A validated ultra-performance liquid chromatography with diode array detection coupled to electrospray ionization and triple quadrupole mass spectrometry method to simultaneously quantify taurine, homotaurine, hypotaurine and amino acids in macro- and microalgae

Abstract

A fast and reliable method for the simultaneous quantification of Taurine, Homotaurine, Hypotaurine and 19 amino acids in algae samples by Ultra-performance liquid chromatography coupled with diode array and tandem mass spectrometry (UHPLC-DAD-MS/MS) was optimized and validated.Target compounds were chromatographically resolved in less than 15 minutes. (ESI)-MS/MS electrospray ionization and pure analytical standards were used to confirm the identity of all analytes, while quantitation was carried out with diode array detection. Validation parameters of the method were satisfactory: Resolution of peak pairs was always higher than 1.55; all analytical curves showed R 2 > 0.99, with working ranges between 0.04 mg/g to 33.1 mg/g and 9.13 mg/g to 107 mg/g and the Lack-of-fit test was not significant. The intra and inter-day precision of the method (expressed as relative standard deviation) were lower than 6 % and recovery values ranged between 95 % and 105 %. The method was demonstrated to be robust to small deliberate variations of seven variables such sample weight, volume of hydrolysis reagent, hydrolysis time and temperature, derivatization time, column temperature and flow rate.The mean expanded uncertainty for all the target compounds were 0.7 mg/g with a coverage factor of 2.Method Limits of detection and quantification varied from 0.005 * 10 -3 mg/g to 0.11 * 10 -3 mg/g and 0.01* 10 -3 mg/g to 0.22 * 10 -3 mg/g respectively, allowing the routine determination of these bioactive compounds in algae extracts.Therefore, the method was successfully applied for the quantitative determination of the 22 target compounds in five seaweed commercial samples.Relevant compounds were quantified for the first time in the five algae species, namely: i) Taurine in Gracilaria longissima and Chlorella spp., ii) Gamma-aminobutyric acid in G. longissima and L. japonica, iii) Hydroxyproline in G. longissima, Ulva lactuca, Porphyra spp., and L. japonica and iv) Homotaurine and Hypotaurine in the five species studied. Highlights• Simultaneous analysis of Taurine, Homotaurine, Hypotaurine and 19 amino acids • The method was validated and applied for the analysis of algae extracts • Quantitation was performed with DAD to support routine analyses • Homotaurine and Hypotaurine were quantified for the first time in commercial samples Keywords Homotaurine, Hypotaurine, Taurine, amino acids, Algae, UPLC-DAD-MS/MS * u(RSD R ) = standard uncertainty for within lab reproducibility ** u(rec) = standard uncertainty for recovery *** u (Pse) = Expanded uncertainty

Identification of protein-polysaccharide nanoparticles carrying hepatoprotective bioactives in freshwater clam (Corbicula fluminea Muller) soup

“…Terriente, et al [20]. Briefly, it was a function of retention time of the two peaks, tR1 and tR2, and the peak widths at half height, W11/2 and W21/2, of the analyte and another peak Eq.…”

Taurine, homotaurine, GABA and hydrophobic amino acids content influences “in vitro” antioxidant and SIRT1 modulation activities of enzymatic protein hydrolysates from algae

“…Total amino acids were quantified as previously described 70 . Briefly, 10 mg of EPH fractions were weighted and added with 1 mL of 8 M per chloric acid and maintained 24 h at 110 °C (Precisiterm, J.P. Selecta, BCN, Spain).…”

Identification of taurine biomarker in human biofluids using plasmonic patterns of silver nanostructure

“…11 As the understanding of the signicance of taurine in different diseases grows, there is a greater demand for a quick and straightforward test to regularly measure this compound in plasma and urine. 12 Conventional techniques to detect the taurine concentration usually include chromatography with spectrometric detection, 13 capillary electrophoresis, 14 and electrochemical assay. 15 Despite the excellent limit of detection, these methods involve timeconsuming sample preparation, costly and hazardous solvents, and require skilled personnel to run the complex procedures.…”

“…Conventional techniques to detect the taurine concentration usually include chromatography with spectrometric detection, 13 capillary electrophoresis, 14 and electrochemical assay. 15 Despite the excellent limit of detection, these methods involve time-consuming sample preparation, costly and hazardous solvents, and require skilled personnel to run the complex procedures.…”