Comparing the high-throughput efficiency

of microextraction techniques, MEPS and µSPEed,

for the isolation of polyphenols from food matrices
José S. 1,2,*
Câmara , Natalia 3
Casado , Jorge 2
Pereira , Catarina L. 2
Silva , Rosa 2
Perestrelo , Isabel Sierra3
1CQM
– Centro de Química da Madeira, Universidade da Madeira, Campus da Penteada, 9020-105
2Faculdade de Ciências Exatas e Engenharia da Universidade da Madeira, Campus da Penteada, 9020-105 Funchal, Portugal
3Departamento de Tecnología Química y Energética, Tecnología Química y Ambiental, Tecnología Mecánica y Química Analítica, E.S.C.E.T, Universidad Rey Juan Carlos, C/ Tulipán s/n, 28933 Móstoles, Spain

INTRODUCTION RESULTS
In this study, microextraction by packed sorbent (MEPS) or micro solid Optimization of extraction parameters
phase extraction (μSPEed) coupled to UHPLC-PDA, were evaluated and
compared regarding their analytical performance or the extraction of
polyphenols from baby foods.
To achieve the highest extraction efficiency, the influential parameters type
of sorbent, the number of extraction cycles, the pH, the elution solvent
and the elution volume, were systematically optimized.
Method selectivity, linear dynamic range, limits of detection (LODs) and
quantification (LOQs), accuracy, precision, and extraction yields, were
determined and discussed for both techniques.

Experimental
EXPERIMENTAL
Design

Figure 2. Optimization of the best experimental conditions for MEPS and SPEed: selection of the best sorbent (A), optimal sample loading condition (B), best sample volume (C), optimum sample pH
0
(D) and best elution solvent (E) and respective optimal volume (F).

Step #1
Sorbent Step #2 Sealing MEPS
5 Activation activation Solvent
discarding
ring
Single
direction
flow
FLOW Check FLOW Check
Valve Valve
Frit MEPS
Open Closed
packed
bed
Step #3
Sorbent Step #4
10
Conditioning conditioning Solvent
discarding End plug
Figure 3. Representative chromatogram of real
Needle sample upon SPEed extraction and UPLC analysis
FLOW FLOW Analytical method validation
µSPEed Table 1. Validation parameters for the MEPS/UHPL-PDA and µSPEed/UHPLC-PDA analysis of the selected phenolic compounds..

