This study aims to establish an ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS) method for the determination of emodin-8-O-β-D-glucoside(EG) and its metabolites in plasma, and to investigate the toxicokinetics(TK) behavior of them in rats. To be specific, the TK of EG and its metabolites from the first to the last administration in the repeated dose toxicity study was determined, and the kinetic parameters were calculated. The exposure of EG prototype and metabolites in rat plasma after oral administration of different doses of EG was evaluated. The result showed that the prototype of EG and its metabolites aloe-emodin-8-O-β-D-glucoside, emodin, aloe-emodin, and hydroxyemodin could be detected in rats after oral administration of high-, medium-, and low-dose EG. The area under the curve(AUC) of the prototype and metabolites after the first and last administration was in positive correlation with the dose. The time to the maximum concentration(T_(max)) of EG and metabolites in the three administration groups was <6 h, and the longest in vivo residence time was 12 h. The T_(max) and in vivo residence time of EG were prolonged with the increase in the dose. The metabolites emodin, aloe-emodin, and hydroxyemodin all had two peaks. Both hydroxyemodin and aloe-emodin exhibited increased plasma exposure, slow metabolism, and accumulation in vivo. In addition, aloe-emodin-8-O-β-D-glucoside and emodin disappeared with the increase in dose, suggesting the change of the metabolic pathway of EG in vivo in the case of high-dose administration. The mechanism of high-dose EG in vivo needs to be further explored. This study preliminarily elucidates the TK behavior of EG in rats, which is expected to support clinical drug use.
Animals ; Anthraquinones ; Chromatography, High Pressure Liquid/methods* ; Emodin/toxicity* ; Glucosides/toxicity* ; Mass Spectrometry ; Rats ; Toxicokinetics

Bones are stained into red color with feeding madder, but we do not know whether the fed madder can change the bone biomechanical properties and bone mineral contents in animals. In this research, we established a rat model with feeding madder. The bone biomechanical properties were detected by universal material mechanics, bone mineral contents were detected by inductively coupled plasma mass spectrometry and spectrometer, and red color material in bone was analyzed by high performance liquid chromatography. The results showed that bone biomechanical parameters in femur diaphysis in the 10% and 15% group rats were significantly higher than those in the control group after feeding madder for 6 months. The level of calcium, magnesium and zinc in femur diaphysis in 10% and 15% group rats were higher than those in the control group after feeding madder for 6 months. However, it was shown that the kidney congestion and hyperemia and the level of blood urea nitrogen and creatinine in the 15% group rats were significantly different compared to those in the control group rats after feeding madder for 6 months. The red colored material in bone is related to alizarin analyzed with high-performance liquid chromatography. The conclusion could be drawn that feeding 10% madder in diet was not toxic to the rats fed for 6 months, and it could improve bone biomechanical properties and increase bone mineral elements.
Animals ; Anthraquinones ; toxicity ; Biomechanical Phenomena ; Bone Density ; Bone and Bones ; drug effects ; physiology ; Calcium ; Femur ; Magnesium ; Rats ; Zinc

The bilirubin metabolism mediated by the phase Ⅱ metabolizing enzyme UGT1A1 in the liver was evaluated to study the potential hepatotoxicity risk based on investigation on the inhibitory effect of rhein and its metabolites on the UGT1A1 enzyme in Rhei Radix et Rhizoma. Firstly, in vitro liver microsomes incubation was used to initiate the phase Ⅱ metabolic reaction to investigate the inhibitory effect of rheinon UGT1A1 enzyme. Secondly, the phase Ⅰ and phase Ⅱ metabolic reactions were initiated to investigate the hepatotoxicity risk of rhein metabolites. It was found that the rhein and its phase Ⅱ metabolites had no significant inhibitory effect on UGT1A1 enzyme, but its phase Ⅰ metabolites significantly reduced UGT1A1 enzyme activity. Based on the metabolites analysis, it is speculated that the rhein phase Ⅰ metabolite rheinhydroxylate and its tautomers have certain hepatotoxicity risks, while the toxicity risk induced by the prototype and phase Ⅱ metabolites of rheinglucoside, rheinglucuronic acid and rhein sulfate is small.
Anthraquinones/toxicity* ; Chemical and Drug Induced Liver Injury ; Drugs, Chinese Herbal/toxicity* ; Glucuronosyltransferase/metabolism* ; Humans ; Liver/enzymology* ; Microsomes, Liver/drug effects* ; Rhizome

