BACKGROUND/AIMS: Our previous studies of ionization and solubility of unconjugated bilirubin (UCB) yielded inappropriately large differences between the two carboxylic pK'a values of UCB. These data, however, were not ideal due to crystal effects, matastability, impurities of the bilirubin, and imprecision of analyses at low UCB. METHODS: The sodium salt of taurocholate (TC) was purified and dissolved in water to 100 mM. Chloroform (CHCl3) was purified by vacuum distillation. Buffers used were: citrate from pH 4 to 6, phosphate from pH 6 to 8, and borate above pH 8. All had an ionic strength of 0.10. The problems were minimized by rapid solvent partition of UCB from CHCl3 into buffered aqueous NaCl, and a new, accurate assay of low UCB in the aqueous phase which was achieved by concentrating the UCB through back extraction into small volumes of CHCl3. RESULTS: In contrast with the crystal dissolution studies, the two pK'a value were similar. H2B0, not HB-, was the dominant UCB species in the pH range of bile (6.0 to 8.0). The aqueous solubilities of UCB were 90 to 98% less. Less than 0.01% of the bile salt partitioned into the CHCl3 phase and self-association of B= was negligible. UCB solubilities in 50 mM TC were 2 to 10% of those obtained by crystal dissolution, and, up to pH 7.9, were below the maximum UCB concentration in normal human bile. CONCLUSIONS: We suggest that the markedly increased binding of UCB with each ionization step is due to the disruption of the internal hydrogen bonds of the ionized carboxyl groups on interaction with the bile salt. We propose to extend the study of partition to determine the activity and the degradation products of calcium salts of unbound bilirubin fractions.
Bilirubin/*chemistry ; Chloroform ; English Abstract ; Hydrogen-Ion Concentration ; In Vitro ; Solubility ; Solvents ; Taurocholic Acid/*chemistry

OBJECTIVE: To investigate concentration changes of amylase in rabbits vitreous humor of acute pancreatitis associated with postmortem different interval. METHODS: To induce animal model of acute pancreatitis with sodium taurocholate, observed concentration changes of amylase after different postmortem interval, then compared with normal. RESULTS: The concentration of amylase in vitreous humor of rabbit 24h after death were related to postmortem interval significantly; The concluded formulae of the relationship between postmortem interval and amylase concentration is y=8.7420+0.7699x-0.0083x2 (R2=92.62792, F=14.89734, P=0.001). CONCLUSION Concentration changes of amylase in vitreous humor of acute pancreatitis associated with postmortem interval may provide a new sensitive and objective method for the forensic early injury time estimation.
Amylases/analysis* ; Animals ; Disease Models, Animal ; Female ; Male ; Pancreatitis/enzymology* ; Postmortem Changes ; Rabbits ; Spectrophotometry/methods* ; Taurocholic Acid ; Time Factors ; Vitreous Body/chemistry*

OBJECTIVETo provide evidence for three-level prevention of cholelithiasis by means of observing the effects of some choleretics on bile compositions drained from patients with pigment gallstone.

METHODSTwenty-seven patients suffering from primary pigment gallstones and having received treatment of choledochostomies plus T-tube or endoscopic nasal bile drainage (ENBD) were divided equally into three groups, and administered respectively with Lidanling (the LDL group), ursodesoxycholic acid (the UDA group) and combination of LDL and UDA (the LDL + UDA group) through oral intake (7 patients in each group). Besides, 6 post-operational patients got no treatment with any drug were allocated in the control group. Bile of all the patients was collected before treatment and on the 1, 3, 5, 7 th day after the treatment started to detect levels of total bile acid (TBA), glycocholic acid (GCA), taurocholic acid (TCA), glycocholic cheno-desoxycholic acid (GCDCA), total bilirubin (TBIL), uncombined bilirubin (UCB), concentration of calcium ion (Ca(2+)) as well as the bacterio-genetic and endogenous beta-glucuronidase activity for comparing.

RESULTSLevels of TBA, GCA, TCA and GCDCA got gradually increased in the UDA group and the LDL + UDA group after treatment (P < 0.05), while those in the LDL group remained unchanged, showing an insignificant difference as compared with those in the control group. In the LDL group and the LDL + UDA group, TBIL gradually increased while UCB gradually decreased in the course of treatment (P < 0.05). Moreover, levels of Ca(2+) and endogenous beta-glucuronidase activity got significantly lowered (P < 0.05).

