Quantitative Analysis of Twelve Active Components Combined With Chromatographic Fingerprint for Comprehensive Evaluation of Qinma Prescription by Ultra-Performance Liquid Chromatography Coupled With Diode Array Detection.

A combination method of ultra-performance liquid chromatography (UPLC) coupled with diode array detection has been developed for quality evaluation of Qinma prescription (QMP), based on chromatographic fingerprint technology with the similarity analysis (SA) and the quantitative analysis of 12 components by hierarchical cluster analysis (HCA). The established method has been validated by linearity, precision, repeatability, stability and recovery tests. The UPLC fingerprints with 17 common peaks of 5 QMP samples prepared by different extraction methods including water decoction extraction, water extraction-ethanol precipitation method, ethanol reflux extraction, ethanol extraction-water precipitation method and methanol ultrasonic extraction were obtained, and the SA results indicated that similarity index was greatly influenced by the large peak. The similarity index ranged from 0.816 to 0.999 basing on 17 peaks, which has been decreased to 0.683-0.999 basing on 16 peaks without the large peak of baicalin (BA). The results of simultaneous quantification of 12 components in these 5 QMP samples proved that BA, gallic acid (GA), wogonoside (WOG) and gentiopicroside (GEN) were the major ingredients in QMP with high contents >1.44 (mg/g), indicating that ethanol reflux was the most effective extraction method. Integrating fingerprint analysis, simultaneous determination and HCA, the established method is rapid, sensitive, accurate and readily applicable. All the results indicated that the combination method can control the quality of QMP and its related traditional Chinese medicinal compounds more comprehensively and scientifically.

pH-sensitive and biocompatible quercetin-loaded GO-PEA-HA carrier improved antitumour efficiency and specificity.

A novel drug carrier was designed based on a new biomaterial, that is, graphene oxide (GO), to improve the efficiency and specificity of anticancer drug. In this study, GO was successively modified with polyetheramine (PEA) and hyaluronic acid (HA). The carrier was utilized to load an antitumor component, that is, quercetin (Que), which was derived from traditional Chinese medicine, namely the pagoda tree flower bud. This drug delivery system (DDS) exhibited pH sensibility under subacid condition and good biocompatibility even when GO concentration reached 350 µg/mL. Moreover, the antitumor efficacy was doubly improved and more long-acting compared with Que alone. Results show that the GO-based material has potential clinical applications for antitumor drug delivery.

Chemical Fingerprinting and Quantification of Chinese Cinnamomi Cortex by Ultra High Performance Liquid Chromatography Coupled with Chemometrics Methods.

To rapidly clarify and quantify the chemical profiling of Cinnamomi cortex a reliable and feasible strategy of chromatographic fingerprinting with a suite of chemometrics methods was developed and validated by ultra-high performance liquid chromatography coupled with diode array detection. Furthermore, to identify more meaningful chemical markers, the chemometrics methods including hierarchical cluster analysis (HCA), principal component analysis (PCA) and similarity, which all generate quality evaluations and correlation classifications of Cinnamomi cortex, were used to improve the Cinnamomi cortex quality control standards. A total of 12 characteristic peaks were confirmed, seven of which were identified by comparing their retention times, UV and MS spectra with authentic compounds. Moreover, 11 analytes were accurately determined, as a complementary quantification method of chromatographic fingerprinting. For quantitative analyses, selective detection was performed at 254, 280 and 340 nm. The tested samples were separated and determined using UPLC and a series of methodologies including linearity, precision, accuracy, limit of detection and quantification and extraction recoveries were validated. Meanwhile the method bias for all the analytes did not exceed 5%. A total of 42 samples were acquired in China and analyzed. The results demonstrated that chromatographic fingerprinting in combination with chemometrics methods provides a promising and practical method to more effectively and comprehensively control the quality of Cinnamomi cortex from various sources, which would be a useful reference for the development and further study of Cinnamomi cortex and related formulations.

