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目的:通过构建高效液相色谱-四极杆-时间飞行(HPLC-Q-TOF)高分辨质谱方法定性地鉴定虎杖醇提物的主要组分,对其中的活性成分白藜芦醇含量进行定量分析,并用电喷雾质谱法(ESI-MS)首次研究白藜芦醇与β-乳球蛋白的相互作用。方法:定性分析采用高效液相色谱-四极杆-时间飞行(HPLC-Q-TOF)高分辨质谱方法,使用Agilent ZORBAX Eclipse XDB-C18柱(2.1×150 mm,5-Micron),流动相为甲醇-水溶液,梯度洗脱:0~30 min 10%~90%甲醇,30~40 min 90%甲醇,40~60 min 90%~10%甲醇,柱温:40℃,进样量:2μL,流速:0.2 m L/min-1,虎杖醇提物采用HPLC-Q-TOF负离子检测模式。结果:由高效液相分离和质谱特征负离子的精确质量解析,共鉴别出虎杖中13种化合物,测定出实验所用虎杖中白藜芦醇的含量为1.181 mg/g,观测到白藜芦醇与β-乳球蛋白能形成稳定的蛋白复合物。结论:该方法定性分析了虎杖醇提物中主要药用成分,首次利用ESI-MS法研究了β-乳球蛋白与白藜芦醇的相互作用,检测到两者能有效形成稳定的蛋白复合物,有助于开发一种新的途径以提高白藜芦醇的稳定性和生物利用率。
OBJECTIVE: To characterize the main components of Polygonum cuspidatum by HPLC-Q-TOF (High Performance Liquid Chromatography-Quadrupole-Time of Flight) method and quantify the content of resveratrol The interaction between resveratrol and β-lactoglobulin was studied for the first time by electrospray ionization mass spectrometry (ESI-MS). METHODS: Qualitative analysis was performed on a HPLC-Q-TOF high resolution mass spectrometry column using an Agilent ZORBAX Eclipse XDB-C18 column (2.1 × 150 mm, 5-Micron) with a mobile phase of Methanol-water, gradient elution: 0-30 min 10% -90% methanol 30-40 min 90% methanol 40-60 min 90% -10% methanol, column temperature 40 ℃, injection volume 2 μL, Flow rate: 0.2 m L / min-1, Polygonum cuspidatum extract HPLC-Q-TOF negative ion detection mode. Results: 13 kinds of compounds were identified from the high quality liquid phase separation and mass spectrometry negative ions. The results showed that the content of resveratrol in Polygonum cuspidatum was 1.181 mg / g, Beta-lactoglobulin can form stable protein complexes. CONCLUSION: This method qualitatively analyzed the main medicinal components in the ethanol extract of Polygonum cuspidatum. The interaction between β-lactoglobulin and resveratrol was studied by ESI-MS for the first time. It was found that both of them could effectively form a stable protein complex It helps to develop a new way to improve the stability and bioavailability of resveratrol.