CHEMISTRY SUPPORT SERVICES TO THE ENVIRONMENTAL TOXICOLOGY PROGRAM

为了支持国家毒理学计划（NTP），在其毒物毒理学评价工作中，我们在2009财政年度完成了近100项化学任务。牙科复合材料是一种复杂的混合物，含有有机树脂基质，增强无机填料和硅烷偶联剂。已经表明，在放置特定的牙科密封剂后，诸如双酚A（BPA）和bis-GMA的化合物存在于唾液中，这些化合物显示出雌激素活性。在这方面，从三个不同的地方接收生物（人尿液和唾液）样本，用于分析物的测定：BPA及其可能的衍生物和代谢物双酚A甘油二甲基丙烯酸酯（bis-GMA）、双酚A二甲基丙烯酸酯（BPADM）、双酚A二缩水甘油醚（BADGE）、双酚A双（2，3-二羟丙基）醚（BPAHPE）和三（乙二醇）二甲基丙烯酸酯（TEGDMA）]。开发方法包括采用液相色谱-质谱检测（LC/MS）测定最低可定量基质浓度和评价基质干扰。在对NTP的新倡议的持续支持中，称为毒性测试的高通量筛选（HTS）计划，不同的任务正在进行中。 在本财政年度，对数百种化学品进行了溶解度测试，制备了10/20 mM溶液，并运送到不同的实验室进行高通量筛选实验。化合物三（2-氯异丙基）磷酸盐（TCPP：四种异构体的混合物），一种添加到家庭家具中使用的柔性聚氨酯泡沫中的阻燃剂，包括。对该化合物进行了以下研究：散装化学品有限分析，NTP-2000和NIH-07饲料和0.5%中的给药制剂开发研究，50，000; 40，000; 30，000 ppm下的14天适口性研究NTP-2000和NIH-07饲料中TCPP的浓度（μ g/g饲料）; Wistar Han大鼠和B6 C3 F1小鼠的nih 07。在本年度进行的离子液体（IL）的持续研究中，在水中进行了给药制剂开发研究，并对以下IL进行了散装化学再分析：1-丁基-1-甲基-吡咯烷鎓氯化物、1-丁基-3-甲基-咪唑鎓氯化物、1-乙基-3-甲基-咪唑鎓氯化物和N-丁基-吡啶鎓氯化物。为支持正丁基缩水甘油醚（BGE）的致癌性评价，进行了全面的化学分析和给药制剂开发研究。为了支持对天然产物的研究，gum guggul提取物（GGE）是从印度原产的植物Commiphora mukul（gum guggul）中分离出的几种类固醇-脂质的混合物，含有guggulsterone的Z-和E-异构体作为主要成分。为支持NTP研究，开发并验证了一种用于测定玉米油制剂中GGE的方法。为支持LUMI-CELL？ER检测国际验证研究的内部员工支持计划，采购、编码并运输化学品至日本、意大利和美国的实验室。根据上述工作人员支持计划，根据内部工作人员的要求，将不同化合物运送至实验室进行MCF-7雌激素受体细胞增殖试验。此外，根据GenTox测试计划，化学品被运送用于Salmonella测试。此外，还采购了25种化合物，根据要求制备溶液，并运送至实验室进行血液毒性试验。此外，为了支持内部人员支持计划，进行了该研究以测定4-苯基-3-丁烯酸（PBA）处理的前列腺癌细胞（PC-3）裂解物的可溶性和膜级分中的肽基-β-酰胺化单加氧酶（PAM）活性。以TNP-三肽为底物，用HPLC-UV法测定了甘氨酸延伸的中间体（TNP-D-Tyr-Val-β-hydroxy-Gly）和酰胺化的TNP-二肽（TNP-D-Tyr-Val-NH_2）。 根据产品浓度结果和细胞裂解物蛋白浓度，测定比活性。 每年进行动物房水检测。每三个月对正丁基缩水甘油醚进行一次过氧化物检测。每年都对化学和生物样品进行处理。 2009年15-19日在马里兰州巴尔的摩举行的第48届毒理学学会会议上展示了以下海报： ¿ NTP-2000和NIH-07啮齿动物饲料中磷酸三（氯丙基）酯混合物的分析方法验证 ¿ NTP-2000和NIH-07啮齿动物饲料中2-甲氧基-4-硝基苯胺的分析方法验证 ¿ 磷酸三氯丙酯（TCPP）在Wistar Han大鼠和B6 C3 F1小鼠中的初步适口性试验 ¿ 磷酸三（氯丙基）酯混合物、1-氯丙酮和N-乙酰基-L-半胱氨酸的提取和分析的分析方法开发

In support of National Toxicology Program (NTP), in their efforts of toxicological evaluations of toxicants, we have completed nearly 100 chemistry tasks were performed during FY2009. Dental composites are complex mixtures containing an organic resin matrix, reinforcing inorganic filler and a silane-coupling agent. It has been shown that the compound like bisphenol A (BPA) and bis-GMA were present in saliva after placement of a specific dental sealant that are shown to posses estrogenic activity. In this regard biological (human urine and saliva) samples were received from three different places for the assay of analytes: BPA and its possible derivatives and metabolites bisphenol A glycerolate dimethacrylate (bis-GMA), bisphenol A dimethacrylate (BPADM), bisphenol A diglycidyl ether (BADGE), bisphenol A bis (2,3-dihydroxypropyl) ether (BPAHPE), and tri (ethylene glycol) dimethacrylate (TEGDMA)]. The development method included the determination of the lowest quantifiable matrix concentration using liquid chromatography with mass spectrometric detection (LC/MS) and evaluation of matrix interferences. In an ongoing support of NTP¿s new initiative called ¿High Throughput Screening (HTS) program¿ for the toxicity testing different tasks are in progress. During the Fiscal Year hundreds of chemicals were tested for solubility, prepared 10/20mM solutions and shipped to different labs for high through-put screening experiments. The compound tris(2-Chloroisopropyl)phosphate (TCPP: mixture of four isomers), a flame retardant added to flexible polyurethane foam used in home furnishings includes. Following studies were performed for this compound: a bulk chemical limited analysis, dose formulation development study in NTP-2000 & NIH-07 feeds and in 0.5%,a 14-Day palatability study on at 50,000; 40,000; 30,000 ppm (¿g/g of feed) concentration of TCPP in NTP-2000 and NIH-07 feeds; nih07 for both Wistar Han rats and B6C3F1 mice. In continues studies on Ionic Liquids (IL) in the current year conducted Dose Formulation Development study in water and Bulk Chemical Reanalysis for the following IL¿s: 1-butyl-1-methyl-pyrrolidinium chloride, 1-butyl-3-methyl-imidazonium chloride, 1-ethyl-3-methyl-imidazonium chloride, and N-butyl-pyridinium chloride. To support the carcinogenic evaluation of n n-Butyl glycidyl ether (BGE), a comprehensive chemical analysis and dose formulation development study were performed. In support of studies on natural produces, gum guggul extract (GGE), a mixture of several steroid-lipids isolated from the plant Commiphora mukul (gum guggul) of Indian origin, containing Z- and E- isomers of guggulsterone as major constituents. In support of NTP studies a method was developed and validated for the assay of GGE in corn oil formulations. In support of the internal staff support program for LUMI-CELL¿ ER assay international validation study chemicals were purchased, coded, and shipped to labs in Japan, Italy and USA. Under above staff support program different compounds were shipped to the lab for MCF-7 estrogen receptor cell proliferation assay as requested by the internal staff. Also under GenTox Testing program chemicals were shipped for salmonella testing. Additionally 25 compounds are procured, prepared solutions as per the requirement and shipped to the lab for hemotoxicity assay. Additionally in support of the of the internal staff support program the study was conducted to determine the peptidyl-α-amidating monooxygenase (PAM) activity in soluble and membrane fractions of 4-phenyl-3-butenoic acid (PBA) treated prostate cancer cell (PC-3) lysates. With the TNP-tripeptide as substrate, the two reaction products, glycine-extended intermediate (TNP-D-Tyr-Val-α-hydroxy-Gly) and the amidated TNP-dipeptide (TNP-D-Tyr-Val-NH2) were determined using HPLC-UV. From the product concentration results and the cell lysate protein concentrations, the specific activity, was determined. Animal room water testing was conducted annually. Peroxide test on n-Butylglycidylether was performed every three months. Every year disposal of chemical and of biological samples was conducted. Following posters were presented in the 48th Society of Toxicology meeting, held in Baltimore, MD, during 15-19, 2009: ¿ Analytical Method Validation of Tris(chloropropyl) phosphate, Mixture in NTP-2000 and NIH-07 Rodent Diets ¿ Analytical Method Validation of 2-Methoxy-4-nitroaniline in NTP-2000 and NIH-07 Rodent Diets ¿ Preliminary Palatability Testing of Tris (chloropropyl) phosphate (TCPP) in Wistar Han Rats and B6C3F1 Mice ¿ Analytical Method Development for the Extraction and Analysis of Tris(chloropropyl)phosphate, mixture, 1-Chloroacetone and N-Acetyl-L-Cysteine