Liquid chromatography-electrospray ionization tandem mass spectrometry for the quantification of styraxlignolide A in rat plasma.

Styraxlignolide A is a pharmacologically active ingredient isolated from Styrax japonica Sieb. et Zucc. A rapid, selective, and sensitive liquid chromatographic method with electrospray ionization tandem mass spectrometry was developed for use in the quantification of styraxlignolide A in rat plasma. Styraxlignolide A was extracted from rat plasma using ethyl acetate at neutral pH. The analytes were separated on an Atlantis dC18 column using a mixture of methanol and ammonium formate (10 mM, pH 3.0) (70:30, v/v) and detected by tandem mass spectrometry in multiple reaction monitoring mode. The standard curve was linear (r(2) =0.9978) over the concentration range of 100-10000 ng/mL. The lower limit of quantification was 100 ng/mL using 50 µL of plasma sample. The coefficient of variation and relative error for intra- and inter-assays at four QC levels were 1.6-8.3% and from -12.0 to -1.7%, respectively. The present method was applied successfully to the pharmacokinetic study of styraxlignolide A after intravenous administration of styraxlignolide A at a dose of 10 mg/kg in male Sprague-Dawley rats.

Anticancer activity of 2α, 3α, 19ß, 23ß-Tetrahydroxyurs-12-en-28-oic acid (THA), a novel triterpenoid isolated from Sinojackia sarcocarpa.

BACKGROUND: Natural products represent an important source for agents of cancer prevention and cancer treatment. More than 60% of conventional anticancer drugs are derived from natural sources, particularly from plant-derived materials. In this study, 2α, 3α, 19ß, 23ß-tetrahydroxyurs-12-en-28-oic acid (THA), a novel triterpenoid from the leaves of Sinojackia sarcocarpa, was isolated, and its anticancer activity was investigated both in vitro and in vivo. PRINCIPAL FINDINGS: THA possessed potent tumor selected toxicity in vitro. It exhibited significantly higher cytotoxicity to the cancer cell lines A2780 and HepG2 than to IOSE144 and QSG7701, two noncancerous cell lines derived from ovary epithelium and liver, respectively. Moreover, THA showed a dose-dependent inhibitory effect on A2780 ovary tumor growth in vivo in nude mice. THA induced a dose-dependent apoptosis and G2/M cell cycle arrest in A2780 and HepG2 cells. The THA-induced cell cycle arrest was accompanied by a downregulation of Cdc2. The apoptosis induced by THA was evident by induction of DNA fragmentation, release of cytoplasmic Cytochrome c from mitochondria, activation of caspases, downregulation of Bcl-2 and upregulation of Bax. CONCLUSION: The primary data indicated that THA exhibit a high toxicity toward two cancer cells than their respective non-cancerous counterparts and has a significant anticancer activity both in vitro and in vivo. Thus, THA and/or its derivatives may have great potential in the prevention and treatment of human ovary tumors and other malignancies.

Purification and anticancer activity investigation of pentacyclic triterpenoids from the leaves of Sinojackia sarcocarpa L.Q. Luo by high-speed counter-current chromatography.

Sinojackia sarcocarpa L.Q. Luo has high aesthetic value, which is clinically used to treat diseases such as the variations of arthritis, thromboangiitis obliterans, angina pectoris as well as many other diseases. Pentacyclic triterpenoids are the main pharmacological effective compounds. High-speed counter-current chromatography method was performed on a Midi-DE centrifuge at 25°C. The solvent system was n-hexane-ethylacetate-methanol-water (10:5:3:1, v/v). Peak fractions were collected according to the elution profile for subsequent high-performance liquid chromatography analysis. The structure identification was performed by ultraviolet, infrared, mass spectrometry, (1)H-NMR and (13)C-NMR. Compound 2α,3α,19ß,23ß-tetrahydroxyurs-12-en-28-oic acid and 2α,3α,23ß-trihydroxyurs-12-en-28-oic acid were purified from the plant of Styracaceaen genius for the first time, whose purities were 96.57% and 97.33%, respectively. Compared with the same dose of oral 5-fluorouracil with 57.6% inhibition rate, the S(180) tumour inhibition rates of 20 mg kg(-1)d(-1) two compounds were 59.5% and 48.9%, respectively.