App ID: FLM-C220001

蛋白中殘留的TFA于HPLC上UV檢測器下的定量檢測分析

（此應(yīng)用同樣適用于多肽中TFA殘留量的測定)

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殘留量的測定。多肽的出峰時(shí)間將于蛋白相似或更早， 都不會干擾TFA的定量。此外，UV下的分析，也可采用Comixsil HAX 3u 100*4.6mm

(貨號：FMD-HAX3-EONU)，流速為1ml/min，其它色譜條件一樣。