Pharmacokinetics and metabolism of trans-emodin dianthrones in rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Polygonum multiflorum Thunb. (PM) is a common traditional Chinese medicine with diverse biological activities of resolving toxins, nourishing livers and promoting hairs. Nevertheless, in recent years hepatotoxic adverse reactions caused by the administration of PM have raised worldwide concerns. In our previous study, we found that emodin dianthrones showed hepatotoxicity and may be potential toxicity markers. However, the metabolic transformation and pharmacokinetic behavior of emodin dianthrones in vivo have still not been elucidated. AIM OF THE STUDY: Taking trans-emodin dianthrones (TED) as an example, the present study was conducted to investigate the pharmacokinetics and bioavailability of TED in rats and characterized its metabolic transformation in the plasma, urine and feces of rats. MATERIALS AND METHODS: A rapid and sensitive UPLC-qqq-MS/MS method was developed for accurate quantification of TED in plasma and successfully applied to the pharmacokinetic evaluation of TED in rats after intravenous and oral administration. A reliable UFLC-Q-TOF-MS high resolution mass spectrometry combined with a scientific metabolite identification strategy was used to comprehensively characterize the metabolic transformation of TED in plasma, urine and feces in rats. RESULTS: The established UPLC-qqq-MS/MS method had a linear range of 1-500 ng/mL, and the method was accurate and reliable to meet the quantitative requirements. When 20 mg/kg TED was given by gavage rats, it was rapidly absorbed into the circulatory system and had a long half-life time of 6.44 h and wide tissue distribution in vivo. While intravenous injection of 0.4 mg/kg TED in rats, it was rapidly metabolized and eliminated with a half-life time of 1.82 h. The oral absorption bioavailability of TED was only 2.83%. Furthermore with a sensitive UFLC-Q-TOF-MS technique and metabolite identification strategy, 21 metabolites were successfully identified, including 11 in plasma, 12 in urine and 18 in feces. The main Ⅰ and Ⅱ phase metabolic processes involved glucuronidation, oxidation, carbonylation, (de)methylation, sulfation and hydrogenation. CONCLUSION: TED could be rapidly absorbed into the blood circulation and widely distributed and slowly metabolized in the body and underwent extensive cleavage and metabolic transformation in vivo. The study provided a basis for in-depth elucidation of the toxicology and mechanism research of TED, but also laid the foundation for further research on the material basis of hepatotoxicity of PM.

Use of liquid chromatography hybrid triple-quadrupole mass spectrometry for the detection of emodin metabolites in rat bile and urine.

Emodin is the representative form of rhubarb, which is widely used in traditional Chinese medicine for the treatment of purgative, anti-inflammatory, antioxidative and antiviral, etc. Previous reports demonstrated that emodin glucuronide was the major metabolite in plasma. Owing to the extensive conjugation reactions of polyphenols, the aim of this study was to identify the metabolites of emodin in rat bile and urine. Neutral loss and precursor ion scan methods of triple-quadrupole mass spectrometer revealed 13 conjugated metabolites in rat bile and 22 metabolites in rat urine, which included four phase I and 18 phase II metabolites. The major metabolites in rat biosamples were emodin glucuronoconjugates. Moreover, rhein monoglucuronide, chrysophanol monoglucuronide and rhein sulfate were proposed for the first time after oral administration of emodin. Overall, liquid chromatography hybrid triple-quadrupole mass spectrometry analysis leads to the discovery of several novel emodin metabolites in rat bile and urine and underscores that conjugated with glucuronic acid is the main metabolic pathway.

[Analysis of emodin and its metabolites based on hollow fiber liquid phase microextraction].

