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Reliable, sensitive, rapid and quantitative enzyme-based assay for gamma-hydroxybutyric acid (GHB) 总被引:1,自引:0,他引:1
Several assays for gamma-hydroxybutyrate (4-hydroxybutyrate, GHB) have been developed based on the enzyme gamma-hydroxybutyrate dehydrogenase (GHB-DH). Enzymatic oxidation of GHB by NAD+ is coupled to diaphorase-mediated reduction of pro-dye to yield colored product. GHB-DH from Ralstonia eutropha was cloned and expressed as a stable fusion protein easily purified by affinity chromatography. Quantitative initial velocity and endpoint versions of the assay in solution are described. Michaelis-Menten parameters for oxidation of GHB and ethanol were estimated. A semi-quantitative "dipstick" version of the assay on paper also is described. Both solution endpoint and "dipstick" assays are sensitive to about 0.05 mg GHB/mL using 10 microL of sample. Ethanol at concentrations possible in urine and agents used to stabilize physiological fluids for forensics analysis do not interfere significantly. The "dipstick" assay also allows detection of GHB in alcoholic beverages after evaporation of about one-fourth drop of beverage before testing. The enzymatic assay for GHB is reliable, sensitive, inexpensive and rapid. 相似文献
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Genipin, the hydrolytic product of geniposide, which is extracted from gardenia fruit, shows good potential as a fingerprint reagent. It develops latent fingerprints on paper as blue impressions with good contrast and resolution. Even very faint impressions that are barely visible in ambient light will fluoresce brightly upon illumination at ca. 590 nm and are best viewed with a barrier filter above 630 nm. Potential advantages of genipin are the combination of colorimetric and fluorogenic activity in one reagent as well as its being a safe and environmentally friendly natural product. 相似文献
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Deana Dorman Logan Judith A. Calder Betty L. Cohen 《Journal of youth and adolescence》1980,9(3):263-269
A cultural tradition to acknowledge a girl's first menstrual period is proposed in order to overcome the negative connotation of the event. Three basic questions regarding such a tradition are addressed: Who would participate? What would it mean? What type of activity should it be? A research team consisting of a psychologist, a nurse, and a social worker suggests answers based on their interviews with mothers and daughters, as well as their collection of menarche anecdotes from women psychologists. 相似文献