以QuEChERS萃取法搭配 Biphenyl靜相與超效能液相層析串聯式質譜系統定量葡萄中花青素

Abstract

花青素(anthocyanosides)廣泛地分佈於植物組織中，對於人體生理作用而言，以抗氧化與抗發炎特性而被受到重視。於富含多種天然成份的葡萄中，亦能發現花青素的存在，能夠為人類健康提供必需之營養。本研究開發了利用QuEChERS (quick, easy, cheap, effective, rugged and safe method)搭配聯苯(biphenyl)靜相-液相層析串聯質譜技術之分析平台，並將其應用於快速分離葡萄樣品中的十二種常見花青素。來自葡萄中所被偵測的單醣(mono-)與非醣基化(un-glycosylation)的花青素而言，聯苯靜相能夠快速達到有效地層析分離，且運用優化之QuEChERS樣品前處理與超高效能液相層析串聯式質譜儀，開發了一種高靈敏度且精確的方法並克服複雜的葡萄基質干擾。針對此分析方法進行驗證，其動態定量範圍為0.5至500 ng mL-1，日內與日間性精密度(%RSD)皆低於11.8%。同時成功地檢測來自不同國家及地區的葡萄樣品，其所測定含量範圍為3.31至23.97 mg/100 g之間。藉由已開發之QuEChERS搭配液相層析串聯式質譜技術，為花青素之定量分析提供了一種更有價值與吸引力的新穎方法。Anthocyanosides (AnCS), which are widely distributed in plant tissues, have important physiological roles and are known for their antioxidant and anti-inflammatory effects. As a source of nutrition in the human diet, grapes contain a variety of natural compounds, including anthocyanosides. In this study, a sensitive and accurate method was developed using quick, easy, cheap, effective, rugged, and safe method (QuEChERS) and ultra-high performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) system for the determination of anthocyanosides in grapes. A liquid chromatography method using a biphenyl stationary phase was developed and applied for the rapid separation of 12 anthocyanosides. Efficient separation was achieved for a wide range of mono- and un-glycosylated anthocyanosides and specific glycosylation patterns were found in grapes. The linear ranges of the target anthocyanosides were 0.5–500 ng mL−1. The intra- and interday precisions (%RSD) were both below 11.8%. Anthocyanosides in grapes from different sources were successfully quantified, with concentrations ranging from 3.31 to 23.97 mg/100 g. Thus, the developed QuEChERS and UHPLC-MS/MS method provides an attractive alternative for the quantification of anthocyanosides in grapes.