15 Step #5 eVOL®
Analytical Method Validation Method Application
Sample Filtered
sample
Step #6
Unretained
XCHANGE® Phenolic compounds Extraction LDR
R 2 LOD LOQ Recovery Intra-day Inter-day Banana Apple
Multi-fruits Chicken, beef
with cereals & vegetables
syringe
Loading loading analytes
discarding
interface
technique (µg/mL) (µg/kg) (µg/kg) (%) precision precision
Conc. RSD Conc. RSD Conc. RSD Conc. RSD
MEPS 0.994 12.8 42.5 63 2.3 4.2 99 5.3 71 2.4 133 4.6 112 1.8
Gallic acid 0.25 - 2
μSPEed 0.997 2.1 6.9 67 2.8 4.5 138 7.3 169 3.8 154 2.7 125 1.1
FLOW
FLOW
MEPS 0.998 1.0 3.3 77 3.3 3.1 48 6.8 48 5.2 - - - -
High Protocatechuic acid 0.10 - 1.5
μSPEed 0.993 2.3 7.6 85 1.6 3.4 109 2.8 50 3.9 - - - -
Pressure
MEPS 0.995 1.7 5.6 53 2.4 4.1 - - 815 2.2 569 2.6 194 3.1
Connection Chlorogenic acid 0.03 - 0.75
20 Step #7 μSPEed 0.990 2.5 8.3 77 2.6 3.2 - - 1790 1.4 1353 2.9 284 4.1
MeOH Sorbent MEPS 0.992 5.3 17.6 48 0.8 1.0 31.5 1.7 - - 43 3.1 31 5.1
Step #8 4-Hydroxybenzoic acid 0.10 - 2
Time
(min)
Elution loading Target Bed μSPEed
MEPS
0.999
0.995
5.1
7.1
17.1
23.8
52
81
1.5
3.2
3.3
3.3
37 4.1
153 3.7
-
469 1.8
- 44.7
293
1.9
3.3
30 4.1
-
analytes High Epicatechin 0.05 - 1.5
Micro μSPEed 0.993 3.8 12.6 93 2.3 3.7 294 2.9 565 2.4 416 3.6 -
elution Pressure
One-Way MEPS 0.992 2.1 7.2 71 2.1 2.5 67 2.6 32.6 2.3 64 3.8 65 3.3
Seal p-Coumaric acid 0.03 - 0.75
FLOW
FLOW
Valve μSPEed 0.996 1.4 4.6 67 2.7 4.8 63 5.5 31 2.6 60.3 1.4 71.5 1.3
Cartridge
MEPS 0.993 1.6 5.4 63 1.0 1.8 236 2.5 - - 365 2.2 112 3.7
Ferulic acid 0.01 - 0.95
μSPEed 0.992 1.6 5.2 86 1.1 1.9 307 3.8 - - 409 1.6 165 1.3
Figure 1. Experimental layout of the MEPS and SPEed extraction procedures using the semi-automatic eVol® syringe. MEPS 0.992 9.6 31.9 70 1.6 1.9 340 3.4 169 2.5 360 2.3 < LOQ -
Rutin 0.05 - 2
μSPEed 0.984 2.5 8.4 77 2.1 2.8 387 3.2 192 4.3 367 1.7 36.3 1.3
MEPS 0.997 1.8 5.9 83 2.6 2.8 - - - - - - - -
Parameters MEPS µSPEed Resveratrol
μSPEed
0.03 - 0.75
0.992 1.4 4.7 84 2.4 1.7 - - - - - - - -
MEPS 0.992 3.3 11.2 79 1.5 3.1 108 5.1 52 2.1 98 2.2 < LOQ -
C2, C8, C18, Sil, M1, PEP, PS-DVB/RP-NP, C18 Myricetin
μSPEed
0.03 - 0.75
0.998 2.8 9.2 96 2.2 4.8 179 3.1 74 2.9 161 3.3 < LOQ -
Sorbent type R-CX, DVB/HLB, SCX, R- PS-DVB/RP, PS- Naringenin
MEPS
0.05 - 1.5
0.992 10.0 33.3 93 1.0 3.5 128 3.2 61 3.0 285 5.7 148 5.4
μSPEed 0.992 7.8 26.1 91 1.4 4.1 104 4.3 65 4.9 307 4.4 147 1.6
AX, PGC, HyDRC DVB/SCX, PS-DVB/SAX MEPS 0.985 7.4 24.6 98 2.2 4.2 < LOQ - < LOQ - - < LOQ 110 6.4
Kaempferol 0.05 - 1.5
Sample cycles 3×, 5×, 10× μSPEed 0.980 13.6 45.2 97 1.8 3.1 < LOQ - 52 4.3 - < LOQ 71 3.1
Legend: LDR – linear dynamic range; R2 – correlation coefficient; LOD – limits of detection; LOQ – limits of quantification; ME – Matrix effect; RSD – relative standard deviation.
Sample volume (µL) 100, 250, 500 25, 50, 100
Elution volume (µL) 100, 150, 200 25, 50, 100
Elution solvent
pH
ACN, MeOH
2, 7, 10
CONCLUSIONS
 MEPS and μSPEed, combined with UHPLC-PDA, were evaluated and compared to determine the most suitable
Chromatographic conditions (UPLC®-PDA)
extraction approach for the simultaneous quantification of 12 polyphenols in baby food products.
Mobile phase 0.1% Formic Acid/MeOH  The proposed analytical approaches were successfully optimized and validated in terms of selectivity, linearity,
Column HSS T3 LOD, LOQ, accuracy and precision.
Flow rate 250 µL/min  Nevertheless, the best performance was achieved with the μSPEed procedure since it exhibited higher extraction
potential towards the target analytes providing, in general, high recovery values and very low LODs and LOQs and
ACKNOWLEDGEMENTS requiring less amount of sample and solvents.
Natalia Casado would like to thank the Rey Juan Carlos University for providing her a mobility grant to perform a pre-doctoral stay in the
CQM. This work was supported by FCT-Fundação para a Ciência e a Tecnologia (project PEst-OE/ QUI/UI0674/2013, CQM;  Both extraction methods were used for the qualitative and quantitative determination of polyphenols from baby
SFRH/BD/97039/2013), and through Madeira 14-20 Program, project PROEQUIPRAM - Reforço do Investimento em Equipamentos e
food samples and constitute a promising tool to evaluate and improve the knowledge of the nutritional quality of
http://cqm.uma.pt
Infraestruturas Científicas na RAM (M1420-01-0145-FEDER-000008) and by ARDITI-Agência Regional para o Desenvolvimento da
Investigação Tecnologia e Inovação, through the project M1420-01-0145- FEDER-000005 -Centro de Química da Madeira -
CQM+(Madeira 14-20) and Project M1420-09-5369-FSE-000001 for the JAMP Post-Doctoral fellowship this kind of products

UID/QUI/
00674/2013