1. J Chromatogr B Biomed Sci Appl. 2001 Feb 25;751(2):377-82.

New sensitive assay of vancomycin in human plasma using high-performance liquid

chromatography and electrochemical detection.

Favetta P(1), Guitto J, Bleyzac N, Dufresne C, Bureau J.

Author information:
(1)Service de Pharmacie, H pital Lyon-Sud--Secteur Ste Eugnie, Pierre-Bnite,
France.

A method using reversed-phase high-performance liquid chromatography with

electrochemical detection for the analysis of vancomycin in human plasma was
developed. Chromatographic conditions included an octadecyl column, a mobile
phase of acetonitrile-sodium phosphate buffer (pH 7) (12:88), a total run time of
12 min, and coulometric electrochemical detection at +700 mV. Linear detector
response was found in the range 5-100 microg ml(-1) after a 1:80 dilution or from
0.5 to 50 microg ml(-1) after a 1:20 dilution of the samples. In both cases the
correlation coefficient (r) of the calibration curve standard was better than
0.995. Vancomycin determination was based on a denaturation of plasma proteins
with methanol, then a dilution with mobile phase was performed. Recovery of
vancomycin from plasma was 103.1+/-3.9%, and no interference from commonly used
drugs or endogenous compounds was observed. A significant correlation was shown

with the EMIT assay (r=0.92, P<0.001) using clinical samples from children. This
HPLC technique is simple, sensitive, rapid, precise, selective and requires only
100 microl of plasma for completion.

PMID: 11236095 [PubMed - indexed for MEDLINE]

Innovative Electrochemical Sensor Based on Electroactive Nanoparticles of Molecularly

Imprinted Polymers for the Indirect Detection of Vancomicyn
1

Antonio Turco, 1 Elisabetta Mazzotta, 1 Cosimino Malitesta, 2 Iva Chianella, 2 Sergey A.

Piletsky
1

University of Salento, Dipartimento di scienze e tecnologie biologiche ed ambientali, Via

Monteroni, 1 Lecce, Italy
2
Cranfield University, Cranfield Health Vincent Building, MK43 0AL, Bedfordshire, Cranfield,
UK
elisabetta.mazzotta@unisalento.it
In recent years, Molecularly Imprinted Polymers (MIPs) have been applied as a useful tool in
molecular recognition processes and in the development of sensing systems. The imprinting
technique gives the possibility to easily prepare low cost materials containing cavities able to
selectively recognize analytes ranging from biologically relevant markers, drugs and
agrochemicals [1], [2], [3]. MIPs represent an alternative to natural receptors (enzymes,
antibodies, etc.) with the main advantages to be robust, versatile and cost effective systems [4].
For the successful application of MIPs in sensors it is necessary to improve their binding
kinetics, shorten analysis times and achieve complete template removal [5]. The possibility of
design MIPs at nanodimensional scale has been demonstrated to have a key effect on these issues
by enhancing surface-to-volume ratio and thus making binding sites more accessible to analytes
[4]. Different MIP nanostructures have been prepared [4] and successfully applied in the design
of sensing devices. Among these, MIP nanoparticles received particular attention being prepared
by different chemical and electrochemical strategies [5] and being applied in different analytical
fields ranging from chromatographic [6] to sensing applications [7]. In the latter case, their
integration with the transducer surface represents a key aspect strongly influencing the
performance of the whole sensing systems. Among possible integration schemes, their
entrapment in an electrosynthesized film has been proposed [7]. In this work we developed an
easy way to prepare electrochemical sensor based on electroactive MIP nanoparticles deposited
on an electropolymerized film for the indirect electrochemical detection of vancomicyn, a not
electroactive antibiotic widely used in farm. Electroactive MIP nanoparticles specific for
vancomicyn were synthesized by solid-phase synthesis [8] adding vinyl-ferrocene in different

amount to polymerization mixture. MIP nanoparticles were studied by cyclic voltammetry to

identify the amount of vinyl-ferrocene determining the highest electrochemical activity. Then
nanoparticles were deposited by casting on glassy carbon electrode surface before and after the
electrochemical deposition of different conducting and not conducting polymers. It was verified
that the presence of a polymeric layer on the electrode surface increases MIP nanoparticles
adhesion preserving their electrochemical activity thus promoting a higher stability of the
system. Finally the electrochemical detection of vancomicyn was studied both by cyclic
voltammetry and amperometrically being based on the modification of the electrochemical signal
due to MIP electroactivity after increasing vancomycin concentration. The observed behavior
could be ascribed to the interaction between MIP cavities and the antibiotic influencing the
electron transfer process of the ferrocene redox sites within nanoparticles. References [1] X.
Kan, H. Zhan, C. Li, A. Zhu, Z. Xiang, Z. Zhao, Electrochim Acta, 63, 2012, 69-75 [2] D. Zang,
M. Yan, S. Ge, L. Ge, J. Yu, Analyst 138, 2013, 2704-2711 [3] E. Mazzotta, R.A. Picca,
C.Malitesta, S.A. Piletsky, E.V. Piletska, Biosens Bioelectron, 23, 2008, 1152-1156 [4] C.
Malitesta, E. Mazzotta, R.A.Picca, A. Poma, I. Chianella, S.A. Piletsky, Anal Bional Chem, 402,
2012, 1827-1846 [5] S. Tokonami, H. Shigi, T. Nagaoka, Anal Chim Acta 641, 2009, 7-13 [6] E.
Turiel, M. Esteban, Anal Bional Chem 378, 2004, 1876-1886 [7] E. Mazzotta, R.A. Picca, C.
Malitesta, S.A. Piletsky, E.V. Piletska, Biosens Bioelectron 23, 2008, 1152-1156 [8] A. Poma, A.
guerreiro, M.J. Whitcombe, E.V. Piletska, A.P.F. Turner, S. Piletsky, Adv Func Mat, 23, 2013,
2821-2827