A novel assessment technique of reproductive and genetic toxicity in domestic animals using a nuclear magnetic resonance instrument

在体内和体外磷-31和质子核磁共振（NMR）光谱的卵巢，肝脏，骨骼肌和胚胎的实验和农场动物获得使用自制的表面线圈，传统的圆形线圈，和/或成像线圈。在体内观察了代表高能态的β-ATP/无机磷酸盐比值和细胞内pH等。许多环境毒物的鉴定和定量没有任何提取过程，从这些器官和胚胎使用NMR光谱。质子NMR实验通过常规技术的改进来进行，并且31 P NMR实验如下进行：在NMR检查期间，将样品保持在25 ℃的受控空气温度下的探针室中。C.In.vivo完整动物体内稳态器官的31 P NMR光谱是使用配备有89-mm孔超导磁体（9.20特斯拉; Oxford Inst）的JNM-alpha-400 FT-NMR光谱仪（JEOL，Tokyo，Japan）获得的 ...更多信息 仪器，贝德福德，MA，USA）。将动物安装在表面线圈探针（两圈，2.0cm直径）上，该探针在161.7MHz下调谐到31 P，并与由计算机胶带条组成的铁氧体屏幕（Scotch 700 ; Three M Co.，St.Louis，MO）以消除由浅表肌肉产生的31 P NMR信号（7）。通过使用源自器官样本的水1H信号调整匀场参数来控制场均匀性。采集条件设置如下以获得完全弛豫信号：脉冲宽度为20微秒，脉冲重复时间为3.0秒，累积200次扫描。因此，每次测量花费5-25分钟。在指数乘法之后，对累积的自由感应衰减进行傅里叶变换。采用JEOL软件（JEOL）通过计算机积分计算单个光谱峰面积，并将磷酸肌酸（CP）峰用作位置参考（0 ppm）。通过与球形球中外部参比品（20 mM亚甲基二膦酸盐水溶液）的数据进行比较，将每种化合物的相对量转换为绝对浓度。本研究表明，NMR光谱是一种新的评估工具，用于评估实验动物和家畜的生殖和遗传毒性。少

In vivo and in vitro phosphorus-31 and proton nuclear magnetic resonance (NMR) spectra of the ovary, liver, skeletal muscle and embryo of experimental and farm animals were acquired using a house-made surface coil, conventional round coil, and/or imaging coils. Beta-ATP/inorganic phosphate ratio which represents the high energy state and intracellular pH etc.were observed in vivo. Many environmental toxicants were identified and quantified without any extraction processes from these organs and embryos using a NMR spectroscopy. Proton NMR experiments were done by a modification of conventional techniques, and 31P NMR experiments were performed as follows : During the NMR examination, the sample was maintained in a probe chamber at controlled air temperature of 25. C.In.vivo 31P NMR spectra of the steady-state organs within intact animals were obtained using a JNM-alpha-400 FT-NMR spectromenter (JEOL,Tokyo, Japan) equipped with 89-mm bore super-conducting magnet (9.20 Tesla ; Oxford Inst … More ruments, Bedford, MA,USA). The animal was mounted on a surface coil probe (two-turns, 2.0cm diameter) with was tuned to 31P at 161.7MHz and used in conjunction with a ferrite screen composed of strips of computer tape (Scotch 700 ; Three M Co., St.Louis, MO) to eliminate 31P NMR signals arising from superficial muscles (7). Field homogeneity was controlled by adjusting shim parameters using a water 1H signal derived from the organ sample. Acquisition conditions were set as follows to obtain fully relaxd signals : the pulse width was 20 micro-seconds, the pulse repetition time was 3.0s, and 200 scans were accumulated. Thus, it took 5-25 min for each measurement. After exponential multiplication, the accumulated free induction decays were subjected to Fourier transformation. Individual spectral peak areas were calculated by computer integration with JEOL software (JEOL), and the creatine phosphate (CP) peak was used as a positional reference (0 ppm). The relative amount of each compound was converted to absolute concentration by comparison with the data of an external reference (20mM methylene diphosphonate in water) in a spherical bulb. The present study showed that NMR spectroscopy is a novel assessment tools for assessment of reproductive and genetic toxicity in experimental and domestic animals. Less