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Review
Chiral Drug Analysis in Forensic Chemistry:
An Overview
Cláudia Ribeiro 1,2 ID , Cristiana Santos 1 ID
, Valter Gonçalves 3 ID
, Ana Ramos 4 , Carlos Afonso 2,3
and Maria Elizabeth Tiritan 1,2,3, * ID
1 Institute of Research and Advanced Training in Health Sciences and Technologies, Cooperativa de Ensino
Superior Politécnico e Universitário (CESPU), Rua Central de Gandra, 1317, 4585-116 Gandra PRD, Portugal;
claudia.ribeiro@iucs.cespu.pt (C.R.); cristiana.s@live.com.pt (C.S.)
2 Interdisciplinary Centre of Marine and Environmental Research (CIIMAR/CIMAR), University of Porto,
Edifício do Terminal de Cruzeiros do Porto de Leixões, Av. General Norton de Matos s/n,
4050-208 Matosinhos, Portugal; cafonso@ff.up.pt
3 Laboratory of Organic and Pharmaceutical Chemistry, Department of Chemical Sciences,
Faculty of Pharmacy, University of Porto , Rua de Jorge Viterbo Ferreira, 228, 4050-313 Porto, Portugal;
valter.cma@gmail.com
4 Institute of Science and Innovation in Mechanical and Industrial Engineering (INEGI),
Faculty of Engineering of the University of Porto, Rua Dr. Roberto Frias, 400, 4200-465 Porto, Portugal;
shortinha.sa@gmail.com
* Correspondence: elizabeth.tiritan@iucs.cespu.pt; Tel.: +351-22-415-7178; Fax: +351-22-415-7102
Received: 30 December 2017; Accepted: 25 January 2018; Published: 28 January 2018
Abstract: Many substances of forensic interest are chiral and available either as racemates or
pure enantiomers. Application of chiral analysis in biological samples can be useful for the
determination of legal or illicit drugs consumption or interpretation of unexpected toxicological
effects. Chiral substances can also be found in environmental samples and revealed to be useful
for determination of community drug usage (sewage epidemiology), identification of illicit drug
manufacturing locations, illegal discharge of sewage and in environmental risk assessment. Thus,
the purpose of this paper is to provide an overview of the application of chiral analysis in
biological and environmental samples and their relevance in the forensic field. Most frequently
analytical methods used to quantify the enantiomers are liquid and gas chromatography using both
indirect, with enantiomerically pure derivatizing reagents, and direct methods recurring to chiral
stationary phases.
Keywords: chiral drugs; forensic chemistry; enantiomers; pharmaceuticals; illicit drugs
1. Introduction
Chiral compounds are asymmetric three dimensional molecules with one or more stereogenic
centers or asymmetry originated by planes or axis that gives two non-superimposable mirror images
molecules, called enantiomers [1]. In an achiral environment, a pair of enantiomers shares similar
physical and chemical properties, however, in a chiral environment such as living organisms,
enantiomers may exhibit different biological activities and/or toxicity due to enantioselective
interactions [2–4]. Separation of enantiomers has gained relevance in forensic chemistry and has
been applied in the analysis of biological fluids, environmental samples and in the control of illicit
drug preparations [5–9]. Figure 1 summarizes the applications of chiral analysis in forensic chemistry.
Molecules 2018, 23, 262; doi:10.3390/molecules23020262 www.mdpi.com/journal/molecules

synthesized by a clandestine laboratory [5,15]. Chiral analysis in biological fluids can give
information regarding consumption and differentiation between illegal drugs or legal
pharmaceuticals containing only a single enantiomer [7,16]. For example, the ingestion of dexedrine
(S-(+)-amphetamine (AM)) used in the treatment of narcolepsy, attention deficit disorders and
hyperactivity in children results only in serum concentrations of S-(+)-AM in contrast to the
Molecules 2018, 23, 262 2 of 47
ingestion of the illegal AM that leads to both enantiomers [7,8,16].
Figure 1. Application of chiral drug analysis in forensic chemistry.

chemistry.