Yinchenhao Decoction Ameliorates Alpha-Naphthylisothiocyanate Induced Intrahepatic Cholestasis in Rats by Regulating Phase II Metabolic Enzymes and Transporters.

Yinchenhao Decoction (YCHD), a famous traditional Chinese formula, has been used for treating cholestasis for 1000s of years. The cholagogic effect of YCHD has been widely reported, but its pharmacodynamic material and underlying therapeutic mechanism remain unclear. By using ultra-high-performance liquid chromatography (UHPLC)-quadrupole time-of-flight mass spectrometry, 11 original active components and eight phase II metabolites were detected in rats after oral administration of YCHD, including three new phase II metabolites. And it indicated that phase II metabolism was one of the major metabolic pathway for most active components in YCHD, which was similar to the metabolism process of bilirubin. It arouses our curiosity that whether the metabolism process of YCHD has any relationship with its cholagogic effects. So, a new method for simultaneous quantitation of eight active components and four phase II metabolites of rhein, emodin, genipin, and capillarisin has been developed and applied for their pharmacokinetic study in both normal and alpha-naphthylisothiocyanate (ANIT)-induced intrahepatic cholestasis rats. The results indicated the pharmacokinetic behaviors of most components of YCHD were inhibited, which was hypothesized to be related to different levels of metabolic enzymes and transporters in rat liver. So dynamic changes of intrahepatic enzyme expression in cholestasis and YCHD treated rats have been monitored by an UHPLC-tandem mass spectrometry method. The results showed expression levels of UDP-glucuronosyltransferase 1-1 (UGT1A1), organic anion-transporting polypeptide 1A4 (OATP1A4), multidrug resistance-associated protein 2 (MRP2), multidrug resistance protein 1, sodium-dependent taurocholate cotransporter, and organic anion-transporting polypeptide 1A2 were significantly inhibited in cholestasis rats, which would account for reducing the drug absorption and the metabolic process of YCHD in cholestatic rats. A high dose (12 g/kg) of YCHD remarkably increased the expression of UGT1A1, bile salt export pump, MRP2, OATP1A4 in cholestasis rats presented it exhibited the greatest ameliorative effect on cholestasis, also particularly in histopathological examination and reducing levels of alanine transaminase, aspartate transaminase, total bilirubin, direct bilirubin, and total bile acid. Considering the metabolic process of bilirubin in vivo, the choleretic effect of YCHD is proven to be related to its regulatory action on expression of metabolic enzymes and transporters in cholestatic liver.

Simultaneous quantitation of 14 active components in Yinchenhao decoction with an ultrahigh performance liquid chromatography-diode array detector: Method development and ingredient analysis of different commonly prepared samples.

J. Sep. Sci. 2016, 39, 4147-4157 DOI: 10.1002/jssc.201600284 Yinchenhao decoction (YCHD) is a famous Chinese herbal formula recorded in the Shang Han Lun which was prescribed by Zhongjing Zhang during 150-219 AD. A novel quantitative analysis method was developed, based on ultrahigh performance liquid chromatography coupled with a diode array detector for the simultaneous determination of 14 main active components in Yinchenhao decoction. Furthermore, the method has been applied for compositional difference analysis of the 14 components in eight normal extraction samples of Yinchenhao decoction, with the aid of hierarchical clustering analysis and similarity analysis. The present research could help hospital, factory and lab choose the best way to make Yinchenhao decoction with better efficacy.

Simultaneous quantitation of 14 active components in Yinchenhao decoction by using ultra high performance liquid chromatography with diode array detection: Method development and ingredient analysis of different commonly prepared samples.

