公开(公告)号：US20080199848A1

公开(公告)日：2008-08-21

申请号：US11915828

申请日：2006-05-13

申请人： Stefanie M. Bode-Boger ,  Jens Martens-Lobenhoffer

发明人： Stefanie M. Bode-Boger ,  Jens Martens-Lobenhoffer

IPC分类号： C12N9/00 ,  G01N33/68

CPC分类号： G01N33/6812

摘要： The invention relates to a method for determining the concentration of asymmetric dimethylarginine (ADMA) simultaneously with arginine and with symmetric dimethylarginine (SDMA) in biological samples by means of HPLC-MS-MS. The sample preparation consists exclusively of adding a solution of isotope-labeled internal standards (13C6-argininc and D6-ADMA) and of adding a mixture consisting of acetonitrile/propionic acid/trifluoroacetic acid for precipitating high-molecular proteins, the quantity and composition being realized in such a manner that a bringing of the sample composition in line with the composition of the mobile phase in the HPLC separation is achieved without requiring a derivitization or extraction of the analytes. The chromatographic separation of the analytes ensues on a silica normal phase HPLC column while using a mobile phase consisting of water/acetonitrile/propionic acid/trifluoroacetic acid, the volume ratio of water to acetonitrile ranging from 2 to 98 to 30 to 70, the volume percent of trifluoroacetic acid ranging from 0.01 to 0.5, the volume percent of propionic acid being 10 to 100 times higher than trifluoroacetic acid, and the optimized composition of the mobile phase consists of acetonitrile/propionic acid/trifluoroacetic acid with 10/90/1/0.025 volume percents. The detection and quantification ensue by means of tandem mass spectrometry with the following fragmentations being observed: 175.2 m/z→70.1 m/z for arginine, 181.2 m/z→74.1 m/z for 13C6-arginine, 203.2 m/z→172.1 m/z for SDMA, 203.2 m/z→46.1 m/z for ADMA and 209.2 m/z→70.1 m/z D6-ADMA.

摘要翻译： 本发明涉及通过HPLC-MS-MS在生物样品中与精氨酸和对称二甲基精氨酸（SDMA）同时测定不对称二甲基精氨酸（ADMA）的浓度的方法。 样品制备仅仅包括加入同位素标记的内部标准溶液（ C 6+金属和D 6 -ADMA）和 加入由乙腈/丙酸/三氟乙酸组成的混合物用于沉淀高分子蛋白质，其数量和组成按照在HPLC分离中使流动相组成与样品组合物一致的方式实现 实现而不需要衍生或提取分析物。 分析物的色谱分离在二氧化硅正相HPLC柱上进行，同时使用由水/乙腈/丙酸/三氟乙酸组成的流动相，水与乙腈的体积比为2至98至30至70，体积 三氟乙酸的百分比范围为0.01至0.5，丙酸的体积百分比为三氟乙酸的10至100倍，流动相的最佳组成由乙腈/丙酸/三氟乙酸与10/90/1 / 0.025体积百分比。 通过串联质谱法进行检测和定量，观察到以下碎裂：对于精氨酸，为175.2m / z-> 70.1m / z，对于13精氨酸为181.2m / z-> 74.1m / z 对于SDMA为203.2m / z-> 172.1m / z，对于ADMA为203.2m / z-> 46.1m / z，对于ADMA为203.2m / z-> 46.1m / z，对于ADMA为209.2m / z-> 70.1m / z D < SUB> 6 -ADMA。

