OBJECTIVETo establish a RP-HPLC method for simultaneous determination of thymine, hypoxanthine and uracil contents in medical pipefish.

METHODSamples were extracted with distilled water by ultrasonic wave and separated on Waters C18 column eluted with a mobile phase of 0.05 mol x L(-1) KFI2PO4-acetonitrile (97:3). The flow rate was 0.6 mL x min(-1). The determination wavelength was 260 nm and the column temperature was set at 40 degrees C.

RESULTThe method had good linearity in the range of 0.033-0.660 (r = 0.9996), 0.620-12.400 (r = 0.9999), 0.048-0.960 microg (r = 0.9995), with average recoveries of 98.67% (RSD 1.6%), 99.03% (RSD 0.74%), 98.65% (RSD 1.8%), for thymine, hypoxanthine and uracil respectively.

CONCLUSIONThe simultaneous determination method of thymine, hypoxanthine and uracil in medical pipefish is established by RP-HPLC for the first time. The contents of the three constituents in different kinds of medical pipefish are significantly different. The method is simple, rapid and sensitive, and can be used for control the quality of medical pipefish.

Animals ; Chromatography, High Pressure Liquid ; methods ; Hypoxanthine ; analysis ; Medicine, Chinese Traditional ; Smegmamorpha ; Thymine ; analysis ; Uracil ; analysis

OBJECTIVETo examine the quality of Whitmania pigra from the different populations to provide the basis for new species selection.

METHODThe contents of the moisture, alcohol extractive, total ash, acid-insoluble ash, and the activity of antiplatelet aggregation enzyme of wild and the breeding W. pigra were determined by the methods were determined in Chinese Pharmacopoeia (2005 edition). The contents xanthine and hypoxanthine by HPLC.

RESULTIt showed that the contents of the alcohol extractive, total ash, acid-insoluble ash, antiplatelet aggregation enzyme, and xanthine and hypoxanthine content of the breeding population in Nanjing are lations. Many quality indexes of the breeding in Zhejiang Tongxiang base were beffer than the wild.

CONCLUSIONThe breeding W. pigra are good in quality, It suggested large-scale promotion of aquaculture.

Animals ; Breeding ; Female ; Hypoxanthine ; analysis ; Leeches ; chemistry ; physiology ; Male ; Xanthine ; analysis

This study is to establish a method for simultaneously determination of five nucleosides and nucleobases, including hypoxanthine, uridine, adenine, guanosine and adenosine in Rehmannia glutinosa Libosch. which was collected from different regions in China. A Diamonsil C18 column (250 mm x 4.6 mm, 5 microm) was used. Acetonitrile and 0.04 mol L(-1) potassium dihydrogen phosphate solution were adopted as mobile phase with gradient elution. The flow rate was 1 mL min(-1) and column temperature was 30 degrees C. The detection wavelength was at 254 nm. The method had good linearity over the range of 1.0 - 16.0 microg mL(-1) (r2 = 0.999 8), 5.0 - 80.0 microg mL(-1) (r2 = 0.999 8), 1.0 - 16.0 microg mL(-1) (r2 = 0.999 5), 1.25 - 20.0 microg mL(-1) (r2 = 0.999 8) and 1.0 - 16.0 microg mL(-1) (r2 = 0.999 8) for hypoxanthine, uridine, adenine, guanosine and adenosine, respectively. The average recoveries were between 98.8% and 100.7%. The content of hypoxanthine, uridine, adenine, guanosine and adenosine in Rehmannia glutinosa Libosch. from different regions was significantly different. This established method was sensitive and reliable for the quantification of five chemical constituents in Rehmannia glutinosa Libosch.
Adenine ; analysis ; Adenosine ; analysis ; Chromatography, High Pressure Liquid ; Guanosine ; analysis ; Hypoxanthine ; analysis ; Nucleosides ; analysis ; Plants, Medicinal ; chemistry ; Rehmannia ; chemistry ; Uridine ; analysis

