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Simple, rapid, and cost-effective microextraction by the packed sorbent method for quantifying of urinary free catecholamines and metanephrines using liquid chromatography-tandem mass spectrometry and its application in clinical analysis

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Abstract

Pheochromocytoma and paraganglioma (PPGL) are rare neuroendocrine tumors arising from adrenal and extra-adrenal chromaffin cells. They produce excessive amounts of catecholamines and their metabolites. A newly analytical procedure based on the semi-automated microextraction by packed sorbent (MEPS) technique, using a digitally controlled syringe (eVol) combined with liquid chromatography-tandem mass spectrometry (LC-MS/MS), was developed to quantify free urinary catecholamines and metanephrines. The important parameters affecting MEPS performance, namely the type of sorbent material (porous graphitized carbon (PGC), polar enhanced polymer (PEP), cation-exchange (CX) and C18), number of extraction cycles, and elution solvent system, were evaluated. The optimal experimental conditions involved the loading of sample mixture in seven extraction cycles through a C18 sorbent in a MEPS syringe, followed by using elution solutions (water/acetonitrile/formic acid, 95/4.75/0.25). The entire sample preparation took about 4 min. Chromatographic separation was well achieved with an HSS PFP column using the gradient elution. The linearity range of the method was 0.167–33.4 ng/mL for epinephrine, 0.650–130 ng/mL for norepinephrine, 1.53–306 ng/mL for dopamine, 1.34–268 ng/mL for metanephrine, 3.43–686 ng/mL for normetanephrine, and 1.33–265 ng/mL for 3-methoxytyramine. The intra- and interassay precisions were ≤ 12.8%, and the respective accuracies were 88.4–112.0% and 89.0–109.5%. The carryover and sample stability without acidification were also investigated. Validation using clinical urine specimens showed that the proposed method had higher sensitivity compared with other urinary biochemical tests. The developed MEPS-LC-MS/MS method was simple, fast, and cost-effective; it helped to obtain information about multiple metabolites. It is applicable in routine clinical laboratories for the screening of PPGL.

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Abbreviations

APS:

Amino-propyl silane

CX:

Cation-exchange

DA:

Dopamine

DPB:

2-Aminoethyldiphenylborinate

EPI:

Epinephrine

ESI:

Electrospray ionization

IS:

Internal standard

LOQ:

Limit of quantification

LOD:

Limit of detection

ME:

Matrix effect

MEPS:

Microextraction by packed sorbent

MN:

Metanephrine

MRM:

Multiple reaction monitoring

NE:

Norepinephrine

NMN:

Normetanephrine

PPGL:

Pheochromocytoma and paraganglioma

PGC:

Porous graphitized carbon

PEP:

Polar enhanced polymer

QC:

Quality control

SPE:

Solid phase extraction

SD:

Standard deviation

3-MT:

3-Methoxytyramine

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Funding

This study received financial support from the National Natural Science Foundation of China NSFC-81672107 and NSFC-21405007.

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Authors and Affiliations

  1. Department of Pharmacy, Peking University Third Hospital, Beijing, 100191, China

    Xin Xiong & Yuanyuan Zhang

  2. Therapeutic Drug Monitoring and Clinical Toxicology Center of Peking University, Beijing, 100191, China

    Xin Xiong & Yuanyuan Zhang

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  1. Xin Xiong
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  2. Yuanyuan Zhang
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Correspondence to Xin Xiong.

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The ethical committee of Peking University Third Hospital approved the study.

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Xiong, X., Zhang, Y. Simple, rapid, and cost-effective microextraction by the packed sorbent method for quantifying of urinary free catecholamines and metanephrines using liquid chromatography-tandem mass spectrometry and its application in clinical analysis. Anal Bioanal Chem 412, 2763–2775 (2020). https://doi.org/10.1007/s00216-020-02436-8

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