Mobilized plasma lead as an index of lead body burden and its relation to the heme-related indices.

Plasma lead (Pb-P) from workers were distributed in two main fractions: a protein bound fraction and low molecular weight fractions. Lead mobilized into plasma by CaEDTA was mainly observed in the low molecular weight fraction corresponding to lead disodium ethylenediamine tetraacetic acid (PbEDTA). The peak levels of Pb-P was attained around 1.5 and 2.5 hours after the start of CaEDTA injection. Pb-P and blood lead levels (Pb-B) at 2 h after the injection were 4.26 (+/- 2.84) and 0.96 (+/- 0.27) fold of the initial levels just before the injection. Pb-P concentrations at 2 hours after the start of CaEDTA injection (MPb-P) were well correlated (r = 0.740) with amounts of lead excreted in urine for 24 h thereafter (MPb-U). log MPb-P as well as log MPb-U were correlated with Pb-B (r = 0.765 and 0.817, respectively). Correlation coefficients of lead body burden (MPb-P or MPb-U) vs the logarithms of the effect indices (delta-aminolevulinic acid (ALA) dehydratase, ALA in urine, coproporphyrin in urine, and erythrocyte zinc protoporphyrin) were higher than the correlation coefficients of exposure indices (Pb-B or Pb-U) vs the logarithms of the effect indices. Thus the biological effect monitoring is significant and reliable for evaluating the functional components of lead body burden (MPb-P or MPb-U).

Comparative study of Scutellaria planipes and Scutellaria baicalensis.

Scutellaria planipes, a species of Scutellaria, was explored by comparing to Scutellaria baicalensis, a pharmacopoeia species. Four principle flavonoids in both the plant roots were analyzed by using a reversed-phase chromatographic system with a chemically bonded ODS silica gel column and phosphate buffer:methanol (68:32 and 1:1) as mobile phase. Their contents were similar in both plant roots. Antiallergic and antibacterial activities in vitro and acute toxicity were compared. The results provided valuable data for S. planipes as a potential medicinal resource.

Studies on the constituents of roots of Scutellaria planipes.

From the roots of Scutellaria planipes (L.), three new flavonoids, 5,7,3',6'-tetrahydroxy-6,8,2'-trimethoxyflavone, (2 R,3 R)-3,5,7,2',5'-pentahydroxyflavanone and 7,2'-dihydroxy-5-methoxyflavone were isolated, together with ten known flavonoids, skullcapflavone I, wogonin, baicalein, norwogonin, viscidulin III, 5,7,2',6'-tetrahydroxy-flavone, 3,5,7,2',6'-pentahydroxyflavone, wogonin 7- O-beta- D-glucuronide, baicalin, baicalein 7- O-beta- D-glucopyranoside, and three phenethyl alcohol glycosides, acteoside, martynoside, and jionoside D. The structures were elucidated by spectroscopic methods.

Determination of schizandrin in human plasma by gas chromatography-mass spectrometry.

Schizandrin (SZ) is one of the lignan components from Schisandra fruits. A highly sensitive and precise method for the determination of SZ in human plasma was developed involving selected-ion monitoring with gas chromatography-mass spectrometry using a fused-silica capillary column. A 0.1-ml plasma sample was used for solid-phase extraction. A good linear relationship was obtained in the concentration range studied (2.0-500 ng/ml) and the method was sufficiently accurate and precise to support clinical pharmacokinetic studies. After oral administration of SZ at a dose of 15 mg to healthy male subjects, the average value of the maximum plasma concentration of SZ was 96.1 +/- 14.1 ng/ml. The plasma concentration of this substance could be monitored for 8 h after administration.

Three acyclic bis-phenylpropane lignanamides from fruits of Cannabis sativa.

Three new acyclic bis-phenylpropane lignanamides, named cannabisin E, F and G were isolated from the fruits of Cannabis sativa. Their structures have been elucidated based on spectral and chemical evidence.

Structure determination of biliary metabolites of schizandrin in rat and dog.