Furthermore, the presence of these compounds in wastewater has been shown to be a tool for
Substances of forensic interest include pharmaceuticals and various classes of illicit drugs misused
the monitoring drug consumption at a community level (sewage epidemiology) [17–19]. In fact,
to improve sports performance or due to their psychotropic effects. The consumption of these
once excreted, residues of chiral drugs reach the aquatic environment mainly through the sewage
substances can cause toxicological effects and/or increased risk of death [10–13]. These substances
system as parent compounds and metabolites [20,21]. Concentration of target pharmaceuticals or
can be consumed by medical prescription or illicit practice and are available either as a racemates,
or as a single enantiomer. Data from chiral analysis in both biological samples and illicit drugs
preparations can be important in the control of manufacturing, consumption of illicit drugs or linking
between illicit drug preparations, consumers and traffickers [5,6,14]. Besides, both impurity and
chiral profile may provide a link between starting materials and the illicit drugs synthesized by
a clandestine laboratory [5,15]. Chiral analysis in biological fluids can give information regarding
consumption and differentiation between illegal drugs or legal pharmaceuticals containing only
a single enantiomer [7,16]. For example, the ingestion of dexedrine (S-(+)-amphetamine (AM)) used
in the treatment of narcolepsy, attention deficit disorders and hyperactivity in children results only
in serum concentrations of S-(+)-AM in contrast to the ingestion of the illegal AM that leads to both
enantiomers [7,8,16].
Furthermore, the presence of these compounds in wastewater has been shown to be a tool for
the monitoring drug consumption at a community level (sewage epidemiology) [17–19]. In fact,
Molecules 2018, 23, 262 3 of 47
once excreted, residues of chiral drugs reach the aquatic environment mainly through the sewage
system as parent compounds and metabolites [20,21]. Concentration of target pharmaceuticals or
illicit drug residues in wastewater influent may be used to backcalculate drug consumption for local
communities (sewage forensics) [17,22–24]; an approach to provide direct quantitative estimates,
in a non-invasive manner and in almost real-time [17,24,25]. Since most of these compounds are chiral,
the determination of the enantiomeric fraction (EF) can give further information about the use of
legal and illegal substances. Furthermore, once in the sewage system these compounds are subject
to biotic processes that causes changes in the enantiomeric composition. This information may be
used to evaluate the efficiency of the wastewater treatment plant (WWTP) or illegal discharges of
sewage since it may be expected that their EF in untreated sewage would differ from the one observed
in treated effluents [9,18,26]. Also, information about environmental occurrence and distribution
of chiral pharmaceuticals in the environment is important for evaluation of enantio-(eco)toxicity in
particular for aquatic organisms [27–29]. In this sense, chiral analysis applied to drugs preparations,
biological fluids and environmental samples may give information about: distinction between legal
and illicit drugs; linking between samples, illegal laboratories, consumers and trafickers; estimation of
consumption patterns at community level (sewage epidemiology); identification of manufacturing
locations of illicit drugs; illegal discharge of sewage and information about ecotoxicity (Figure 1).
Concerning the importance of chiral drug analyses in various forensic contexts, the present work
aims to critical discuss the applicability of chiral drug analyses concerning pharmaceuticals and illicit
drugs in forensic chemistry regarding biological and environmental matrices. The references search
were based in ScienceDirect and ISI Web of Knowledge databases considering articles up to 2017 that
comprise biological matrices such as urine, plasma, serum, blood and hair and environmental samples
as surface waters, influents and effluents from WWTPs as aquatic environmental matrices.
2. Chromatography in Chiral Analyses

The separation of enantiomers is currently carried out using liquid chromatography (LC),
gas chromatography (GC), capillary electrophoresis (CE) and supercritical fluids chromatography [15,
20,30–44]. Chromatographic methods for resolution of enantiomers include indirect and direct methods.
The indirect method is based on the reaction of the enantiomers with chiral derivatization reagents
(CDRs) and the formation of diastereoisomers with different physico–chemical properties that can be
separated by conventional means such as chromatography. The direct method can be achieved by chiral
stationary phases (CSPs), mostly applied in LC, or using chiral mobile phase additives. Both GC and LC
methods are available in indirect and direct [35,45–49]. However, for GC only few chiral columns are
available. Thus, most works with GC used indirect approaches with CDRs, which are then separated
by achiral columns. Examples of most used CDRs are S-(−)-N-(trifluoroacetyl)propyl chloride (S-TPC),
R-(−)-α-methoxy-α-(trifluoromethyl)phenylacetyl (R-MTPCl) and S-(−)-N-(heptafluorobutyryl)propyl
chloride (S-HFBPrCl) [46,50]. Regarding direct methods, many types of CSP are available, however
the cyclodextrin (CD), Pirkle-type, polysaccharide derivatives, antibiotics-based and polymeric-based
are most used [20,51–53].
