OBJECTIVE: To determinate triptolide and wilforlide A in biological samples by liquid chromatography-tandem mass spectrometry (LC-MS/MS) method and to verify the method. METHODS: After 0.4 mL blood, urine or 0.4 g hepatic tissues with internal standard were extracted by ethyl acetate, they were separated on a Allure PFP Propyl (100 mm x 2.1 mm, 5 µm) with a mobile phase of methanol-20 mmol/L ammonium acetate using gradient elution. For mass spectrometric detection, electrospray ionization (ESI⁺) in positive mode was elected and the data was collected using multiple-reaction monitoring (MRM). RESULTS: The linearity was good (r > 0.995 0) and the limit of detection was 2 ng/mL or 2 ng/g for triptolide and wilforlide A. The recovery was 61.08%-102.98%. The intra-day and inter-day precision was less than 12.58% for each biological sample, and the accuracy was 90.61%-105.80%. CONCLUSION This method is simple, convenient and good selective, and could be applied to analysis of triptolide and wilforlide A in different biological samples. And the method may provide technical support for forensic medicine identification, clinical diagnosis and treatment of tripterygium wilfordii Hook. f. poisoning.
Chromatography, High Pressure Liquid ; Diterpenes/urine* ; Epoxy Compounds/urine* ; Humans ; Oleanolic Acid/urine* ; Phenanthrenes/urine* ; Sensitivity and Specificity ; Tandem Mass Spectrometry

OBJECTIVETo analyze the relationship between metabolites of polycyclic aromatic hydrocarbons (PAHs) and lung function in coke oven workers, and to provide scientific basis for further exploring the potential mechanism and developing the preventing strategies of the workers' early lung damage.

METHODSWe measured carbon monoxide, sulfur dioxide, benzene soluble matter, particulate matters, and PAHs at different workplaces of a coke oven plant. Detailed information on demography and occupational health condition of 912 workers were collected. We divided these workers into control group and coke oven group according to their workplaces and the different concentrations of COEs in the environment. We detected 10 urinary PAH metabolites and lung function using gas chromatography-mass spectrometry and spirometric tests, respectively.

RESULTSFEV(1.0) (91.12 ± 13.31) and FEV(1.0)/FVC (108.61 ± 20.37) of the coke oven group is significantly lower than the control group (94.16 ± 15.57, 113.45 ± 19.70). In the coke oven group, the hydroxyphenanthrene and 1-hydroxypyrene are negatively correlated with FEV(1.0)/FVC (β = -0.136, β = -0.100), Ptrend < 0.05 for all.

CONCLUSIONThe dose response decrease of lung function is associated with the urinary PAH metabolites in coke oven workers. Indicated that the long exposure to PAHs may cause the early lung damage in coke oven workers, phenanthrene and pyrene may be the main factors.

Adult ; Air Pollutants, Occupational ; urine ; Coke ; Humans ; Lung ; physiopathology ; Male ; Middle Aged ; Occupational Exposure ; analysis ; Phenanthrenes ; urine ; Polycyclic Aromatic Hydrocarbons ; urine ; Pyrenes ; urine ; Respiratory Function Tests

OBJECTIVETo observe the change of urinary monocyte chemottractant protein-1 (MCP-1) in patients with diabetic nephropathy (DN), and to explore the therapeutic effect and mechanism of triptolide (TL) in treating DN.

METHODSThirty-five patients in the treated group were treated with TL plus benazepril and thirty two patients in the control group were treated with benazepril alone for six months. The change of urinary MCP-1 was measured before and after treatment.

RESULTSLevel of urinary MCP-1 in DN patients was significantly higher than that in healthy subjects (P < 0.01), but it could be significantly decreased after TL treatment, showing significant difference as compared with that in the control group (P < 0.05).

CONCLUSIONDetermination of urinary MCP-1 level is beneficial to know the degree of kidney inflammation in DN patients. TL can inhibit inflammatory reaction to decrease the level of urinary MCP-1, and thus improve the renal function.

