To better detect cardiovascular diseases in people, researchers used high-performance liquid chromatography–tandem mass spectrometry (HPLC–MS/MS) to simultaneously quantify metabolites in human plasma.
Scientists from Sechenov First Moscow State Medical University tested a high-performance liquid chromatography–tandem mass spectrometry (HPLC–MS/MS) assay for cardiovascular diseases. Their findings were published in the Journal of Chromatography B (1).
Metabolomes represent sets of low molecular weight compounds that reflect the physiological and pathological state of the body. Compared to genes, metabolomic biomarkers more rapidly reflect the influence of external and internal factors on the body and may serve as novel indicators of the phenotype changes. Metabolomic approaches may be useful in personalized medicine, offering new diagnostic and prognostic markers. As a consequence, identifying associations between changes in metabolites and (patho)physiological processes is a significant challenge, as metabolites are often involved in numerous biochemical reactions and metabolic pathways. Metabolites may function as either end products or substrates of metabolic processes that occur within the context of several competitive or non-competitive biochemical reactions.
Significant discoveries have been made in the past few decades regarding relations between myocardial metabolism and cardiac activity regulation. Given that the heart’s function depends on essential compounds being transferred through the body, metabolic dysfunction represents a primary etiological factor in a range of cardiovascular pathologies. These factors include imbalances in general metabolism, abnormalities in signaling pathways, or changes in the biosynthesis and degradation. The metabolomic profile of blood plasma directly reflects the state of the heart and blood vessels.
Deeper understanding of metabolic alterations in the body may facilitate more profound grasps of the fundamental mechanisms that occur during pathophysiological processes. Further, the obtained data may be utilized to distinguish the initial stages of cardiovascular disease.
Cardiovascular diseases are a series of disorders, such as coronary heart disease or congenital heart disease, that affect the heart and blood vessels (2). Diseases in this category are the leading cause of death globally; in 2022, an estimated 19.8 million people died from cardiovascular diseases, representing approximately 32% of all global deaths. Of these deaths, 85% were due to heart attacks or strokes. It is important to detect cardiovascular diseases as early as possible, so that proper management can begin.
In this study, the researchers optimized and validated a high-throughput high-performance liquid chromatography–tandem mass spectrometry (HPLC–MS/MS) for simultaneous quantification of 98 metabolites in human plasma. This includes amino acids and its derivatives, tryptophan pathway metabolites, nucleosides, water-soluble vitamins, acylcarnitines, and others. This method utilizes chemical derivatization to enhance retention and sensitivity of polar metabolites, providing accurate analysis across diverse physicochemical properties.
The presented method was validated in accordance with European Medicines Agency (EMA) guidelines and included assessment of linearity, accuracy, precision, matrix effects, recovery, and stability. Parallelism testing confirmed the suitability of a surrogate matrix for calibration. From there, the method was applied for the analysis of plasma samples from 399 patients with cardiovascular diseases and 75 healthy controls. This revealed significant metabolic alterations in pathways associated with inflammation, nitric oxide metabolism, and mitochondrial function.
With the comprehensive approach presented here, this technique could be used as a rapid screening method for identifying selective cardiovascular disease biomarkers using targeted metabolomic profiling.
(1) Baskhanova, S. N.; Moskaleva, N. E.; Shestakova, K. M.; et al. Targeted Metabolomics for Cardiovascular Disease: Validation of a High-Throughput HPLC-MS/MS Assay. J. Chromatogr. B 2025, 1264, 124732. DOI:
(2) Cardiovascular Diseases (CVDs). World Health Organization 2025.
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