In this article, canonical correlation analysis was used to explore the relationship between the toxicity-attenuating effect and the variation of chemical contents in rhubarb caused by processing. With quasi-acute toxicity test, the difference of hepatic and renal toxicity to mice with the processed materials of rhubarb was researched. The chemical contents of anthraquinones and tannins in rhubarb were measured by UV-vis spectrophotometry and high performance liquid chromatography. The results showed that there were toxic effects to liver and kidney in mice after repeated intragastric administration of rhubarb and its processed materials for 14 days at a dosage of 76 g x kg(-1). The toxic effect of processed materials was much lower than crude drug. With canonical correlation analysis, the sequence of the hepatic and renal toxicity of chemical contents in rhubarb were found as follows: total anthraquinone glycosides (AQGs) > tannins (Tns) > total anthraquinones (AQs); aloe-emodin (AE) > physcione (Ph) > rhein (Rn) > emodin (Ed) > chrysophanol (Ch) and AEG > PhG > ChG > EdG > RnG of glycosyl-anthraquinone. It could be concluded that processing would attenuate the toxicity of crude drug of rhubarb. The toxicity-attenuating effect might be correlated to the decline of the contents of both anthraquinone glycosides and tannins, especially the aloe-emodin glycoside and physcione glycoside. The results also suggested that the serum alanine aminotransferase (ALT) and creatine (CREA) would be useful to monitor the hepatic and renal toxicity of rhubarb.
Alanine Transaminase ; blood ; Animals ; Anthraquinones ; analysis ; Creatinine ; blood ; Kidney ; drug effects ; Liver ; drug effects ; Mice ; Plant Roots ; chemistry ; Proanthocyanidins ; analysis ; Rheum ; chemistry ; toxicity ; Technology, Pharmaceutical ; methods

OBJECTIVETo compare the bidirectional effect of rhubarb total anthraquinone (TA) and total tannins (TT) on rats' liver.

METHODSOne hundred rats were randomly divided into 10 groups, i.e., the blank group, the model group, the blank + high dose TA group, the blank +low dose TA group, the blank + high dose TT group, the blank + low dose TT group, the model + high dose TA group, the model + low dose TA group, the model +high dose TT group, and the model + low dose TT group, 10 in each group. The carbon tetrachloride (CCI4) was used to prepare the acute liver injury rat model. TA and TT of rhubarb (at 5.40 g crude drugs/kg and 14.69 g crude drugs/kg) were intragastrically administrated to rats in all groups except the blank group and the model group, once daily for 6 successive days.The general state of rats, biochemical indices such as alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), laminin (LN), hyaluronic acid (HA), transforming growth factor beta1 (TGF-beta1), as well pathological results of rat liver tissues. Finally the protection laws of TA and TT for rats' liver were analyzed using factor analysis.

RESULTSCompared with the blank control group, all biochemical indices increased in the blank group (P < 0.05, P < 0.01). HA also increased in the blank + high dose TA group; AST, ALT, and HA also increased in the blank +high dose TT group (P < 0.05). Compared with the model group, AST, ALT, ALP, HA, and TGF-beta1 significantly decreased in the model + low dose TA group, the model + high dose TA group, the model + low dose TT group (P < 0.05, P < 0.01). Serum AST, ALT, and ALP also decreased in the model + high dose TT group (P < 0.05, P < 0.01). Pathological results showed that mild swollen liver cells in the model + high dose TA group. Fatty degeneration and fragmental necrosis around the central veins occurred in the blank + high dose TA group. The pathological injury was inproved in the model +low dose TA group. Two common factors, liver fibrosis and liver cell injury, were extracted by using factor analysis. TA showed stronger improvement of the two common factors than TT.

CONCLUSIONSRhubarb TA and TT showed protective and harmful effects on rats' liver. At an equivalent dosage, TA had better liver protection than TT. High dose TT played a role in liver injury to some extent.

Animals ; Anthraquinones ; adverse effects ; pharmacology ; Carbon Tetrachloride ; toxicity ; Chemical and Drug Induced Liver Injury ; drug therapy ; pathology ; Dose-Response Relationship, Drug ; Female ; Liver ; drug effects ; Male ; Rats ; Rats, Sprague-Dawley ; Rheum ; chemistry ; Tannins ; adverse effects ; pharmacology