GC and LC methods have been widely used for the enantioselective analysis of various classes
of illicit drugs in biological fluids though LC methods are most used concerning environmental
samples [20,54]. This is probably due to the high number of commercial columns available for LC and
the limit of quantification that LC with mass spectrometer (MS) can be achieved. For GC methods
most work uses MS while LC use different detectors as MS, ultraviolet-visible (UV/Vis), diode array
(DAD) and fluorescence detectors (FD) (Tables 1 and 2).
LC/MS and LC/MS/MS are the most applied techniques to quantify chiral compounds in the
environment. Nevertheless, MS detection presents some limitations in the type of elution mode and
the additives that can be used. Capillary electrophoresis (CE) has also been used for the separation
of enantiomers of toxicological, doping and forensic interest due to its simplicity and inexpensive
Molecules 2018, 23, 262 4 of 47
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illicit drugs
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pharmaceuticals
Chiral
ChiralChiral Compounds
Compounds
Compounds
Table
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Chiral
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Compounds
chiral illicitdrugs
Compounds drugs and pharmaceuticals
and pharmaceuticals.
Stimulants
Stimulants
Stimulants
Molecules 2018, 23, × FOR PEER REVIEW Stimulants and and
and
theirtheir
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and
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their metabolites
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4 of 47
Chiral
Stimulants
Stimulants Compounds
Chiral
Chiral
andand
Chiral
Chiral Compounds
Compounds
theirtheir 3metabolites
metabolites
Compounds
ChiralCompounds
NH2 NHNH
2 2 NHCHNHCH
NHCH
3 3
NH
NH NH
NH22 NH2 2
NH
Compounds NHCH
NHCH
NHCH NHCH
3 3
33NHCH
NHCH O OO NHCHNHCH
NHCH
3 33
22 33
NH2 NH2
Stimulants
Stimulants
Table 1. Structures
Stimulantsand
of the
Stimulants
Stimulants
Stimulants and
and their
and
and
and
their metabolites
their
chiral NHCH
their
their
their metabolites
illicit3 drugs
NHCH and pharmaceuticals
metabolites
metabolites
metabolites
3
metabolites
O O
O O
OO
NHCH
NHCHNHCH
NHCH
NHCH
NHCH
NHCH
3
3 33 3
3
CH3 CH CH
3 3 CH CH CH
3 3
O OOO O CHNHCH
3 CH
CH33 3 3
3
CH33 CH
NH32 3NH
CH CH
NHCH
3 3NHCH
CH O OO CH33CH
CH CH
33 3
CH
CH 3
CH CH
NH
NH
CH3 CH32
323NH
CH NH
2 2
NH
NH 22 2 Chiral CH
Compounds
CH33 CH
NHCH
NHCH
CH3 CH3 3
NHCH
3NHCH
3 3NHCH
CH NHCH
3
3 3
33 3 O OO
OO 3CHNHCH
CHNHCH 3NHCH
O O
O NHCHNHCH 3
NHCH
NHCH 3 3