Adult ; Aged ; Chemokine CCL2 ; urine ; Diabetic Nephropathies ; drug therapy ; urine ; Diterpenes ; therapeutic use ; Epoxy Compounds ; Female ; Humans ; Immunosuppressive Agents ; therapeutic use ; Male ; Middle Aged ; Phenanthrenes ; therapeutic use ; Phytotherapy ; Prospective Studies

Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous environmental contaminants present in air and food. Among PAHs, benzo(a)pyrene(BaP), phenanthrene (PH) and pyrene (PY) are considered to be important for their toxicity or abundance. To investigate the changes of biomarkers after PAH exposure, rats were treated with BaP (150 microgram/kg) alone or with PH (4,300 microgram/kg) and PY (2,700 microgram/kg) (BPP group) by oral gavage once per day for 30 days. 7-ethoxyresorufin-O-deethylase activity in liver microsomal fraction was increased in only BaP groups. The highest concentration (34.5 ng/g) of BaP, was found in muscle of rats treated with BaP alone at 20 days of treatment; it was 23.6 ng/g in BPP treated rats at 30 days of treatment. The highest PH concentration was 47.1 ng/g in muscle and 118.8 ng/g in fat, and for PY it was 29.7 ng/g in muscle and 219.9 ng/g in fat, in BPP groups. In urine, 114-161 ng/ml 3-OH-PH was found, while PH was 41-69 ng/ml during treatment. 201-263 ng/ml 1-OH-PY was found, while PH was 9-17 ng/ml in urine. The level of PY, PH and their metabolites in urine was rapidly decreased after withdrawal of treatment. This study suggest that 1-OH-PY in urine is a sensitive biomarker for PAHs; it was the most highly detected marker among the three PAHs and their metabolites evaluated during the exposure period and for 14 days after withdrawal.
Adipose Tissue/chemistry/drug effects ; Animals ; Benzo(a)pyrene/analysis/metabolism/*toxicity ; Biological Markers/metabolism/urine ; Blood Chemical Analysis ; Body Weight/drug effects ; Cytochrome P-450 CYP1A1/metabolism ; Environmental Pollutants/blood/metabolism/*toxicity/urine ; Female ; Liver/drug effects/enzymology ; Lymphocytes/drug effects/metabolism ; Muscle, Skeletal/drug effects/metabolism ; Organ Size/drug effects ; Phenanthrenes/blood/metabolism/*toxicity/urine ; Pyrenes/analysis/metabolism/*toxicity ; Rats ; Rats, Sprague-Dawley ; Time Factors

AIMInvestigations on reducing the toxicity of triptolide through poly(D, L-lactic acid) nanoparticles as a drug carrier by oral administration to Wistar rats.

METHODSTriptolide-loaded poly (D, L-lactic acid) nanoparticles (TP-PLA-NPs) were prepared by modified spontaneous emulsification solvent diffusion (modified-SESD). The shape of nanoparticles was observed by transmission electron microscope (TEM). The size distribution and mean diameter were measured by laser light scattering technique. The entrapment efficiency and contents of drug loading were determined by RP-HPLC. The physical state of drug loaded in nanopartiles were primarily investigated by X-ray powder diffractometry. TP-PLA-NPs release behavior in vitro was carried out. After oral administration of the nanoparticles to Wistar rats in 15d, the toxicity for liver and kidney were studied by determining aspartate transaminase (AST), alanine transaminase (ALT) and blood urea nitrogen in serum and concentration of protein in urine.

RESULTSThe preparation process adapted to the formulation was as follows: the volume ratio of the aqueous and organic phases was 40/15; the surfactant concentration was 1%; the drug concentration was 0.3%; triptolide-PLA was 1:15 (w/w). The mean diameter was 149.7 nm and the polydispersity index was 0. 088 for the nanoparticles prepared by above conditions. The entrapment efficiency and content of drug loading were 74.27% and 1.36%, respectively. The release behavior of drug in vitro showed an initial burst effect, subsequently a slower rate stage. The results indicated that the liver toxicity (P < 0.01) and kidney toxicity (P < 0.05) caused by triptolide could be decreased significantly by nanoparticles carrier.

CONCLUSIONPLA-NPs might be used as a new oral carrier for triptolide.

Alanine Transaminase ; blood ; Animals ; Aspartate Aminotransferases ; blood ; Blood Urea Nitrogen ; Delayed-Action Preparations ; Diterpenes ; administration & dosage ; isolation & purification ; toxicity ; Drug Carriers ; Drug Delivery Systems ; Epoxy Compounds ; Lactic Acid ; Male ; Nanotechnology ; Particle Size ; Phenanthrenes ; administration & dosage ; isolation & purification ; toxicity ; Polyesters ; Polymers ; Proteinuria ; urine ; Rats ; Rats, Wistar ; Tripterygium ; chemistry