Metabolite profiles of ginsenosides Rk1 and Rg5 in zebrafish using ultraperformance liquid chromatography/quadrupole–time-of-flight MS

AbstractBackgroundIn the present study, metabolite profiles of ginsenosides Rk1 and Rg5 from red ginseng or red notoginseng in zebrafish were qualitatively analyzed with ultraperformance liquid chromatography/quadrupole–time-of-flight MS, and the possible metabolic were pathways proposed.MethodsAfter exposing to zebrafish for 24 h, we determined the metabolites of ginsenosides Rk1 and Rg5. The chromatography was accomplished on UPLC BEH C18 column using a binary gradient elution of 0.1% formic acetonitrile–0.1% formic acid water. The quasimolecular ions of compounds were analyzed in the negative mode. With reference to quasimolecular ions and MS2 spectra, by comparing with reference standards and matching the empirical molecular formula with that of known published compounds, and then the potential structures of metabolites of ginsenosides Rk1 and Rg5 were acquired.ResultsFour and seven metabolites of ginsenoside Rk1 and ginsenoside Rg5, respectively, were identified in zebrafish. The mechanisms involved were further deduced to be desugarization, glucuronidation, sulfation, and dehydroxymethylation pathways. Dehydroxylation and loss of C-17 residue were also metabolic pathways of ginsenoside Rg5 in zebrafish.ConclusionLoss of glucose at position C-3 and glucuronidation at position C-12 in zebrafish were regarded as the primary physiological processes of ginsenosides Rk1 and Rg5

Comparative Study on the Pharmacokinetics of Rutin and Quercetin in Diabetic and Normal Rats by HPLC-DAD

Diabetes mellitus (DM) is a serious health problem affecting millions of individuals worldwide. It is showed that some changes of many enzymes and transporters concerned with metabolism and disposal of drug have taken place in organism under pathologic state of DM. The pharmacokinetic of drug should be different between diabetic and normal animals. Rutin and quercetin can also be used to treatment of diabetic mellitus. So, this paper investigated the difference of pharmacokinetic profiles of rutin and quercetin in diabetic and normal rats in vivo by HPLC-DAD method. The pharmacokinetic parameters were analyzed by double-compartmental method (DAS2.0). The pharmacokinetic parameters of rutin in normal and diabetic rats were: (22.203 ± 2.6) and (36.174 ± 7.5) mg · h/L for AUC(0-4); (0.726 ± 0.13) and (1.069 ± 0.17) h for MRT(0-4), (5.413 ± 0.57) and (6.595 ± 0.38) h for t1/2; (0.424 ± 0.071) and (0.226 ± 0.072) L/h/kg for Cl, respectively. The pharmacokinetic parameters of quercetin in normal and diabetic rats were: (5.243 ± 0.82) and (2.376 ± 0.61) mg h/L for AUC(0-4); (2.556 ± 0.52) and (1.616 ± 0.35) h for MRT(0-4), (1.216 ± 0.17) and (0.992 ± 0.12) h for t1/2; (1.918 ± 0.32) and (4.342 ± 0.99) L/h/kg for Cl, respectively. Those results indicate that the pharmacokinetic profiles of rutin and quercetin were changed by DM.Colegio de Farmacéuticos de la Provincia de Buenos Aire

Pharmacokinetics of lansoprazole injection in peptic ulcer and healthy volunteers

The pharmacokinetics of lansoprazole after a single intravenous dose of 30 mg was determined in 10 healthy volunteers and 10 peptic ulcers patients. In this work, a liquid-liquid extraction and enrichment method with RP-HPLC determination route was taken with high sensitivity and low limit detection of 5 ng/mL. The concentration-time curves in the two groups were best fitted to a two-compartment model, but their main kinetic parameters were remarkably different between healthy and ulcers volunteers. The mean maximum plasma concentration (Cmax ) and area under the curve (AUC0t ) were increased from 975.8 ng/mL to 1298.7 ng/mL and from 1439 ng·h/mL to 2301 ng·h/mL, respectively, and peak time (tmax ) decreased from 0.36 h to 0.26 h. Meanwhile, the half life (t1/2 ) prolonged from 2.25 h to 2.91 h and the clearance (CL) reduced from 20.04 L/h to 13.96 L/h. That variability of lansoprazole pharmakinetic parameter indicates that ulcers have significant effect on its metabolic process.Colegio de Farmacéuticos de la Provincia de Buenos Aire