We developed a novel quantitative analysis method based on ultra high performance liquid chromatography coupled with diode array detection for the simultaneous determination of the 14 main active components in Yinchenhao decoction. All components were separated on an Agilent SB-C18 column by using a gradient solvent system of acetonitrile/0.1% phosphoric acid solution at a flow rate of 0.4 mL/min for 35 min. Subsequently, linearity, precision, repeatability, and accuracy tests were implemented to validate the method. Furthermore, the method has been applied for compositional difference analysis of 14 components in eight normal-extraction Yinchenhao decoction samples, accompanied by hierarchical clustering analysis and similarity analysis. The result that all samples were divided into three groups based on different contents of components demonstrated that extraction methods of decocting, refluxing, ultrasonication and extraction solvents of water or ethanol affected component differentiation, and should be related to its clinical applications. The results also indicated that the sample prepared by patients in the family by using water extraction employing a casserole was almost same to that prepared using a stainless-steel kettle, which is mostly used in pharmaceutical factories. This research would help patients to select the best and most convenient method for preparing Yinchenhao decoction.

Metabolism profiles of icariin in rats using ultra-high performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry and in vitro enzymatic study.

Icariin (ICA), the major constituent of Epimedium brevicornu Maxim, is recognized as an effective agent for the treatment of osteoporosis. In our previous report, the relatively short half-life (74min) and poor bioavailability (approximately 0.1%) of ICA in rats suggested that not only ICA itself but also its metabolites could be responsible for the observed osteoporosis treatment effect. Therefore, the present study aimed at identifying the metabolites of ICA in rat plasma, bile, urine, and feces after the administration of a single oral dose of ICA (150mg/kg). In this work, ultra-high performance liquid chromatography coupled with electrospray ionization quadrupole time-of-flight tandem mass spectrometry (UHPLC/Q-TOF-MS) method was established to identify the metabolic profiles following intragastric administration of pure ICA in rats. The blood, bile, urine, and feces samples of the rats were investigated to explore the complete metabolic pathway of ICA in vivo. A total of 14 metabolites were detected in the bile, revealing that the bile is the main excretion pathway for ICA and its metabolites. The conjugated metabolites as observed in vivo, was further confirmed by the in vitro enzymatic study. Five metabolites of ICA, including demethylicariin, icariside I-3-O-glucuronide, demethylicariside II, demethylicariside II-7-O-glucuronide, and dehydroxyicaritin-glucuronide, were reported for the first time in the literature. In addition, the results revealed that the principal metabolism pathways of ICA in rat were deglycosylation and glucuronidation after deglycosylation.

Stereoselective pharmacokinetic and metabolism studies of 20(S)- and 20(R)-ginsenoside Rg3 epimers in rat plasma by liquid chromatography-electrospray ionization mass spectrometry.

20(S)- and 20(R)-ginsenoside Rg3 are a pair of epimers which could be deglycosylated to ginsenoside Rh2 and protopanaxadiol (PPD) in vivo. To better understand the differences of pharmacokinetic parameters and metabolism behaviors of Rg3 epimers in rat plasma, a sensitive and specific liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) method was developed and fully validated. This chromatographic method separate 20(S)-/20(R)-Rg3, 20(S)-/20(R)-Rh2 and 20(S)-/20(R)-PPD by gradient elution of 10 mM ammonium acetate solution (pH 5.0) and acetonitrile on a C18 column with a total run time of 15 min. 20(S)-protopanaxatriol (PPT) was used as internal standard, and multiple reaction monitoring (MRM) mode with negative electrospray ionization were performed. The lower limit of quantitations (LLOQs) were between 4.2 and 4.8 ng/ml, and the accuracies were between 91.7% and 112.2% with intra- and inter-day precisions less than 11.6%. This method was successfully applied to a pharmacokinetic study of intravenous and intra-gastric administration of 20(S)-Rg3 and 20(R)-Rg3 to rats. It has been found that both epimers can be deglycosylated to their corresponding chiral metabolites, i.e., Rh2 and PPD, with different extents. However, 20(R)-Rg3 underwent single direction chiral inversion to 20(S)-Rg3 in rats. Stereoselective pharmacokinetic parameters, metabolic degrees and chiral inversion extents of Rg3 epimers in rats were also discussed for the first time.