Candidate Reference Measurement Procedures for Serum 25-Hydroxyvitamin D3 and 25-Hydroxyvitamin D2 by Using Isotope-Dilution Liquid Chromatography–Tandem Mass Spectrometry

Stepman, Hedwig; Vanderroost, An; Uytfanghe, Katleen Van; Thienpont, Linda M.

doi:10.1373/clinchem.2010.152553

Cited by 179 publications

(135 citation statements)

References 27 publications

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“…LC-MS/MS-based reference methods [10] usually involve stable isotope labeled compounds for internal standardization. Since vancomycin is a bio-product, production of a respective material is hardly possible.…”

Section: Introductionmentioning

confidence: 99%

Quantification of vancomycin in human serum by LC-MS/MS

König¹,

Kobold²,

Fink³

et al. 2013

Clinical Chemistry and Laboratory Medicine

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Background: The aim of our work was to develop and validate a reliable LC-MS/MS-based measurement procedure for the quantification of vancomycin in serum, to be applied in the context of efforts to standardize and harmonize therapeutic drug monitoring of this compound using routine assays. Methods: Sample preparation was based on protein precipitation followed by ultrafiltration. In order to minimize differential modulation of ionization by matrix constituents extended chromatographic separation was applied leading to a retention time of 9.8 min for the analyte. Measurement was done by HPLC-ESI-MS/MS. For internal standardization the derivative vancomycin-glycin (ISTD) prepared by chemical synthesis was used, HPLC conditions ensured coelution of ISTD with the analyte. Results: In a bi-center validation total CVs of < 4% were observed for quality control material ranging from 5.3 mg/L to 79.4 mg/L; accuracy was ± 4%. No relevant ion suppression was observed. Comparative measurement of aliquots from 70 samples at the two validation sites demonstrated close agreement. Conclusions: Employing a closely related homologue molecule for internal standardization and the use of MS/ MS following highly efficient sample pre-fractionation by HPLC, the method described here can be considered to offer the highest level of analytical reliability realized so far for the quantification of vancomycin in human serum. Thus, the method is suitable to be used in a comprehensive reference measurement system for vancomycin.

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Section: Introductionmentioning

confidence: 99%

Quantification of vancomycin in human serum by LC-MS/MS

König¹,

Kobold²,

Fink³

et al. 2013

Clinical Chemistry and Laboratory Medicine

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“…This study adds to a growing literature [9][10][11][12][13][14][15][16][17][22][23][24][25][26][27][28] [13,15,23]. In the accuracy performance testing using Labquality reference serum panel, both Roche and Abbott displayed a similar bias at higher 25(OH)D concentrations as with study serum samples.…”

Section: Discussionmentioning

confidence: 65%

Analytical and clinical performance of the new Fujirebio 25-OH vitamin D assay, a comparison with liquid chromatography-tandem mass spectrometry (LC-MS/MS) and three other automated assays

Saleh

Müller

Eckardstein

2016

Clinical Chemistry and Laboratory Medicine (CCLM)

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Results: Intra-and inter-day imprecision of the Fujirebio 25(OH)D assay was < 5%. Fujirebio 25(OH)D assay showed the highest correlation among the assays tested with the LC-MS/MS method (R = 0.986). The mean relative bias obtained was -15.6% (Fujirebio), -12.7% (Beckman), -2.1% (Abbott) and 9.7% (Roche) as compared to LC-MS/ MS. Comparison with the Labquality certified reference serum panel yielded a mean bias of -11.8% (Fujirebio), -14.1% (Beckman), 4.4% (Abbott) and 3.2% (Roche), respectively. Compared to LC-MS/MS, the sensitivity of different methods in detecting vitamin D insufficiency ( < 50 nmol/L) varied from 100% for the Fujirebio assay to 72.7% for Roche, and specificity ranged from 94.4% for Roche to 87.6% for Beckman.

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“…The main limitation of immunochemical methods is the cross reactivity of the antibodies used in the assay. Another restriction is related to their inability to discriminate the D 2 and D 3 forms of the 25OH metabolite [6]. The chromatographic methods are too sophisticated and require high-qualified laboratory staff.…”

Section: Introductionmentioning

confidence: 99%

Untitled

2014

Integr Food Nutr Metab

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Twenty five-hydroxy-vitamin D (25OHD) levels are accepted as an indicator for vitamin D status and their measurement is still a challenge. The aim of the present study was to develop a robust and easy to use HPLC-method for simultaneous analysis of 25OHD 2 and 25OHD 3 in human plasma and to assess its analytical performance, Protein precipitation followed by liquid-liquid extraction with n-hexane was used for separation from the matrix substances. A HPLC-system with UV detector (Thermo Scientific Spectra System, USA) was used. HPLC-UV analysis was performed on a Synergy hydro-RP column 4,6x250 mm, 4.0 µm particle size with mobile phase consisting of methanol/acetonitrile/water (70/25/5) under isocratic elution at a flow rate of 1.2 mL/min and UV-detection at 265 nm. Our HPLC-method was compared with a validated ID-LC-MS/MS method and with a commercial immunochemical method. Linear regression was used for data analysis. Linearity of the HPLC-assay was between 5-160 ng/ml for each analyte (r 2 =0.99) with lower limit of detection 4 nmol/L, and limit of quantification 10 nmol/L. Intra-assay precision was determined by extracting and quantifying 10 replicates of 4 QC levels (RSD ≤ 9%). Inter-assay precision was determined over 21 consecutive days analyzing 10 replicates of all QC samples (RSD<10%). All results were within 14.2% deviation of the 25OHD LC-MS/MS method mean. Method comparison demonstrated good agreement between HPLC and LC-MS/MS (r=0.85) and disparity with the commercial immunoassay (r=0.48). This assay demonstrates excellent linearity, acceptable accuracy and precision, a good agreement with a validated ID-LC-MS/MS method. The simple sample preparation and ease of determination make it useful for the routine clinical laboratory.

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Candidate Reference Measurement Procedures for Serum 25-Hydroxyvitamin D3 and 25-Hydroxyvitamin D2 by Using Isotope-Dilution Liquid Chromatography–Tandem Mass Spectrometry

Cited by 179 publications

References 27 publications

Quantification of vancomycin in human serum by LC-MS/MS

Quantification of vancomycin in human serum by LC-MS/MS

Analytical and clinical performance of the new Fujirebio 25-OH vitamin D assay, a comparison with liquid chromatography-tandem mass spectrometry (LC-MS/MS) and three other automated assays

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