ABSTRACT
A reversed-phase high-performance liquid chromatography with diode-array detection (HPLC-DAD) was developedand validated to estimate the phenolic acids (gallic acid, caffeic acid, syringic acid, p-coumaric acid, sinapic acid, andferrulic acid), flavonoids (catechin rutin, myricetin, quercetin, apigenin, and kaempferol), ascorbic acid, and eugenol.The chromatogram condition was set in suitable wavelength 272 nm and run flow rate 0.7 µl/minutes using HPLCAgilent Technologies 1260 Infinity, a reversed-phase Zorbax SB-C18 column (3.5 µm particle size, i.d. 4.6 mm × 250mm) with the mobile phase solution (1:9, HPLC-grade acetonitrile:1% acetic acid). The linearity, precision, limit ofdetection, limit of detection, and accuracy were R2 > 0.9907, relative standard deviation < 1%, 0.005 µg/ml, 0.015 µg/ml, and 96%–102%, respectively. As a result, all the selective compounds were successfully separated, identified, andquantified. The enormous contents were found in quercetin and eugenol, expressing crude content (mean, 5.989 mg/g)and residue content (mean, 1.934 mg/g) for quercetin, while crude content (mean, 3.209 mg/g) and residue content(mean, 0.184 mg/g) for eugenol. Consequently, this method could be applied, repeated, and developed for the laterobservation, especially in commercially inclination of Piper betle analysis
ABSTRACT
OBJECTIVE: To study the chemical constituents of different solvent extracts from Fructus Trichosanthis, and to study their antioxidant activity and protection of myocardial ischemia reperfusion injury in rats. METHODS: The chemical constituents in different solvent extracts were determined by UV-Vis spectrophotometry. Myocardial ischemia reperfusion model was made by ligation of left coronary artery in rats. Rats were randomly divided into 6 groups, with 6 rats in each group. The sham operation group and the model groups were respectively given normal saline, Fructus Trichosanthis (equivalent to crude drug 22.5 g·kg-1), compound Danshen dripping pill group(0.085 g·kg-1). Medicines were given once a day for 7 d. After the last drug 1 h, left coronary artery was ligated for 30 min and then reperfusion was established for 120 min by removing the ligation. During this time, ECG was recorded. At the end, animals were euthanized. Blood was collected to evaluate the contents of CK-MB, MYO-MB, cTnT. The heart was removed and fixed to observe the changes of myocardial tissue by optical microscope. RESULTS: Compared with the water extract group and the alcohol extract group of Fructus Trichosanthis, the total amino acids content of Fructus Trichosanthis dichloromethane extract was not detected, but the content of total flavonoids is higher (P < 0.01). Compared with the water extraction liquid of Fructus Trichosanthis, the antioxidant properties on DPPH radical of Fructus Trichosanthis dichloromethane extract group is lower (P < 0.01), and the antioxidant activity on-OH free radical of the alcohol extract of Fructus Trichosanthis group is lower (P < 0.01). Compared with model group, the elevation of ST segment of electrocardiogram was significantly suppressed in each group during reperfusion (P < 0.01 or P < 0.05). The plasma CK-MB, cTnT and MYO-MB in water extract group were significant lowered (P < 0.05). CONCLUSION: The extraction of Fructus Trichosanthis is able to decrease the production of oxidants. The water extract of Fructus Trichosanthis could ameliorate myocardial ischemia reperfusion injury.
ABSTRACT
A reversed-phase high-performance liquid chromatographic method using photodiode array detector with gradient elution has been developed and validated for the simultaneous estimation of ascorbic acid, free phenolic acids and flavonoids (catechin, rutin, quercetin, myrecetin, apigenin and Kaempferol) in four different solvent extracts of two wild edible leaves of viz. Sonchus arvensis and Oenanthe linearis, collected from North-eastern region in India . The chromatographic separation was carried out on Acclaim C 18 column (5 μm particle size,250 x 4.6 mm), Dionex Ultimate 3000 liquid chromatograph and detection was carried out at three different wave lengths (272, 280 and 310 nm) using a mobile phase of acetonitrile and 1% aqueous acetic acid solution with gradient elution. The experimental results showed high amount of ascorbic acid in S. arvensis and O. Linearis (1.2% and 2.3 % respectively) and gallic acid (0.02% and 0.06% respectively) in 1% aq. acetic acid extract of these two plants. The high percentage of recovery (96-103%), low coefficient of variation ( R2 > 0.99) and low limit of detection (LOD) and limit of quantitation (LOQ) confirm the suitability of the method for simultaneous quantification of ascorbic acid and all phenolic compounds in the two plants under investigation.