Hollow fiber liquid phase microextraction (HFLPME) coupled with high performance liquid chromatography (HPLC) has been developed to analyze the emodin and its metabolites in plasma and urine samples. The abilities of the absorption and metabolism for the active components in traditional Chinese medicines between the male and female rats were compared, and the biological metabolism and transmutation of the analyte were detailed discussed. Emodin and its metabolites in plasma and urine samples were extracted into n-octanol (acceptor) in hollow fiber. The acceptor phase was dried and dissolved by 50 microL methanol and then analyzed by HPLC. Under the optimal conditions, the linearities of the analytes were all very good in biological samples (r > 0.9960), the detection limits of the analytes were within the ranges of 0.1-3.0 microg/L. The enrichment factors were 12.2 to 26.3. The relative standard deviations for intra-day and inter-day precision were lower than 11.0%. The average recoveries of the analytes in plasma and urine samples were all in the range of 97.9% to 103%. HFLPME-HPLC can eliminate interference from complex biological samples, improve the sensitivity and reduce the detection limit, thus this method is suitable for the determination of trace compounds in complex sample.

Use of cetyltrimethylammonium bromide as a simple probe for rapid determination of emodin by resonance light scattering technique.

A new resonance light scattering (RLS) method for emodin determination with cationic surfactant cetyltrimethylammonium bromide (CTAB) as probe has been developed. In Britton-Robinson buffer (pH 6.5) medium, emodin reacted with cationic surfactant CTAB and formed the emodin-CTAB complex. The complex aggregated together through hydrophobic forces and causing great enhancement of RLS signals with the maximum peak located at about 350 nm. The enhanced RLS intensities were found to be proportional to the concentration of emodin in the range of 0.54-9.72 µg ml(-1) with the detection limit (3σ) of 10.3 ng ml(-1). In this work, the characteristics of RLS, absorption, fluorescence spectra of the system were studied. The optimum reaction condition and the influencing factors on the RLS signal were investigated in detail. The proposed method was applied to the analysis of emodin in synthetic samples and human urine with satisfactory results. Furthermore, the forms of the substances under the experimental condition and the mechanism of the reaction were discussed in detail.

Comparison of dispersive liquid-liquid microextraction based on organic solvent and ionic liquid combined with high-performance liquid chromatography for the analysis of emodin and its metabolites in urine samples.

In this paper, two methods based on organic solvent dispersive liquid-liquid microextraction (OS-DLLME) and ionic liquid dispersive liquid-liquid microextraction (IL-DLLME) coupled with high-performance liquid chromatography have been critically compared for analyzing emodin and its metabolites (aloe-emodin, anthraquinone-2-carboxylic acid, rhein, danthron, chrysophanol and physcion) in urine samples. Several important parameters influencing the extraction recoveries of DLLME were carefully optimized. Under optimal conditions, the enrichment factors (EFs) for emodin and its metabolites by OS-DLLME and IL-DLLME were within the range of 90-295 and 63-192 respectively; the relative standard deviations (RSDs, n=3) for intra-day and inter-day precision were lower than 7.2 and 8.7% by OS-DLLME, and lower than 5.7 and 6.4% by IL-DLLME; the recoveries of emodin and its metabolites were from 87.1 to 105% for OS-DLLME and from 94.8 to 103% for IL-DLLME, respectively. There were no significant deviations between the two methods for the determination of emodin and its metabolites. From the results of HPLC/UV of urine sample after DLLME, the metabolites aloe-emodin, rhein, chrysophanol and physcion were identified by comparing the retention times with the standards. From the results of HPLC/MS, anthraquinone-2-carboxylic acid and danthron as unreported metabolites of emodin were found.

Mixed hemimicelles SPE based on CTAB-coated Fe3O4/SiO2 NPs for the determination of herbal bioactive constituents from biological samples.

In this paper, a solid-phase extraction (SPE) method based on mixed hemimicelles of cetyltrimethyl ammonium bromide (CTAB) on silica-coated magnetic nanoparticles (MNPs) is developed for extraction and preconcentration of compounds from the biological samples. We selected rhein and emodin which are the major active anthraquinones of rhubarb as model analytes. A high performance liquid chromatography-fluorescence detection (HPLC/FLD) method was developed for the determination of rhein and emodin in urine and serum samples. The main factors influencing the extraction efficiency including the amount of surfactant, the concentration of MNPs, the shaking time and the desorption ability of organic solvents were investigated and optimized. No interferences were caused by proteins or endogenous compounds in urine and serum samples. Good linearities (r(2)>0.9995) for all calibration curves were obtained, and the limits of detection (LODs) for rhein and emodin were 0.2 and 0.5 ng/mL in urine samples and 7 and 10 ng/mL in serum samples, respectively. Satisfactory recoveries (92.76-109.90% and 97.53-107.72% for rhein and emodin) in the biological matrices were achieved.

