See

discussions, stats, and author profiles for this publication at: https://www.researchgate.net/publication/236919433

Hydrophilic interaction chromatography

(HILIC) in the analysis of antibiotics

Article in Journal of pharmaceutical and biomedical analysis · April 2013

DOI: 10.1016/j.jpba.2013.04.015 · Source: PubMed

CITATIONS READS

44 682

5 authors, including:

Getu Kahsay Huiying Song

Mekelle University University of Leuven
24 PUBLICATIONS 118 CITATIONS 9 PUBLICATIONS 63 CITATIONS

SEE PROFILE SEE PROFILE

Ann Van Schepdael Deirdre Cabooter

University of Leuven University of Leuven
254 PUBLICATIONS 3,822 CITATIONS 76 PUBLICATIONS 1,097 CITATIONS

SEE PROFILE SEE PROFILE

Some of the authors of this publication are also working on these related projects:

- To develop a new sampling technique combined with gas chromatography for the analysis of
residual solvents from specific pharmaceutical formulations View project

All content following this page was uploaded by Getu Kahsay on 09 November 2017.

The user has requested enhancement of the downloaded file.

 Journal of Pharmaceutical and Biomedical Analysis 87 (2014) 142–154

Contents lists available at ScienceDirect

Journal of Pharmaceutical and Biomedical Analysis

journal homepage: www.elsevier.com/locate/jpba

Review

Hydrophilic interaction chromatography (HILIC) in the analysis of antibiotics夽

Getu Kahsay, Huiying Song, Ann Van Schepdael, Deirdre Cabooter, Erwin Adams ∗
Laboratorium voor Farmaceutische Analyse, Faculteit Farmaceutische Wetenschappen, KU Leuven, O&N2, PB-923, Herestraat 49, B-3000 Leuven, Belgium

a r t i c l e i n f o a b s t r a c t

Article history: This paper presents a general overview of the application of hydrophilic interaction chromatography
Received 28 February 2013 (HILIC) in the analysis of antibiotics in different sample matrices including pharmaceutical, plasma,
Received in revised form 10 April 2013 serum, fermentation broths, environmental water, animal origin, plant origin, etc. Specific applications of
Accepted 13 April 2013
HILIC for analysis of aminoglycosides, ␤-lactams, tetracyclines and other antibiotics are reviewed. HILIC
Available online 22 April 2013
can be used as a valuable alternative LC mode for separating small polar compounds. Polar samples usu-
ally show good solubility in the mobile phase containing some water used in HILIC, which overcomes the
Keywords:
drawbacks of the poor solubility often encountered in normal phase LC. HILIC is suitable for analyzing
Antibiotics
HILIC
compounds in complex systems that elute near the void in reversed-phase chromatography. Ion-pair
Retention mechanism reagents are not required in HILIC which makes it convenient to couple with MS hence its increased
popularity in recent years. In this review, the retention mechanism in HILIC is briefly discussed and a list
of important applications is provided including main experimental conditions and a brief summary of
the results. The references provide a comprehensive overview and insight into the application of HILIC
in antibiotics analysis.
© 2013 Elsevier B.V. All rights reserved.

Contents

1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 142
2. Stationary and mobile phases for HILIC applications . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 143
2.1. Stationary phases in HILIC . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 143
2.2. Mobile phases in HILIC . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 146
3. Use of HILIC for analysis of antibiotics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 146
3.1. Aminoglycoside antibiotics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 146
3.1.1. Analysis of aminoglycosides . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 146
3.1.2. HILIC in the analysis of aminoglycosides . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 147
3.2. ␤-lactam antibiotics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 147
3.3. Tetracycline antibiotics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 150
3.4. Other antibiotics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 151
4. Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 152
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 152

1. Introduction phase, however, exhibited heterogeneity that resulted in peak

tailing and non-linear retention factors with varying analyte
Looking back into the history of liquid chromatography (LC), concentrations [1]. Elution in NPLC has been accomplished by
the pioneer systems were operated in normal phase mode (NPLC) non-polar organic solvents. Some variants like the use of isohy-
which implied that the stationary phase was polar. Such stationary dric solvents were tried, but this approach was found to be rather
tedious and failed to yield good performance over a longer period
[2–4]. The development of stationary phases for chromatography
夽 This is an open-access article distributed under the terms of the Creative Com- driven by pharmaceutical studies of water/octanol partitioning,
mons Attribution-NonCommercial-No Derivative Works License, which permits led to the now well-established reversed-phase (RP) systems [5,6].
non-commercial use, distribution, and reproduction in any medium, provided the
original author and source are credited.
Bonded octadecyl stationary phases allowed efficient separation of
∗ Corresponding author. Tel.: +32 16323444; fax: +32 16323448. analytes within a broad range of polarity and fast column equil-
E-mail address: Erwin.Adams@pharm.kuleuven.be (E. Adams). ibration. The lack of retention of highly hydrophilic compounds

0731-7085/$ – see front matter © 2013 Elsevier B.V. All rights reserved.
http://dx.doi.org/10.1016/j.jpba.2013.04.015
 G. Kahsay et al. / Journal of Pharmaceutical and Biomedical Analysis 87 (2014) 142–154
High
sensitivity
HILIC
ESI-MS
sensitivity Reversed Phase
Normal Phase
Low
sensitivity
Polar
Analyte Polarity
Fig. 1. HILIC as a complement to normal phase and reversed-phase LC separations.
with ionizable functional groups has been supplemented by ion-
exchange chromatography [7] or ion pairing on RP columns [8].
However, for a large group of strongly hydrophilic compounds that
could not be ionized in solution, it was impossible to obtain reten-
tion on either stationary phase. The problem has been overcome
sometimes in GC by derivatization, and recently, in LC by develop-
ing hydrophilic interaction chromatography (HILIC) [9].
The term “hydrophilic” refers to the affinity to water. The con-
cept of HILIC is as follows: a polar stationary phase retains polar
analytes that are eluted by a mixture of organic solvent (usually ace-
tonitrile (ACN)) and water. HILIC is an alternative high-performance
liquid chromatography (HPLC) mode for separating polar com-
pounds. HILIC has been reported as a variant of NPLC, but the
separation mechanism in HILIC is more complicated than that
in NPLC [1,10,11]. While the acronym HILIC was first suggested
by Alpert in 1990 [9], the number of publications on HILIC has
increased substantially since 2003 as indicated elsewhere [11] and
as outlined in the review by Hemström and Irgum [12]. Alpert noted
that charged basic groups in a solute led to pronounced hydrophilic-
ity and retention. He considered that the retention mechanism
consisted mostly of partitioning between the bulk mobile phase and
a layer of mobile phase enriched with water, partially immobilized
on the stationary phase [9]. Hemström and Irgum [12] concluded
that both adsorption and the partitioning mechanism contribute
to retention in HILIC. Models describing the retention mechanism
in HILIC can be found in references [10] and [13]. Like NPLC, HILIC
employs traditional polar stationary phases such as silica, amino or
cyano, but the mobile phase used is similar to those employed in
the RPLC mode [9,11,12].
Originally, HILIC was applied mainly for the determination of
carbohydrates, amino acids and peptides [10]. Since then, HILIC
has been successfully applied to all types of liquid chromatographic
separations, including small molecules [14], pharmaceutical com-
pounds [15], metabolites [16], drugs of abuse [17], toxins [18],
carbohydrates [19], oligosaccharides [20], amino acids [21], pep-
tides and proteins [9,22]. The interest in the HILIC technique in the
last years has been promoted by growing demands for the analysis
of polar drugs, metabolites and biologically important compounds
in proteomics, glycomics and clinical analysis [23]. HILIC also allows
the analysis of charged substances, as in ion chromatography (IC).
Another reason for the increasing popularity of HILIC is its excel-
lent suitability for coupling to mass spectrometry (MS). Fig. 1
shows how HILIC complements other areas of chromatography and
extends the range of separation options [11].
In general, advantages of HILIC have been summarized [24] as
follows: (i) good peak shapes can be obtained for bases, (ii) MS
143
sensitivity is enhanced due to the high organic content in the mobile
phase and the high efficiency of spraying and desolvation tech-
niques, (iii) direct injection can often be made of extracts eluted
from C18 solid-phase extraction columns with solvents of high
organic content, as these are weak solvents in HILIC, (iv) the order
of elution of solutes is different and generally the opposite of that
found in RP separations, (v) good retention of polar compounds is
obtained in HILIC, whereas very poor retention is often obtained
in RPLC and (vi) higher flow rates are possible due to the high
organic content and hence lower viscosity of typical mobile phases.
In addition, polar samples always show good solubility in the water
containing mobile phase used in HILIC, which overcomes the draw-
backs of the poor solubility often encountered in NPLC. Ion-pair
reagents are not required in HILIC, and it can be conveniently cou-
pled to MS, especially in the electrospray ionization (ESI) mode. In
contrast to RPLC, gradient elution HILIC begins with a low-polarity
Apolar
organic solvent and elutes polar analytes by increasing the polar
aqueous content [25].
Other advantages of HILIC that have emerged more recently,
include the use of long columns to achieve highly efficient sepa-
rations, a superior loading capacity to RP for charged basic solutes
and the potential of very fast analysis due to the good mass transfer
characteristics of the columns operated in mobile phases of much
lower viscosity than typically used in RPLC [24,26]. Nevertheless,
HILIC also has some drawbacks [27] compared to RP separations
which include: (i) the separation mechanism is at present less well
understood than that of RPLC. Thus, it may be difficult to predict the
effect of a change in conditions on the separation outcome, (ii) the
technique does not have the broad applicability of RPLC. Analytes
that are neutral and non-polar generally show very little retention.
In addition, ionized acidic analytes (negatively charged ions) can
show little retention due to repulsion of the ion from negatively
charged column silanol groups on some silica-based columns and
(iii) HILIC is potentially an environmentally less friendly technique
than RPLC as it consumes much larger volumes of organic solvents.
2. Stationary and mobile phases for HILIC applications
2.1. Stationary phases in HILIC
The chemical modification of silica gels using chemical reac-
tions began in the 1960s. Reaction between alkylsilyl chlorides
and silanol groups (Si–OH) is the most common method and was
reported in 1970 [28]. Approximately at the same time as Alpert [9]
introduced the HILIC technique, Huber et al. [29] noted that the cor-
rect selection of a suitable adsorbent is a crucial step for the success
of so-called “solvent generated” liquid–liquid chromatography, the
category to which the HILIC mechanism can be attributed [30].
The family of HILIC stationary phases with various support mate-
rials and surface chemistries has continuously enlarged since, to
suit specific separation problems. The basic types of HILIC columns
include plain silica, neutral, polar chemically bonded, ion-exchange
and zwitterionic stationary phases. NP separations on bare sil-
ica and amino-silica columns in aqueous-organic mobile phases
were reported as early as the mid-seventies [31]. Using neutral
stationary phases (e.g. diol and amide phases) there will be no
electrostatic interactions, whereas with charged phases (e.g. plain
silica and aminopropyl phases) strong electrostatic interactions can
take place between the analytes and the stationary phase. Zwitteri-
onic phases (e.g. sulfobetaine silica) exhibit only weak electrostatic
interactions with analytes [1].
The first separations in HILIC mode were reported in 1975.
Linden and Lawhead [31] separated carbohydrates using an amino-
silica phase, Bondapak (Waters, Milford, MA, USA) in a mixture
of ACN and water (75:25 v/v). The next generation of stationary
144 G. Kahsay et al. / Journal of Pharmaceutical and Biomedical Analysis 87 (2014) 142–154

