Quantification of Histidine-Containing Dipeptides in Dolphin Serum Using a Reversed-Phase Ion-Pair High-Performance Liquid Chromatography Method

Abstract

The quantification of histidine-containing dipeptides (anserine, carnosine, and balenine) in serum might be a diagnostic tool to assess the health condition of animals. In this study, an existing reversed-phase ion-pair high-performance liquid chromatography (HPLC)–ultraviolet detection method was improved and validated to quantify serum anserine, carnosine, and balenine levels in the dolphin. The serum was deproteinized with trichloroacetic acid and directly injected into the HPLC system. Chromatographic separation of the three histidine-containing dipeptides was achieved on a TSK–gel ODS-80Ts (4.6 mm × 150 mm, 5 µm) analytical column using a mobile phase of 50 mmol/L potassium dihydrogen phosphate (pH 3.4) containing 6 mmol/L 1-heptanesulfonic acid and acetonitrile (96:4). The standard curve ranged from 0.1 µmol/L to 250 µmol/L. The average accuracy of the intra- and inter-analysis of anserine, carnosine, and balenine was 97–106%. The relative standard deviations of total precision (RSDr) of anserine, carnosine, and balenine in dolphin serum were 5.9%, 4.1%, and 2.6%, respectively. The lower limit of quantification of these compounds was 0.11–0.21 µmol/L. These results indicate that the improved method is reliable and concise for the simultaneous determination of anserine, carnosine, and balenine in dolphin serum, and may be useful for evaluation of health conditions in dolphins. Furthermore, this method can also be applied to other biological samples.

Determination of Free Histidine in Complex Hair Care Products with Minimum Sample Preparation Using Cation-Exchange Chromatography and Post Column Derivatization

“…Due to the complexity of the samples, it was important to investigate the potential postextraction matrix effects [ 21 , 22 ]. The postextraction matrix effect was studied individually for the shampoo and conditioner products, using samples that did not contain endogenous histidine (according to the manufacturers’ labels).…”

Enhanced capillary zone electrophoresis in cyclic olefin copolymer microchannels using the combination of dynamic and static coatings for rapid analysis of carnosine and niacinamide in cosmetics

“…The HPLC method was used to verify the reliability of the developed method as described previously [34]. The sample preparation method is the same as described in Section 2.5.…”

Unveiling Novel Chaotropic Chromatography Method for Determination of Pralidoxime in Nerve Agent Antidote Autoinjectors

“…For the HPLC separation of highly polar molecules, analytical methods such as the derivatization of the corresponding compounds, using a hydrophilic interaction chromatography (HILIC) column as a stationary phase, or adding ion-pairing reagents to the mobile phase are typically employed [19][20][21][22][23][24]. To solve these problems, cation-exchange chromatography, size exclusion chromatography, ion-pair chromatography (IPC), reverse-phase chromatography, normal-phase chromatography, capillary electrophoresis, and several other analytical methods have been reported [25][26][27][28][29]. Due to the highly polar nature of the analytes, IPC methods using various ion-pairing reagents are the most frequent and common chromatographic techniques for pralidoxime separation, covering more than 50% of all column chromatography methods.…”