Dried Blood Spot Testing for Seven Steroids Using Liquid Chromatography-Tandem Mass Spectrometry With Reference Interval Determination in the Korean Population

Kim, Borahm; Lee, Mi Na; Park, Hyung-Doo; Kim, Jong Won; Chang, Yun Sil; Park, Won Soon; Lee, Soo-Youn

doi:10.3343/alm.2015.35.6.578

Cited by 25 publications

(27 citation statements)

References 21 publications

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“…Of the 1,107 metabolites, 65% of the features with MSMS spectral data ( n = 176) were annotated by at least compound class, which included phospholipids, hormones and steroids, and saturated and unsaturated fatty acids. Steroid concentrations in newborns can be as low as 10 ng/mL (Kim et al 2015), while polyunsaturated fatty acid concentrations (PUFA) in cord blood can be as high as 1 mg/mL (Niinivirta et al 2011), highlighting the dynamic range of the method.…”

Section: Resultsmentioning

confidence: 99%

An untargeted metabolomics method for archived newborn dried blood spots in epidemiologic studies

et al. 2017

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Introduction For pediatric diseases like childhood leukemia, a short latency period points to in-utero exposures as potentially important risk factors. Untargeted metabolomics of small molecules in archived newborn dried blood spots (DBS) offers an avenue for discovering early-life exposures that contribute to disease risks. Objectives The purpose of this study was to develop a quantitative method for untargeted analysis of archived newborn DBS for use in an epidemiological study (California Childhood Leukemia Study, CCLS). Methods Using experimental DBS from the blood of an adult volunteer, we optimized extraction of small molecules and integrated measurement of potassium as a proxy for blood hematocrit. We then applied this extraction method to 4.7-mm punches from 106 control DBS samples from the CCLS. Sample extracts were analyzed with liquid chromatography high resolution mass spectrometry (LC-HRMS) and an untargeted workflow was used to screen for metabolites that discriminate population characteristics such as sex, ethnicity, and birth weight. Results Thousands of small molecules were measured in extracts of archived DBS. Normalizing for potassium levels removed variability related to varying hematocrit across DBS punches. Of the roughly 1,000 prevalent small molecules that were tested, multivariate linear regression detected significant associations with ethnicity (3 metabolites) and birth weight (15 metabolites) after adjusting for multiple testing. Conclusions This untargeted workflow can be used for analysis of small molecules in archived DBS to discover novel biomarkers, to provide insights into the initiation and progression of diseases, and to provide guidance for disease prevention.

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Section: Resultsmentioning

confidence: 99%

An untargeted metabolomics method for archived newborn dried blood spots in epidemiologic studies

et al. 2017

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“…LC-MS/MS is the most accurate method currently available for measuring small molecules, and it can measure multiple steroid hormones simultaneously [6]. Steroid profiling with LC-MS/MS allows for evaluation of the status of enzymes involved in adrenal steroid biosynthetic pathways; thus, it is a better diagnostic tool than the evaluation of a single steroid [6]. A specific LC-MS/MS method with simultaneous detection of multiple steroids has been introduced to minimize unnecessary follow-up analyses [36].…”

Section: Introductionmentioning

confidence: 99%

“…Steroid profiling with LC-MS/MS allows for evaluation of the status of enzymes involved in adrenal steroid biosynthetic pathways; thus, it is a better diagnostic tool than the evaluation of a single steroid [6]. A specific LC-MS/MS method with simultaneous detection of multiple steroids has been introduced to minimize unnecessary follow-up analyses [36]. Although previous studies examined multiple steroid hormonal analyses using LC-MS/MS in Korea, they included limited numbers of steroids [67] or did not evaluate clinical screening for CAH using patient samples [6].…”

Section: Introductionmentioning

confidence: 99%

“…A specific LC-MS/MS method with simultaneous detection of multiple steroids has been introduced to minimize unnecessary follow-up analyses [36]. Although previous studies examined multiple steroid hormonal analyses using LC-MS/MS in Korea, they included limited numbers of steroids [67] or did not evaluate clinical screening for CAH using patient samples [6]. Testosterone and progesterone, steroid hormones useful for the screening and diagnosis of CAH, have not been studied using dried blood spots (DBS) in Korean neonates [67].…”

