23 advantages of supported liquid extraction (sle+) and the new evolute express for sample...

© 2012 Biotage 1

5 Minutes of Fame- Advantages of the Evolute Express and Supported Liquid Extraction (SLE+)Solvent Evaporation Technology

Sung [email protected]

Chris Sochacki704.654.4882 [email protected]

© 2012 Biotage 2

Do less with NO loss in performance

New Benefits EVOLUTE EXPRESSLoad-Wash-Elute & Minimum elution volume

Load-Wash-EluteVs.

Cond-Equil-Load-Wash-Elute

½ Elution Volume=Less Dry Down Times

© 2012 Biotage 3

Less CloggingComparison with main competitors and Original Evolute

Even Flow Less Channeling Increased capacity

Pore Size optimized Selectively Reduce Protein Interferences

Water Wettable, Reduced Drying

Additional Benefits

© 2012 Biotage 4

ISOLUTE SLE+ :- Supported Liquid Extraction

Page 74-79 in the 2012 Catalog

© 2012 Biotage 5

Traditional LLE

Collection plate well or vial

Typical bioanalytical LLE procedure:

1. Dispense sample into well

2. Dispense solvent into well

3. Seal vial or plate

4. Mix (often manual)

5. Centrifuge to separate layers (manual)

6. Remove seal

7. Remove unwanted aqueous layer (top or bottom)

8. Sample for analysis

3. to 6. Cap, mix, centrifuge, de-cap

1. and 2.

7.

8.

© 2012 Biotage 6

Supported Liquid Extraction: how does it work?

Collect organic extract

containing drug

Whole sample is absorbed onto

bed

(unlike SPE – no ‘flow though’)

Step 1:

Apply aqueous sample

Step 2:

Wait for 5 min.

Step 3:

Add organic solvent

Individual cartridge or

96-well plate

© 2012 Biotage 7

IMPROVED RECOVERIES WITH ISOLUTE SLE+

Isolute SLE+ Liquid-Liquid Extraction

Analyte Analyte Recovery %RSD Analyte Recovery %RSD

Imipramine 97 4 65 4

Trimipramine 96 2 57 4

Nortriptyline 91 4 62 5

© 2012 Biotage 8

Processing Time

Technique SLE+ LLE

Off line Steps None 4

Total Time 12.5 minutes 22.5 minutes

Productivity 4 plates/hour 2 plates/hour

© 2012 Biotage 9

How clean are the extracts? Protein Assay

10

30

70

120

220

40

20

50

Raw

Seru

m

Mol.WtMarker

SLE

+

HM-N

© 2012 Biotage 10

MRM TIC’s comparing phospholipid removal between various extraction conditions

2.00 4.00 6.00 8.00

%

0

100

2: MRM of 11 Channels ES+TIC

5.63e77.56

6.556.11

8.15

2.00 4.00 6.00 8.00

%

0

100


5.63e7

2.00 4.00 6.00 8.00

%

0

100


5.63e7

2.00 4.00 6.00 8.00

%

0

100


5.63e7

7.66

Time2.00 4.00 6.00 8.00

%

0

100


3.27e73.58

3.44

4.22

Time2.00 4.00 6.00 8.00

%

0

100


3.27e7

Time2.00 4.00 6.00 8.00

%

0

100


3.27e7

Time2.00 4.00 6.00 8.00

%

0

100


3.27e7

Phospholipids

Lyso phospholipids

Plasma extracted with PPT+ Plasma pre-treated 1:1 0.5M NH4OHExtraction solvent MTBE

Plasma pre-treated 1:1 1% formic acidExtraction solvent DCM

Plasma pre-treated 1:1 1% formic acidExtraction solvent EtOAc

© 2012 Biotage 12

Patented-Gas Vortex

• No moving parts = Robust• ~12 min Evaporation times (500uL

MeOH, TV96)• Most Published Evaporation Unit

on the Market