Table 1
Summary of stationary phases used in HILIC separations [11].

Packing materials Structure of stationary phase Application

Underivatized silica stationary [12,23]

phases that contain
functional groups such as
siloxanes and silanols with
(or without) a small quantity
of metals

Polymeric structures of poly [23]

(succinimide) derivatives

Amino-bonded phases [23,31]

Diol bonded phases [32]

Amide bonded phases [23,33]

Alkylamide [34]

Mix-mode [35]

Cyano bonded phases [45]

 G. Kahsay et al. / Journal of Pharmaceutical and Biomedical Analysis 87 (2014) 142–154 145

Table 1 (Continued)

Packing materials Structure of stationary phase Application

Polyethylene glycol/silica (HS [46]

PEG)

“Click” ␤-cyclodextrin [47]

“Click” saccharides (“click” [48]

maltose)

“Click” dipeptide [49]

Zwitterionic sulfobetaine [50]

bonded phases (ZIC-HILIC)

Cationic exchangers bonded [51]

phases

Mix-mode RP/anionic [50,52]

exchangers bonded phases

phases for HILIC used diol- and amide-silica. The diol-silica col-
umn has mainly been used for the separation of proteins [32].
Amide-silica columns have been available since the mid-eighties.
This particular phase is described as consisting of non-ionic car-
bamoyl groups that are chemically bonded to the silica gel, but is
commonly known as an amide bonded silica. After Yoshida [33]
applied these phases for the separation of peptides, the amide-
silica phase soon found common usage in HILIC. Chemically bonded
stationary phases with specific structural properties have been pre-
pared by Buszewski et al. [34,35]. Since then silica hydride, hybrid
146 G. Kahsay et al. / Journal of Pharmaceutical and Biomedical Analysis 87 (2014) 142–154
silica-organic phases and a large variety of silica-based or polymer
HILIC columns have been designed. Amide-, cyano-, diol-, polyeth-
ylene glycol-, poly(succinimide)-, sulfoalkylbetaine-, cyclodextrin,
pentafluorophenylpropyl, polyvinylalcohol, polypeptide and other
polar chemically bonded stationary phases have become avail-
able for HILIC separations, some of them showing both HILIC and
RP application possibilities [10]. Hence, the structural variations
of HILIC-type stationary phases are wider than those found in
reversed-phase systems. Table 1 shows a summary of the different
structures of stationary phases that are applicable to HILIC-mode
separation [11].
Chirita et al. [36] suggested a good scheme for column selec-
tion and applied it to the analysis of neurotransmitters. In terms of
the retention characteristics and selectivity, their column selection
method seems to be acceptable but the separation efficiency must
be considered too. The characterization of HILIC stationary phases
has been reported, but there is no test scheme to describe the struc-
ture selectivity relationships for HILIC phases, in contradistinction
with the well-accepted column tests for RP stationary phases
[37–40]. Kawachi et al. [41] suggested an inclusive test scheme
of HILIC stationary phases using trimethylphenylammonium chlo-
ride, sodium p-toluenesulfonate, nucleosides, saccharides, and
xanthines to describe the degree of hydrophilicity, the selectiv-
ity for hydrophilic and hydrophobic groups, positional selectivity
and the configuration of hydrophilic groups, the anion and cation
exchange properties, the local pH conditions on the stationary
phases and the shape selectivity. In this test scheme, it was possible
to divide the HILIC phases into several groups with varying selec-
tivity. This can be helpful to select stationary phases in combination
with the structural characteristics of the target analyte.
2.2. Mobile phases in HILIC
A typical mobile phase for HILIC chromatography includes
water-miscible polar organic solvents with a small amount of water
[9]. Although the retention mechanism in HILIC is not completely
understood, it is supposed to be caused by partitioning. This mode
is based on the differential distribution of the injected analyte
molecules between the organic-rich mobile phase and a water-
enriched layer adsorbed onto the hydrophilic stationary phase.
The more hydrophilic the analyte, the more the partitioning equi-
librium is shifted towards the immobilized water layer on the
stationary phase, and thus, the more the analyte is retained [9,12].
Usually, ACN is used as organic solvent, but any aprotic solvent
that is miscible with water (e.g., tetrahydrofuran, THF) can be used.
Alcohols can also be adopted, although a higher concentration is
needed to achieve the same degree of retention of the analyte rel-
ative to an aprotic solvent–water combination [12]. HILIC on polar
stationary phases employs organic-rich mobile phases, usually con-
taining 5–40% water or a volatile buffer if used in combination
with MS or charged aerosol detection. Mobile phase pH and ionic
strength significantly affect the retention and separation selectivity
of ionizable analytes in HILIC separations which can be detrimen-
tal for the robustness. Buffer salts and mobile phase additives such
as ammonium acetate, ammonium formate, ammonium phosphate
and trifluoroacetic acid are commonly used to improve peak shape
and to control analyte polarity so as to effect differential changes
in retention. However the positive impact of mobile phase pH and
ionic strength on a separation also depends on the properties of
the stationary phase [10,11]. It is recommended to perform initial
experiments at a relatively high concentration (e.g., 40%) of aqueous
buffer in ACN, to ensure the elution of all sample compounds. Then
the elution strength is adjusted by increasing the concentration of
ACN, until acceptable sample retention is achieved. Alternatively,
a scouting gradient of decreasing ACN concentration can be run.
Gradients with high initial concentration of organic modifier and
Fig. 2. Schematic picture of an adsorbed diffuse water layer at the surface of a polar
stationary phase in highly organic environment.
Figure adapted from [10].
increasing concentrations of water were successfully applied for
HILIC separations of various biological samples, especially with
bare silica columns. The gradient usually starts from 95% ACN
containing 5% aqueous ammonium acetate or ammonium formate
buffer, with gradually decreasing ACN concentration. Running the
gradient up to a high water concentration (up to 90%) may help
removing strongly retained sample interferences [10].
In HILIC water acts as the strong solvent, hence the higher the
organic content the greater the retention of the polar analytes will
be [42]. For this reason, solvents typically used in HILIC contain
60–90% of organics and are thus suitable for LC/MS analysis using
ESI, without significant changes in ionization efficiency during gra-
dient elution [1,42]. In mobile phases containing some water, the
preferential adsorption causes formation of a water-rich adsorbed
liquid multi-layer (Fig. 2). In aqueous-organic NP containing more
than 0.5–1% water, the layer of adsorbed water is thick enough to
induce liquid–liquid partition between the bulk mobile phase and
the adsorbed aqueous liquid layer. However, sample adsorption on
polar surface centers may significantly contribute to the retention
[10]. The NPLC drawback of insolubility of hydrophobic compounds
is largely solved in HILIC because of its mobile phase properties. It
often provides sufficient retention of strongly polar compounds,
for which it offers different selectivity compared to traditional RP
chromatography [9,43].
The selection of the organic solvent in HILIC has a strong effect on
the retention. The elution strength of organic solvents in the HILIC
mode increases generally in the order of increasing solvent polarity
and ability to participate in proton–donor/proton–acceptor inter-
actions: methanol > ethanol > 2-propanol > THF > ACN [43]. ACN is
strongly preferred as the organic compound because mobile phases
containing other solvents often provide insufficient sample reten-
tion and broad or non-symmetrical peak shapes [10]. Another
interesting organic solvent to be used in the mobile phase is THF,
which is a strong hydrogen bond acceptor which may result in
a different analyte elution order than ACN. Polar protic solvents,
such as methanol, ethanol or isopropanol can act as both hydrogen
bond donors and acceptors and can compete for active polar sites
on the HILIC surface. Therefore analytes, whose retention is based
on strong hydrogen bonding, are poorly retained. Approaches for
method development in HILIC have been extensively discussed in