Simultaneous quantification of nine flavonoids in Ginkgo biloba extract tablets by HPLC-DAD

A new HPLC-DAD method has been developed and validated for the simultaneous analysis of nine flavonoids (rutin, myricetin, quercitrin, quercetin, luteolin, genistein, kaempferol, apigenin, and isorhamnetin) in Ginkgo biloba tablets. The analytes were separated on a kromasil C18 column and recorded at 254 nm. The greatest resolution was achieved with methanol-0.1 % formic acid gradient at a flow rate of 1.0 mL min-1 For all the analytes, the correlation coefficients for all the calibration plots (R2<0.999) showed good linearity over the range tested. The method was validated for repeatability, precision, stability, accuracy, selectivity, and robustness. The validated method has been successfully applied to simultaneous analysis of these active components in Ginkgo biloba tablets from different manufacturers.Colegio de Farmacéuticos de la Provincia de Buenos Aire

Structure and Evolution of Glycogen Branching Enzyme N-Termini From Bacteria

In bacteria, glycogen plays important roles in carbon and energy storage. Its structure has recently been linked with bacterial environmental durability. Among the essential genes for bacterial glycogen metabolism, the glgB-encoded branching enzyme GBE plays an essential role in forming α-1,6-glycosidic branching points, and determines the unique branching patterns in glycogen. Previously, evolutionary analysis of a small sets of GBEs based on their N-terminal domain organization revealed that two types of GBEs might exist: (1) Type 1 GBE with both N1 and N2 (also known as CBM48) domains and (2) Type 2 GBE with only the N2 domain. In this study, we initially analyzed N-terminal domains of 169 manually reviewed bacterial GBEs based on hidden Markov models. A previously unreported group of GBEs (Type 3) with around 100 amino acids ahead of the N1 domains was identified. Phylogenetic analysis found clustered patterns of GBE types in certain bacterial phyla, with the shorter, Type 2 GBEs predominantly found in Gram-positive species, while the longer Type 1 GBEs are found in Gram-negative species. Several in vitro studies have linked N1 domain with transfer of short oligosaccharide chains during glycogen formation, which could lead to small and compact glycogen structures. Compact glycogen degrades more slowly and, as a result, may serve as a durable energy reserve, contributing to the enhanced environmental persistence for bacteria. We were therefore interested in classifying GBEs based on their N-terminal domain via large-scale sequence analysis. In addition, we set to understand the evolutionary patterns of different GBEs through phylogenetic analysis at species and sequence levels. Three-dimensional modeling of GBE N-termini was also performed for structural comparisons. A further study of 9,387 GBE sequences identified 147 GBEs that might belong to a possibly novel group of Type 3 GBE, most of which fall into the phylum of Actinobacteria. We also attempted to correlate glycogen average chain length (ACL) with GBE types. However, no significant conclusions were drawn due to limited data availability. In sum, our study systematically investigated bacterial GBEs in terms of domain organizations from evolutionary point of view, which provides guidance for further experimental study of GBE N-terminal functions in glycogen structure and bacterial physiology

Comparative study on the pharmacokinetic of lansoprazole in gastric ulcer and normal rabbits by HPLC-DAD

