DNS氨基酸的双向聚酰胺薄膜层析

1、1会计学DNS氨基酸的双向聚酰胺薄膜层析氨基酸的双向聚酰胺薄膜层析是是类化学纤维原料，即锦纶类化学纤维原料，即锦纶(又称尼龙又称尼龙)。由己二酸与己二胺聚合而成。由己二酸与己二胺聚合而成。因为在这类物质分子中都含有大量酰胺基团因为在这类物质分子中都含有大量酰胺基团，故统称聚酰胺。，故统称聚酰胺。DNS-Cl在在pH过高时，水解产生副产物过高时，水解产生副产物DNS-OH，即：，即：在在DNS-Cl过量时，会过量时，会产生产生DNS-NH2，即：，即：pH9.840,30min反应的副产物：反应的副产物：DNS-NH2黄色荧光和黄色荧光和DNS-OH蓝色荧光蓝色荧光颉颉亮亮苯苯丙丙赖赖甘甘丝丝天

2、天脯脯谷谷1、DNS氨基酸的制备氨基酸的制备2、混合、混合DNS氨基酸的双向层析氨基酸的双向层析（1）点样。距）点样。距右右下角距两边各下角距两边各0.5cm，直径控制在，直径控制在23mm之间，点在无光泽面之间，点在无光泽面,可重复几次点样。可重复几次点样。（2）苯冰醋酸层析：展层剂前缘距薄膜顶端）苯冰醋酸层析：展层剂前缘距薄膜顶端3mm处处即可停止即可停止,冷冷风吹干，紫外观察。风吹干，紫外观察。（3）甲酸水层析：调转）甲酸水层析：调转90度与第一相垂直，度与第一相垂直，热热风吹风吹干，紫外灯下观察，用铅笔干，紫外灯下观察，用铅笔轻轻轻轻标记。标记。（1 1）严格控制点样位置以及点样直径。

3、）严格控制点样位置以及点样直径。（2 2）展层时勿将点样浸入溶剂系统。）展层时勿将点样浸入溶剂系统。（3 3）展层后必须经电吹风将膜吹干。）展层后必须经电吹风将膜吹干。（4 4）使用紫外照射时要注意使用时间短）使用紫外照射时要注意使用时间短. .1. Prepare the developing container.The developing container for TLC can be a specially designed chamber, a jar with a lid, or a beaker with a watch glass on the top: Pour solve

4、nt into the beaker to a depth of just less than 0.5 cm. To aid in the saturation of the TLC chamber with solvent vapors, line part of the inside of the beaker with filter paper Cover the beaker with a watch glass, swirl it gently, and allow it to stand while you prepare your TLC plate. TLC plates us

5、ed in the organic chem teaching labs are purchased as 5 cm x 20 cm sheets. Each large sheet is cut horizontally into plates which are 5 cm tall by various widths; the more samples you plan to run on a plate, the wider it needs to be.Prepare the TLC plate.Shown in the photo to the left is a box of TL

6、C plates, a large un-cut TLC sheet, and a small TLC plate which has been cut to a convenient size. Plates will usually be cut and ready for you when you come to lab.Handle the plates carefully so that you do not disturb the coating of adsorbent or get them dirty.Measure 0.5 cm from the bottom of the

7、 plate. Take care not to press so hard with the pencil that you disturb the adsorbent. Using a pencil, draw a line across the plate at the 0.5 cm mark. This is the origin: the line on which you will spot the plate. Its kind of hard to see the pencil line in the above photos, so here is a close-up of

8、 how the plate looks after the line has been drawn. Under the line, mark lightly the name of the samples you will spot on the plate, or mark numbers for time points. Leave enough space between the samples so that they do not run together, about 4 samples on a 5 cm wide plate is advised. Use a pencil

9、 and do not press down so hard that you disturb the surface of the plate. A close-up of a plate labeled 1 2 3 is shown to the right. . . swirl until dissolvedadd a few drops of solvent . . . dip the microcap into solution - the arrow points to the microcap, it is tiny and hard to see make sure it is

10、 filled - hold it up to the light if necessary touch the filled microcap to TLC plate to spot it - make sure you watch to see that all the liquid has drained from the microcap rinse the microcap with clean solvent by first filling it . . . . . . and then draining it by touching it to a paper towel h

11、eres the TLC plate, spotted and ready to be developed place the TLC plate in the developing container - make sure the solvent is not too deepThe solvent will rise up the TLC plate by capillary action. In this photo, it is not quite halfway up the plate.In this photo, it is about 3/4 of the way up th

12、e plate.Remove the plate from the beaker.quickly mark a line across the plate at the solvent front with a pencilAllow the solvent to evaporate completely from the plate. If the spots are colored, simply mark them with a pencil.Most samples are not colored and need to be visualized with a UV lamp. Ho

13、ld a UV lamp over the plate and mark any spots which you see lightly with a pencil.this is a UV lamp here are two proper sized spots, viewed under a UV lamp (you would circle these while viewing them)The plate shows three compounds run at three different concentrations. The middle and right plate sh

14、ow reasonable spots; the left plate is run too concentrated and the spots are running together, making it difficult to get a good and accurate Rf reading. Heres what overloaded plates look like compared to well-spotted plates. The plate on the left has a large yellow smear; this smear contains the s

15、ame two compounds which are nicely resolved on the plate next to it. The plate to the far right is a UV visualization of the same overloaded plate.The retention factor, or Rf, is defined as the distance traveled by the compound divided by the distance traveled by the solvent.反应的副产物：反应的副产物：DNS-NH2黄色荧

16、光和黄色荧光和DNS-OH蓝色荧光蓝色荧光1. Prepare the developing container.The developing container for TLC can be a specially designed chamber, a jar with a lid, or a beaker with a watch glass on the top: Pour solvent into the beaker to a depth of just less than 0.5 cm. TLC plates used in the organic chem teaching l

17、abs are purchased as 5 cm x 20 cm sheets. Each large sheet is cut horizontally into plates which are 5 cm tall by various widths; the more samples you plan to run on a plate, the wider it needs to be.Prepare the TLC plate.Shown in the photo to the left is a box of TLC plates, a large un-cut TLC sheet, and a small TLC plate which has been cut to a convenient size. Plates will usu