a publication by Dejaegher et al. [44].
3. Use of HILIC for analysis of antibiotics
3.1. Aminoglycoside antibiotics
3.1.1. Analysis of aminoglycosides
Aminoglycosides (AGs) are very polar and lack chromophores
(neomycin B is shown as an example in Fig. 3), which makes them
 G. Kahsay et al. / Journal of Pharmaceutical and Biomedical Analysis 87 (2014) 142–154
Fig. 3. Chemical structure of neomycin B.
difficult to separate using conventional RPLC with UV detection.
Researchers have overcome this problem by employing derivatiza-
tion agents and ion-pair reagents for detection [53–63]. However,
derivatization steps make the analytical process more time con-
suming and often give problems with quantitation. HILIC combined
with MS has been receiving great attention for AGs analysis as there
is no need for derivatization or addition of ion-pairing reagents.
3.1.2. HILIC in the analysis of aminoglycosides
HILIC has been widely used to analyze aminoglycosides in differ-
ent matrices (Table 2), including muscle and kidney [64,65], plasma
[66], serum [67,68], manure [69], apples [70], milk [71], honey [72]
and bulk samples [73]. Kumar et al. [72] reported a HILIC method for
the analysis and investigation of the chromatographic behavior of
several AGs (streptomycin, dihydrostreptomycin, spectinomycin,
apramycin, neomycin, paramomycin, kanamycin A and gentam-
icins). Stationary phases with three different charge states, such
as bare silica (negative), aminopropyl (positive), amide (neutral)
and ZIC-HILIC (zwitterionic), were chosen for the screening exper-
iments. Mobile phase constituents (pH and ionic strength) and
column temperature were also investigated. The zwitterionic HILIC
provided the most satisfactory separation and sensitivity. The
method was directly applicable for the analysis of AG residues in
honey. Based on these research results, Kumar et al. [74] continued
to study AG residues in kidney samples from food-producing ani-
mals and in honey samples by zwitterionic HILIC in combination
with a triple quadrupole mass analyzer. The LOQ ranged from 2 to
125 ␮g/kg in honey and 25 to 264 ␮g/kg in kidney samples, which
is well below the maximum residue limits (MRLs) established.
Also in other studies zwitterionic HILIC stationary phases were
found to perform well [64,67,68,72,74]. Six AGs (amikacin, genta-
micin, kanamycin, neomycin, paromomycin and tobramycin) were
simultaneously quantified by ZIC-HILIC combined with ESI-MS/MS
detection in human serum [67]. Similarly, neomycin was quanti-
tated by LC–MS/MS using HILIC in human serum [68]. Analysis of
neomycin, apramycin and kanamycin was possible by zwitterionic
HILIC with MS detection and the same column was also successfully
employed in a retention mechanism study (Fig. 4) [75]. A crucial
part in the separation mechanism is a water layer, which is built up
when equilibrating the stationary phase with the mobile phase. A
polar analyte will experience hydrophilic partitioning between the
water layer and the mobile phase. Besides, a charged analyte may
have weak and/or strong electrostatic interactions (both attraction
and repulsion) by the zwitterionic groups, which greatly contribute
to the separation [10]. Since the AGs have strong electrostatic inter-
action with the zwitterionic stationary phase, it is necessary to use
formic acid and a high buffer concentration in the mobile phase to
achieve good chromatographic performance.
Multi-residue AG antibiotics in bovine and porcine muscle
and kidney were quantitated using LC–MS/MS [64]. A ZIC-HILIC
147
Fig. 4. Retention of polar/hydrophilic compounds on a zwitterionic HILIC stationary
phase.
Figure adapted from [75].
column was employed for the chromatographic separation. The
method was found to be sensitive, robust and highly specific for
the simultaneous quantification of the AGs with LOQs of 25 ng/g
for gentamicin, 50 ng/g for spectinomycin, dihydrostreptomycin,
kanamycin and apramycin; and 100 ng/g for streptomycin and
neomycin, which are all well below the maximum residue limits
set by the Codex Alimentarius Commission [64].
Streptomycin (STR) and dihydrostreptomycin (DHSTR) are two
of the most commonly used AG antibiotics in veterinary medicine.
Gremilogianni et al. [71] compared HILIC with ion-pair chromatog-
raphy (IPC) for the determination of STR and DHSTR residues in
milk. Comparison of the validation parameters (sensitivity, pre-
cision and recovery) of these two methods revealed superiority
for the HILIC method (Table 3). HILIC separation was performed
using a silica-based HILIC Fortis column. The sensitivity of the HILIC
method was greater than that of the IPC method for both STR and
DHSTR.
A HILIC–MS/MS method for detection and quantification of
spectinomycin and lincomycin from liquid manure and rainfall
run-off was described by Peru et al. [69]. The chromatographic
separation was performed on a silica-based Altima HP HILIC col-
umn. MS detection was carried out using an atmospheric pressure
chemical ionization (APCI) interface in multiple reaction monitor-
ing (MRM) positive ion mode. They found that the method was
sensitive for the quantification of the antibiotics and that HILIC
provided excellent retention and separation of spectinomycin and
lincomycin from interfering matrix components without the need
for ion-pairing reagents.
An automated LC–MS/MS method using a CAPCELL PAK ST
HILIC column for the simultaneous quantification of 15 AGs
residues in muscle, liver (pigs, chicken and cattle), kidney (pigs
and cattle), cow milk and hen eggs was reported elsewhere
[65]. The 15 AGs consist of streptomycin (STREP), amikacin
(AMIKA), hygromycin B (HYGRO), dihydrostreptomycin (DIHY),
netilmicin (NETIL), kasugamycin (KASUG), kanamycin B (KANA),
sisomicin (SISO), spectinomycin (SPECT), gentamicin C1 (GENT),
apramycin (APRA), paromomycin (PAROM), tobramycin (TOBRA),
ribostamycin (RIBOS) and neomycin B (NEOM). The detection capa-
bilities in this method ranged from 10 to 50 ␮g/kg. The MRM
LC–MS/MS chromatograms of the extractions of blank bovine liver
spiked with these AGs are shown in Fig. 5. The report concluded
that the LC–MS/MS method could be applied to trace analysis of
multi-component AG contaminants in complex sample matrices.
The HILIC method gave satisfactory retention and separation of the
AGs residues.
3.2. ˇ-lactam antibiotics
Penicillins and cephalosporins have been thoroughly inves-
tigated in RP-HPLC systems coupled with different methods of
 148
Table 2

G. Kahsay et al. / Journal of Pharmaceutical and Biomedical Analysis 87 (2014) 142–154

HILIC applications in the analysis of aminoglycosides.

Matrix Compound (s) of interest Stationary phase Mobile phase Detection Refs

Kidney and muscle tissues Gentamicin/spectinomycin/dihydrostreptomycin/ ZIC-HILIC (SeQuant) 150 mM ammonium acetate + 1% ESI-MS [64]
kanamycin/apramycin/streptomycin/neomycin (100 mm × 2.1 mm, 5 ␮m) FA-ACN; gradient
Muscle, liver, kidney, cow milk and Streptomycin/amikacin/hygromycin CAPCELL PAK ST ACN-0.1% TFA; gradient ESI-MS/MS [65]
hen eggs B/dihydrostreptomycin/netilmicin/kasugamycin/ (150 mm × 2.0 mm, 3 ␮m)
kanamycin B/sisomicin/spectinomycin/gentamicin
C1/ apramycin/paromomycin/tobramycin/
ribostamycin/neomycin B
Mouse, rat and guinea pig plasma Amikacin/streptomycin/spectinomycin/gentamicin ZIC-HILIC (SeQuant) 5 or 25 mM ammonium formate, ESI-MS [66]
(50 mm × 2.0 mm, 5 ␮m) pH 2.5-ACN – 1% FA; gradient
Human serum Amikacin/gentamicin/kanamycin/neomycin/ ZIC-HILIC ACN-2 mM ammonium acetate – ESI-MS/MS [67]
paromomycin/tobramycin (100 mm × 2.1 mm) FA; gradient
Human serum Neomycin ZIC-HILIC ACN-10 mM ammonium acetate – ESI-MS/MS [68]
(100 mm × 2.1 mm) FA; gradient
Manure Spectinomycin/lincomycin Altima HP HILIC ACN-water-0.1% FA; isocratic APCI-MS/MS [69]
(150 mm × 2.1 mm, 3 ␮m)
Apples Streptomycin Atlantis HILIC silica A: water + 0.05% FA/B: ACN + 0.05% ESI-MS/MS [70]
(150 mm × 2.1 mm, 3 ␮m) FA; gradient
Milk Streptomycin/dihydrostreptomycin HILIC Fortis 150 mM ammonium formate-ACN, ESI-MS [71]
(100 mm × 3.0 mm, 3 ␮m) pH 4.5; gradient
Honey Streptomycin/spectinomycin/dihydrostreptomycin/ ZIC-HILIC 175 mM ammonium formate, pH ESI-MS/MS [72]
gentamicin C1/gentamicin C1a/gentamicin C2/C2a/ (150 mm × 2.1 mm, 3.5 ␮m) 4.5 – 0.2% FA in ACN; gradient
Apramycin/paromomycin/kanamycin A/neomycin B
Bulk samples Impurities in streptomycin and dihydrostreptomycin Halo HILIC 100–200 mM ammonium formate, ESI-QIT/TOFMS [73]
(100 mm × 2.1 mm, 2.7 ␮m) pH: 3.0–4.5-ACN; isocratic

