Journal Article

Simultaneous extraction and determination of monoamine neurotransmitters in human urine for clinical routine testing based on a dual functional solid phase extraction assisted by phenylboronic acid coupled with liquid chromatography-tandem mass spectrometry

Li, Shu, Kellermann, Gottfried, Li, Xiaoguang (Sunny)

2017

Overview

The major monoamine neurotransmitters, serotonin (5-HT) and catecholamines (i.e., norepinephrine (NE), epinephrine (E), and dopamine (DA)), are critical to the nervous system function, and imbalances of the neurotransmitters have been connected to a variety of diseases, making their measurement useful in a clinical setting. A simple, rapid, robust, sensitive, and specific LC-MS/MS method has been developed and validated for the simultaneous quantitation of urinary serotonin and catecholamines with low cost, which is ideal for routine clinical applications. A simple extraction from complex urine was accomplished using tailored solid phase extraction incorporating phenylboronic acid complexation on a 96-well HLB microplate for the sample extraction and resulted in significantly improved throughput, selectivity, and extraction recovery. Compared to 1–10 mL of urine typically used, this method required only 10 μL. A rapid chromatographic elution with a total cycle time of 6 min per sample compared to reported run times of 19–75 min was achieved on a PFP column. The sensitivity of l and 2 ng mL −1 for the detection of low abundant E and NE combined with the high coverage of 1024 ng mL −1 for DA enabled the multi-analyte detection of these biogenic amines in a single run. Good linearity (2.0–512, 1.0–512, 4.0–1024, and 4.0–1024 ng mL −1 for NE, E, DA, and 5-HT, respectively), accuracy (87.6–104.0%), precision (≤8.0%), extraction recovery (69.6–103.7%), and matrix effect (87.1–113.1% for catecholamines and 63.6–71.4% for 5-HT) were obtained. No autosampler carryover was observed. The analytes were stable for 5 days at 20 °C, 14 days at 4 °C, and 30 days at −20 °C and five freeze–thaw cycles. The easy sample preparation, rapid LC, and multi-analyte MS detection allow two 96-well plates of samples to be extracted within 2 h and analyzed on an LC-MS/MS system within 24 h. The applicability and reliability of the assay were demonstrated by assessment of the reference interval for authentic urine specimens from 90 healthy individuals. Graphical abstract A simple, rapid, robust, sensitive and specific LC-MS/MS method combined with a dual functional solid phase extraction has been developed and validated for the simultaneous extraction and quantitation of monoamine neurotransmitters in human urine with low cost

• Publisher: Springer Berlin Heidelberg,Springer,Springer Nature B.V
• Subject: Adolescent / Adult / Aged / Amines / Analytical Chemistry / Biochemistry / Biogenic amines / Biogenic Monoamines - chemistry / Biogenic Monoamines - urine / Boronic Acids - chemistry / Boronic Acids - urine / Catecholamine / Catecholamines / Characterization and Evaluation of Materials / Chemical properties / Chemistry / Chemistry and Materials Science / Chromatography / Chromatography, Liquid / Complex Mixtures - urine / Cycle time / Dopamine / Elution / Epinephrine / Extraction / Extraction (Chemistry) / Female / Food Science / freeze-thaw cycles / Freeze-thawing / Human wastes / Humans / Laboratory Medicine / Linearity / Liquid chromatography / Low cost / Male / Mass Spectrometry / Mass spectroscopy / Measurement / Methods / Middle Aged / Monitoring/Environmental Analysis / Monoamines / Nervous system / Neurotransmitter Agents - urine / Neurotransmitters / Norepinephrine / Quantitation / Recovery / Reliability analysis / Reproducibility of Results / Research Paper / Routines / Sample preparation / Selectivity / Sensitivity and Specificity / Serotonin / Solid Phase Extraction / Solid phases / tandem mass spectrometry / Therapeutic applications / Urinalysis / Urinalysis - methods / Urine / Young Adult