CONCLUSIONCombined use of LDL and UDA could elevate levels of TBA, GCA, TCA, GCDCA, enhance the excretion of TBIL in patients with pigment gallstone after bile drainage, lower levels of UCB and Ca(2+) and the activity of endogenous beta-glucuronidase in the bile, so as to reduce the possibility of stone formation of bile, and therefore, it could be used to prevent the production of pigment gallstone, especially to prevent post-operative recurrence of stones.

Adult ; Bile ; chemistry ; Bile Acids and Salts ; analysis ; Bilirubin ; analysis ; Calcium ; analysis ; Cholagogues and Choleretics ; pharmacology ; Choledochostomy ; Cysteic Acid ; analogs & derivatives ; pharmacology ; Drainage ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Female ; Gallstones ; metabolism ; Glucuronidase ; analysis ; Glycocholic Acid ; analysis ; Humans ; Male ; Middle Aged ; Taurocholic Acid ; analysis ; Ursodeoxycholic Acid ; analogs & derivatives ; pharmacology

Tanreqing (TRQ), a traditional Chinese medicine (TCM) formula, can alleviate liver injury and improve liver function. Its pharmacological mechanisms of actions are still unclear due to its complex components and multi-target natures. Metabolomic study is an effective approach to investigating drug pharmacological actions, new diagnostic markers, and potential mechanisms of actions. In the present study, a new strategy was used to evaluate the protective effect of TRQ capsule against carbon tetrachloride (CCl)-induced hepatotoxicity in rats, by analyzing metabolic profiling of endogenous bile acids (BAs) along with biochemical and histological analyses. BAs concentrations were determined by ultra-performance liquid chromatography coupled with quadrupole mass spectrometry (UPLC-MS). Principal component analysis and partial least squares discriminant analysis were then employed to analyze the UPLC-MS results and compare the hepatoprotective effect of TRQ capsule in different groups at the doses of 0.36, 1.44, and 2.88 g·kg body weight, respectively. Moreover, our results suggested that taurocholic acid (TCA) and taurohyodesoxycholic acid (THDCA) were the most important biochemical markers, which were indicative of CCl-induced acute hepatic damage and hepatoprotective effect of TRQ capsule. Therefore, this new strategy would be an excellent alternative method for evaluating hepatoprotective effect and proposing potential mechanisms of action for other drugs as well.
Alanine Transaminase ; blood ; Animals ; Aspartate Aminotransferases ; blood ; Bile Acids and Salts ; blood ; metabolism ; Biomarkers ; blood ; Carbon Tetrachloride ; pharmacology ; Chemical and Drug Induced Liver Injury ; drug therapy ; metabolism ; pathology ; Chromatography, Liquid ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; therapeutic use ; Female ; Liver ; drug effects ; pathology ; Male ; Mass Spectrometry ; Metabolome ; drug effects ; Metabolomics ; Rats ; Rats, Wistar ; Taurocholic Acid ; blood ; Taurodeoxycholic Acid ; analogs & derivatives ; blood

Phyllanthus Urinaria L. (PUL) is a traditional Chinese medicine used to treat hepatic and renal disorders. However, the mechanism of its hepatoprotective action is not fully understood. In the present study, blood biochemical indexes and liver histopathological changes were used to estimate the extent of hepatic injury. GC/MS and LC/MS-based untargeted metabolomics were used in combination to characterize the potential biomarkers associated with the protective activity of PUL against CCl-induced liver injury in rats. PUL treatment could reverse the increase in ALT, AST and ALP induced by CCl and attenuate the pathological changes in rat liver. Significant changes in liver metabolic profiling were observed in PUL-treated group compared with liver injury model group. Seventeen biomarkers related to the hepatoprotective effects of PUL against CCl-induced liver injury were screened out using nonparametric test and Pearson's correlation analysis (OPLS-DA). The results suggested that the potential hepatoprotective effects of PUL in attenuating CCl-induced hepatotoxicity could be partially attributed to regulating L-carnitine, taurocholic acid, and amino acids metabolism, which may become promising targets for treatment of liver toxicity. In conclusion, this study provides new insights into the mechanism of the hepatoprotection of Phyllanthus Urinaria.
Amino Acids ; metabolism ; Animals ; Carbon Tetrachloride ; adverse effects ; Chemical and Drug Induced Liver Injury ; metabolism ; prevention & control ; Drugs, Chinese Herbal ; administration & dosage ; Gas Chromatography-Mass Spectrometry ; Humans ; Liver ; drug effects ; enzymology ; metabolism ; Male ; Metabolomics ; Phyllanthus ; chemistry ; Rats ; Rats, Sprague-Dawley ; Taurocholic Acid ; metabolism