FA: formic acid; TFA: trifluoroacetic acid; QIT: quadrupole ion trap.
 G. Kahsay et al. / Journal of Pharmaceutical and Biomedical Analysis 87 (2014) 142–154
Table 3
Comparison of the validation parameters of HILIC versus IPC method [71].
HILIC method
STR
LOQ 13.9 ␮g/kg
Recovery 85.5%
Inter-day precision 6.7–8.1%
detection such as UV detection [76,77], MS [78,79] or chemi-
luminescence detection [80]. Since these compounds are rather
small polar molecules, they could hardly be analyzed in RP-HPLC
mode. Ion-pairing reagents, buffers and acids are often used as
additives [81–83] to improve the retention and peak shape [84].
In addition, multidimensional [85] and capillary HPLC systems
have been developed to analyze cephalosporins at lower concen-
trations in different sample matrices [86]. Some cephalosporins
were used as model substances in mixtures with other drugs in
HILIC mode, but their particular chromatographic retention was
not explained [23,87]. It is reported that HILIC was also used to
analyze cephalosporin C [88,89].
The simple mobile phase of HILIC and its compatibility with
MS opened a new door for the analysis of cephalosporins. Liu
et al. [88] developed a HILIC method to separate seven commonly
used cephalosporins (cefotaxime sodium, cefpiramide, cefazolin
Fig. 5. MRM LC–MS/MS chromatograms of the extractions of blank bovine liver spiked with AGs equivalent to detection capabilities of 20 ␮g/kg for APRA, STREP, DIHY,
SPECT, GENT, TOBRA, PAROM, HYGRO, RIBOS, KASUG, AMIKA, NEOM, KANA, SISO, NETIL.
Figure adapted from [65].
149
IPC method
DHSTR STR DHSTR
14 ␮g/kg 109 ␮g/kg 31 ␮g/kg
72.3% 69.3% 56.5%
9.8–11.2% 8.9–12.1% 9.7–13.4%
sodium, cefepime hydrochloride, cefixime, ceftazidime and cef-
triaxone sodium). Chromatograms for the seven cephalosporins
on the Click ␤-CD column and Atlantis HILIC silica column are
shown in Fig. 6. The orthogonality between HILIC and RPLC for
cephalosporins was also investigated. The authors further devel-
oped a successful HILIC method to analyze cefotaxime sodium and
its degradation products. A HILIC–MS/MS method for the simul-
taneous determination of three polar, non-structurally related
compounds – (1) a carbapenem antibiotic, imipenem (IMP), (2) a
renal dehydropeptidase inhibitor, cilastatin (CIL) and (3) an investi-
gational ␤-lactamase inhibitor MK-4698 (BLI) – in biological fluids
was described elsewhere [89]. A HILIC-ESI-MS/MS method for the
multitarget quantitative analysis of the hydrophilic metabolites
of penicillins and cephalosporins has also been described [90].
Jovanović et al. [91] reported the chromatographic behavior of
mixture of ␤-lactam antibiotics (cefotaxime, cefalexin, cefaclor,
150 G. Kahsay et al. / Journal of Pharmaceutical and Biomedical Analysis 87 (2014) 142–154

Fig. 6. Chromatograms for seven cephalosporins on the Click ␤-CD column (a) and Atlantis HILIC silica column (b, c). Mobile phase: A, 10 mM ammonium formate at pH 6.8;
B, ACN-100 mM ammonium formate = 90:10 at pH 6.8. Gradient for (a) and (b) was 88–65% B in 20 min and 65% B in the next 10 min; gradient for (c) was 100–75% B in 20 min
and 75% B in the next 15 min. (1) cefotaxime sodium, (2) cefpiramide, (3) cefazolin sodium, (4) cefepime hydrochloride, (5) cefixime, (6) ceftazidime and (7) ceftriaxone
sodium.
Figure adapted from [88].

cefuroxime, cefuroxime axetil, ampicillin and amoxicillin) using In order to avoid this problem, adding chelating agents, such as
HILIC-UV and their retention prediction models were designed by oxalic acid and EDTA salts to the mobile phase and RP end-capped
applying Box–Behnken Design. ␤-lactam antibiotics analyzed using columns have been employed. Using HILIC silica with high-purity
HILIC with different detection systems are summarized in Table 4. silica columns, non-selective reactions such as complex formation

3.3. Tetracycline antibiotics

Tetracycline antibiotics (TCs) are a large class of antibiotics that

characteristically contain an octahydronaphthacene ring skeleton
with four fused rings. The various TCs mainly differ in their substi-
tution groups at the C5, C6 and C7 positions and include tetracycline
(TC), chlortetracycline (CTC), doxycycline (DC) and oxytetracycline
(OTC) (Fig. 7).
Analysis methods of TCs over the past 20 years have been R1 R2 R3 R4
reviewed [92–98]. Among them, LC using a C18 or C8 column, with
Teteracycline (TC) H OH CH3 H
UV, fluorometry and mass spectrometry detection are generally
preferred. However, owing to the presence of two ketone groups Chlortetracyclin (CTC) H OH CH3 Cl
in positions 1 and 11, and the enols around C10 and C12, TCs can
readily chelate with trace metal ions present in silica packing mate- Oxytetracycline (OTC) OH OH CH3 H
rial or from the equipment. Most of them have a strong tendency to Doxycycline (DC) H H CH3 H
irreversibly bind to the silanol groups in a silica-based packing col-
umn. As a result, this causes severe tailing of TCs peaks [99–102]. Fig. 7. Structure of some tetracyclines.
 G. Kahsay et al. / Journal of Pharmaceutical and Biomedical Analysis 87 (2014) 142–154 151

Table 4
Summary of ␤-lactam antibiotics analyzed using HILIC with different detection systems.

Matrix Compound (s) of interest Stationary phase Mobile phase Detection Refs

Fermentation broths
Raw materials
Plasma, blood
Reference substances
2-Aminoadipic acid, ZIC-HILIC (and A: 10% (v/v) 200 mM ESI-MS/MS [88]
2-amino-5-(4-carboxy-2-thiazolyl)-valeric ZIC-HILIC Guard FA, pH 4.0 in water/B:
acid, 6-aminopenicillanic acid, 20 mm × 2.1 mm; 10% (v/v) 200 mM FA,
6-aminopenicilloic acid, 5 ␮m) pH 4.0 in ACN; gradient
7-aminocephalosporanic acid,
8-hydroxypenillic acid, cephalosporin C,
cephalosporin C lactone,
deacetoxycephalosporin,
deacetylcephalosporin, penicillamine
disulfide, phenoxyacetic acid,
phenoxymethylpenicilloic acid,
phenoxymethylpenillic acid,
p-hydroxyphenoxyacetic acid,
␦-(l-␣-Aminoadipoyl)-l-Cys-d-Val
Ceftazidime, cefixime, cefpiramide, Atlantis HILIC silica A: 10 mM ammonium UV/vis ESI-MS/MS [89]
ceftriaxone sodium, cefotaxime sodium, (100 mm × 2.1 mm, formate, pH 6.8/B:
cefazolin sodium, cefepime hydrochloride 5 ␮m) ACN-100 mM
and degradation products of cefotaxime Click ␤-CD column ammonium formate,
sodium (150 mm × 2.1 mm, pH 6.8; gradient
5 ␮m)
Imipenem (IMP), cilastatin (CIL), MK-4698 Atlantis HILIC silica 15 mM ammonium TIS-MS/MS [90]
␤-lactamase inhibitor (BLI) (50 mm × 2.1 mm, formate, pH 3 in 80%
3 ␮m) ACN; isocratic
Cefotaxime, cefalexin, cefaclor, Alltech Silica ACN-ammonium UV/vis [91]
cefuroxime, cefuroxime axetil, ampicillin (250 mm × 4.6 mm, acetate, pH 4.5–6.5;
and amoxicillin 5 ␮m) isocratic

