DNS氨基酸的雙向聚酰胺薄膜層析

1、1會計學DNS氨基酸的雙向聚酰胺薄膜層析氨基酸的雙向聚酰胺薄膜層析是是類化學纖維原料，即錦綸類化學纖維原料，即錦綸(又稱尼龍又稱尼龍)。由己二酸與己二胺聚合而成。由己二酸與己二胺聚合而成。因為在這類物質分子中都含有大量酰胺基團因為在這類物質分子中都含有大量酰胺基團，故統(tǒng)稱聚酰胺。，故統(tǒng)稱聚酰胺。DNS-Cl在在pH過高時，水解產生副產物過高時，水解產生副產物DNS-OH，即：，即：在在DNS-Cl過量時，會過量時，會產生產生DNS-NH2，即：，即：pH9.840,30min反應的副產物：反應的副產物：DNS-NH2黃色熒光和黃色熒光和DNS-OH藍色熒光藍色熒光頡頡亮亮苯苯丙丙賴賴甘甘絲絲天

2、天脯脯谷谷1、DNS氨基酸的制備氨基酸的制備2、混合、混合DNS氨基酸的雙向層析氨基酸的雙向層析（1）點樣。距）點樣。距右右下角距兩邊各下角距兩邊各0.5cm，直徑控制在，直徑控制在23mm之間，點在無光澤面之間，點在無光澤面,可重復幾次點樣?？芍貜蛶状吸c樣。（2）苯冰醋酸層析：展層劑前緣距薄膜頂端）苯冰醋酸層析：展層劑前緣距薄膜頂端3mm處處即可停止即可停止,冷冷風吹干，紫外觀察。風吹干，紫外觀察。（3）甲酸水層析：調轉）甲酸水層析：調轉90度與第一相垂直，度與第一相垂直，熱熱風吹風吹干，紫外燈下觀察，用鉛筆干，紫外燈下觀察，用鉛筆輕輕輕輕標記。標記。（1 1）嚴格控制點樣位置以及點樣直徑。

3、）嚴格控制點樣位置以及點樣直徑。（2 2）展層時勿將點樣浸入溶劑系統(tǒng)。）展層時勿將點樣浸入溶劑系統(tǒng)。（3 3）展層后必須經電吹風將膜吹干。）展層后必須經電吹風將膜吹干。（4 4）使用紫外照射時要注意使用時間短）使用紫外照射時要注意使用時間短. .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.反應的副產物：反應的副產物：DNS-NH2黃色熒

16、光和黃色熒光和DNS-OH藍色熒光藍色熒光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