After oral administration of schizandrin (1) to rat, the bile was collected and treated with beta-glucuronidase and arylsulfatase. Eleven metabolites, SZ-M0 (3), SZ-M1 (4), SZ-M2 (5), SZ-M3 (6), SZ-M4 (7), SZ-M5 (8), SZ-M6 (9), SZ-M7 (10), SZ-M8 (11), SZ-M9 (12) and SZ-M10 (13) were isolated from the bile treated with the enzymes. The bile after oral administration of 1 to dog was collected and treated with enzymes in the same way, and eight metabolites, 3-8, 10 and SZ-MD2 (14) were isolated from the bile treated with enzymes. The structures of these metabolites were determined on the basis of chemical and spectral studies. The major metabolite in the bile of rat was 7 and the major metabolites in the bile of dog were 7 and 14.

Xanthone C-glycoside and acylated sugar from Polygala tenuifolia.

A new xanthone C-glycoside, polygalaxanthone III (1), and a new acylated sugar, tenuifoliside E (2) were isolated from the roots of Polygala tenuifolia. Their structures were characterized as 4-C-[beta-D-apiofuranosyl-(1-->6)-beta-D-glucopyranosyl]-1, 3,6-trihydroxy-7-methoxyxanthone (1) and beta-D-(1-O-acetyl-3-O-feruloyl-6-O-sinapoyl)-fructofuranosy l-alpha-D-(2,4,6- O-triacetyl)glucopyranoside (2), respectively, on the basis of chemical and spectral evidence including two dimensional nuclear magnetic resonance (2D-NMR) studies.

Studies on anti-allergic components in the roots of Asiasarum sieboldi.

The methanolic extract of the roots of Asiasarum sieboldi has been evaluated for anti-allergic effects on various experimental models in vitro and in vivo. Bioassay-guided fractionation of the methanolic extract resulted in isolation of suppressive compounds in the PCA test: methyleugenol (1), elemicin (2), gamma-asarone (3), (-)-asarinin (4), and (-)-sesamin (5). Furthermore, elemicin (2) and (2E,4E,8Z,10E)-N-isobutyl-2,4,8,10-dodecatetraenami de (6) were found to exhibit an inhibitory action on 5-lipoxygenase (5-LOX) from RBL-1 cells. 3',4'-Dimethoxycinnamaldehyde (7) and xanthoxylol (8) potently reduced the contractile response of guinea pig ileal strips to LTD4.

[Effect of hydrolysis conditions on the determination of urinary 2,5-hexanedione in workers exposed or not exposed to N-hexane].

In order to determine the optimal conditions of acid hydrolysis for urinary 2,5-hexanedione (HD) measurement, the effects of urine pH or volume of HCl added to urine and hydrolysis period were studied using gas chromatography-mass spectrometry (GC-MS). When 0.3 ml of concentrated HCl was added to 3 ml of urine, complete hydrolysis to liberate HD was attained 2 h after the start of heating at 100 degrees C for urine from non-exposed subjects. On the contrary, in urine from exposed subjects most of HD was liberated in 30 min at 100 degrees C, suggesting that the urinary substrates converted to HD by acid hydrolysis were different between exposed and non-exposed subjects. It was confirmed that hydrolysis for 2 h at 100 degrees C with 0.3 ml of concentrated HCl added to 3 ml of urine gave the most reliable levels and the greatest amounts of HD in both urine from exposed and non-exposed subjects. Reference values were determined by the hydrolysis conditions presented here. In 84 males, 52 females, and 136 with sexes combined the urinary HD levels were 0.35, 0.49, and 0.39 mg/l (geometric mean), respectively. Urinary HD levels determined without hydrolysis (free HD) were less than 0.006 mg/l in 31 control subjects examined. In urine from exposed subjects the amount of free HD was about one-fifth of the total HD (free plus conjugated HD) as determined with acid hydrolysates, although the percentage of free to total HD varied from 0.4 to 28%.(ABSTRACT TRUNCATED AT 250 WORDS)

Gomisin A inhibits tumor promotion by 12-O-tetradecanoylphorbol-13-acetate in two-stage carcinogenesis in mouse skin.