Gastric ulcer is one of ulcerous diseases and may result in some changes of many enzymes and transporters concerned with metabolism and disposal of drug. The pharmacokinetic of drug should be different between peptic ulcer and normal animals. Lansoprazole has been one of important medicine for treatment of ulcerous diseases. So, this paper investigated the difference of pharmacokinetic profiles of lansoprazole in gastric ulcer and normal rabbits in vivo by HPLC-DAD method. In this work, a liquid-liquid extraction and enrichment method with RP-HPLC determination route was taken. The pharmacokinetic parameters were analyzed by double-compartmental method (DAS2.0). The pharmacokinetic parameters of lansoprazole in normal and ulcer rabbits were as follows: (614.42 ± 152.25) and (875.73 ± 316.34) mg h/L for AUC(0-6.5); (0.68 ± 0.12) and (0.83 ± 0.22) h for MRT(0-6.5), (0.52 ± 0.23) and (0.87 ± 0.42) h for t1/2 ; (6.13 ± 2.11) and (2.54 ± 1.65) L/h/kg for CL, respectivelyGastric ulcer is one of ulcerous diseases and may result in some changes of many enzymes and transporters concerned with metabolism and disposal of drug. The pharmacokinetic of drug should be different between peptic ulcer and normal animals. Lansoprazole has been one of important medicine for treatment of ulcerous diseases. So, this paper investigated the difference of pharmacokinetic profiles of lansoprazole in gastric ulcer and normal rabbits in vivo by HPLC-DAD method. In this work, a liquid-liquid extraction and enrichment method with RP-HPLC determination route was taken. The pharmacokinetic parameters were analyzed by double-compartmental method (DAS2.0). The pharmacokinetic parameters of lansoprazole in normal and ulcer rabbits were as follows: (614.42 ± 152.25) and (875.73 ± 316.34) mg h/L for AUC(0-6.5); (0.68 ± 0.12) and (0.83 ± 0.22) h for MRT(0-6.5 , (0.52 ± 0.23) and (0.87 ± 0.42) h for t1/2 ; (6.13 ± 2.11) and (2.54 ± 1.65) L/h/kg for CL, respectivelyColegio de Farmacéuticos de la Provincia de Buenos Aire

Gradient HPLC-DAD determination and pharmacokinetic study of Ginkgo biloba extract in rabbits

HPLC-DAD was used and validated for the simultaneous determination of five flavonoids (rutin, quercitrin, quercetin, kaempferol and isorahamnetin) in rabbit plasma. Chromatographic separation was performed on an Aglient Zorbax SB-C18 column (5 μm particle size, 250 mm × 4 . 6 mm i.d.) maintained at 35 ºC. The mobile phase was a mixture of methanol and 0.1 % formic acid water solution with a step linear gradient. At 1.0 ml/min flow rate, the eluent of five flavonoids were detected simultaneously at 350 nm with good separation. For all the analytes, the correlation coefficients for all the calibration plots (r > 0.999) showed good linearity over the range tested. The method was validated for precision, stability, accuracy, and selectivity. The validated method has been successfully applied to determine drug concentrations in plasma samples from rabbit that had been intravenously administrated Ginkgo biloba extract. The main pharmacokinetic parameters of rutin, quercitrin, quercetin, kaempferol and isorahamnetin in rabbit after intravenously administration of 80 mg/kg EGb were as follows, t1/2: (2.134 ± 0.594), (3.408 ± 0.917), (1.919 ± 0.62), (1.171 ± 0.261), (1.829 ± 1.756) h; AUC0-∞: (3.661 ± 0.518), (1.584 ± 0.17), (9.951 ± 1.253), (1.002 ± 0.164), (0.373 ± 0.037) μg·h·L-1 ; MRT(0-t): (0.929 ± 0.132), (1.256 ± 0.038), (1.174 ± 0.065), (0.989 ± 0.099), (1.041 ± 0.117) h; Cl: (5.559 ± 0.814), (12.743 ± 1.304), (2.034 ± 0.224), (20.382 ± 3.165), (54.068 ± 5.474) L/h·kg.Colegio de Farmacéuticos de la Provincia de Buenos Aire

Multiple compounds determination and fingerprint analysis of herbal preparation Shuang-Huang-Lian capsule by HPLC-DAD

