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. Author manuscript; available in PMC: 2010 Jun 15.
Published in final edited form as: J Chromatogr B Analyt Technol Biomed Life Sci. 2009 Apr 24;877(18-19):1709–1715. doi: 10.1016/j.jchromb.2009.04.025

Table 1.

LC method for the focus mode HTLC online extraction method

Step1 Start Sec2 Loading pump Flow (mL/min) Grad %A %B %C %D Tee3 Loop4 Eluting pump Flow (mL/min) Grad %A %B
1 0.00 20 3.00 Step 100 - - - - Out 0.37 Step 98 2.0
2 0.33 120 0.07 Step 100 - - - T In 0.3 Step 98 2.0
3 2.33 60 2.00 Step - 100 - - - In 0.36 Ramp 65 35
4 3.33 45 2.00 Step - 100 - - - Out 0.37 Step 65 35
5 4.08 20 2.00 Step - - 80 20 - In 0.38 Ramp 25 75
6 4.42 25 2.00 Step - - 80 20 - Out 0.39 Step 2.0 98
7 4.83 25 2.00 Step - - 80 20 - In 0.38 Step 98 2.0
8 5.25 90 2.00 Step 100 - - - - Out 0.37 Step 98 2.0
1

At each step, the mobile phases for both loading and eluting pump were introduced by step gradient

2

time as second

3

the loop connected to analytical column

4

the transfer loop connected to HT column