ABSTRACT
Objective To develop an HPLC method for simultaneous determination of the eleven constituents (agaric-alcohol, naringin, hesperidin, neohesperidin, honokiol, emodin, magnolol, costunolide, dehydrocostus, chrysophanol, and physcion) in Chenxiang Huazhi Pills (CHP) by HPLC with gradient elution. Methods The chromatographic separation was performed on an Thermo Syncronis C18 column (4.6 mm × 250 mm, 5 μm) which was operated at 30 ℃. The mobile phase was a linear gradient prepared from water (A) and acetonitrile (B). The linear gradient elution program was programmed as follows: 0—10 min, 20% acetonitrile; 10—20 min, 20%—40% acetonitrile; 20—24 min, 40% acetonitrile; 24—26 min, 40%—52% acetonitrile; 26—30 min, 52% acetonitrile; 30—31 min, 52%—90% acetonitrile; 31—35 min, 90% acetonitrile; 35—40 min, 90%—100% acetonitrile; 40—43 min, 100% acetonitrile; 43—45 min, 100%—20% acetonitrile. The flow rate was 1 mL/min and the detection wavelength was 215 nm. Results The analysis permitted very good separation of eleven constituents within 43 min. A good linear relationship between the peak area and the injection volume was obtained. The ranges of the eleven constituents were 1.4—13.6, 10.0—200.0, 31.5—315.0,1.0—120.1, 1.8—50.6, 0.93—10.1, 1.8—30.0, 0.2—40.3, 1.8—18.1, 1.7—25.0, and 0.45—10.70 μg/mL. The average recoveries of eleven constituents in the samples were in the range of 98.90%—100.87%. The precision RSD of the peak areas of the 11 components ranged from 0.55%—1.54%; Eleven components had good stability within 30 h, and the concentration RSD of each component ranged from 0.75% to 1.94%; The repeatability RSD of each component ranged from 0.39% to 1.73%. The content of agaric-alcohol, naringin, hesperidin, neohesperidin, honokiol, emodin, magnolol, costunolide, dehydrocostus, chrysophanol, and physcion in six batches were 92.0—201.0, 511.5—9 033.0, 5 475.0—12 635.5, 54.5—5 095.5, 192.0—2 137.5, 117.0—391.5, 106.5—1 281.5, 13.0—136.5, 93.5—199.0, 177.0—1 207.0, and 33.5—251.5 μg/g, respectively. Conclusion The method is accurate, rapid and simple with high sensitivity, precision and repeatability, which has been successfully applied as an effective tool for the multicomponent analysis of CHP.
ABSTRACT
Objective To study the chemical constituents and bioactivities of leaves of Torreya grandis. Methods The chemical constituents were isolated by MCI-Gel CHP-20, Diaion HP-20, Toyopearl HW-40, Sephadex LH-20, RP-18 and silica gel column chromatographic methods. Their structures were identified on the basis of physicochemical and spectroscopic analysis. The cytotoxic activity and antitumor activity of compounds 1-4 were investigated by lethal-to-prawn larva bioactivity determining method and MTT assay. Results Ten compounds were isolated from EtOAc extract and n-BuOH of leaves of T. grandis. Their structures were identified as torreyagrandate (1), hinokiol (2), 4-epiagathadial (3), 3,4-dihydroxybenzoic acid 3-O-β-D-glucoside (4), dehydroabietic acid (5), trans-communic acid (6), cis-communic acid (7), (2-methoxy-1,4-phenylene) dimethanol (8), pinoresinol (9), and β-sitosterol (10). The bioactivity experiment indicated that compounds 1-4 possessed certain cytotoxic activity towards brine shrimp, and LC50 value were 7.7, 8.0, 8.8, and 4.2 μg/mL, respectively. In addition, it was found that compound 4 presented remarkable inhibitory effect on two kinds of cells of human liver cancer cells of Huh7 (67%) and HepG2 (69%) at a dose of 10 μg/mL. Conclusion All compounds are isolated from leaves of T. grandis for the first time expect compound 10. Compounds 1-4 exhibite certain cytotoxic activity, and compound 4 displays stronger antitumor activity towards Huh7 and HepG2 liver cancer cells.
ABSTRACT
Aim To explore the alterations of blood corticosterone (CORT) level and adrenal steroidogenic function,as well as its sex specificity in intrauterine growth retardation (IUGR) rats induced by prenatal nicotine exposure (PNE) with high-fat diet (HFD) after birth,and to make clear its mechanism through insulin-like growth factor-1 (IGF-1) signaling pathway.Methods IUGR model was established by PNE (2.0mL · kg-1 · d-1),and the offspring rats were administered with HFD until postnatal week (PW) 24 after weaning.Blood CORT concentration,adrenal steroidogenesis enzymes,expressions of IGF-1 signaling pathway and 11β-HSDs/CR system were tested.Results Compared with HFD control group,the CORT concentration in male offspring of PNE group represented a decrease trend,while an increase trend in female;the expressions of adrenal steroidogenesis enzymes (such as StAR,3β-HSD and P450cll) in male offspring decreased,while increased in female offspring (such as SF-1 and P450c21);the expressions of IGF-1 signalling pathway (IGF-1 and IGF-1R) in male offspring increased,and they significantly increased in female offspring;the expression levels of 11 β-HSD2 and GR decreased,but 11β-HSD1/11β-HSD2 ratio was enhanced in male PNE group,while in female PNE group,the corresponding gene expressions increased.Conclusions PNE could induce abnormal alterations of adrenal steroidogenic function,and exhibit apparent gender differences.The potential mechanism is related to low adrenal steroidogenesis function programming induced by nicotine and catch-up growth mediated by IGF-1 after birth.