公开(公告)号：US07537704B2

公开(公告)日：2009-05-26

申请号：US11915828

申请日：2006-05-13

申请人： Stefanie M. Bode-Böger ,  Jens Martens-Lobenhoffer

发明人： Stefanie M. Bode-Böger ,  Jens Martens-Lobenhoffer

IPC分类号： B01D15/08

CPC分类号： G01N33/6812

摘要： The invention relates to a method for determining the concentration of asymmetric dimethylarginine (ADMA) simultaneously with arginine and with symmetric dimethylarginine (SDMA) in biological samples by means of HPLC-MS-MS. The sample preparation consists exclusively of adding a solution of isotope-labeled internal standards (13C6-arginine and D6-ADMA) and of adding a mixture consisting of acetonitrile/propionic acid/trifluoroacetic acid for precipitating high-molecular proteins, the quantity and composition being realized in such a manner that a bringing of the sample composition in line with the composition of the mobile phase in the HPLC separation is achieved without requiring a derivitization or extraction of the analytes. The chromatographic separation of the analytes ensues on a silica normal phase HPLC column while using a mobile phase consisting of water/acetonitrile/propionic acid/trifluoroacetic acid, the volume ratio of water to acetonitrile ranging from 2 to 98 to 30 to 70, the volume percent of trifluoroacetic acid ranging from 0.01 to 0.5, the volume percent of propionic acid being 10 to 100 times higher than trifluoroacetic acid, and the optimized composition of the mobile phase consists of acetonitrile/propionic acid/trifluoroacetic acid with 10/90/1/0.025 volume percents. The detection and quantification ensue by means of tandem mass spectrometry with the following fragmentations being observed: 175.2 m/z→70.1 m/z for arginine, 181.2 m/z→74.1 m/z for 13C6-arginine, 203.2 m/z→172.1 m/z for SDMA, 203.2 m/z→46.1 m/z for ADMA and 209.2 m/z→70.1 m/z D6-ADMA.

摘要翻译： 本发明涉及通过HPLC-MS-MS在生物样品中与精氨酸和对称二甲基精氨酸（SDMA）同时测定不对称二甲基精氨酸（ADMA）的浓度的方法。 样品制备仅包括加入同位素标记的内标（13C6-精氨酸和D6-ADMA）溶液，并加入由乙腈/丙酸/三氟乙酸组成的混合物用于沉淀高分子蛋白质，其数量和组成为 以如下方式实现：在HPLC分离中使样品组合物与流动相的组成一致，而不需要衍生或提取分析物。 分析物的色谱分离在二氧化硅正相HPLC柱上进行，同时使用由水/乙腈/丙酸/三氟乙酸组成的流动相，水与乙腈的体积比为2至98至30至70，体积 三氟乙酸的百分比范围为0.01至0.5，丙酸的体积百分比为三氟乙酸的10至100倍，流动相的最佳组成由乙腈/丙酸/三氟乙酸与10/90/1 / 0.025体积百分比。 通过串联质谱法进行检测和定量，观察到以下碎裂：精氨酸为175.2m / z-> 70.1m / z，13C6-精氨酸为181.2m / z-> 74.1m / z，203.2m / z - 对于SDMA为> 172.1m / z，对于ADMA为203.2m / z-> 46.1m / z，对于ADMA为209.2m / z-> 70.1m / z D6-ADMA。

公开(公告)号：US20100173991A1

公开(公告)日：2010-07-08

申请号：US12669913

申请日：2008-06-25

申请人： Peter Lorenz ,  Peter Kreutzmann ,  Fritz Rothe ,  Jens Martens-Lobenhoffer ,  Harry Schmidt ,  Gerald Wolf

发明人： Peter Lorenz ,  Peter Kreutzmann ,  Fritz Rothe ,  Jens Martens-Lobenhoffer ,  Harry Schmidt ,  Gerald Wolf

IPC分类号： A61K31/235 ,  C07C69/94 ,  C07C233/65 ,  A61K31/166 ,  A61P31/04

CPC分类号： C07C231/02 ,  C07C2603/44 ,  C07C237/48

摘要： The aim of the invention to provide a less complex method for the production of A-ring aromatized acetyl minocyclines of the formula (I), wherein R1 to R5=acetyl and/or H, which can also be used on an industrial scale, is achieved in that minocycline hydrochloride is reacted with acetanhydride in the presence of a proton catcher, the reaction product is subjected to chromatographic filtration using a carrier material and an eluant, the eluant is distilled off, and the product is subsequently cleaned by recrystallization.

摘要翻译： 本发明的目的是提供一种不太复杂的制备式（I）芳构化的乙酰基二环胞菊素的方法，其中R1至R5 =也可以工业规模使用的乙酰基和/或H是H为 在质子捕获剂存在下，盐酸米诺环素与乙酸酐反应，使用载体材料和洗脱液对反应产物进行色谱过滤，蒸除洗脱液，随后通过重结晶清洗产物。