Pharmacokinetics and absolute bioavailability of isoschaftoside in rat by LC-MS/MS / 中国药科大学学报

@#The aim of this study was to develop a highly sensitive and specific LC-MS/MS method to explore the pharmacokinetic properties and absolute bioavailability of isoschaftoside in rats. Blood sampling was performed at different time points after intragastric administration of isoschaftoside(1. 5, 3. 0, 6. 0 mg/kg)and 0. 5 mg/kg by intravenous injection. Isoschaftoside was analyzed by a validated LC-MS/MS method in plasma; the pharmacokinetic parameters and absolute bioavailability were evaluated by software DAS 3. 0. The results showed that the linear concentration ranges of isoschaftoside was 1. 0- 500. 0 ng/mL(r=0. 997 6). The precision, accuracy, matrix effect, sensitivity, dilution reliability and stability met the requirements of biological sample analysis. For ig administration of isoschaftoside(1. 5, 3. 0, 6. 0 mg/kg), the pharmacokinetic parameter cmax was(109. 34±22. 87), (259. 84±95. 35)and(499. 26±288. 09)ng/mL; AUC0-t was(310. 57±46. 18), (552. 67±207. 14)and(1 075. 03±371. 19)h ·ng/mL; t1/2 was(2. 36±0. 22), (2. 91±0. 19)and(3. 04±0. 86)h; tmax was(1. 03±0. 25), (1. 18±0. 17)and(1. 5±0. 43)h; MRT0-t was(11. 33±1. 53), (11. 27±1. 09)and(8. 29±0. 76)h, respectively. For iv administration of isoschaftoside(0. 5 mg/kg), the pharmacokinetic parameter AUC0-t was(1 536±421. 3)h ·ng/mL; t1/2 was(2. 57±0. 46)h; MRT0-t was(9. 55±2. 37)h. Furthermore, the absolute bioavailability was 6. 73%, 5. 99%, 5. 80%, respectively. The LC-MS/MS analysis method established in this study was accurate and sensitive, so it can be applied to the pharmacokinetic study of isoschaftoside.

Simultaneous Determination of Caffeic Acid,Isoschaftoside and Schaftoside in Xiaoyan Lidan Tablet by RP-HPLC / 中国药房

ABSTARCT OBJECTIVE:To establish a method for the simultaneous determination of caffeic acid,isoschaftoside and schafto-side in Xiaoyan lidan tablet. METHODS:RP-HPLC was performed on the column of Phenomenex C18 with mobile phase of metha-nol- 0.1% phsphate acid(gradient elution)at flow rate of 0.8 ml/min,the detection wavelength was 334 nm,column temperature was 25 ℃,and the injection volume was 10 μl. RESULTS:The linear range was 0.10-2.81 μg for caffeic acid(r=0.999 9), 0.20-5.63 μg for isoschaftoside(r=0.999 9)and 0.10-2.63 μg for schaftoside(r=0.999 9);RSDs of precision,stability and repro-ducibility tests were lower than 3%;recoveries were 96.3%-100.4%(RSD=1.59%,n=9),97.3%-101.2%(RSD=1.28%,n=9) and 96.6%-100.6%(RSD=1.39%,n=9),respectively. CONCLUSIONS:The method is sensitive,accurate,reliable and reproduc-ible,and can be used for the simultaneous determination of caffeic acid,isoschaftoside and schaftoside in Xiaoyan lidan tablet the quality control of Xiaoyan lidan tablet.

Study on the Content Dynamic Changes of Caffeic Acid,Vicenin-2 and Isoschaftoside in Rabdosia lophanthoi-des / 中国药房

OBJECTIVE:To establish a method for simultaneous determination of caffeic acid,vicenin-2 and isoschaftoside in Rabdosia lophanthoides to assess its optimal harvest period. METHODS:HPLC was performed on the column of Diamonsil C18 with mobile phase of methanol-0.1% phosphoric acid solution (gradient elution) at a flow rate of 0.8 ml/min ,detection wave-length was 334 nm ,column temperature was 25 ℃ and volume injection was 15 μl. RESULTS:The linear range was 0.23-5.68 μg/ml for caffeic acid(r=0.999 9),0.31-7.76 μg/ml for vicenin-2(r=0.999 8)and 0.45-11.30 μg/ml for isoschaftoside(r=0.999 9);RSDs of precision,stability and reproducibility tests were lower than 2.0%,average recoveries were 95.1%-98.9%(RSD=1.8%, n=6),97.4%-101.3%(RSD=1.7%,n=6) and 95.5%-98.8%(RSD=1.4%,n=6),respectively. The contents of above-men-tioned 3 ingredients of R. lophanthoides were the highest harvested in Apr. to May and Jul. to Aug. in a year. CONCLUSIONS:The method is simple,accurate and reproducible,and can be used for the simultaneous determination of caffeic acid,vicenin-2 and isoschaftoside in R. lophanthoides. The optimal harvest period is Apr. to May and Jul. to Aug. in a year.

