App ID: FLM-C220001
蛋白中殘留的TFA于HPLC上UV檢測(cè)器下的定量檢測(cè)分析
（此應(yīng)用同樣適用于多肽中TFA殘留量的測(cè)定)
Equipment
Agilent 1100 HPLC system with DAD Analytical Balance
HPLC Column: Comixsil HAX 3u 3.0x100 mm (P/N:FMD-HAX3-YONU)
Materials:
Protein NS3278
Trifluoracetic acid (Aldrich, catalog #74564) ACN (Tedia, catalog #AS1127)
Distilled water (Aldrich, catalog #270733) Sulfuric acid (Aldrich, catalog #258105) Method Description
HPLC Q-Pump set up: A: DI water, B: Acetonitrile, C: 1% H2SO4 Mobile phase: 60% ACN with 0.1% H2SO4
Mobile phase composition: : A: 30%, B: 60%, C: 10%
|
Concentration, mg/ml |
Average Peak Area, mAU*s |
Concnetration of TFA, mg/ml |
|
0.1 |
12.9 |
- |
|
0.2 |
25.9 |
- |
|
0.5 |
69 |
- |
|
1 |
135.6 |
- |
|
2 |
261 |
- |
|
Protein |
41.8 |
0.31 |
說明：此方法同時(shí)適合多肽中TFA殘留量的測(cè)定。多肽的出峰時(shí)間將于蛋白相似或更早， 都不會(huì)干擾TFA的定量。此外，UV下的分析，也可采用Comixsil HAX 3u 100*4.6mm
(貨號(hào)：FMD-HAX3-EONU)，流速為1ml/min，其它色譜條件一樣。
