超高效液相色谱-线性离子阱-高分辨质谱同步检测果蔬及饮料中花青素

Simultaneous analysis of anthocyanins in fruit,vegetable and beverage by ultra-high performance liquid chromatography coupled to a linear ion trap-orbitrap high resolution mass spectrometer

建立超高效液相色谱联用线性离子阱高分辨质谱(UPLC-LTQ-Orbitrap-HRMS)同时检测果蔬及饮料中6种花色苷和6种花色素。样品经5%(V/V)甲酸甲醇溶液超声提取30 min后,过膜上机。采用Hypersil GOLD C18色谱柱(100 mm×2.1 mm i.d,1.9μm)分离,流动相为1%(V/V)甲酸水溶液和1%(V/V)甲酸乙腈溶液,梯度洗脱,流速0.3 mL/min;电喷雾电离源,正离子模式,扫描范围(m/z):100~1000。考察了方法线性范围、检出限、定量限、重现性和回收率。结果表明:12种花青素在0.0424~19.9μg/mL范围内线性相关系数均≥0.9951,回收率范围在61.9%~112.7%之间。方法检出限(S/N=3)在1.73~8.18 ng/mL之间,定量限(S/N=10)为5.77~27.3 ng/mL。12种花青素质量误差≤2.14×10-6,72 h内相对标准偏差≤7.8%。

A method of ultra-high performance liquid chromatography coupled with a linear ion trap-orbitrap high resolution mass spectrometry(UHPLC-LTQ-Orbitrap HRMS)for the determination of 6 anthocyanins and 6 anthocyanidins in fruits,vegetables and botanical beverage was developed.The samples were determined after extracted by 5%(V/V)formic acid/methanol for 30 min.12 analytes were separated on a Hypersil GOLD C18 column(100 mm×2.1 mm i.d,1.9μm)by gradient elution,with a mobile phase consisting of 1%formic acid/water(V/V)and 1%formic acid/acetonitrile(V/V)at a flow rate of 0.3 mL/min.The detection was performed using a LTQ-Orbitrap MS detector equipped with electrospray ionization(ESI)using full scan in positive mode from(m/z)100~1000.This method was validated according to linearity,limits of detection and quantitation,reproducibility and recoveries.The method showed linear response for concentration in range of 0.0424-19.9μg/mL for 12 analytes,and the correlation coefficients(R2)were greater than 0.9951.The average recoveries of 12 analytes in different kinds of samples ranged from 61.9%~112.7%.The limits of detection(LODs)ranged from 1.73-8.18 ng/mL.The limits of quantification(LOQs)were in range of 5.77-27.3 ng/mL.The mass error for 12 analytes was not more than 2.14×10-6,yielding relative standard deviations(RSDs)within 72 hours not more than 7.8%.