Simultaneous determination by UPLC-ESI-MS of scoparone, capillarisin, rhein, and emodin in rat urine after oral administration of Yin Chen Hao Tang preparation.

A simple, sensitive, and validated method was developed for simultaneous determination of scoparone, capillarisin, rhein, and emodin in rat urine by ultra-performance liquid chromatography/electrospray ionization quadruple time-of-flight mass spectrometry (UPLC-MS). The urinary samples were analyzed on an Acquity UPLC BEH C18 1.7 microm 2.1x50 mm column. Scoparone, capillarisin, rhein, and emodin in rat urine were simultaneously analyzed with good separation. The lower limits of detection were 6.0, 9.0, 7.0, and 3.0 ng/mL, and the lower limits of quantification were 20.0, 33.0, 24.0, and 12.0 ng/mL for scoparone, capillarisin, rhein, and emodin, respectively. The intra- and inter-day precisions (RSD) were less than 9%. The intra- and inter-accuracies were found to be in the range of 94.14-104.54% for scoparone, 101.72-107.34% for capillarisin, 95.24-103.59% for rhein, and 101.32-107.82% for emodin at three concentration levels. The absolute recoveries for scoparone, capillarisin, rhein, and emodin were not less than 77.0%. The developed method has been applied to determine scoparone, capillarisin, rhein, and emodin in rat urine after oral administration of Yin Chen Hao Tang preparation, a traditional Chinese medicine formulation widely used in China for treatment of jaundice and liver disorders.

Metabolism of 1,8-dihydroxy 3-hydroxy methyl anthraquinone (aloe-emodin) isolated from the leaves of Cassia tora in albino rats.

Cassia tora is a well known plant of India. Aloe-emodin was isolated from the leaves of this plant and its metabolism pattern was studied. The results showed that about 15.4% of the administered aloe-emodin was excreted and the rest was probably bound or metabolized in the system.

Polarographic and absorptive stripping voltammetric determination of mitoxantrone, emodin and chrysarobin using mercury and glassy carbon electrodes.

Polarographic and voltammetric methods were used to study drugs of anthrone derivatives: mitoxandrone (antineoplastic), emodin (cathartic) and chrysarobin (antipsoriatic) on mercury and glassy carbon electrodes. The surface-active properties were exploited for developing a highly sensitive adsorptive stripping voltammetric method for determination of trace amounts of these compounds. Effective pre-concentration was obtained at glassy carbon electrode, with the surface species being measured via its reduction or oxidation, respectively. It was concluded that observed cathodic and anodic currents had the diffusive character. A simple, quick and accurate methods for the determination of all studied compounds in pure form and mitoxantrone in vials have been developed. The acetate buffer pH 4.6 containing 5 to 30% ethanol was used as a supporting electrolyte. The linear calibration range was 0.5 = 100 microg cm-3 on mercury and 25 - 250 ng cm-3 on glassy carbon electrode.

A rapid high-performance thin-layer chromatographic urine screen for laxative abuse.

Laxative abuse, if unrecognized, can lead to unnecessary and costly investigations. The stimulants and cathartics are the most commonly abused laxatives and have the potential for causing the most long-term damage. We have optimized a solid-phase extraction coupled with high-performance thin-layer chromatography (HPTLC) to provide rapid, simple, and sensitive detection of phenolphthalein, danthron, rhein, aloin, and bisacodyl and its metabolites in urine. Positive screens have revealed laxative abuse in 3 of 42 samples from patients presenting with diarrhea of unknown origin.