or ion effects leading to less favorable separations could be pre-
vented [103,104].
TCs are not only widely used as veterinary medicines, but also as
growth additives in animal feeds or water [92]. It has been reported
that only a small portion of the applied TCs (veterinary or food
additives) is metabolized or absorbed in animals and most of the
unmetabolized form is released in excreta [105], which enters into
the environment and disrupts the indigenous microbial population.
Therefore, many researchers focus on the development of a rapid
and sensitive method for the determination of TCs in the environ-
ment. Li et al. [106] reported the environmental fate and transport
of four zwitterionic TCs (oxytetracycline, doxycycline, chlortetracy-
cline and tetracycline) and developed a HILIC–MS/MS method for
detection and quantification of these antibiotics in environmen-
tal waters. The chromatographic separation was achieved on an
amino-bonded silica HILIC column under isocratic conditions. Dur-
ing method development, the effects of mobile phase components
(buffer type, pH and type and concentration of organic modifier)
on retention and separation of TCs on the column was investigated
and a mixed-mode retention mechanism composed of partitioning,
adsorption and ion-exchange interaction was proposed.
Determination of OTC, TC and CTC in different sample matri-
ces using a HILIC or mixed HILIC–ion-exchange mode were also
described elsewhere [107,108]. The methods gave rapid and robust
separations of the TCs. HILIC using high-purity silica as station-
ary phase has been applied for the separation of epirubicin and its
analogues [109]. Several parameters affecting the chromatographic
behavior such as organic modifier, buffer pH and ion strength have
been investigated. Of utmost importance for successful separation
of the analogues (doxorubicin, daunorubicin and epidaunorubicin)
is the choice of organic modifier. ACN was shown to offer superior
separation to methanol, isopropanol or THF. Table 5 summarizes
the application of HILIC for TCs in different sample matrices.
3.4. Other antibiotics
Besides ␤-lactams, AGs and TCs, HILIC has been widely
employed for the analysis of other antibiotics (Table 6). Although
RPLC is the predominant LC mode applied to peptide separation,
there is a growing awareness of the potential of HILIC as a comple-
mentary mode in situations where RPLC does not offer the required
retention and overall separation efficiency. Determination of the
glycopeptide antibiotic avoparcin in kidney using a 200 Å, 5 ␮m
HILIC column was reported by Curren and King [110]. Van Dorpe
et al. [111] reported HILIC analysis of peptide antibiotics (van-
comycin and polymyxin) using Alltima HP and ZIC-HILIC columns.
Determination of 13 sulfonamide antibacterials in milk and eggs
using the polymer monolith microextraction (PMME) technique
was also possible [112] on a Luna NH2 HILIC column followed by
MS detection. Quantification of antibacterial drugs like carbadox
and olaquindox with matrix solid-phase dispersion (MSPD) extrac-
tion in swine feed using a BEH HILIC column [113] and bicozamycin

Table 5
HILIC in the analysis of tetracylines antibiotics.

Matrix Compound (s) of Stationary phase Mobile phase Detection Refs

interest

Environmental water
Environmental waters
Drug substances
Epirubicin raw material
OTC, TC, CTC, Kromasil 100-5 NH2 ACN-6.7 mM ammonium citrate, UV–vis [106]
doxycycline (250 mm × 4.6 mm, 5 ␮m) pH 4.0 (85:15, v/v); isocratic
OTC Kromasil KR100-5SIL ACN-10 mM oxalate, pH 2.5 (90:10, UV–vis [107]
(250 mm × 4.6 mm, 5 ␮m) v/v); isocratic
TC, CTC, OTC ThermoHypersil APS2 ACN-6.7 mM citrate, pH 5.0 (85:15, UV–vis [108]
(50 mm × 4.6 mm, 3 ␮m) v/v); isocratic
Epidaunorubicin, Kromasil KR100-5SIL 30 mM sodium formate, pH UV–vis [109]
daunorubicin, (250 mm × 4.6 mm, 5 ␮m) 2.9-ACN, (90:10, v/v); isocratic
epirubicin, doxorubicin
152 G. Kahsay et al. / Journal of Pharmaceutical and Biomedical Analysis 87 (2014) 142–154

Table 6
HILIC in the analysis of other antibiotics.

Matrix Compounds of interest Stationary phase Mobile phase Detection Refs

Kidney Avoparcin Polyhydroxyethyl aspartamide 47% 15 mM triethyl ammonium UV [110]

HILIC phosphate in ACN; isocratic
(200 mm × 4.6 mm, 5 ␮m)
Bulk substances Vancomycin and polymyxin Alltima HP HILIC A: 0.1% w/v FA in ACN Photodiode array detector [111]
(250 mm × 2.1 mm, 5 ␮m) B: 0.1% w/v FA in water
Milk and eggs Sulfonamides antibacterial Luna HILIC ACN + 0.05% FA-water; gradient ESI-MS [112]
residues (150 mm × 2.0 mm, 3 ␮m)
Swine feed Carbadox and olaquindox BEH HILIC 10 mM ammonium acetate in Photodiode array detector [113]
(100 mm × 2.1 mm, 1.7 ␮m) ACN-water (95:5, v/v); isocratic
Milk Bicozamycin TSK-Gel Amide 80 50 mM ammonium acetate in ESI-MS/MS [114]
(150 mm × 2.0 mm, 5 ␮m) water (pH 4)-ACN; gradient
Human plasma Levofloxacin Atlantis HILIC silica ACN-100 mM ammonium formate, MS/MS [115]
(50 mm × 3 mm, 5 ␮m) pH 6.5 (82:18 v/v); gradient
Rat blood Gemifloxacin ZIC-HILIC-C18 ACN-10 mM ammonium acetate Fluorescence [116]
(100 mm × 4.6 mm, 5 ␮m) (pH 3.5, 80:20, v/v); isocratic
Distillers grains Ampicillin, Penicillin G, OTC, Atlantic HILIC silica 0.1% aqueous FA – ACN; gradient ESI-MS [117]
TC, CTC, Bacitracin A, (100 mm × 2.1 mm, 5 ␮m)
Virginiamycin M1,
Erythromycin A, Tylosin A,
Clarithromycin, Monensin A,
STR
Chicken muscle Aminoglycosides (3), ZIC-HILIC A: 50 mM ammonium formate, pH ESI-MS/MS [118]
amprolium, ␤-lactams (3), (2.1 mm × 100 mm, 3.5 ␮m) 2.5; B: ACN; gradient
lincosamides (2), macrolides
(4), quinolones (4),
sulfonamides (4) and
tetracyclines (3)