3,4-Methylenedioxy-
OO 3
3,4-Methylenedioxy-
O 3
3,4-Methylenedioxy-
3 33
Stimulants and their metabolites
Amphetamine
Amphetamine
Amphetamine (AM) (AM)
(AM) Metamphetamine
Metamphetamine
Metamphetamine (MA) (MA)
(MA) O 3,4-Methylenedioxy-
3,4-Methylenedioxy-
3,4-Methylenedioxy-
O3,4-Methylenedioxy-
3,4-Methylenedioxy-
CH CH CH
3 3 NH2 3 3 NHCH3
Amphetamine (AM) Metamphetamine 33 3(MA) methamphetamine
CH333CH
methamphetamine (MDMA)
(MDMA)
3
Amphetamine (AM) Metamphetamine (MA) methamphetamine
O
(MDMA)
CH
CH CH3 CH
CH333 CH
3,4-Methylenedioxy-
CH
3,4-Methylenedioxy-
O 3
Amphetamine (AM) Metamphetamine (MA)
CH CH 3
Amphetamine
Amphetamine (AM)
(AM) Metamphetamine
Metamphetamine (MA)
(MA) O
CH CH
CHCH CH33 CH
CH CH
CH O O 33
33 3
NHCH
Amphetamine
Amphetamine (AM) (AM) Metamphetamine
Metamphetamine (MA) (MA) methamphetamine
methamphetamine
methamphetamine
O
methamphetamine
(MDMA)(MDMA)
(MDMA)
(MDMA)
3
NH methamphetamine
(MDMA)
O OO NHCH 2 NH 22 NH NH
NH 3,4-Methylenedioxy-
3,4-Methylenedioxy-
3,4-Methylenedioxy-
NH CH
NH
3,4-Methylenedioxy-
NH3
Amphetamine
O Amphetamine
Amphetamine(AM)
NH(AM)
2NH NH
3
(AM)
O OO
Metamphetamine
O Metamphetamine(MA)
Metamphetamine (MA)
(MA)
CHCH
3 3
CH
CH
33 3,4-Methylenedioxy-
3,4-Methylenedioxy-
O
3,4-Methylenedioxy-methamphetamine
Amphetamine
O O
Amphetamine
Amphetamine
OO
Amphetamine (AM) (AM)
NH
NH22(AM)
(AM) NH
NH22 2
22
O Metamphetamine
O
OOO NH NH
Metamphetamine
Metamphetamine
NH NH
MetamphetamineNH (MA)
NHCH
(MA)
CH(MA)
3CH
(MA) CH
3 3
CH
CH methamphetamine
methamphetamine
NH NH
NH NH3 CH
CH
NH
NH
NH(MDMA)
CH
(MDMA)
33
O O CH3CH CH 33 O O
O OO CHNH
CH
3
NH
CH
3 3
3
CH CH3 3
3 3 methamphetamine
methamphetamine
methamphetamine
methamphetamine
HO (MDMA)(MDMA)
NH
CH
CH (MDMA)
(MDMA)
(MDMA)
3CH
3,4-Methylenedioxy-
HO 3
CH
CH3 3
CH33
O OO CH33CH CH CH33CH
CH CH
33 33
CH
CH
HO CH CH 3 3
O Amphetamine CHCH(AM) O OMetamphetamine (MA)
O O 33 33 O OO
O HO
HO HO HO
HO HO
OO CH NHCH NH NH CH CH methamphetamine
O OO NH
3 2NH
2 3NH
22 22 OO NH
3
NH NH
3NHCH CH HO HO NH
NH NH NH(MDMA)
O 3,4-Methylenedioxy
OO
3,4-Methylenedioxy
3,4-Methylenedioxy NH2
2 Methylenedioxy-N-ethylamphetamine
O O
Methylenedioxy-N-ethylamphetamine
O OOO NH
NH CH
NHCH33CH33 3
CH
3 CH 33 p-Hydroxymethamphetamine
p-Hydroxymethamphetamine
NH
p-HydroxymethamphetamineNH
NH
CH CH
3,4-Methylenedioxy
3,4-Methylenedioxy
O
3,4-Methylenedioxy
3,4-Methylenedioxy
3,4-Methylenedioxy Methylenedioxy-N-ethylamphetamine
NH
O CH p-Hydroxymethamphetamine
CH CH
NH333 CH
CH
CH 33 3
33
O amphetamine
amphetamine
amphetamine
O (MDA)
CH 3(MDA)
3,4-Methylenedioxy
CH
3,4-Methylenedioxy CH
