Identification and determination of isoflavones in germinated black soybean sprouts by UHPLC−Q-TOF-MS mass spectrometry and HPLC-DAD

Ultra-performance liquid chromatography coupled with electrospray ionization tandem quadrupole time-of-flight mass spectrometry (UHPLC?Q-TOF-MS) and HPLC-DAD with an ultrasonic extraction method was developed for qualitative and quantitative compositions and contents in black soybean sprouts. Operational conditions of ultrasonic extraction were optimized by single-factor experiment. Phytochemical compositions of black soybean sprouts were identified by UHPLC?Q-TOF-MS in positive ion mode. A total of 23 compounds were tentatively characterized and identified by means of accurate mass and characteristic fragment ions. Among of them, the six isoflavones in different germinated black soybean sprouts were further quantified by HPLC-DAD. The results indicated that the calibration curves showed good linearity (r² > 0.9994). The intra-day and inter-day precision variations were less than 1.65% and 1.73%, respectively. The recoveries were in the range of 94.95–106.45%. The results indicated that the maximum amounts of isoflavones were produced on the fourth to fifth day germinated black soybean sprouts.     

Solid lipid nanoparticles of anticancer drug andrographolide: formulation,in vitro and in vivo studies

Diterpenoidal anti-cancer drug andrographolide (AD) was encapsulated into solid lipid nanoparticle (SLN) because of poor aqueous solubility and high lipophilicity. AD-SLNs were prepared by solvent injection method and characterized for droplet size, surface morphology, zeta potential, etc. In vitro drug release was carried out by dialysis-membrane method. A pharmacokinetic study was performed by UPLC/Q-TOF-MS method to determine the maximum plasma concentration (C_max), area under the curve (AUC), etc. There was an improvement in C_max and AUC of AD-SLNs when compared with AD, thereby enhancing the bioavailability of AD. The t_max was increased than that of AD suspension, indicating the sustained release pattern of AD-SLNs. The antitumor activity was carried out on Balb/c mice showing better results with AD-SLNs as compared to AD. Thus, the AD-loaded SLNs would be useful for delivering poorly water-soluble AD with enhanced bioavailability and improved antitumor activity.     

基于UPLC／Q—TOF—MS技术的毛茛生物碱类成分快速分析鉴定

应用液质联用技术uPLc／Q—TOF—MS建立一种有效快速分析鉴定毛茛生物碱类成分的方法并研究其质谱裂解规律。采用液质联用技术UPLC／Q—TOF—MS,根据毛莨生物碱类成分的一级质谱数据,二级特征裂解碎片及标准品比对鉴定毛莨生物碱类成分。共鉴定了12种生物碱类成分,并探讨了其裂解规律。首次从毛茛中分离鉴定了12种生物碱类物质,建立了一种有效的基于UPLC／Q—TOF-MS鉴定毛茛生物碱类成分的方法。     

急性放射损伤小鼠血清蛋白质组分析

为寻找急性放射病早期诊断指标和新的治疗靶点，探讨辐射损伤的发病机理，采用蛋白质组学的双向电泳和蛋白质氨基酸序列分析技术研究了8Gy γ射线照射后24h小鼠血清蛋白质的变化，鉴定有差异的蛋白质，并用蛋白质印迹(Western blotting)方法进行验证。结果显示，双向电泳发现8Gy照射后24h小鼠血清有一显著变化蛋白点，分子量约为15ku，经氨基酸序列分析发现为结合珠蛋白(Haptoglobin，Hp)的α亚单位。Western blotting方法进一步验证了双向电泳的结果。结果表明，经8Gy γ射线照射后24h小鼠血清中结合珠蛋白发生显著增加，可能在辐射损伤中发挥着一定的作用，此结果为探索急性放射损伤的发病机理提供了新的线索。     

基于UPLC/Q-TOF-MS/MS分析鳖血柴胡皂苷类化学成分

以鳖血柴胡(Bupleuri radix with turtle blood)炮制品为原料,基于超高效液相色谱-飞行时间质谱(UPLC/Q-TOF-MS/MS)分析手段,建立皂苷类成分快速分析方法。采用Waters ACQUITY UPLC BEH C18色谱柱(100 mm×2.1 mm,1.7μm),以0.01%甲酸水溶液(A)-乙腈(B)为流动相进行梯度洗脱,体积流量0.3 mL/min,进样量2μL,柱温40℃,ESI采用负离子扫描模式采集数据。结果表明:利用已建立的筛查数据库将满足质量误差小于5×10-6、同位素分布正确且含有二级质谱裂解碎片的离子作为分析目标化合物,结合软件Formula Finder,Mass Calculators等功能、在线数据库(Human Metabolome Database、Pub Chem、Mass Bank、Chem Spider等)及二级碎片裂解规律,从鳖血柴胡中共分离和鉴定出44个皂苷类成分,主要为环氧醚型、异环双烯型、12-烯型、12-烯-28-羧酸型柴胡皂苷。综上分析,UPLC/Q-TOF-MS/MS方法能快捷、准确、较全面地鉴定鳖血柴胡中皂苷类成分,为鳖血柴胡的质量评价指标选择和药效物质基础的研究提供可行方法。     

