ABSTRACT
Effective supplementation of probiotics can be beneficial to intestinal health, but in situ analysis of probiotics activity has rarely been reported. In this study, by coupling fluorescein 5-isothiocyanate (FITC) and 5(6)-carboxytetramethylrhodamine N-succinimidyl ester (5(6)-TAMRA-SE) with D-lysine, two fluorescent D-amino acids (FDAAs) probes were obtained: green probe (fluorescein-D-lysine, FDL) and red probe (TAMRA-D-lysine, TDL). Then, we tried to label the three kinds of probiotics, Lactobacillus acidophilus (LA), Lactobacillus casei (LC) and Veillonella atypica (VA) in vitro. FDAAs was applied to the labeling of intestinal flora in mice, and a method was established to investigate the oral survival rate of three commonly used probiotics. All animal experiments were approved by the Animal Ethics Committee of Guangzhou University of Chinese Medicine. The results show that the two synthetic FDAAs can be non-toxic and 100% for the in vitro labeling of the three probiotics. Known by FDAAs two-step labeling of oral probiotics, the high survival rate of LA was 92.30% ± 1.67%. The survival rates of VA and LC are similar, 84.13% ± 4.06% and 82.27% ± 2.43%, respectively. This study can quickly compare the changes of colonization survival rate of different probiotics in vivo, provide theoretical support for the in situ colonization activity of probiotics in the intestine, and guide the rational drug use of clinical probiotics.
ABSTRACT
The present study is to establish the fingerprints for the quality evaluation of Ilicis Pubescentis Radix by HPLC-UV. The chromatographic conditions were defined as Phenomenex Luna C₁₈(4.6 mm × 250 mm, 5 μm). Mobile phase was acetonitrile-0.05% phosphoric acid in gradient elution, and the flow rate was 0.8 mL·min⁻¹.Column temperature was 30 °C and the injection volume was 10 μL．The detection wavelength was 210 nm. According to the similarity evaluation, the chemometric method was used to assess the quality of Ilicis Pubescentis Radix. The fingerprints of 16 batches of Ilicis Pubescentis Radix were established. There were 29 common peaks in the fingerprints and 12 common peaks were identified by reference substances. Fingerprints similarity of samples were greater than 0.92. The samples were classified into three groups by hierarchical cluster analysis combined with principal component analysis (PCA) and orthogonal partial least squares-discriminant analysis (OPLS-DA), and seven components were the main markers that cause differences in the different batches of samples. By comparing the on-line UV spectra of chromatographic peaks, the chromatographic fingerprint was divided into three regions: region A showed seventeen main peaks (mainly lignans and phenolic acids); region B showed eight main peaks, which were proved as saponins; region C showed four main peaks, which were proved as other components. The established HPLC-UV fingerprint is highly specific, and can be used to evaluate the quality consistency of different batches of Ilicis Pubescentis Radix.
ABSTRACT
A comprehensive HPLC-DAD-MS method was developed to study the chemical components of semi-bionic extract of Coptis-Evodia herb couple. The extract was isolated on a Hypersil BDS C18 column (4.6 mm x 200 mm, 5 microm) using acetonitrile-ammonium formic buffer as mobile phase by gradient elution. Detection was performed on DAD and MS equipped with an electrospray ionization (ESI) source by full scan and product full scan on positive mode. The chromatogram of Coptis-Evodia showed seventeen main peaks, eight of which were from Evodia while the others were from Coptis. By comparison of the retention time, the on-line UV spectra and MS spectra, four peaks were identified as jatrorrhizine, hydroxevodiamine, palmatine and berberine, and three peaks were deduced as epiberberine, columbamine and coptisine. In addition, berberine and palmatine were quantitatively determined. No new component was created in the semi-bionic extract of the herb couple, yet the solubilities of berberine and palmatine decreased.
Subject(s)
Berberine , Chemistry , Berberine Alkaloids , Chemistry , Chromatography, High Pressure Liquid , Methods , Coptis , Chemistry , Drugs, Chinese Herbal , Evodia , Chemistry , Plant Extracts , Chemistry , Spectrometry, Mass, Electrospray IonizationABSTRACT
The progress in research on the active ingredients of Sambucus Chinensis and pharmacological activities was reviewed. It is important to study the chemical constituents and phamacological activities of Sambucus chinensis.