2026-08-18 北海道大学
本研究の概念図
＜関連情報＞
- https://www.hokudai.ac.jp/news/2026/08/post-2415.html
- https://www.sciencedirect.com/science/article/abs/pii/S0963996926009506
血漿中ヒアルロン酸オリゴ糖の液体クロマトグラフィー・タンデム質量分析による同時定量法の確立 Development of a method of liquid chromatography coupled with tandem mass spectrometry for simultaneous determination of hyaluronan oligosaccharide in plasma
Nao Takabayashi, Yuki Sato, Kuma Aoyagi, Shunsuke Nashimoto, Hitoshi Kashiwagi, Yoh Takekuma, Mitsuru Sugawara
Food Research International Available online: 24 April 2026
DOI:https://doi.org/10.1016/j.foodres.2026.119273
Highlights
- A one-pot derivatization/extraction method was applied.
- DMT-MM, GP and d5-GP were used to HA 4, 6 and 8-mer labeling.
- Simultaneous determination method of HA 4, 6 and 8-mer by LC-MS/MS was established.
- This method enabled the quantification of HA 4, 6 and 8-mers in plasma.
- HA 4, 6 and 8-mer were detected even though 8 h passed after oral administration.
Abstract
Hyaluronan (HA) exists in the body with various molecular weights ranging from several hundred to several million. Recently the physiological functions of HA oligosaccharides, in addition to high-molecular weight HA, have been reported. It is important to clarify the relationship between physiological function and pharmacokinetics. When HA is ingested orally, most of it is broken down into HA oligosaccharides less than 8-mers by the gut microbiota and absorbed. However, there is no method to simultaneously determine these HA oligosaccharides. This study aimed to elucidate the pharmacokinetics of exogenous HA, in plasma samples, by developing a simple pretreatment and highly sensitive simultaneous determination method for HA less than 8-mers. First, carboxy groups of HA oligosaccharides were derivatized with a condensing reagent (DMT-MM) and Girard’s reagent P. After pretreatment by solid-phase extraction using hydrophilic interaction, peaks of HA 2, 4, 6 and 8-mer were detected by LC-MS/MS. Validation studies were conducted for this method. Additionally, plasma concentrations of HA were determined after oral administration of HA formulation to rats. We confirmed by MS scan that the derivatized HA oligosaccharides could be analyzed by the developed quantitative method and identified the peak of derivatized HA oligosaccharides. The MS/MS conditions were optimized, and a calibration curve was generated, which showed good linearity for HA 4, 6, and 8-mer. This method was valid and enabled measurement of HA 4, 6, and 8-mer concentrations in plasma. This simultaneous analytical method can reduce experiment time compared to conventional methods.
