New Technique For Sample Preparation in Analytical Chemistry PDF

Division for Chemistry
Department of Analytical Chemistry
Zeki Altun
New Techniques for

Sample Preparation in
Analytical Chemistry
Karlstad University Studies

2005:24
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H CH3 O H CH3 O
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Ropivacaine, Mw 274 g/mol PPX, Mw 232 g/mol
HO CH3 O H CH3 O
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CH3 CH3
3-OH-ropivacaine, Mw 290 g/mol 2H -ropivacaine, Mw 281 g/mol

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Lidocaine, Mw 234 g/mol Bupivacaine, Mw 288 g/mol
Figure 6. Structures and molecular weights of local anaesthetics utilized in this work.

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Figure 7. The structures and molecular weights of roscovitine and olomoucine (internal
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Sorbents

Figure 8. Effect of type of sorbents on the recovery of ropivacaine.

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2 mg 1 mg 0.5 mg

Figure 9. Effect of amount of sorbent, C2, on the recovery of ropivacaine and its
metabolites PPX and 3-OH-ropivacaine compared to direct injection of pure standard
solutions.
Ropivacaine
80%
PPX
3-OH-ropivacaine
Internal Standard
60%
Recovery
40%
20%
0%
0.5 mg 1 mg 2 mg

Figure 10. Effect of amount sorbent, benzenesulphonic acid, on the recovery of
ropivacaine and its metabolites PPX and 3-OH-ropivacaine compared to direct injection of
pure standard solutions.

21

7$4$4,! 5!
#

* 66+ +. 4
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0,9
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at
r1
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ate
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Figure 11. Effect of the washing solution on the response of ropivacaine.

66 ++
+.
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+

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23
80% C8
C2
60%
Recovery
40%
20%
0%
M eOH/ Wat er 8:2 M eOH/ Wat er 9:1(v/ v) M eOH/ Wat er/ ACN M eOH/Water 95:5
(v/ v) (0.25%NH4OH) (0.25%NH4OH) 8:1:1(v/v) (0.25% (v/v) (0.25%NH4OH)
NH4OH)
Figure 12. Effect of different elution solvents on the recovery of ropivacaine.

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Figure 13. A photographic image of UV-polymerized monolith inside a disposable micro
pipette tip. The polymerization mixture contained GMA 20%, EGDMA 15.5%, BMA 3.5%,
AIBN 1% (with respect to monomers), cyclohexanol 30% and 1-dodecanol 30%.
Polymerization was allowed to proceed for 85 minutes using 254 nm light.
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26

Figure 14. Packed 96-tips with monolithic methacrylate polymer.
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27
041005024 MRM of 2 Channels ES+
355.3 > 233
8.58e4
100
Roscovitine 125 nM in plasma sample A
Blank plasma
0 Time
1.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00
041005086 MRM of 2 Channels ES+
299 > 91
4.87 2.96e5
100
IS B
Blank plasma
0 Time
1.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00
Figure 15. Chromatograms of (A) blank plasma and plasma spiked with 125 nM
roscovitine and (B) blank plasma and plasma spiked with olomoucine (I.S.).
Chromatographic conditions: column Zorbax 50 x 2.1 mm I.D., SB-C8, 3,5 ȝm connected
to a guard column, an Optiguard 10 x 1 mm I.D., C8; mobile phase (A) 0.1 % formic acid in
ACN-Water (10:90, v/v), (B) 0.1% formic acid in ACN-Water (80-20, v/v); linear gradient
0% B for 1 min, 0-80% B in 4 min, 80% B in 1 min; flow rate 150 ȝL/min.
28
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34
New Techniques for
Sample Preparation in
Analytical Chemistry
Sample preparation is often a bottleneck in systems for chemical analysis. The aim
of this work was to investigate and develop new techniques to address some of the
shortcomings of current sample preparation methods. The goal has been to provide
full automation, on-line coupling to detection systems, short sample preparation
times and high-throughput.
A new technique for sample preparation that can be connected on-line to liquid
chromatography (LC) and gas chromatography (GC) has been developed. Microex-
traction in packed syringe (MEPS) is a new solid-phase extraction (SPE) technique
that is miniaturized and can be fully automated. In MEPS approximately 1 mg of
sorbent material is inserted into a gas tight syringe (100-250 µL) as a plug. Sample
preparation takes place on the packed bed. Evaluation of the technique was done
by the determination of local anaesthetics in human plasma samples using MEPS
on-line with LC and tandem mass spectrometry (MS-MS). MEPS connected to an
autosampler was fully automated and clean-up of the samples took one minute. In
addition, in the case of plasma samples the same plug of sorbent could be used for
about 100 extractions before it was discarded.
A further aim of this work was to increase sample preparation throughput. To do
that disposable pipette tips were packed with a plug of porous polymer monoliths
as sample adsorbent and were then used in connection with 96-well plates and LC-
MS-MS. When roscovitine in human plasma and water samples was used as model
substance, a 96-plate was handled in two minutes.
Karlstad University Studies

ISSN 1403-8099
ISBN 91-85335-65-7

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New Techniques for

Karlstad University Studies

+ 46+0 +<

B7 "

6$;$6 # !

Ropivacaine, Mw 274 g/mol PPX, Mw 232 g/mol

3-OH-ropivacaine, Mw 290 g/mol 2H -ropivacaine, Mw 281 g/mol

Lidocaine, Mw 234 g/mol Bupivacaine, Mw 288 g/mol

Roscovitine, Mw 354 g/mol Olomoucine (I.S), Mw 298 g/mol

7$4 !& #"

Figure 12. Effect of different elution solvents on the recovery of ropivacaine.

6\QWKHVLVRI3RO\0$('0$%0$0RQROLWKVIRU3LSHWWH7LSV

Roscovitine 125 nM in plasma sample A

+, 5 (

, 0J+, &

/&8 #B7+,B7+

Karlstad University Studies

You might also like

New Technique For Sample Preparation in Analytical Chemistry PDF

Uploaded by

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Original Title

Copyright

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New Technique For Sample Preparation in Analytical Chemistry PDF

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Copyright:

Available Formats

Division for Chemistry

Department of Analytical Chemistry

New Techniques for

Karlstad University Studies

 + 4 6+ 0  +<

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Ropivacaine, Mw 274 g/mol PPX, Mw 232 g/mol

3-OH-ropivacaine, Mw 290 g/mol 2H -ropivacaine, Mw 281 g/mol

Lidocaine, Mw 234 g/mol Bupivacaine, Mw 288 g/mol

Roscovitine, Mw 354 g/mol Olomoucine (I.S), Mw 298 g/mol

7$4 ! &  # "  

Figure 12. Effect of different elution solvents on the recovery of ropivacaine.

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Karlstad University Studies

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