Section: Introductionmentioning

confidence: 99%

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Dried Blood Spot Multiplexed Steroid Profiling Using Liquid Chromatography Tandem Mass Spectrometry in Korean Neonates

et al. 2019

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Background Screening for congenital adrenal hyperplasia (CAH) using immunoassays for 17α-hydroxyprogesterone generates many false-positive results. We developed and validated a liquid chromatography-tandem mass spectrometry (LC-MS/MS) assay for simultaneous quantification of nine steroid hormones in dried blood spot (DBS) samples, and established reference intervals for these hormones. Methods We examined our method for linearity, precision, accuracy, extraction recovery, and matrix effects and determined the reference intervals of cortisol, 17α-hydroxyproges-terone, 11-deoxycortisol, 21-deoxycortisol, androstenedione, corticosterone, 11-deoxycorticosterone, testosterone, and progesterone in 1,146 DBS samples (from 272 preterm and 874 full-term neonates). Immunoassay and LC-MS/MS methods were compared for 17α-hydroxyprogesterone. Fourteen additional samples were tested to validate the clinical applicability of the LC-MS/MS method. Results The linearity range was 2.8–828.0 nmol/L for cortisol and 0.9–40.0 nmol/L for the other steroids (R 2 >0.99). Intra-day and inter-day precision CVs were 2.52–12.26% and 3.53–17.12%, respectively. Accuracy was 80.81–99.94%, and extraction recovery and matrix effects were 88.0–125.4% and 61.7–74.2%, respectively. There was a negative bias, with higher values measured by immunoassay compared with LC-MS/MS ( r =0.8104, P <0.0001). The LC-MS/MS method was successfully applied to the analysis of nine steroids in DBS for screening and diagnosis of CAH using the 14 additional samples. Conclusions Our method enables highly sensitive and specific assessment of nine steroids from DBS and is a promising tool for clinical analysis of CAH.

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“…Standard whole-blood with six drug concentrations (streptomycin, kanamycin, cycloserine, and PAS at 0, 5, 10, 20, 40, and 100 µg/mL; clarithromycin and prothionamide at 0, 0.5, 1, 2, 4, and 10 µg/mL; and moxifloxacin, levofloxacin, and linezolid at 0, 1, 2, 4, 8, and 20 µg/mL) were prepared by mixing equal amounts of standard plasma and blank packed red blood cells. For standard DBSs, 100 µL of six standard whole-blood was added dropwise onto Whatman 903 filter paper by using an adjustable pipette [ 10 11 ]. After samples were dried for 12 hr at room temperature, they were stored in a sealed bag at -20℃.…”

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confidence: 99%

Multiplex Assay of Second-Line Anti-Tuberculosis Drugs in Dried Blood Spots Using Ultra-Performance Liquid Chromatography-Tandem Mass Spectrometry

et al. 2016

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As dried blood spots (DBSs) have various advantages over conventional venous blood sampling, some assays for detection of one or two anti-tuberculosis (TB) drugs in DBSs have been developed. However, there are no assays currently available for the simultaneous measurement of three or more anti-TB drugs in DBSs. In this study, we developed and evaluated a multiplex method for detecting nine anti-TB drugs including streptomycin, kanamycin, clarithromycin, cycloserine, moxifloxacin, levofloxacin, para-aminosalicylic acid, prothionamide, and linezolid in DBSs by using ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). Seventy-nine patient samples of DBS were analyzed on the UPLC-MS/MS system. All drug concentrations were determined within 4 min, and assay performance was evaluated. All drugs were clearly separated without ion suppression. Within-run and between-run precisions were 1.7-13.0% and 5.7-17.0%, respectively, at concentrations representing low and high levels for the nine drugs. Lower limits of detection and quantification were 0.06-0.6 and 0.5-5.0 µg/mL, respectively. Linearity was acceptable at five level concentrations for each drug. Correlations between drug concentrations in plasma and DBSs by using Passing-Bablock regression and Pearson's rho (ρ, 0.798-0.989) were acceptable. In conclusion, we developed a multiplex assay to measure nine second-line anti-TB drugs in DBSs successfully. This assay provided convenient and rapid drug quantification and could have applications in drug monitoring during treatment.

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Dried Blood Spot Testing for Seven Steroids Using Liquid Chromatography-Tandem Mass Spectrometry With Reference Interval Determination in the Korean Population

Cited by 25 publications

References 21 publications

An untargeted metabolomics method for archived newborn dried blood spots in epidemiologic studies

An untargeted metabolomics method for archived newborn dried blood spots in epidemiologic studies

Dried Blood Spot Multiplexed Steroid Profiling Using Liquid Chromatography Tandem Mass Spectrometry in Korean Neonates

Multiplex Assay of Second-Line Anti-Tuberculosis Drugs in Dried Blood Spots Using Ultra-Performance Liquid Chromatography-Tandem Mass Spectrometry

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