ABSTRACT
Objective: To study the chemical constituents from the root bark of Litsea glutinosa. Methods: Silica gel and Sephedex LH-20 column chromatographies as well as semi-preparative HPLC were applied to isolate and purify the compounds. Their structures were elucidated based on the spectrum analysis. Results: Fifteen compounds were obtained and identi-fied as schizandriside (1), lyonside (2), coclaurine (3), alangisesquin A (4), alangisesquin B (5), dihydrobuddlenol (6), ssioriside (7), dendranthemoside B (8), epi-anhydrocinnzeylanol (9), isolariciresinol-5’-methoxy-iso-larixa-9’-O-β-D-xylopyranoside (10), benzyl alcohol-β-D- glucopyranoside (11), phenylethyl-β-D-glucopyranoside (12), n-butyl-β-D-fructoside (13), N-cis-feruloyl tyramine (14), and N-trans-sphingoyl tyramine (15). Conclusion: Compounds 1-13 are isolated from this plant for the first time.
ABSTRACT
Objective: The aim of this work was to investigate the quality markers (Q-markers) of Ziziphi Spinosae Semen (ZSS) based on biotransformation by human intestinal microbiota. Methods: In this study, in vitro biotransformation of ZSS aqueous extract by normal human intestinal microbiota was analyzed using UPLC-Q-Orbitrap-MS. Furthermore, the time course of the biotransformation was studied to probe into the biotransformation mechanism of compounds in ZSS by human intestinal flora. The change rules of flavonoids, saponins and alkaloids in the incubation solution at different time points were plotted based on the percentage of peak area of compounds. Results: A total of 31 original ingredients and four metabolites were characterized in transformed ZSS aqueous extract by human intestinal microbiota. No obvious degradation was observed for benzylisoquinoline alkaloids within 24 h. As far as flavones concerned, a wide range of metabolic reactions as well as significant reaction were shown. Meanwhile, these flavonoids were completely degraded during 24 h. In addition, both jujuboside A and jujuboside B were metabolized to their saponins by deglycosylation reactions. Thus, coclaurine, zizyphusine, kaempferol-3-O-rutinoside, spinosin, vicenin II, jujuboside A, and jujuboside B were referred as prospective Q-markers. Conclusion: The results indicated that the chemical compounds in ZSS were obviously affected by transformation. Intestinal transformation studies play an important role for the elucidation of therapeutic material basis of ZSS and it should be taken into account during the process of the investigation of Q-marker.
ABSTRACT
Objective: A high performance liquid chromatography coupled to triple quadrupole mass spectrometry method (HPLC- MS/MS) was established to simultaneously determine the content of 25 characteristic components (gallic acid, tanshinol, catechin, chlorogenic acid, caffeic acid, epicatechin, rutin, polydatin, hyperin, astragalin, naringin, hesperidin, rosmarinic acid, resveratrol, salvianolic acid B, quercetin, emodin-8-O-β-D-glucoside, isorhamnetin, emodin, coclaurine, nuciferine, cryptotanshinone, tanshinone I, dehydronuciferine, tanshinone IIA) in Danhe Granules (DG), and the consistency between different batches was investigated. Methods The analysis was conducted on Agilent Rapid Resolution HD C18 column (50 mm × 2.1 mm, 1.8 μm) with the mobile phase of 0.1% formic acid water-0.1% formic acid acetonitrile. The dynamic multi-response detection (dMRM) scanning mode was used in the mass spectrometry. Results: Based on the established HPLC-MS/MS method, the simultaneous quantitative analysis of 25 characteristic components could be completed within 10 min, with the quantitative limits of isorhamnetin, coclaurine, and dehydronuciferine of 0.025 ng/mL; emodin and tanshinone I of 0.050 ng/mL; emodin-8-O-β-D-glucoside of 0.200 ng/ml; gallic acid, chlorogenic acid, caffeic acid, epicatechin, rutin, hyperin, astragalin, resveratrol, quercetin, nuciferine, tanshinone IIA of 0.250 ng/ml; catechin, rosmarinic acid, and cryptotanshinone of 0.500 ng/mL; tanshinol, hesperidin, and salvianolic acid B of 1.000 ng/mL; polydatin of 2.000 ng/mL; naringin of 5.000 ng/mL, respectively. The linear relationships of the 25 constituents within their respective mass concentrations were good, with the average recovery of 85.16%-113.46% and the RSD of 2.01%-8.80%. Furthermore, this method also included the main components named monarch, minister, assistant and guide of herbs in a relatively comprehensive way. The total content of the 25 components was 31.49 mg/g, among which the content of salvianolic acid B (9.44 mg/g) and hesperidin (7.60 mg/g) was the highest, and the content of isorhamnetin (0.79 μg/g) was the lowest. According to boxplot analysis, the content of 25 components in 10 different batches of DG fluctuated (P value) within 75% < P < 125%; and the RSD value of 25 components ranged from 2.58% to 13.10% by statistical analysis. The above results showed that the consistency of the component content among 10 batches of DG was acceptable. Conclusion: The analytical method established in this study is fast and sensitive. Furthermore, the results of this study are reliable and can provide scientific methods and basis for quality control and consistency analysis of DG.
ABSTRACT
Objective To establish a quality evaluation system for slice decoction of Ziziphi Spinosae Semen (SZR). Methods Ten batches of SZR slice were collected and prepared into decoction, and then the decoctions of SZR were prepared as freeze-dried powders. The extraction rate and the content of the index components were used as indicators to optimize the extraction process. Multi-dimensional chemical fingerprints were established by HPLC coupled with UV detector and ELSD detector. The similarity, RSD of relative retention time, and RSD of relative peak area were analyzed to comprehensively evaluate the quality of ten batches of SZR slice decoction. A sensitive method based on ultra-HPLC coupled with triple quadrupole mass spectrometry (UPLC-MS/MS) was established and validated for quantitative analysis of the nine compounds in SZR slice decoction, including coclaurine, magnoflorine, vicenin II, spinosin, swertisin, kaempferol-3-O-rutinoside, 6'''-feruloylspinosin, jujuboside A, and jujuboside B. Consequently, the developed methods were successfully applied to the quality assessment of SZR slice decoction. Results The average dry extraction rate of ten batches of SZR slice decoction was 20.77%. Three, four, and two common peaks were identified in the HPLC-UV 227 nm, 335 nm, and HPLC-ELSD fingerprints. The similarity of the fingerprints at different detection wavelengths was in the range of 0.942-0.988, relative retention time and relative peak area RSD% were less than 3%. The method validation results indicated that the methods were simple, specific and reliable. All the compounds showed good linearity (r > 0.999 0) with a relatively wide concentration range, acceptable recovery at 87.40%-110.42%, and RSD% less than 4.12%. The content of nine chemical compounds in ten batches of SZR slice decoction were as follows: 0.25-0.27 mg/g of coclaurine, 3.2-3.6 mg/g of magnoflorine, 0.077-0.084 mg/g of vicenin II, 0.82-0.89 mg/g of spinosin, 0.007 6-0.008 5 mg/g of swertisin, 0.057-0.060 mg/g of kaempferol-3-O-rutinoside, 0.32-0.35 mg/g of 6'''-feruloylspinosin, 0.83-0.91 mg/g of jujuboside A, and 0.095-0.110 mg/g of jujuboside B. Conclusion A stable and reliable quality evaluation method of SZR slice decoction was established, which provides a stable material basis for the follow-up pharmacokinetic and pharmacodynamic experiments and provides a reference for the quality control of the prescription containing SZR at the same time.