using a TSK-GEL NH2 HILIC column and MS detection [114] showed
to be successful with greater separation efficiency. Analysis of levo-
floxacin in human plasma using an Atlantis HILIC silica column
and determination of gemifloxacin on dried blood samples using
a ZIC-HILIC C18 column have also been reported [115,116] and the
methods were successfully applied for pharmacokinetic studies of
these drugs.
4. Conclusion
The use of HILIC for the analysis of antibiotics is reviewed. In
recent years, the HILIC separations have received greater atten-
tion mainly due to their versatility for the analysis of polar drugs,
metabolites, biologically important compounds, etc. in complex
sample matrices of different origin. Due to the very hydrophilic
nature of most antibiotics, HILIC is more suitable for the analysis
of antibiotics compared to traditional RP chromatography. There
are many kinds of HILIC columns available on the market with
diverse stationary phases. However, bare silica, zwitterionic and
aminopropyl based stationary phases are most commonly used.
ESI-MS/MS is the most popular detection method employed in
HILIC separation of most antibiotics. The on-going developments of
new HILIC methods in conjunction with more powerful detection
systems will probably increase its use and popularity in antibiotics
analysis in the future.
References
[1] J. Bernal, A.M. Ares, J. Pól, S.K. Wiedmer, Hydrophilic interaction liquid chro-
matography in food analysis, J. Chromatogr. A 1218 (2011) 7438–7452.
[2] J.-P. Thomas, A. Brun, J.-P. Bounine, Isohydric solvents in liquid-solid column
chromatography: importance for the reproducibility of chromatographic sep-
arations and application to the experimental determination of mobile phase
polarity, J. Chromatogr. A 139 (1977) 21–43.
[3] G.A. Bens, E. Crombez, P. De Moerloose, Applications of a modified “Isohydric
solvent system” in HPLC on silica gel for the analysis of the macrolide antibi-
otics turimycins and spiramycins, J. Liq. Chromatogr. 5 (1982) 1449–1465.
[4] P. Jandera, J. Churáček, Gradient Elution in Column Liquid Chromatography:
Theory and Practice, J. Chromatogr. Library, Vol. 31, Elsevier, Amsterdam,
1985.
[5] K. Valkó, L.R. Snyder, J.L. Glajch, Retention in reversed-phase liquid chro-
matography as a function of mobile-phase composition, J. Chromatogr. A 656
(1993) 501–520.
[6] J.G. Dorsey, M.G. Khaledi, Hydrophobicity estimations by reversed-phase liq-
uid chromatography, J. Chromatogr. A 656 (1993) 485–499.
[7] A. Nordborg, E.F. Hilder, Recent advances in polymer monoliths for ion-
exchange chromatography, Anal. Bioanal. Chem. 394 (2009) 71–84.
[8] J. Dai, P.W. Carr, Effect of mobile phase anionic additives on selectivity, effi-
ciency, and sample loading capacity of cationic drugs in reversed-phase liquid
chromatography, J. Chromatogr. A 1216 (2009) 6695–6705.
[9] A.J. Alpert, Hydrophilic-interaction chromatography for the separation of
peptides, nucleic acids and other polar compounds, J. Chromatogr. A 499
(1990) 177–196.
[10] P. Jandera, Stationary and mobile phases in hydrophilic interaction chro-
matography: a review, Anal. Chim. Acta 692 (2011) 1–25.
[11] B. Buszewski, S. Noga, Hydrophilic interaction liquid chromatography
(HILIC)—a powerful separation technique, Anal. Bioanal. Chem. 402 (2012)
231–247.
[12] P. Hemström, K. Irgum, Hydrophilic interaction chromatography, J. Sep. Sci.
29 (2006) 1784–1821.
[13] Y. Guo, S. Gaiki, Retention and selectivity of stationary phases for hydrophilic
interaction chromatography, J. Chromatogr. A 1218 (2011) 5920–5938.
[14] Y. Hsieh, J. Chen, Simultaneous determination of nicotinic acid and its
metabolites using hydrophilic interaction chromatography with tandem
mass spectrometry, Rapid Commun. Mass Spectrom. 19 (2005) 3031–3036.
[15] B.A. Olsen, Hydrophilic interaction chromatography using amino and silica
columns for the determination of polar pharmaceuticals and impurities, J.
Chromatogr. A 913 (2001) 113–122.
[16] Y. Hsieh, Potential of HILIC-MS in quantitative bioanalysis of drugs and
metabolites, J. Sep. Sci. 31 (2008) 1481–1491.
[17] A. Gheorghe, A. van Nuijs, B. Pecceu, L. Bervoets, P.G. Jorens, R. Blust, H. Neels,
A. Covaci, Analysis of cocaine and its principal metabolites in waste and
surface water using solid-phase extraction and liquid chromatography–ion
trap tandem mass spectrometry, Anal. Bioanal. Chem. 391 (2008)
1309–1319.
[18] C. Dell‘Aversano, P. Hess, M.A. Quilliam, Hydrophilic interaction liquid
chromatography–mass spectrometry for the analysis of paralytic shellfish
poisoning (PSP) toxins, J. Chromatogr. A 1081 (2005) 190–201.
[19] S.C. Churms, Recent progress in carbohydrate separation by high-
performance liquid chromatography based on hydrophilic interaction, J.
Chromatogr. A 720 (1996) 75–91.
[20] G.O. Staples, M.J. Bowman, C.E. Costello, A.M. Hitchcock, J.M. Lau, N. Leymarie,
C. Miller, H. Naimy, X. Shi, J. Zaia, A chip-based amide-HILIC LC/MS platform
for glycosaminoglycan glycomics profiling, Proteomics 9 (2009) 686–695.
[21] T. Langrock, P. Czihal, R. Hoffmann, Amino acid analysis by hydrophilic
interaction chromatography coupled on-line to electrospray ionization mass
spectrometry, Amino Acids 30 (2006) 291–297.
[22] P.J. Boersema, N. Divecha, A.J. Heck, S. Mohammed, Evaluation and optimiza-
tion of ZIC-HILIC-RP as an alternative MudPIT strategy, J. Proteome. Res. 6
(2007) 937–946.
 G. Kahsay et al. / Journal of Pharmaceutical and Biomedical Analysis 87 (2014) 142–154
[23] M.A. Strege, Hydrophilic interaction chromatography-electrospray mass
spectrometry analysis of polar compounds for natural product drug discovery,
Anal. Chem. 70 (1998) 2439–2445.
[24] D.V. McCalley, Is hydrophilic interaction chromatography with silica
columns a viable alternative to reversed-phase liquid chromatography
for the analysis of ionisable compounds? J. Chromatogr. A 1171 (2007)
46–55.
[25] S. Cubbon, T. Bradbury, J. Wilson, J. Thomas-Oates, Hydrophilic interaction
chromatography for mass spectrometric metabonomic studies of urine, Anal.
Chem. 79 (2007) 8911–8918.
[26] D.V. McCalley, Evaluation of the properties of a superficially porous silica
stationary phase in hydrophilic interaction chromatography, J. Chromatogr.
A 1193 (2008) 85–91.
[27] D.V. McCalley, The challenges of the analysis of basic compounds by high
performance liquid chromatography: some possible approaches for improved
separations, J. Chromatogr. A 1217 (2010) 858–880.
[28] T. Ikegami, K. Tomomatsu, H. Takubo, K. Horie, N. Tanaka, Separation efficien-
cies in hydrophilic interaction chromatography, J. Chromatogr. A 1184 (2008)
474–503.
[29] J.F.K. Huber, M. Pawlowska, P. Markl, Selection of the solid support—a crucial
step for the realization of solvent generated liquid-liquid chromatography, J.
Chromatogr. A 500 (1990) 257–280.
[30] J.F.K. Huber, M. Pawlowska, P. Markl, Solvent-generated liquid-liquid chro-
matography with aqueous ternary systems, Chromatographia 19 (1984)
19–28.
[31] J.C. Linden, C.L. Lawhead, Liquid chromatography of saccharides, J. Chro-
matogr. A 105 (1975) 125–133.
[32] M. Rubinstein, Preparative high-performance liquid partition chromatogra-
phy of proteins, Anal. Biochem. 98 (1979) 1–7.
[33] T. Yoshida, Prediction of peptide retention time in normal-phase liquid chro-
matography, J. Chromatogr. A 811 (1998) 61–67.
[34] B. Buszewski, M. Jezierska-Świtała, S. Kowalska, Stationary phase with spe-
cific surface properties for the separation of estradiol diastereoisomers, J.
Chromatogr. B 792 (2003) 279–286.
[35] S. Kowalska, K. Krupczyńska, B. Buszewski, The influence of the mobile phase
pH and the stationary phase type on the selectivity tuning in high perfor-
mance liquid chromatography nucleosides separation, J. Sep. Sci. 28 (2005)
1502–1511.
[36] R.I. Chirita, C. West, A.L. Finaru, C. Elfakir, Approach to hydrophilic interaction
chromatography column selection: application to neurotransmitters analysis,
J. Chromatogr. A 1217 (2010) 3091–3104.
[37] E. Cruz, M.R. Euerby, C.M. Johnson, C.A. Hackett, Chromatographic
classification of commercially available reverse-phase HPLC columns, Chro-
matographia 44 (1997) 151–161.
[38] U.D. Neue, K. Van Tran, P.C. Iraneta, B.A. Alden, Characterization of HPLC
packings, J. Sep. Sci. 26 (2003) 174–186.
[39] D. Visky, E. Haghedooren, P. Dehouck, Z. Kovács, K. Koczian, B. Noszál, J. Hoog-
martens, E. Adams, Faccilitated column selection in pharmaceutical analyses
using a simple column classification system, J. Chromatogr. A 1101 (2006)
103–114.
[40] K. Kóczián, E. Haghedooren, S. Dragovic, B. Noszál, J. Hoogmartens, E. Adams,