OBJECTIVE: To investigate the relationship between postmortem interval (PMI) and concentration changes of components in swine vitreous humor. METHODS: Ninety-six porcine eyes from swine dying from acute massive hemorrhage, being randomly divided into 24 groups, were stored in dark situation, at temperature of (15 +/- 2) degrees C and humidity of (50 +/- 5)% for 2-96 hours separately. The vitreous humor was collected. Concentrations of K+, Na+, Cl- and hypoxanthine (Hx) were analyzed by automatic biochemical analyzer and ultra performance liquid chromatograph (UPLC). The data were statistically analyzed by SPSS software. RESULTS: Linear regression analysis showed that concentrations of vitreous K+ and Hx were positively correlated with PMI(R2=0.767 and R2 = 0.793, respectively). Binary linear regression showed a higher correlation for K+ and Hx with PMI estimation (R2 = 0.866). PMI was not significantly correlated with vitreous Na+ and Cl- concentrations. CONCLUSION Vitreous K+ and Hx concentrations can be used as the objective markers for PMI estimation. The binary linear regression functions of vitreous K+ and Hx concentrations with PMI are more accurate for estimating the PMI.
Animals ; Chromatography, High Pressure Liquid/methods* ; Female ; Forensic Pathology ; Hypoxanthine/analysis* ; Male ; Postmortem Changes ; Potassium/analysis* ; Regression Analysis ; Sodium/analysis* ; Swine ; Temperature ; Time Factors ; Vitreous Body/chemistry*

A new method based on high performance liquid chromatography-electrospray ionization time of flight-mass spectrometry (HPLC-ESI-TOF/MS) was developed for the rapid identification of active compounds in Styela clava and the development of its specific chromatograms. Samples were extracted by ultrasonic-assisted extraction, and the extraction conditions were optimized. The developed HPLC-ESI-TOF/MS method was used to identify the components in Styela clava extract, and a specific chromatogram based on HPLC analysis was established. Ten compounds in Styela clava extract have been primary identified by HPLC-ESI-TOF/MS on-line detection combined with literature review. The result of similarity evaluation for specific chromatograms indicated that the quality of different Styela clava samples was not entirely consistent. This method has the advantages of simple operation, rapid measurement and it is a powerful tool for identification of active components in Styela clava and its quality control.
Animals ; Chromatography, High Pressure Liquid ; methods ; Hypoxanthine ; analysis ; Quality Control ; Spectrometry, Mass, Electrospray Ionization ; methods ; Tyrosine ; analysis ; Uridine ; analysis ; Urochordata ; chemistry

OBJECTIVETo evaluate the effects of processing methods of Hirudo.

METHODBoth water and alcohol extracts of Hirudo were studied according to Chinese Pharmacopeia (Edition 2005). The content of hypoxanthine in Hirudo was measured by high performance liquid chromatography (HPLC) and Hirudin was determined by thrombin.

RESULTThe contents of water, water soluble extraction, ethanol soluble extraction and hirudin in crude Hirudo are higher than those in processed Hirudo. But the contents of hypoxanthine in processed Hirudo is higher than in crude Hirudo.

CONCLUSIONHirudo fried by French chalk can decrease the active components with high intensive drug property, accordingly the toxicity of Hirudo was decreased. As a result, the effects of Hirudo as invigorate the circulation of blood and stimulate the menstrual flow were abated.

Animals ; Blood Circulation ; drug effects ; Ethanol ; chemistry ; Hirudins ; analysis ; Hirudo medicinalis ; chemistry ; Hypoxanthine ; analysis ; Menstrual Cycle ; drug effects ; Solubility ; Water ; chemistry

OBJECTIVETo study the chemical constituents of the leaves of Aquilaria sinensis.

METHODThe compounds were isolated and purified by the methods of solvent extraction and chromatographic technique, and their structures were identified on the basis of the analyses of spectral data.

RESULTThirty-three compounds were obtained. Among them, twelve compounds were identified as 5-hydroxyl-7,4'-dimethoxyflavone (1), acacetin (2), luteolin (3), genkwanin (4), yuankanin (genkwanin-5-O-beta-D-primeveroside, 5), adenosine (6), genkwanin-5-O-beta-D-glucopyranoside (7), hypolaetin-7-O-beta-D-glucopyranoside (8), hypoxanthine (9), uracil (10), 8-C-beta-D-galactopyranosylisovitexin (11), and 4-(1,2,3-trihydroxypropyl) -2,6-dimethoxyphenyl-1-O-beta-D-glucopyranoside (12), respectively.

CONCLUSIONAll compounds except for 1, 3 and 4 were isolated from the leaves of A. sinensis for the first time.

Adenosine ; analysis ; isolation & purification ; Chromatography ; methods ; Flavones ; analysis ; isolation & purification ; Glycosides ; analysis ; isolation & purification ; Hypoxanthine ; analysis ; isolation & purification ; Luteolin ; analysis ; isolation & purification ; Plant Extracts ; analysis ; isolation & purification ; Plant Leaves ; chemistry ; Thymelaeaceae ; chemistry

OBJECTIVETo evaluate the genotoxic and nongenotoxic effects of short-term exposure to glycidyl mathacrylate (GMA) on human lung fibroblast cells (2BS cells) in vitro.

