Leveraging supported liquid extraction and automated workflow for high efficiency and high quality clinical LC-MS/MS method for quantification of urinary 5-hydroxyindoleacetic acid (5-HIAA), vanillylmandelic acid (VMA) and homovanillic acid (HVA)
Difei Sun , Dawn-Marie Murphy McLean , Jan Palaty , Danijela Konforte
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Abstract
Background and Aims
5-hydroxyindoleacetic acid (5-HIAA), vanillylmandelic acid (VMA) and homovanillic acid (HVA) are neurotransmitter metabolites used for the diagnosis and monitoring of neuroendocrine tumors. This study’s objective was to apply automation to improve efficiency and quality of a liquid chromatography tandem mass spectrometry (LC-MS/MS) procedure for quantification of 5-HIAA, VMA and HVA in human urine.
Methods
5-HIAA, VMA and HVA in urine samples were extracted with automated supported liquid extraction (SLE) and quantified by LC-MS/MS utilizing scheduled multiple reaction monitoring (MRM). An in-house developed middleware was used to automate data analysis and reporting.
Results
The average recovery for 5-HIAA, VMA and HVA in different types of urine matrix varied between 92.5 % and 99.0 %, while the average matrix factor varied between 98.6 % and 103.4 %. The results of other validation studies: stability, sensitivity, analytical measurement range, selectivity, accuracy, precision, dilution verification, carryover, and method comparison all met the performance criteria.
Conclusions
Automated SLE is a suitable sample preparation technique for quantification of 5-HIAA, VMA and HVA in human urine. Workflow automation described in this study resulted in 54 % of overall time savings and 80 % of hands-on time savings compared to all manual approach.
期刊介绍：
The Official Journal of the International Federation of Clinical Chemistry and Laboratory Medicine (IFCC)
Clinica Chimica Acta is a high-quality journal which publishes original Research Communications in the field of clinical chemistry and laboratory medicine, defined as the diagnostic application of chemistry, biochemistry, immunochemistry, biochemical aspects of hematology, toxicology, and molecular biology to the study of human disease in body fluids and cells.
The objective of the journal is to publish novel information leading to a better understanding of biological mechanisms of human diseases, their prevention, diagnosis, and patient management. Reports of an applied clinical character are also welcome. Papers concerned with normal metabolic processes or with constituents of normal cells or body fluids, such as reports of experimental or clinical studies in animals, are only considered when they are clearly and directly relevant to human disease. Evaluation of commercial products have a low priority for publication, unless they are novel or represent a technological breakthrough. Studies dealing with effects of drugs and natural products and studies dealing with the redox status in various diseases are not within the journal''s scope. Development and evaluation of novel analytical methodologies where applicable to diagnostic clinical chemistry and laboratory medicine, including point-of-care testing, and topics on laboratory management and informatics will also be considered. Studies focused on emerging diagnostic technologies and (big) data analysis procedures including digitalization, mobile Health, and artificial Intelligence applied to Laboratory Medicine are also of interest.
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