J. Pharm. Biomed. Anal. 44 (2007) 894–905.
[41] Y. Kawachi, T. Ikegami, H. Takubo, Y. Ikegami, M. Miyamoto, N. Tanaka,
Chromatographic characterization of hydrophilic interaction liquid chro-
matography stationary phases: hydrophilicity, charge effects, structural
selectivity, and separation efficiency, J. Chromatogr. A 1218 (2011)
5903–5919.
[42] Z. Hao, B. Xiao, N. Weng, Impact of column temperature and mobile
phase components on selectivity of hydrophilic interaction chromatography
(HILIC), J. Sep. Sci. 31 (2008) 1449–1464.
[43] N.S. Quiming, N.L. Denola, A.B. Soliev, Y. Saito, K. Jinno, Retention behav-
ior of ginsenosides on a poly(vinyl alcohol)-bonded stationary phase in
hydrophilic interaction chromatography, Anal. Bioanal. Chem. 389 (2007)
1477–1488.
[44] B. Dejaegher, D. Mangelings, Y. Vander Heyden, Method development for
HILIC assays, J. Sep. Sci. 31 (2008) 1438–1448.
[45] K. Kaczmarski, W. Prus, T. Kowalska, Adsorption/partition model of liquid
chromatography for chemically bonded stationary phases of the aliphatic
cyano, reversed-phase C8 and reversed-phase C18 types, J. Chromatogr. A
869 (2000) 57–64.
[46] L. Rong, T. Takeuchi, Determination of iodide in seawater and edible salt by
microcolumn liquid chromatography with poly(ethylene glycol) stationary
phase, J. Chromatogr. A 1042 (2004) 131–135.
[47] Z. Guo, Y. Jin, T. Liang, Y. Liu, Q. Xu, X. Liang, A. Lei, Synthesis, chromatographic
evaluation and hydrophilic interaction/reversed-phase mixed-mode behav-
ior of a “Click beta-cyclodextrin” stationary phase, J. Chromatogr. A 1216
(2009) 257–263.
[48] Q. Fu, T. Liang, X. Zhang, Y. Du, Z. Guo, X. Liang, Carbohydrate separation by
hydrophilic interaction liquid chromatography on a ‘click’ maltose column,
Carbohydr. Res. 345 (2010) 2690–2697.
[49] M. Xue, H. Huang, Y. Ke, C. Chu, Y. Jin, X. Liang, Click dipeptide”: a novel station-
ary phase applied in two-dimensional liquid chromatography, J. Chromatogr.
A 1216 (2009) 8623–8629.
[50] O. Núñez, K. Nakanishi, N. Tanaka, Preparation of monolithic silica columns
for high-performance liquid chromatography, J. Chromatogr. A 1191 (2008)
231–252.
153
[51] H. Lindner, B. Sarg, C. Meraner, W. Helliger, Separation of acetylated core
histones by hydrophilic-interaction liquid chromatography, J. Chromatogr. A
743 (1996) 137–144.
[52] M. Lämmerhofer, M. Richter, J. Wu, R. Nogueira, W. Bicker, W. Lindner,
Mixed-mode ion-exchangers and their comparative chromatographic char-
acterization in reversed-phase and hydrophilic interaction chromatography
elution modes, J. Sep. Sci. 31 (2008) 2572–2588.
[53] R.E. Hornish, J.R. Wiest, Quantitation of spectinomycin residues in
bovine tissues by ion-exchange high-performance liquid chromatogra-
phy with post-column derivatization and confirmation by reversed-phase
high-performance liquid chromatography-atmospheric pressure chemi-
cal ionization tandem mass spectrometry, J. Chromatogr. A 812 (1998)
123–133.
[54] S. He, Q.Y. Chen, Y. Sun, Y.C. Zhu, L.X. Luo, J.Q. Li, Y.S. Cao, Determination
of tobramycin in soil by HPLC with ultrasonic-assisted extraction and solid-
phase extraction, J. Chromatogr. B 879 (2011) 901–907.
[55] M. Mashat, H. Chrystyn, B.J. Clark, K.H. Assi, Development and validation of
HPLC method for the determination of tobramycin in urine samples post-
inhalation using pre-column derivatization with fluorescein isothiocyanate,
J. Chromatogr. B 869 (2008) 59–66.
[56] A. Posyniak, J. Zmudzki, J. Niedzielska, Sample preparation for residue
determination of gentamicin and neomycin by liquid chromatography, J.
Chromatogr. A 914 (2001) 59–66.
[57] P. Vinas, N. Balsalobre, M. Hernández-Córdoba, Liquid chromatography on
an amide stationary phase with post-column derivatization and fluorimetric
detection for the determination of streptomycin and dihydrostreptomycin in
foods, Talanta 72 (2007) 808–812.
[58] V. Coquart, M.C. Hennion, Trace-level determination of polar phenolic com-
pounds in aqueous samples by high-performance liquid chromatography and
on-line pre-concentration on porous graphitic carbon, J. Chromatogr. A 591
(1992) 195–201.
[59] D.N. Heller, J.O. Peggins, C.B. Nochetto, M.L. Smith, O.A. Chiesa, K. Moulton,
LC-MS/MS measurement of gentamicin in bovine plasma, urine, milk, and
biopsy samples taken from kidneys of standing animals, J. Chromatogr. B 821
(2005) 22–30.
[60] M. Cherlet, S. De Baere, P. De Backer, Quantitative determination of
dihydrostreptomycin in bovine tissues and milk by liquid chromatography-
electrospray ionization-tandem mass spectrometry, J. Mass Spectrom. 42
(2007) 647–656.
[61] R.H. Granja, A.M. Niño, R.A. Zucchetti, R. Patel, A.G. Salerno, Determination
of streptomycin residues in honey by liquid chromatography–tandem mass
spectrometry, Anal. Chim. Acta 637 (2009) 64–67.
[62] F.L. van Holthoon, M.L. Essers, P.J. Mulder, S.L. Stead, M. Caldow, H.M. Ashwin,
M. Sharman, A generic method for the quantitative analysis of aminogly-
cosides (and spectinomycin) in animal tissue using methylated internal
standards and liquid chromatography tandem mass spectrometry, Anal.
Chim. Acta 637 (2009) 135–143.
[63] H. Berrada, J.C. Moltó, J. Mañes, G. Font, Determination of aminoglycoside and
macrolide antibiotics in meat by pressurized liquid extraction and LC-ESI-MS,
J. Sep. Sci. 33 (2010) 522–529.
[64] R. Ishii, M. Horie, W. Chan, J. MacNeil, Multi-residue quantitation of aminogly-
coside antibiotics in kidney and meat by liquid chromatography with tandem
mass spectrometry, Food Addit. Contam. 25 (2008) 1509–1519.
[65] Y.F. Tao, D.M. Chen, H. Yu, L.L. Huang, Z.Y. Liu, X.Q. Cao, C.X. Yan, Y.H. Pan,
Z.L. Liu, Z.H. Yuan, Simultaneous determination of 15 aminoglycoside(s)
residues in animal derived foods by automated solid-phase extraction and
liquid chromatography–tandem mass spectrometry, Food Chem. 135 (2012)
676–683.
[66] A. Shen, L. Morgan, M.L. Barroso, X. Zhang, Method Development of LC-MS/MS
Analysis of Aminoglycosides Drugs: Challenges and Solutions, Tandem Labs-
Tuyen Nguyen; Sepracor Inc., 2008, pp. 1–8.
[67] R. Oertel, V. Neumeister, W. Kirch, Hydrophilic interaction chromatography
combined with tandem-mass spectrometry to determine six aminoglycosides
in serum, J. Chromatogr. A 1058 (2004) 197–201.
[68] R. Oertel, U. Renner, W. Kirch, Determination of neomycin by LC–tandem
mass spectrometry using hydrophilic interaction chromatography, J. Pharm.
Biomed. Anal. 35 (2004) 633–638.
[69] K.M. Peru, S.L. Kuchta, J.V. Headley, A.J. Cessna, Development of a hydrophilic
interaction chromatography–mass spectrometry assay for spectinomycin
and lincomycin in liquid hog manure supernatant and run-off from cropland,
J. Chromatogr. A 1107 (2006) 152–158.
[70] D.A. Bohm, C.S. Stachel, P. Gowik, Confirmatory method for the determination
of streptomycin in apples by LC–MS/MS, Anal. Chim. Acta 672 (2010) 103–106.
[71] A.M. Gremilogianni, N.C. Megoulas, M.A. Koupparis, Hydrophilic interaction
vs ion pair liquid chromatography for the determination of streptomycin and
dihydrostreptomycin residues in milk based on mass spectrometric detection,
J. Chromatogr. A 1217 (2010) 6646–6651.
[72] P. Kumar, A. Rubies, R. Companyó, F. Centrich, Hydrophilic interaction
chromatography for the analysis of aminoglycosides, J. Sep. Sci. 35 (2012)
498–504.
[73] S.I. Kawano, Analysis of impurities in streptomycin and dihydrostrepto-
mycin by hydrophilic interaction chromatography/electrospray ionization
quadrupole ion trap/time-of-flight mass spectrometry, Rapid Commun. Mass
Spectrom. 23 (2009) 907–914.
[74] P. Kumar, A. Rúbies, R. Companyó, F. Centrich, Determination of
aminoglycoside residues in kidney and honey samples by hydrophilic
154 G. Kahsay et al. / Journal of Pharmaceutical and Biomedical Analysis 87 (2014) 142–154
interaction chromatography-tandem mass spectrometry, J. Sep. Sci. 35 (2012)
2710–2717.
[75] W. Jiang, P. Appelblad, T. Jonsson, P. Hemström, Analysis of aminoglycosides
with a Zwitterionic HILIC stationary phase and mass spectrometry detection,
Chromatography Today 4 (2011) 26–28.
[76] A.C. Moffat, M.D. Osselton, B. Widdop, J. Watts, Clarke’s Analysis of Drugs and
Poisons, Pharmaceutical Press, London, 2004.
[77] B.C. McWhinney, S.C. Wallis, T. Hillister, J.A. Roberts, J. Lipman, J.P. Ungerer,
Analysis of 12 beta-lactam antibiotics in human plasma by HPLC with ultra-
violet detection, J. Chromatogr. B 878 (2010) 2039–2043.
[78] H. Liu, H. Wang, V.B. Sunderland, An isocratic ion exchange HPLC method for
the simultaneous determination of flucloxacillin and amoxicillin in a pharma-
