The anti-tumor effects of catechins and gallic acid(GA)in-vitro was investigated in this paper. Fluo-3AM, Calcium-AM(Ca-AM), 2′, 7′-dichlorofluorescein-diacetate(DCFH-DA), 4′, 6-diamidino-2- phenylindole(DAPI) and Ca-AM plus colbat were used to characterize intracellular calcium, labile iron pool(LIP), reactive oxygen species(ROS), nuclei morphology and mitochondrial permeability transition pore(mPTP) opening, respectively. High performance liquid chromatography(HPLC) was used to quantitate catechins and GA in the cultural medium. The results indicated that each of them showed dose response inhibition of cell growth, provoking nuclei condensation, intracellular calcium elevation, mPTP opening, LIP reduction, and cytochrome c(Cyt-C) to release into cytosol. The caspase inhibitors, 2-aminoethoxydiphenol borate(APB) or Fe3+ could inhibit lethal effects of GA and(-)-epigallocatechin(EGC), but failed to affect(-)-epigallocatechin gallate(EGCG) and(-)-epicatechin gallate(ECG). Level of ROS presented negative growth while their concentration decreased in the medium. In conclusion, our findings suggest that viability of RKO decreased because of their good correlation with elevation of calcium and loss of LIP and ROS in cytosol.
First order interaction of antioxidant capability of epicatechin was investigated in vitro by methods of response surface and central composite general rotary design.The results indicated that based on molar concentration,the ability to scavenge free radicals were in the order:ECG > EC > EGCG > ECG × EGCG > EGC.This suggests that the antioxidant ability of individual catechin had positively interacted effects under physiological condition.The free radical scavenging ability changed during tea processing.As compared with the fresh tea leaves,the leaves after indoor-spreading and Yao-Qing had a higher IC50of for DPPH free radical scavenge,being increased by 11.01%.It is deduced that the process technology of indoor-spreading might improve the quality of manufactured green tea.
对茶叶中的茶氨酸和γ-氨基丁酸(GABA)建立了柱前衍生方法,并利用高效液相色谱法(HPLC)对衍生物进行了测定。采用邻苯二甲醛(OPA)及N-乙酰-L-半胱氨酸(NAC)为衍生试剂,考察了脯氨酸(Pro)、茶多酚(Tp)及维生素C(Vc)等对衍生体系的影响,并对色谱分离条件进行了优化。结果表明,茶氨酸与GABA衍生物在色谱柱上的保留行为与分离效率主要受流动相pH及缓冲盐浓度的影响;Pro、Tp及Vc对茶氨酸的衍生产率影响较小。而Vc可以提高GABA检测的灵敏度,并能稳定衍生用储备液。GABA与茶氨酸衍生后的方法检出限(LOD)分别为3.01×10-5 mmol/L和7.98×10-5 mmol/L;定量限(LOQ)分别为9.99×10-5 mmol/L和2.658×10-4m m o l/L;线性范围分别为0.01～0.4 m m o l/L(相关系数为0.996)和0.05～0.8 m m o l/L(相关系数为0.995);方法的回收率分别为99.29%～119.60%和93.18%～141.06%(除本身含茶氨酸量高的绿茶中回收率为62.88%外)。这说明经优化的柱前衍生-高效液相色谱体系可用于茶叶中两种特征性氨基酸的测定。
