Hydrophilic interaction liquid chromatography coupled with tandem mass spectrometry method for the simultaneous determination of l -valine, l -l eucine, l -isoleucine, l -phenylalanine, and l -tyrosine in human serum

Abstract

l-Valine, l-leucine, l-isoleucine, l-phenylalanine, and l-tyrosine are important proposed biomarkers for the early detection and diagnosis of type 2 diabetes. A simple and selective hydrophilic interaction chromatography with tandem mass spectrometry method was developed for the simultaneous determination of these amino acids in human serum, using stable isotope-labeled amino acids as internal standards. Chromatographic separation was carried out on a Syncronis HILIC column (150 mm × 2.1 mm, 5 μm) with the column temperature of 35°C and a mobile phase consisted of acetonitrile/120 mM ammonium acetate (89:11, v/v), and the run time was 11.0 min. The mass spectrometric analysis was performed using a QTRAP 5500 mass spectrometer coupled with an electrospray ionization source in positive ion mode. As these five amino acids are endogenous compounds in serum, we used the corresponding stable isotope-labeled amino acids to evaluate the matrix effect and recovery in serum. The matrix effect was 98.7-107.3%, and the recovery was 92.7-102.3%. Calibration curves spiked unlabeled amino acids in water were linear over the range of 0.200-100 μg/mL. The accuracy, inter-, and intraday precision were below 10.2%. Analytes were stable during the study. This assay method has been validated and applied to the early diagnosis research of type 2 diabetes.

Relationship between Branched-Chain Amino Acids, Metabolic Syndrome, and Cardiovascular Risk Profile in a Chinese Population: A Cross-Sectional Study

“…BCAAs (L-Leu, L-Ile, and L-Val) were detected by a simple, robust, and sensitive hydrophilic interaction chromatography-tandem mass spectrometric (HILIC-MS/MS) method using stable isotope-labeled amino acids as internal standard [ 15 ]. Chromatographic separation was carried out on a Syncronis HILIC column (150 mm × 2.1 mm, 5 μ m) with the column temperature of 35°C and a mobile phase consisted of 120 mmol/L ammonium acetate buffer-acetonitrile containing 20 mmol/L ammonium acetate (89 : 11, v/v).…”

Amino acid analyses of the exosome-eluted fractions from human serum by HPLC with fluorescence detection

“…Amino acid levels in the serum or plasma have been used for early detection and diagnosis of type 2 diabetes, cervical cancer, pancreatic cancer, and post-stroke cognitive impairment [1] , [2] , [3] , [4] . Recently, exosomes (30–200 nm diameter), phospholipid bilayer-enclosed extracellular vesicles (EVs), have become the focus of disease diagnosis because they are released by exocytosis [5] into serum, urine, cerebrospinal fluid, saliva, and breast milk and may contain cellular information.…”

Analysis of amino acids in human tears by hydrophilic interaction liquid chromatography and quadrupole orbitrap mass spectrometry

“…More recently, liquid chromatography-tandem mass spectrometry (LC-MS/MS) has been adopted for amino acid analysis in physiological samples due to its excellent selectivity and sensitivity. [13][14][15][16][17][18] However, co-eluting endogenous compounds in the matrix interferes with ionization efficiency and reproducibility of the ionization source. To enhance the detection sensitivity and chromatographic retention, derivatization reagents such as o-phthaldialdehyde and 9-uorenylmethyl chloroformate were applied.…”

“…To enhance the detection sensitivity and chromatographic retention, derivatization reagents such as o-phthaldialdehyde and 9-uorenylmethyl chloroformate were applied. 15 However, the methods with chemical derivatization may suffer from laborious sample preparations as well as long testing time. 19,20 With the development of mass spectrometry (MS) technology, such as orbitrap MS and time of ight (ToF) MS, high resolution MS has become an acceptable method for the direct analysis of amino acids (AAs).…”