Study on Content Dynamic Changes of Three Water-soluble Flavonoids in Isodon Lophanthoides / 中国药师

Objective:To establish an HPLC method for the simultaneous determination of viceninⅡ, isoschaftoside and schafto-side in Isodon lophanthoides, and analyze their content dynamic changes. Methods:The HPLC system consisted of a Phenomenex luna C18(250 mm ×4.6 mm, 5 μm)column and a solution system of methanol and 0.5% formic acid with gradient elution, the flow rate was 0. 8 ml·min-1 and the detection wavelength was 334 nm at the column temperature of 25℃and the injection volume was 10 μl. Results:The linear range of viceninⅡ, isoschaftoside and schaftoside was 0.130-3.110 μg(r =0.999 8), 0.180-4.540 μg(r =0.999 9) and 0.080-1.970 μg(r =0. 9999), respectively. The average recovery was 96.8% (RSD =1.9%), 99.2% (RSD =1. 6%) and 97. 1% (RSD=1. 6%) (n=6), respectively. The above three kinds of water-soluble flavonoids from seedling stage to significant accumulation, isoschaftoside and schaftoside reached the highest value in March and August, the content of VitexinⅡ in January and April tends to be stable, in May began to increase gradually, reached the maximum value in August, then began to de-crease. Conclusion:With the contents of water-soluble flavonoids as the indicators, the best harvest time of Isodon lophanthoides is March and August, and the quality of the medicinal materials harvested in August is the best.

Simultaneous Determination of Schaftoside, Isoschaftoside, Deoxyelephantopin and 4, 5-Dicaffeoylquinic Acid in Shennongcha Granules by HPLC / 中国药师

Objective:To establish an HPLC method for determining four constituents ( schaftoside, isoschaftoside, deoxyelephan-topin and 4,5-dicaffeoylquinic acid) in Shennongcha granules. Methods:An HPLC method was performed on a Hypersil C18 column (200 mm × 4. 6 mm,5 μm) with the mobile phase of acetonitrile as the phase A and 0. 025 mol·L-1 phosphoric acid solution as the phase B with gradient elution. The flow rate was 1. 3 ml·min-1 . The detection wavelength was set at 270 nm for schaftoside and isos-chaftoside, 208 nm for deoxyelephantopin and 327 nm for 4, 5-dicaffeoylquinic acid. The column temperature was room temperature. Results:The calibration curve was linear over the concentration range of 5. 850-117. 000 μg·ml-1 for schaftoside, 4. 650-93. 000 μg ·ml-1 for isoschaftoside, 4. 160-83. 200 μg · ml-1 for deoxyelephantopin and 5. 470-109. 400 μg · ml-1 for 4, 5-dicaffeoylquinic acid. The correlation coefficient of all curves was more than 0.999. The average recoverywas 97.70% (RSD=1.40%), 96.87%(RSD=1.13%), 97.53%(RSD =1.69%) and 99.29%(RSD =1.01%) (n =6) , respectively. Conclusion: The developed HPLC method is simple,accurate,and can be used in the content determination of Shennongcha granules.

Water-soluble constituents from Isodon lophanthoides var. gerardianus and their antitumor activities / 中草药

Objective: To investigate the water-soluble constituents of Isodon lophanthoides var. gerardianus. Methods: Compouds were isolated and purified by D-101, Sephadex LH-20, and ODS column chromatographies. The chemical structures were elucidated on the basis of spectral data and physicochemical properties. Results: Twelve compounds were isolated and identified as vitexin II (1), vicenin III (2), isoschaftoside (3), schaftoside (4), vitexin (5), 6, 8-di-C-α-L-arabinosylapigenin (6), apigenin-7-O-glucuronide (7), apigenin 6-C-β-L-arabin-osyl-8-C-α-L-arabinopyranoside (8), apigenin 6-C-β-D-xylopyranosyl-8-C-α-L-arabinopyranoside (9), caffeic acid (10), rosmarinic (11), and rutin (12). Conclusion: Compounds 1-9 are separated from I. lophanthoides var. gerardianus for the first time, and constituents of the water-soluble parts of I. lophanthoides var. gerardianus are studied systematically for the first time. Compounds 1, 6, and 11 exert antiproliferative effects on HepG2 cells.

Studies on reference substance of Guangjinqiancao Zonghuangtong Capsule / 中成药

AIM:To prepare a compound as the chemical reference substance of Guangjinqiancao Zonghuangtong Capsule.METHODS:To apply general column chromatography combined with preparative HPLC to isolate the target compound,to use analytic HPLC to determine the purity,stability and its content in the capsule,and to employ spectroscopic analysis (UV,IR,ESI-MS,1H-NMR,13C-NMR,DEPT,1H-13CCOSY,1H-1HCOSY,1DHOHAHA.1D.NOE,HMBC) to elucidate the structure of the isolated compound.RESULTS:The obtained compound was identified as isoschaftoside with the purity of over 99%, which was stable within 3 months at ambient temperature.As for isosehaftoside solution.it was stable within 8 h at ambient temperature.Its content in the capsule was above 3.0%.CONCLUSION:Isoschaftoside is a qualified reference substance for analytic assay ofGuangjinqiancao Zonghuangtong Capsule,and can be isolated from Desmodium styracifolium(Osb.)Merr.

Study on Identification of Rhizoma Arisaematis and Content Determination of Flavonoids / 中国药房

OBJECTIVE:To establish the identification method of Rhizoma Arisaematis and content determination of flavon-oids.METHODS:TLC was used to identify Rhizoma Arisaematis with schaftoside and ischaftoside as reference substances.The content of flavonoids was determined by HPLC.RESULTS:TLC of test sample and that of control substance had same color dots.The linear range of schaftoside and isoschaftoside were 7.925～126.8 ?g?mL-1(r=0.999 9) and 3.996～63.94 ?g?mL-1(r=0.999 7) respectively.Average recoveries were 99.4% for schaftoside (RSD=2.10%,n=9) and 99.52% for isoschaftoside(RSD=2.42%,n=9).CONCLUSION:The method is simple,accurate and reproducible for the quality evaluation of Rhizoma Arisaematis.