The objective of this paper was to develop a high performance liquid chromatography with diode array detection both for chromatographic fingerprint and simultaneous determination of twelve analytes of Shuang-Huang-Lian (SHL) capsule. The chromatographic separation was performed on an Aglient Zorbax SB-C18 column with a gradient elution program using a mixture of acetonitrile and 0.2 % acetic acid as mobile phase within 110 min detected at 278 nm wavelength. For fingerprint analysis, 50 peaks were selected as the common peaks to evaluate the similarities of different samples collected from different pharmaceutical companies in China, and two kinds of data, relative retention time and relative peak area were used to identify the common peaks in samples for investigation. SHL capsules from different batches of the same manufacturer or different manufacturers showed a close similarity. For quantitative analysis, linear regressions, limit of detection and quantification, intra-day and inter-day precisions, recovery, repeatability and stability were all tested and good results were obtained to simultaneously determine the 12 marker compounds in the samples. The validated method coupled with multiple compounds determination and fingerprint analysis is a powerful and meaningful tool to comprehensively conduct the quality control of TCM.Colegio de Farmacéuticos de la Provincia de Buenos Aire

An Abundant Dysfunctional Apolipoprotein A1 in Human Atheroma

Recent studies have indicated that high-density lipoproteins (HDLs) and their major structural protein, apolipoprotein A1 (apoA1), recovered from human atheroma are dysfunctional and are extensively oxidized by myeloperoxidase (MPO). In vitro oxidation of either apoA1 or HDL particles by MPO impairs their cholesterol acceptor function. Here, using phage display affinity maturation, we developed a high-affinity monoclonal antibody that specifically recognizes both apoA1 and HDL that have been modified by the MPO-H2O2-Cl− system. An oxindolyl alanine (2-OH-Trp) moiety at Trp72 of apoA1 is the immunogenic epitope. Mutagenesis studies confirmed a critical role for apoA1 Trp72 in MPO-mediated inhibition of the ATP-binding cassette transporter A1 (ABCA1)-dependent cholesterol acceptor activity of apoA1 in vitro and in vivo. ApoA1 containing a 2-OH-Trp72 group (oxTrp72-apoA1) is in low abundance within the circulation but accounts for 20% of the apoA1 in atherosclerosis-laden arteries. OxTrp72-apoA1 recovered from human atheroma or plasma is lipid poor, virtually devoid of cholesterol acceptor activity and demonstrated both a potent proinflammatory activity on endothelial cells and an impaired HDL biogenesis activity in vivo. Elevated oxTrp72-apoA1 levels in subjects presenting to a cardiology clinic (n = 627) were associated with increased cardiovascular disease risk. Circulating oxTrp72-apoA1 levels may serve as a way to monitor a proatherogenic process in the artery wall

An Abundant Dysfunctional Apolipoprotein A1 in Human Atheroma

Recent studies have indicated that high-density lipoproteins (HDLs) and their major structural protein, apolipoprotein A1 (apoA1), recovered from human atheroma are dysfunctional and are extensively oxidized by myeloperoxidase (MPO). In vitro oxidation of either apoA1 or HDL particles by MPO impairs their cholesterol acceptor function. Here, using phage display affinity maturation, we developed a high-affinity monoclonal antibody that specifically recognizes both apoA1 and HDL that have been modified by the MPO-H2O2-Cl− system. An oxindolyl alanine (2-OH-Trp) moiety at Trp72 of apoA1 is the immunogenic epitope. Mutagenesis studies confirmed a critical role for apoA1 Trp72 in MPO-mediated inhibition of the ATP-binding cassette transporter A1 (ABCA1)-dependent cholesterol acceptor activity of apoA1 in vitro and in vivo. ApoA1 containing a 2-OH-Trp72 group (oxTrp72-apoA1) is in low abundance within the circulation but accounts for 20% of the apoA1 in atherosclerosis-laden arteries. OxTrp72-apoA1 recovered from human atheroma or plasma is lipid poor, virtually devoid of cholesterol acceptor activity and demonstrated both a potent proinflammatory activity on endothelial cells and an impaired HDL biogenesis activity in vivo. Elevated oxTrp72-apoA1 levels in subjects presenting to a cardiology clinic (n = 627) were associated with increased cardiovascular disease risk. Circulating oxTrp72-apoA1 levels may serve as a way to monitor a proatherogenic process in the artery wall