable over time and independent of the calibrant concentration; comparable results with low variability were obtained regardless of the quantitation model used. Conversely, the highly selective MS responses were found to vary both with time and as a function of the calibrant concentration. A dynamic quantitation model based on polynomial data-fitting was used to reduce the variability in the quantitative results using the MS data.

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Lane C. Sander

National Institute of Standards and Technology

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Stephen A. Wise

National Institute of Standards and Technology

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Katherine E. Sharpless

National Institute of Standards and Technology

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Joseph M. Betz

National Institutes of Health

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James H. Yen

National Institute of Standards and Technology

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Karen W. Phinney

National Institute of Standards and Technology

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Katrice A. Lippa

National Institute of Standards and Technology

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Susan S.-C. Tai

National Institute of Standards and Technology

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David L. Duewer

National Institute of Standards and Technology

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Brian E. Lang

National Institute of Standards and Technology

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