Histamine determination in tears as an index of safety in use of cosmetic products

To assess ocular irritancy caused by chemical and cosmetic products a reliable method based on evaluation of histamine (Hm) in tears is presented. Hm is measured at picomole levels by HPLC and fluorimetric detection after fluorescamine-HM derivatization. In order to avoid any uncontrolled irritation and stimulation of the conjunctiva during sample collection, a procedure of conjunctiva lavage was developed. A balanced salt solution (50 μl) containing a known amount of Hm-fluorophore as reference standard is instilled in the conjunctiva fornix. After a few seconds 20 μl of tear fluid is collected: 10 μl are immediately analysed and 10 μl after derivatization reaction. In this way it is possible to evaluate tear dilution and to assess Hm content in less than 10 minutes.
In a group of 20 normal subjects Hm has been determined in comparison with that of two volunteers after topical application of 50 μ of 0.2% and 0.4% sodium lauryl sulphate solution. A contact of 30 seconds of the cosmetic ingredient caused an immediate dose-dependent Hm release through a direct cytotoxic damage of cell membranes due to the surfactant action.
Évaluation de l'irritation occulaire due aux cosmétiques par la détermination de l'histamine lacrimale     

Validation of a Gas Chromatography-Mass Spectrometry Method for the Measurement of the Redox State Metabolic Ratios Lactate/Pyruvate and β-Hydroxybutyrate/Acetoacetate in Biological Samples

The metabolic ratios lactate/pyruvate and β-hydroxybutyrate/acetoacetate are considered valuable tools to evaluate the in vivo redox cellular state by estimating the free NAD+/NADH in cytoplasm and mitochondria, respectively. The aim of the current study was to validate a gas-chromatography mass spectrometry method for simultaneous determination of the four metabolites in plasma and liver tissue. The procedure included an o-phenylenediamine microwave-assisted derivatization, followed by liquid-liquid extraction with ethyl acetate and silylation with bis(trimethylsilyl)trifluoroacetamide:trimethylchlorosilane 99:1. The calibration curves presented acceptable linearity, with a limit of quantification of 0.001 mM for pyruvate, β-hydroxybutyrate and acetoacetate and of 0.01 mM for lactate. The intra-day and inter-day accuracy and precision were within the European Medicines Agency’s Guideline specifications. No significant differences were observed in the slope coefficient of three-point standard metabolite-spiked curves in plasma or liver and water, and acceptable recoveries were obtained in the metabolite-spiked samples. Applicability of the method was tested in precision-cut liver rat slices and also in HepG2 cells incubated under different experimental conditions challenging the redox state. In conclusion, the validated method presented good sensitivity, specificity and reproducibility in the quantification of lactate/pyruvate and β-hydroxybutyrate/acetate metabolites and may be useful in the evaluation of in vivo redox states.     

Female sex pheromone components ofHeliothis peltigera (Lepidoptera: Noctuidae)

Ten compounds were found in the sex pheromone glands ofHeliothis peltigera (Schiff) and identified as tetradecenal, (Z)-9-tetradecenal, (Z)-9-tetradecenol, (Z)-9-tetradecenyl acetate, hexadecanal, (Z)-7-hiexadecenal, (Z)-9-hexadecenal, (Z)-11-hexadecenal, (Z)-11-hexadecenol, and (Z)-11-hexadecenyl acetate. Behavioral tests in a wind tunnel and subsequent trapping studies conducted in the field indicated that (Z)-11-hexadecenal and (Z)-9-tetradecenal are the main pheromone components ofH. peltigera. Addition of (Z)-11-hexadecenol to the binary blend did not enhance the capture of males ofH. peltigera, but it decreased the number of males of the sympatricH. armigera. Rubber septa impregnated with a mixture of 2 mg (Z)-11-hexadecenal + 1 mg (Z)-9-tetradecenal + 0.6 mg (Z)-11-hexadecenol are recommended for monitoringH. peltigera. Contribution from the Agricultural Research Organization (ARO), No. 2454-E, 1988 series.     

Analysis of epoxidized soybean oil by gas chromatography

A fast and cost-effective procedure to quantitate epoxidized soybean oil by means of an external standard method is reported. This procedure is applicable to commercial epoxidized oils, polymer additive packages and polymers—polyvinyl chloride (PVC)—containing epoxidized oils. The epoxidized soybean oil is converted into fatty acid methyl esters with tetramethylammonium hydroxide, and analyzed by capillary gas chromatography with flame-ionization detection. In PVC samples, the epoxidized soybean oil was extracted with toluene and followed by derivatization prior to analysis. The methyl esters of monoepoxyoctadecanoic, diepoxyoctadecanoic and triepoxyoctadecanoic acid were separated with a short capillary column.     

