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An isotope dilution liquid chromatography-tandem mass spectrometry candidate reference measurement procedure for aldosterone measurement in human plasma

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Abstract

Accurate quantitation of aldosterone is clinically important in standardized testing for primary aldosteronism. The results are often variable when performed by clinical immunoassays. To standardize and ensure the accuracy of clinical systems, reference measurement procedures (RMPs) with higher metrological order are required. A simple and reliable isotope dilution LC-IDMS/MS-based measurement procedure for human plasma aldosterone has been developed. This method involved plasma spiked with a deuterium-labelled internal standard, equilibrated for 0.5 h, and extracted by liquid-liquid extraction (LLE) without derivatization. Aldosterone and its structural analogues were baseline separated with a C18-packed UHPLC column with gradient elution within 7 min. The signal intensity variability and measurement imprecision were reduced by bracketing calibration during plasma aldosterone value assignment. The limit of detection (LoD) was 19.4 pmol/L with a signal-to-noise ratio (S/N) > 3. The lowest limit of quantification (LLoQ) was 27.7 pmol/L (S/N > 10 and CV < 10.0%). LLE was performed with 1 mL of n-hexane/ethyl acetate (3:2, v/v), and the extraction recovery was determined to be 92.15 ± 3.54%. The imprecisions were ≤ 3.18% for samples at 124.8, 867.0, and 2628.5 pmol/L. The recoveries were 98.11–101.61%. The relative bias between this candidate RMP and the established RMP was 2.76–1.89%. The linearity response ranged from 27.7 to 2774.4 pmol/L with R2 = 0.999. The method performance met the requirements of RMPs (≤ 5% total CV and ≤ 3% bias). Furthermore, the developed method was applied to evaluate immunoassays through 41 patient sample comparisons. The calibration and measurement capability (CMC) of this method were also evaluated by measuring these samples. The candidate RMP can serve as an accurate reference baseline for routine methods and can be used for value assignment for reference materials.

Graphical abstract

Selected ion chromatograms by LC-MS/MS using a C18 column for aldosterone and its structural analogues.

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Abbreviations

LC-MS/MS:

Liquid chromatography/tandem mass spectrometry

RMP:

Reference measurement procedure

LoD:

Limit of detection

LLoQ:

Lowest limit of quantification

EQA:

External quality assessment

JCTLM:

Joint Committee for Traceability in Laboratory Medicine

IFCC:

International Federation of Clinical Chemistry and Laboratory Medicine

RELA:

IFCC External Quality assessment scheme for Reference Laboratories in Laboratory Medicine

CMC:

Calibration and measurement capability

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Funding

Funding was provided by The Natural Science Foundation of Guangdong Province (2020A1515010668), the Specific Research Fund for TCM Science and Technology of Guangdong Provincial Hospital of Chinese Medicine (YN2016QJ06, YN2019QL01, and YN2019 MJ04), and Administration of Traditional Chinese Medicine Bureau of Guangdong Province (20211150).

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Authors and Affiliations

  1. Department of Laboratory Medicine, The Second Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou, 510120, Guangdong, China

    Qiaoxuan Zhang, Liqiao Han, Fen Ouyang, Xiaobin Wu, Jun Yan, Min Zhan, Peifeng Ke, Junhua Zhuang & Xianzhang Huang

  2. Second Clinical Medical College, Guangzhou University of Chinese Medicine, Guangzhou, 510120, Guangdong, China

    Qiaoxuan Zhang

  3. Fujian Huayin Medical Laboratory Center, Xiamen, 361101, Fujian, China

    Songbai Zheng

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  1. Qiaoxuan Zhang
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  2. Liqiao Han
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  4. Fen Ouyang
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  5. Xiaobin Wu
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  6. Jun Yan
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  7. Min Zhan
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  10. Xianzhang Huang
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Corresponding authors

Correspondence to Junhua Zhuang or Xianzhang Huang.

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Research involving human participants and/or animals

The research involved plasma samples taken from pregnant women and this study is approved by Ethics Committee of Second Affiliated Hospital of Guangzhou University of Chinese Medicine. The number of this Review is B2016-157-01.

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The ethics committee approved the exemption of informed consent.

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Zhang, Q., Han, L., Zheng, S. et al. An isotope dilution liquid chromatography-tandem mass spectrometry candidate reference measurement procedure for aldosterone measurement in human plasma. Anal Bioanal Chem 413, 4471–4481 (2021). https://doi.org/10.1007/s00216-021-03405-5

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  • DOI: https://doi.org/10.1007/s00216-021-03405-5

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