ceutical formulation for injection, J. Pharm. Biomed. Anal. 37 (2005) 395–398.
[79] M. Dousa, R. Hosmanová, Rapid determination of amoxicillin in premixes by
HPLC, J. Pharm. Biomed. Anal. 37 (2005) 373–377.
[80] E. Benito-Peña, J.L. Urraca, M.C. Moreno-Bondi, Quantitative determination of
penicillin V and amoxicillin in feed samples by pressurized liquid extraction
and liquid chromatography with ultraviolet detection, J. Pharm. Biomed. Anal.
49 (2009) 289–294.
[81] E. Nemutlu, S. Kir, D. Katlan, M.S. Beksac, Simultaneous multiresponse opti-
mization of an HPLC method to separate seven cephalosporins in plasma
and amniotic fluid: application to validation and quantification of cefepime,
cefixime and cefoperazone, Talanta 80 (2009) 117–126.
[82] J.C. Garcia-Gonzalez, R. Méndez, J. Martin-Villacorta, Quantitative deter-
mination of semisynthetic cephamycins in human serum and urine by
ion-exchange, reversed-phase and ion-pair chromatography, J. Chromatogr.
A 812 (1998) 197–204.
[83] C.M. Pistos, A. Tsantili-Kakoulidou, M. Koupparis, The effect of ion pairing
reagents in the retention profile of zwitterionic cephalosporins, J. Liq. Chro-
matogr. Relat. Technol. 26 (2003) 937–952.
[84] M.C. Rouan, F. Abadie, A. Leclerc, F. Juge, Systematic approach to the deter-
mination of cephalosporins in biological fluids by reversed-phase liquid
chromatography, J. Chromatogr. 275 (1983) 133–144.
[85] I. Nishino, H. Fujitomo, T. Umeda, Determination of a new oral cephalosporin,
cefmatilen hydrochloride hydrate and its seven metabolites in human and
animal plasma and urine by coupled systems of ion-exchange and reversed-
phase high-performance liquid chromatography, J. Chromatogr. B 749 (2000)
101–110.
[86] M.I. Bailón-Pérez, A.M. Garcia-Campaña, M. del Olmo-Iruela, L. Gámiz-
Gracia, C. Cruces-Blanco, Trace determination of 10 beta-lactam antibiotics
in environmental and food samples by capillary liquid chromatography, J.
Chromatogr. A 1216 (2009) 8355–8361.
[87] M.A. Strege, S. Stevenson, S.M. Lawrence, Mixed-mode anion-cation
exchange/hydrophilic interaction liquid chromatography-electrospray mass
spectrometry as an alternative to reversed phase for small molecule drug
discovery, Anal. Chem. 72 (2000) 4629–4633.
[88] Q. Liu, L. Xu, Y. Ke, Y. Jin, F. Zhang, X. Liang, Analysis of cephalosporins by
hydrophilic interaction chromatography, J. Pharm. Biomed. Anal. 54 (2011)
623–628.
[89] Y. Xu, W. Xie, C.M. Miller-Stein, E.J. Woolf, Hydrophilic interaction chro-
matography/tandem mass spectrometry for the simultaneous determination
of three polar non-structurally related compounds, imipenem, cilastatin and
an investigational ˇ-lactamase inhibitor, MK-4698, in biological matrices,
Rapid Commun. Mass Spectrom. 23 (2009) 2195–2205.
[90] S. Schiesel, M. Lämmerhofer, W. Lindner, Multitarget quantitative metabolic
profiling of hydrophilic metabolites in fermentation broths of ␤-lactam
antibiotics production by HILIC-ESI-MS/MS, Anal. Bioanal. Chem. 396 (2010)
1655–1679.
[91] M. Jovanović, T. Rakić, B. Jančić-Stojanović, A. Malenović, D. Ivanović,
M. Medenica, Assessment of ␤-lactams retention in hydrophilic interac-
tion chromatography applying Box-Behnken design, J. Sep. Sci. 35 (2012)
1424–1431.
[92] H. Oka, Y. Ito, H. Matsumoto, Chromatographic analysis of tetracycline antibi-
otics in foods, J. Chromatogr. A 882 (2000) 109–133.
[93] A. Marzo, L.D. Bo, Chromatography as an analytical tool for selected antibiotic
classes: a reappraisal addressed to pharmacokinetic applications, J. Chro-
matogr. A 812 (1998) 17–34.
[94] F.J. Schenck, P.S. Callery, Chromatographic methods of analysis of antibiotics
in milk, J. Chromatogr. A 812 (1998) 99–109.
[95] S. O’Connor, D.S. Aga, Analysis of tetracycline antibiotics in soil: advances
in extraction, clean-up, and quantification, Trends Anal. Chem. 26 (2007)
456–465.
View publication stats
[96] M. Seifrtová, L. Novákova, C. Lino, A. Pena, P. Solich, An overview of analytical
methodologies for the determination of antibiotics in environmental waters,
Anal. Chim. Acta 649 (2009) 158–179.
[97] M.S. Díaz-Cruz, D. Barceló, Recent advances in LC-MS residue analysis of
veterinary medicines in the terrestrial environment, Trends Anal. Chem. 26
(2007) 637–646.
[98] R.P. Li, Y. Zhang, Y.P. Huang, Determination of tetracycline antibiotics in the
environmental samples, Prog. Chem. 20 (2008) 2075–2082.
[99] H. Oka, K. Uno, K.I. Harada, K. Yasaka, M. Suzuki, Improvement of chemical
analysis of antibiotics: V. A simple method for the analysis of tetracyclines
using reversed-phase high-performance liquid chromatography, J. Chro-
matogr. A 298 (1984) 435–443.
[100] E.J. Mulders, D.V. de Lagemaat, Determination of residues of tetracycline
antibiotics in animal tissues by high-performance liquid chromatography, J.
Pharm. Biomed. Anal. 7 (1989) 1829–1835.
[101] J.R. Walsh, L.V. Walker, J.J. Webber, Determination of tetracyclines in bovine
and porcine muscle by high-performance liquid chromatography using solid-
phase extraction, J. Chromatogr. A 596 (1992) 211–216.
[102] C.R. Anderson, H.S. Rupp, W.H. Wu, Complexities in tetracycline analysis-
chemistry, matrix extraction, cleanup, and liquid chromatography, J.
Chromatogr. A 1075 (2005) 23–32.
[103] J. Nawrocki, The silanol group and its role in liquid chromatography, J. Chro-
matogr. A 779 (1997) 29–71.
[104] K.K. Unger, R. Skudas, M.M. Schulte, Particle packed columns and monolithic
columns in high-performance liquid chromatography-comparison and criti-
cal appraisal, J. Chromatogr. A 1184 (2008) 393–415.
[105] A.K. Sarmah, M.T. Meyer, A.B. Boxall, A global perspective on the use, sales,
exposure pathways, occurrence, fate and effects of veterinary antibiotics
(VAs) in the environment, Chemosphere 65 (2006) 725–759.
[106] R.P. Li, Y. Zhang, C.C. Lee, Hydrophilic interaction chromatography separation
mechanisms of tetracyclines on amino-bonded silica column, J. Sep. Sci. 34
(2011) 1508–1516.
[107] R. Li, Q. Yuan, Y. Zhang, J. Ling, T. Han, Hydrophilic interaction chromato-
graphic determination of oxytetracycline in the environmental water using
silica column, J. Liq. Chromatogr. Relat. Technol. 34 (2011) 511–520.
[108] J.C. Valette, C. Demesmay, J.L. Rocca, E. Verdon, Separation of tetracycline
antibiotics by hydrophilic interaction chromatography using an amino-
propyl stationary phase, Chromatographia 59 (2004) 55–60.
[109] R. Li, J. Huang, Chromatographic behavior of epirubicin and its analogues on
high-purity silica in hydrophilic interaction chromatography, J. Chromatogr.
A 1041 (2004) 163–169.
[110] M.S.S. Curren, J.W. King, New sample preparation technique for the determi-
nation of avoparcin in pressurized hot water extracts from kidney samples, J.
Chromatogr. A 954 (2002) 41–49.
[111] S. Van Dorpe, V. Vergote, A. Pezeshki, C. Burvenich, K. Peremans, B. De
Spiegeleer, J. Sep. Sci. 33 (2010) 728–739.
[112] M.M. Zheng, M.Y. Zhang, G.Y. Peng, Y.Q. Feng, Monitoring of sulfonamide
antibacterial residues in milk and egg by polymer monolith microextraction
coupled to hydrophilic interaction chromatography/mass spectrometry, Anal.
Chim. Acta 625 (2008) 160–172.
[113] G. Kesiunaite, E. Naujalis, A. Padarauskas, Matrix solid-phase dispersion
extraction of carbadox and olaquindox in feed followed by hydrophilic inter-
action ultra-high-pressure liquid chromatographic analysis, J. Chromatogr. A
1209 (2008) 83–87.
[114] K. Inoue, E. Yamada, T. Hino, H. Oka, Hydrophilic interaction liquid chro-
matography tandem mass spectrometry method for the determination of
bicozamycin in milk, J. Liq. Chromatogr. Relat. Technol. 32 (2009) 1914–1924.
[115] H.Y. Ji, D.W. Jeong, Y.H. Kim, H.H. Kim, D.R. Sohn, H.S. Lee, Hydrophilic
interaction liquid chromatography-tandem mass spectrometry for the deter-
mination of levofloxacin in human plasma, J. Pharm. Biomed. Anal. 41 (2006)
622–627.
[116] R.N. Rao, C.G. Naidu, K.G. Prasad, R. Padiya, S.B. Agwane, Determination of
gemifloxacin on dried blood spots by hydrophilic interaction liquid chro-
matography with fluorescence detector: application to pharmacokinetics in
rats, Biomed. Chromatogr. 26 (2012) 1534–1542.
[117] H.D. Alwis, D.N. Heller, Multiclass, multiresidue method for the detection of
antibiotic residues in distillers grains by liquid chromatography and ion trap
tandem mass spectrometry, J. Chromatogr. A 1217 (2010) 3076–3084.
[118] C. Chiaochan, U. Koesukwiwat, S. Yudthavorasit, N. Leepipatpiboon, Efficient
hydrophilic interaction liquid chromatography-tandem mass spectrometry
for the multiclass analysis of veterinary drugs in chicken muscle, Anal. Chim.
Acta 682 (2010) 117–129.