CH(MDA)
CH
CH 3 3 (MDEA)
(MDEA)
(MDEA)
CH CH
O O O CH 3
CH
3 CH
CH33 3
CH 3
HO HOHO (pOHMA)
(pOHMA) CH3
(pOHMA)
Oamphetamine 3(MDA) O
(MDEA) (pOHMA)
3
amphetamine
O (MDA)
33 3 O OO (MDEA) HO HO (pOHMA)
3 3 HO
OO
amphetamine (MDA) (MDEA)
HO
(pOHMA)
amphetamine
amphetamine
amphetamine (MDA)
(MDA)
CH
(MDA) (MDEA)
(MDEA)
(MDEA)
CH (pOHMA)
(pOHMA)
(pOHMA)
amphetamine
O (MDA) 3 O (MDEA) 3
(pOHMA)
HO
3,4-Methylenedioxy
3,4-Methylenedioxy
3,4-Methylenedioxy
3,4-Methylenedioxy
3,4-Methylenedioxy
3,4-Methylenedioxy
3,4-Methylenedioxy amphetamine Methylenedioxy-N-ethylamphetamine
Methylenedioxy-N-ethylamphetamine p-Hydroxymethamphetamine
Methylenedioxy-N-ethylamphetamine p-Hydroxymethamphetamine
N NN
HO
HO NH NH
3,4-MethylenedioxyNH Methylenedioxy-N-ethylamphetamine p-Hydroxymethamphetamine
N
N N N
NN
amphetamine
HO
HO
amphetamine
amphetamine(MDA)
amphetamine
amphetamine
amphetamine
HO
HO NH(MDA)
(MDA) (MDA) (MDA)
NH3CH
(MDA)
NH
CH
(MDA)
NHNH CH 33 (MDEA)
(MDEA)
(MDEA)
(MDEA)
(MDEA)
(MDEA)
(MDEA)
N NN (pOHMA)
(pOHMA)
(pOHMA)
(pOHMA)
(pOHMA)
(pOHMA)
(pOHMA)
N N NH NHNH 2 2 2
HO
HO
amphetamineNH (MDA)
CH
NH CHCH
CH (MDEA) (pOHMA)
O NH NH
O O NH NH 2 2
HO HO CH
CH3 3CHCH 33CH
3 3CH3
33 33 N N
H HH
N NNN NH
NH
NH O O O NH2
22
2
2 2
O OO CH33CH
CH
CH
CH33 33 NOOH
H
O OO
H NH
HH
O N O ON N
N O N
O N NN
N
O OO
OO 3CHNH
CHNH NH HOOH
O OO
HO HO
HO
HO
HOO HOO NH NH
CH
NH
3NH
CH
O OO
OO NH2 NHNH
HO CH33CH CH
CH33 333
CH3
O OO
N O N
N N NN NH
2 22
NH
NH22 NH
NH
NH22 2
4-Hydroxy-3-methoxy NN OO
4-Hydroxy-3-methoxy
O
4-Hydroxy-3-methoxy H O
CH3CHCH
CH
CH 33 333
H
H H
HH
HH
O
O O
OO
O4-Hydroxy-3-methoxy
4-Hydroxy-3-methoxy
O CH CH
4-Hydroxy-3-methoxy
O 4-Hydroxy-3-methoxy
4-Hydroxy-3-methoxy
O
OO 3 3
methamphetamine
methamphetamine
methamphetamine
O O O O
Methylphenidate
O O
MethylphenidateO
O (MPH)
Methylphenidate
(MPH)
O
(MPH) Aminorex
Aminorex
Aminorex
4-Hydroxy-3-methoxy
4-Hydroxy-3-methoxy O OOO O O
methamphetamine
methamphetamine
methamphetamine
methamphetamine
methamphetamine Methylphenidate
Methylphenidate
Methylphenidate (MPH)
(MPH)
(MPH)
Methylphenidate
(MPH) Aminorex
Aminorex
Aminorex
Aminorex
Aminorex
(HMMA)
(HMMA)
(HMMA)
methamphetamine
methamphetamine
4-Hydroxy-3-methoxy Methylphenidate
(MPH) Aminorex
Aminorex
(HMMA)
(HMMA)
4-Hydroxy-3-methoxy
(HMMA)
4-Hydroxy-3-methoxy
4-Hydroxy-3-methoxy
(HMMA)
(HMMA)
4-Hydroxy-3-methoxy
4-Hydroxy-3-methoxy
4-Hydroxy-3-methoxy Methylphenidate (MPH) Aminorex
4-Hydroxy-3-methoxy
(HMMA)
(HMMA)
methamphetamine (HMMA) Methylphenidate (MPH) Aminorex
methamphetamine
methamphetamine
methamphetamine
methamphetamine (HMMA)
methamphetamine