Gomisin A, isolated from the fruits of Schisandra chinensis, is one of the dibenzocyclooctadiene lignans. Application of 12-O-tetradecanoylphorbol-13-acetate (TPA, 1 microgram/ear), a tumor-promoting agent, to the ears of mice induces inflammation. Among seven dibenzocyclooctadiene lignans assayed, gomisin A, gomisin J, and wuweizisu C inhibited the inflammatory activity induced by TPA in mice. The ED50 of these compounds for TPA-induced inflammation was 1.4-4.4 mumol. Gomisin A, with an ED50 of 1.4 mumol, showed the strongest inhibitory effect. Furthermore, at 5 mumol/mouse, it markedly suppressed the promotion effect of TPA (2.5 micrograms/mouse) on skin tumor formation in mice following initiation with 7,12-dimethylbenz[a]anthracene (50 micrograms/mouse). It is assumed that the inhibition of tumor promotion by gomisin A is due to its anti-inflammatory activity.

[Studies on the metabolic fate of gomisin A (TJN-101). I. Absorption in rats].

Gomisin A (TJN-101) is one of the lignan components isolated from Schisandra Fruits and expected to have some efficacies in clinical treatment of hepatitis. The serum concentrations of TJN-101 and Met. B, which was identified as a demethylenated substance and one of the major metabolites of TJN-101 in rats, were investigated. After intravenous administration at doses of 1.6, 4.0 and 10 mg/kg of body weight, the serum concentration of TJN-101 decreased biphasically, and the terminal elimination half-life at each dose was about 70 min. Dose-dependency was observed for the area under the concentration-time curve (AUC). On the other hand, the serum concentration of TJN-101 increased rapidly and reached maximum within 15 to 30 min when administered orally. This result was supported by the in situ roop method. The Cmax and the AUC values were not exactly dose-dependent, but the values increased with a dose-up of TJN-101. The biotransformation of TJN-101 to Met. B, was very rapid in both intravenous and oral administrations. The AUC value of Met. B after oral administration of TJN-101 at a dose of 1.6 mg/kg was relatively larger than any other dosages. It suggested that TJN-101 was extensively underwent the first pass effect in rats. More than 80% of TJN-101 was bound with rat serum protein in vitro and in vivo. Therefore, it seems to be necessary to pay attention when it was administered concurrently with high protein binding drugs.

Studies on the metabolism of gomisin A (TJN-101). II. Structure determination of biliary and urinary metabolites in rat.

After oral administration of gomisin A (1) to rats, the bile and urine were collected and treated with beta-glucuronidase and arylsulfatase. Seven metabolites, met B (2), met A-III (3), met E (4), met D (5), met F (6), met G (7), and met H (8) were isolated from the bile treated with the enzymes. Eight metabolites 2-8, and met A-II (9) were isolated from the urine treated with the enzymes. A major metabolite 2, and two minor metabolites 3 and 9 were identified as met B, met A-III, and met A-II, respectively, which are oxidative products of 1 formed by rat liver S9 mix. The structures of five new metabolites 4-7, and 8 were determined on the basis of chemical and spectral studies.

[Method for simultaneous determination of six metabolites of toluene, xylene and ethylbenzene, and its application to exposure monitoring of workers in a printing factory with gravure machines].

For the biological monitoring of exposure to solvent composed of toluene, xylene, and ethylbenzene used in a printing factory with gravure machines, we developed a HPLC method for the simultaneous determination of urinary metabolites of this solvent, i.e. hippuric acid, o-, m-, and p-methylhippuric acid, mandelic acid and phenylglyoxylic acid. Except for phenylglyoxylic acid, urinary concentrations of the metabolites determined by the present method correlated well with the air concentrations of the respective solvent components. Hence the present method is useful in monitoring solvent exposure. In 91 workers of the printing factory and 53 control subjects, we also determined the concentrations of some phenolic metabolites and confirmed that o-cresol is a useful indicator for monitoring toluene exposure.

[Automated HPLC method for determining zinc protoporphyrin IX and protoporphyrin IX in erythrocytes of workers exposed to lead].

