Hsu Ying-Yu, Kao Tsai-Hua
Department of Food Science, Fu Jen Catholic University, New Taipei City, 242 Taiwan.
J Food Sci Technol. 2019 Apr;56(4):1939-1953. doi: 10.1007/s13197-019-03660-6. Epub 2019 Mar 8.
This study developed a high performance liquid chromatography with diode array detection (HPLC-DAD) and tandem mass spectrometry (MS-MS) method for determination of prenylflavonoids and hop bitter acids in surplus yeast, a byproduct from beer brewing process. This method enabled the simultaneous separation of 4 prenylflavonoids and 20 hop bitter acids within 30 min by employing a Hypersil-Keystone HyPURITY C18 column and a gradient mobile phase composed of phosphoric acid aqueous solution at pH 1.6 and acetonitrile. For HPLC-DAD analysis, the limits of detection and limits of quantitation ranged from 0.04 to 0.15 µg/mL and from 0.12 to 0.45 µg/mL, respectively, and the recoveries ranged from 82.6 to 99.7%. The intra-day variability and inter-day variability ranged from 1.37 to 8.82% and from 0.68 to 9.74%, respectively. For qualitation by MS-MS, the positive mode was discovered to possess satisfactory collision capacity and high sensitivity for prenylflavonoids, while the negative mode was more suitable for the ionization of hop bitter acids. The content of hop bitter acids in surplus yeast were higher than that of prenylflavonoids, and isomers and oxidation products of hop bitter acids were found. This study has advantages in identifying more components, short separation time, satisfactory resolution, high accuracy and high precision.
本研究开发了一种采用二极管阵列检测的高效液相色谱法(HPLC-DAD)和串联质谱法(MS-MS),用于测定啤酒酿造过程副产物——过剩酵母中的异戊烯基黄酮类化合物和啤酒花苦味酸。该方法使用Hypersil-Keystone HyPURITY C18色谱柱和由pH 1.6的磷酸水溶液与乙腈组成的梯度流动相,能够在30分钟内同时分离4种异戊烯基黄酮类化合物和20种啤酒花苦味酸。对于HPLC-DAD分析,检测限和定量限分别为0.04至0.15μg/mL和0.12至0.45μg/mL,回收率为82.6至99.7%。日内变异和日间变异分别为1.37至8.82%和0.68至9.74%。对于MS-MS定性分析,发现正模式对异戊烯基黄酮类化合物具有令人满意的碰撞能力和高灵敏度,而负模式更适合啤酒花苦味酸的电离。过剩酵母中啤酒花苦味酸的含量高于异戊烯基黄酮类化合物,并且发现了啤酒花苦味酸的异构体和氧化产物。本研究在鉴定更多成分、分离时间短、分辨率令人满意、准确度高和精密度高等方面具有优势。