Görs S, Kucia M, Langhammer M, Junghans P, Metges C C
Nutritional Physiology Oskar-Kellner, Research Institute for the Biology of Farm Animals (FBN), Dummerstorf, Germany.
J Dairy Sci. 2009 Feb;92(2):632-7. doi: 10.3168/jds.2008-1563.
Analysis in individual mouse milk samples is restricted by small sample volumes and hindered by high fat contents. Miniaturized methods were developed for the analysis of dry matter (DM), crude fat, crude protein (CP), and lactose in individual samples of <or=200 microL of fresh or previously frozen mouse milk and used to compare milk from the mouse strain DU6, the largest growth-selected mouse line worldwide, with unselected mice (CON) on lactation d 3, 14, and 18. Individual milk samples were collected by means of a self-constructed milking machine. Aliquots of 10 microL of milk were used to measure DM [coefficient of variation (CV) <2.1%], which was subsequently used to analyze nitrogen for calculation of CP (CV 2.7%). Crude fat was determined in 100 microL via a miniaturized Röse-Gottlieb method (CV 2.8%). An HPLC protocol was used to analyze lactose in 20 microL of diluted whey (CV 5.3%). The miniaturized methods gave similar results compared with conventional approaches. Homogenization was the most important factor affecting milk composition and its reproducibility. Milk storage at -20 degrees C had no effect on composition. Irrespective of the mouse strain, maximum values of 45.5% DM, 29.8% fat, and 12.7% CP were observed at d 14. The greatest lactose contents were found on d 18 (2.41%). Milk lactose concentration at d 3 was lower in DU6 (1.13 +/- 0.10%) than CON (1.67 +/- 0.18%). The method provides an accurate assessment of mouse milk composition.
对单个小鼠乳汁样本的分析受到样本量小的限制,且因高脂肪含量而受阻。已开发出小型化方法,用于分析新鲜或先前冷冻的小于或等于200微升小鼠乳汁的单个样本中的干物质(DM)、粗脂肪、粗蛋白(CP)和乳糖,并用于比较世界上最大的生长选择小鼠品系DU6与未选择小鼠(CON)在哺乳期第3、14和18天的乳汁。通过自行构建的挤奶机收集单个乳汁样本。取10微升乳汁等分试样用于测量DM[变异系数(CV)<2.1%],随后用于分析氮以计算CP(CV 2.7%)。通过小型化的罗斯 - 哥特利布法在100微升中测定粗脂肪(CV 2.8%)。使用高效液相色谱法分析20微升稀释乳清中的乳糖(CV 5.3%)。与传统方法相比,小型化方法给出了相似的结果。均质化是影响乳汁成分及其重现性的最重要因素。在 - 20℃下储存乳汁对成分没有影响。无论小鼠品系如何,在第14天观察到DM的最大值为45.5%、脂肪为29.8%、CP为12.7%。在第18天发现乳糖含量最高(2.41%)。DU6在第3天的乳汁乳糖浓度(1.13±0.10%)低于CON(1.67±0.18%)。该方法可准确评估小鼠乳汁成分。