邻苯二甲醛同步荧光衍生化法测定微量指血中还原型谷胱甘肽

采用微升（μL）级微量指血为检测样品,在未对样品去蛋白的条件下,用邻苯二甲醛（OPA）荧光衍生化法测定了微量静脉血中还原型谷胱甘肽（GSH）的含量.利用标准曲线法和标准加入法进行3次测定的均值分别为4.415μmol/L和5.417 5μmol/L,加标回收率为95.28%～97.18%,检测限为3.6×10-8 mol/L,可基本满足微量成分分析所要求的准确度.本研究为灵敏、简便和快速测定微量血样中GSH的含量提供了实验和方法学的依据.     

Tandem mass spectrometry in the study of fatty acids,bile acids,and steroids

Over the last 50 years, the mass spectrometry of lipids has evolved to become one of the most mature techniques in biomolecule analysis. Many volatile and non-polar lipids are directly amenable to analysis by gas-chromatography-mass spectrometry (GC-MS), a technique that combines the unsurpassed separation properties of gas-chromatography with the sensitivity and selectivity of electron ionization mass spectrometry. Less volatile and/or thermally labile lipids can be analyzed by GC-MS, following appropriate sample derivatization. However, many complex lipids are not readily analyzed by GC-MS, and it is these molecules that are the subject of the current review. Since the early 1970s, there have been three outstanding developments in mass spectrometry that are particularly appropriate in lipid analysis; i.e., the introduction of (i) fast atom bombardment (FAB); (ii) electrospray (ES); and (iii) tandem mass spectrometry (MS/MS). The FAB and ES ionization techniques will be discussed in relation to MS/MS, and examples of their application in biochemical studies will be presented. The review will concentrate on the analysis of fatty acids, bile acids, steroid conjugates, and neutral steroids.     

柱前衍生法在5(6)-羧基罗丹明异构体分离中的应用

利用柱前衍生法对5(6)-羧基-2',7'-二甲基罗丹明6G混合物进行了分离,通过~1HNMR、~(13)CNMR和元素分析对两种异构体的结构进行了表征。荧光光谱测试显示,两种异构体的最大发射波长相差11 nm。该方法操作简单,收率高,纯度好,适用于分离极性较大的5(6)-羧基罗丹明混合物。     

Analysis of triclosan and 4n‐nonylphenol in Colombian reservoir water by gas chromatography‐mass spectrometry

In this work were determined triclosan and 4n‐nonylphenol in water from a reservoir that is used to provide water to a purification plant in an important city in Colombia. The analytical methodology was validated using solid‐phase extraction and analysis by gas chromatography‐mass spectrometry (GC‐MS). The analysis by GC‐MS showed good linearity in the range of 0.05–5 μg/L for both compounds. Recoveries from 79 to 109% and standard deviations of 2.5–7.7 for low concentrations and from 3.8 to 9.6 (n = 5) for high concentrations were obtained for both compounds. In Colombia, this is the first time that these compounds have been analysed in water supplying of a drinking water treatment plant. The validated method was applied to the analysis of 27 samples collected in August 2010 in 11 locations from the reservoir and in the influent and effluent of the drinking water treatment plant. In total, seven samples were found to contain triclosan.     

免疫亲和层析净化高效液相色谱法测定澳洲坚果中黄曲霉毒素B_1

建立了一种测定澳洲坚果中黄曲霉毒素B1的免疫亲和层析净化高效液相色谱法。采用甲醇-水提取,提取液经过滤、水稀释、净化,甲醇洗脱溶解,柱后衍生液相色谱仪荧光检测器测定。实验结果表明,澳洲坚果中黄曲霉毒素B1的检出限为0.2μg/kg,在添加水平为5、10、20μg/kg的回收率实验中,平均回收率为81%~96%,相对标准偏差为0.8%~3.7%。     

柱前衍生-高效液相色谱法检测安立生坦的手性拆分剂L-脯氨酸甲酯

采用4-氯-7-硝基苯-2-（嗯）-1,3-二唑（NBD-Cl）作为柱前衍生试剂,建立了柱前衍生化-高效液相色谱法（HPLC）结合荧光检测器测定安立生坦合成过程中使用的手性拆分剂L-脯氨酸甲酯的方法.考察了衍生反应温度、时间、衍生试剂加入量、缓冲液pH对L-脯氨酸甲酯衍生反应的影响.结果表明,衍生反应优化条件为：反应温度60℃,反应时间60 min,衍生试剂加入量为每毫升反应体系中加入0.3 mg NBD-Cl,衍生反应缓冲液pH为9.0.同时对此方法进行了方法学验证,L-脯氨酸甲酯在0.625～7.5 μg/mL范围内线性关系良好,r=0.999 4;检测限为2.5×10-3μg/mL,定量限为7.5×10-3μg/mL;平均回收率为91.9％～ 106.4％;重复性实验中RSD为0.33％;此方法耐用性良好,L-脯氨酸甲酯样品溶液及衍生产物溶液体系稳定.此方法灵敏、简便、专属性强,能对安立生坦原料药中残留的杂质L-脯氨酸甲酯进行检测及定量分析.