METHODSDNA strand breakage was determined by single cell gel electrophoresis, and DNA ladder formation assay and flow cytometric analysis were carried out to detect apoptic responses of cells to GMA exposure. The HPRT gene mutation assay was used to evaluate the mutagenicity, and the effect of GMA on gap junctional intercellular communication (GJIC) in the exposed cells was examined with the scrape loading/dye transfer technique. The ability of GMA to transform 2BS cells was also tested by an in vitro cell transformation assay.

RESULTSExposure to GMA resulted in a dose-dependent increase in DNA strand breaks but not apoptic responses. GMA was also shown to significantly induce HPRT gene mutations and morphological transformation in 2BS cells in vitro. In contrast, GMA produced a concentration-dependent inhibition of GJIC.

CONCLUSIONSGMA elicits both genotoxic and nongenotoxic effects on 2BS cells in vitro. The induction of DNA damage and gene mutations and inhibition of GJIC by GMA may casually contribute to GMA-induced cell transformation.

Cell Communication ; Cell Differentiation ; Comet Assay ; DNA Damage ; DNA Mutational Analysis ; Epoxy Compounds ; toxicity ; Fibroblasts ; Gap Junctions ; Humans ; Hypoxanthine Phosphoribosyltransferase ; genetics ; Lung ; cytology ; Methacrylates ; toxicity

Potential toxicological interactions of 4-(N-methyl-N-nitrosamino)-1-(3-pyridyl)-1-butanone (NNK) and/or dibuthyl phthalate (DBP) on ozone were investigated after 32- and 52-wk exposures using hprt mutation assay. Male and female B6C3F1 mice exposed to ozone (0.5 ppm), NNK (1.0 mg/kg), DBP (5,000 ppm), and two or three combinations of these toxicants 6 h per day for 32- and 52-wk showed increases in the frequencies of TG rlymphocytes compared to the control groups. Additive interactions were noted from two combination groups compared to the ozone alone in both sexes of 32- and 52-wk studies. The most common specific mutation type in the hprt genes of test materials-treated male and female mice was transversion with very few transition. The results indicate that such dominant transversion may be responsible for toxicity and combined exposure to ozone, NNK, and DBP induces additive genotoxicities compared to ozone alone.
Animals ; Carcinogens/*toxicity ; DNA Mutational Analysis ; Dibutyl Phthalate/*toxicity ; Drug Combinations ; Female ; Hypoxanthine Phosphoribosyltransferase/*genetics ; Male ; Mice ; Mutagenicity Tests ; *Mutation/drug effects ; Nitrosamines/*toxicity ; Ozone/*toxicity ; Reverse Transcriptase Polymerase Chain Reaction ; T-Lymphocytes/drug effects/enzymology

The Lesch-Nyhan syndrome which is caused by the deficiency of hypoxanthine guanine phosphoribosyltransferase is an X-linked recessive disorder characterized by hyperuricemia, choreoathetosis, mental retardation and compulsive self-injurious behavior. Clinical management of the patients with the Lesch-Nyhan syndrome is frustrating and requires burdensome medical treatment since it cripples the patient and shortens the life span by progression of neurological symptoms, but there are no cures or measures for relieving relentless natural course of the disease yet. Therefore, prenatal diagnosis of the affected fetus is important in genetic counselling for the family at high risk. In this study, four different mutations in the HPRT gene of four probands have been identified in four unrelated families; K215X, Q109X, nt.631 A, and nt.289 GT. Two mutations among them altered restriction enzyme sites; SpeI for Q109X and MaeI for nt.289 GT. Based on their molecular defects, prenatal diagnoses of 3 the fetuses were successfully made between ninth and eleventh week of gestation by polymerase chain reaction(PCR), restriction digestion and DNA sequencing using cDNA obtained from chorionic villus samples (CVS). We predicted the outcome of all fetuses prenatally. Among the three fetuses two were male and one was female according to the identification made by PCR amplification of the sex determining region of the Y chromosome(SRY) gene. Each carried a wild type allele for the corresponding mutant allele. They were also tested postnatally for the mutations to be unaffected.
Alleles ; Chorionic Villi ; Digestion ; DNA, Complementary ; Female ; Fetus* ; Humans ; Hyperuricemia ; Hypoxanthine Phosphoribosyltransferase ; Intellectual Disability ; Lesch-Nyhan Syndrome* ; Male ; Polymerase Chain Reaction ; Pregnancy ; Prenatal Diagnosis ; Self-Injurious Behavior ; Sequence Analysis, DNA