超高压液相色谱-飞行时间质谱法分析国产油橄榄叶中酚类化合物

建立国产油橄榄(Olea europaea L.)叶中酚类化合物的超高压液相色谱-飞行时间质谱(ultra-performance liquid chromatography/quadrupole time-of-flight mass spectrometry,UPLC/Q-TOF-MS)定性分析方法。液相色谱条件为Waters BEH-C18(10.0mm×2.1mm,1.7μm)色谱柱,乙腈和甲酸溶液梯度洗脱,流速0.25mL/min,紫外检测波长280nm;质谱条件为Waters ACQUITY Q-TOF-MS质谱仪,电喷雾电离接口,负离子模式检测。采用该方法得到了国产油橄榄叶提取物的紫外检测的色谱图、负离子监测的总离子流图以及一级质谱图,成功鉴定了国产油橄榄叶中的15种酚类化学成分。     

超高压液相色谱-飞行时间质谱法分析牡丹皮化学成分

采用超高压液相色谱-飞行时间质谱(UPLC/Q-TOF-MS)定性分析了牡丹皮化学成分。液相色谱条件为Waters BEH-C18(10.0mm×2.1mm i.d.,1.7μm)色谱柱,乙腈和0.1%甲酸水溶液梯度洗脱,体积流量0.25mL/min,紫外检测波长254nm;质谱条件为Waters ACQUITY Q-TOF-MS质谱仪,电喷雾电离接口,正、负离子模式检测。采用该方法得到了牡丹皮甲醇提取物正、负离子监测的总离子流图,负离子模式监测更适合牡丹皮中化合物的质谱分析,采用分子离子与特征碎片离子提取方式提取色谱图,结合裂解推定及文献数据鉴定了牡丹皮中没食子酸、芍药苷、丹皮酚等13个化合物,芍药苷亚硫酸酯为首次在牡丹皮中发现,总结了芍药苷等单萜类化合物的两种主要裂解途径。     

基于UPLC/Q-TOF-MS不同比例人参配伍藜芦增毒的物质基础及动物毒性关联性研究

通过小鼠的急性毒性实验,探索中药十八反中藜芦与人参药对不同配比时毒性变化趋势,结合超高效液相色谱-四极杆-飞行时间质谱联用技术(UPLC/Q-TOF-MS)对不同配比对应的毒性成分进行分析,采用MassLynx4.1分析软件,结合主成分分析(PCA)及正交偏最小二乘法判别(OPLS-DA)分析生物碱类成分变化。固定藜芦用量时,藜芦与人参在不同配比情况下小鼠死亡率随人参剂量增大呈下降趋势,藜芦酰棋盘花胺、3-当归酰棋盘花胺、红介芬胺、计米亭碱、藜芦胺和3-藜芦酰基计明碱等生物碱含量在不同配比中的变化趋势与动物毒性变化一致。以上几种生物碱在藜芦与人参合用时可能是毒性效应的关键成分,能够反映配伍毒性的变化趋势。     

亲水作用色谱串联质谱结合反相色谱串联质谱的油料磷脂轮廓分析

采用超高效液相色谱串联四极杆-飞行时间质谱(UPLC-Q-TOF-MS)技术,建立了一种亲水作用色谱(HILIC)和反相色谱(RPLC)结合的磷脂轮廓分析方法,并对8种植物油料的磷脂分子种(PE、PC、PI、LPE、LPC)进行了系统分析。结果表明:8种油料的PC和PE分子种组成基本相似; PI和LPC、LPE分子种组成差异显著,油菜籽、长柄扁桃仁和美藤果仁的PI含量均高于大豆,油菜籽中PI组成以16∶0-18∶2和18∶1-18∶2为主,长柄扁桃仁的PI以16∶0-18∶1为主,美藤果仁的PI以16∶0-18∶2和18∶2-18∶2为主。棉籽和亚麻籽的溶血磷脂含量较高,其中棉籽溶血磷脂的脂肪酸组成以亚油酸为主,亚麻籽中则以油酸为主。该检测方法快速、灵敏、准确,能同时分离多种甘油磷脂、溶血磷脂及其同分异构体。