We developed an automated HPLC method for determining erythrocyte protoporphyrin. After a single extraction with N,N-dimethylformamide (DMF), zinc protoporphyrin IX (ZP) and protoporphyrin IX (PP) were separated by the column packed with a new type of reversed phase silica (capcell type) within 5 min. The column life was markedly extended to facilitate sample injection for about 1,000 times. The concentrations of ZP or TP (total protoporphyrin = 0.9 ZP + PP) determined by the present method correlated well with those determined by the conventional methods, i.e. by hematofluorometer (HF) or acid extraction method (FEP). In 150 workers exposed to lead, the ratio of 0.9 ZP/TP varied between 0.43 and 0.99 with the ratio being less than 0.6 in only 5 workers. The correlation coefficient between Pb-B and ZP was significantly higher than that between Pb-B and PP. Both anticoagulants, heparin and EDTA, could be used in the present method as well as in HF and FEP methods. Reference values for TP by HPLC were between 31.2 and 120.9 micrograms/dl RBC. Samples stored at 37 degrees C for 3 days were also used in the present method and the same results were obtained as in the samples stored at 4 degrees C.

[Improved high performance liquid chromatography determination of hippuric acid and methylhippuric acid isomers in urine].

A study was made on the effect of detection wavelength and separation mode of HPLC on determination of urinary hippuric acid (HA) and three isomers of methylhippuric acid (MHA). The interference of other constituents of urine in the determination was effectively decreased by detection at a short wavelength of 227.6 nm. Meta and para MHAs were separated by the addition of beta-cyclodextrine to the mobile phase. Four metabolites were successfully separated from other components of urine by the combination of ODS-silica packed-column and mobile phase (method F). The detection limits were found to be 50 and 5 mg/l for HA and MHAs, respectively. MHAs could not be detected in the non-exposed subjects. Average levels (+/- SD) of HA in non-exposed males and females were 272.2 (+/- 210.8) and 393.0 (+/- 269.8) mg/l, respectively. The urinary levels of HA in females were significantly higher than those in males.

[Pharmacological studies on ginger. V. Pharmacological comparison between (6)-shogaol and capsaicin].

Pharmacological actions of (6)-shogaol and capsaicin were studied. Both (6)-shogaol (0.5 mg/kg, i.v.) and capsaicin (0.1 mg/kg, i.v.) caused a triad such as a rapid fall in blood pressure, bradycardia and aponea in rats. Both drugs-induced marked pressor responses in blood pressure, which occurred after the rapid fall, were markedly reduced by a spinal destruction. In pithed rats, both drugs-induced peripheral pressor responses were markedly reduced with the combined treatment of [D-Arg1, D-Pro2, D-Trp7,9, Leu11]-substance P (0.5 mg/kg, i.v.), phentolamine (10 mg/kg, i.v.) and the section of sciatic nerves. In isolated guinea-pig trachea, (6)-shogaol (100 microM) and capsaicin (10 microM) induced contractile responses which were slightly inhibited by substance P antagonist (10 microM), but exhibited also a tachyphylaxis. Furthermore, although (6)-shogaol (3.6 microM) showed positive inotropic and chronotropic actions on isolated atria in rats, this effect of (6)-shogaol disappeared by repeated injections or pretreatment (100 mg/kg, s.c.) of (6)-shogaol. These results suggest that (6)-shogaol and capsaicin have similar actions, and that both drugs may cause a peripheral action by releasing an unknown active substance from nerve ends.

[Pharmacological studies on ginger. IV. Effect of (6)-shogaol on the arachidonic cascade].

(6)-Shogaol, a pungent component of ginger, which is contained in semi-dried ginger but is rarely found in fresh ginger inhibited carrageenin-induced swelling of hind paw in rats and arachidonic acid (AA)-induced platelet aggregation in rabbits. Moreover, (6)-shogaol prevented prostaglandin I2 (PGI2) release from the aorta of rats when tested as an inhibitor of platelet aggregation. These results suggest that (6)-shogaol may have an inhibitory action on the cyclo-oxygenases in both platelets and aorta. Examination of the effects of (6)-shogaol on cyclo-oxygenases in rabbit platelets and microsome fractions of rat aorta indicated that (6)-shogaol inhibited cyclo-oxygenase activities of both tissues in a concentration-dependent manner. Furthermore, when we examined the effect of (6)-shogaol on 5-lipoxygenase from RBL-1 cells, (6)-shogaol exhibited an inhibitory action on 5-lipoxygenase activity. Therefore, it seems that the inhibitory effects of (6)-shogaol on the carrageenin-induced paw edema, AA-induced platelet aggregation and PGI2 production of aorta may be caused by the inhibition of cyclo-oxygenase activity.