[求助]毛细管电流不出峰有些什么原因? 请指教!

柱长:1米, 内径50, 上样时压力 10psi ,时间 10秒, 分离时电压 20KV, 分离80分钟。
这些是药典上的条件么？
不过1米柱长就太长了，一般用50cm。进样时间也太长。而且，作为一个药典的分析方法，80分钟就太长了，显示不出毛细管电泳的优势。
我没看过欧洲药典（楼主有的话不妨上传），不知道里面的条件和方法是什么，可能说得不对。

多谢回复,我们CE的方法就是下面的方法, 长度是1米,您看看,多谢!

IDENTIFICATION
A. It gives the appropriate response when examined using
the conditions described under Assay.
B. Capillary zone electrophoresis (2.2.47)
Test solution. Dilute the substance to be examined
with water R to obtain a concentration of 1 mg/ml.
Desalt 0.25 ml of the solution by passage through a
micro-concentrator cartridge provided with a membrane
with a molecular mass cut-off of not more than 10 000.
Add 0.2 ml of water R to the sample and desalt again.
Repeat the desalting procedure once more. Dilute the
sample with water R, determine its protein concentration
as described under Tests and adjust to a concentration of
approximately 1 mg/ml with water R.
Reference solution. Dissolve the contents of a vial of
erythropoietin BRP in 0.25 ml of water R. Proceed with
desalting as described for the test solution.
Capillary:
— material: uncoated fused silica,
— size: effective length = about 100 cm, Ø = 50 μm,
Temperature: 35 °C.
CZE buffer concentrate (0.1 M sodium chloride, 0.1 M
tricine, 0.1 M sodium acetate). Dissolve 0.584 g of
sodium chloride R, 1.792 g of tricine R and 0.820 g of
anhydrous sodium acetate R in water R and dilute to
100.0 ml with the same solvent.
1 M putrescine solution. Dissolve 0.882 g of putrescine R
in 10 ml of water R. Distribute in 0.5 ml aliquots.
CZE buffer (0.01 M tricine, 0.01 M sodium chloride,
0.01 M sodium acetate, 7 M urea, 2.5 mM putrescine).
Dissolve 21.0 g of urea R in 25 ml of water R by warming
in a water-bath at 30 °C. Add 5.0 ml of CZE buffer
concentrate and 125 μl of 1 M putrescine solution. Dilute
to 50.0 ml with water R. Using dilute acetic acid R,
adjust to pH 5.55 at 30 °C and filter through a 0.45 μm
membrane filter.
Detection: spectrophotometer at 214 nm.
Set the autosampler to store the samples at 4 °C during
analysis.
Preconditioning of the capillary: rinse the capillary for
60 min with 0.1 M sodium hydroxide filtered through a
0.45 μm membrane filter and for 60 min with CZE buffer.
Apply voltage for 12 h (20 kV).
Between-run rinsing : rinse the capillary for 10 min with
water R, for 5 min with 0.1 M sodium hydroxide filtered
through a 0.45 μm membrane filter and for 10 min with
CZE buffer.
Injection: under pressure or vacuum.
Migration: apply a field strength of 143 V/cm(15.4 kV
for capillaries of 107 cm total length) for 80 min, using
CZE buffer as the electrolyte in both buffer reservoirs.

不好意思，看了相关资料，药典上的条件正如你所列。
没有出峰，原因很多，最主要的是：毛细管没有活化好；溶质在毛细管壁有吸附。
关于不出峰，在中国色谱网论坛上有很多帖子介绍，你可以进去看看。

有道理,这个毛细管不是买的原装的, 我们是从河北永年公司买的毛细管,然后,自己从市场上买了胶管装配起来的,按上面的方法,用氢氢化钠 缓冲液等进行了冲洗,再进行实验, 是不是这个原因, 您看看, 如果要充分活化, 要注意什么地方? 请指教, 谢谢!

另外, 不出峰,连水峰都没有, 后来, 换用其它的物质, 如丙酮来上样, 也是没有峰, 这又是什么原因?

有这种情况吗？没碰到哦

就是不出峰？电流的变化情况怎么样呢？

国产的管子就是永年的了。虽然没有进口的那么好，但是用起来问题不大。
如果活化不够的话，建议先用1M的HCl冲30分钟，水冲5分钟，1M NaOH冲30分钟，水冲5分钟，再用0.1M NaOH冲20分钟，水冲5分钟。这是我师兄的方法。实在不行那就换根管子。
另外，“自己从市场上买了胶管装配起来的”，我不太明白这一步的作用。

我想楼主可能用的Beckman的CE，毛细管外面胶管走冷却液的。