methamphetamine
methamphetamine Drug
DrugDrug precursors
precursors
precursors
Methylphenidate
Methylphenidate
Methylphenidate
Methylphenidate
(MPH)
(MPH)
(MPH)
(MPH) Aminorex
Aminorex
Aminorex
Aminorex
Aminorex
Aminorex
Drug
Drug
Drug
Drug
Drug
Drug precursors
precursors
precursors
precursors
precursors
precursors
(HMMA)
(HMMA)
(HMMA)
(HMMA)
(HMMA)
(HMMA)
OH OH
OH Drug
Drug
Drug precursors
precursors
precursors
OH
OH OH
OH
OHOHOH
OH
OH OH OH
OH
CH3CH CH Drug precursors
Drug precursors
OH OH 33 OHOH
OH
CH3CHCH
Drug precursors
3
Drug precursors
333
CH33CHCHCH
CH
33 33 Drug precursors
OH OH
CH
CH CH
3
3
CH
33
CH
CH 33
CH
NH CH 3 NHCH
NH
NH3CH3
NH
OH NH
OH
OHNH 3
NH3CH
CH CH
CH
OH
OHNH NH3CH
CH
NH
OHNH
NH CH 33 3 OHNH
OH
OHNH
NH
OH
OH
OH
OH NH
NH
CH CHCH
333
NH
NHCHCH CH
CH NHCHCH
NH 3CH
3 CH33
CH 33CH
3
CH
Pseudoephedrine
CH
CH
33 333 33
3CH
CH CH3CH
Ephedrine CH
CH 33
3CH
CH CH3
CH CH 3
CH 3 33
Pseudoephedrine
Pseudoephedrine
Pseudoephedrine
3 3 33 3
Ephedrine
Ephedrine
Ephedrine 3 33
Pseudoephedrine
Pseudoephedrine
Pseudoephedrine
Pseudoephedrine
Pseudoephedrine
NH NH
NH Opioids, Ephedrine
morphine Ephedrine
Ephedrine
Ephedrine
Ephedrine
NH NH
derivatives
NH
NH
NH and their metabolites
NH NH
Pseudoephedrine
Pseudoephedrine
NH CH
Pseudoephedrine
CH Ephedrine
NH NH
EphedrineCH CH
Ephedrine CH
CH33CH CH333CH
CH3
CH 3
CH
Opioids,
Opioids,morphine
morphine derivatives
andand
derivatives andtheir
theirmetabolites
metabolites
33 3 3
CH 3Opioids, morphine derivatives their metabolites
3 33 O
3
CH3
Opioids, Opioids,
Opioids,
Opioids,
morphine
Opioids,
Opioids, morphine
morphine
morphine
derivatives
morphine
morphine derivatives
derivatives
and
derivatives
derivatives their
and
and
N andand
derivatives andtheir
their
their
metabolites
their
their metabolites
metabolites
metabolites
metabolites
metabolites
Pseudoephedrine
Pseudoephedrine Opioids,
Opioids,
O Opioids,
Pseudoephedrine
Pseudoephedrine
Pseudoephedrine
Pseudoephedrine morphine Ephedrine
morphine
morphine Ephedrine
derivatives
derivatives
derivatives
Ephedrine
Ephedrine
Ephedrine
Ephedrine
and
andandtheir
their metabolites
their metabolites
metabolites O OO
H3C N
N NN O CH
O 3CH
O CH 33
HO O OO
O O Opioids,
CH3
morphine
Opioids, morphine derivatives
derivatives and andtheir metabolites
their metabolites
CHO 33CHCH
Opioids, morphine derivatives and their metabolites O CH CH
CH 3 3
H3C H3H
CH3 O Opioids,
Opioids, morphine
morphine Nderivatives
derivatives
N N
N
NN
N and and their
their metabolites
metabolites CH CH
3 N3
3 3CH
3
NC3C N N O O O
CH
H33C H C3C N N
O
O CH
OO
O3 CH CH HO HOHO CH
H H33HNN
C3C NN 3 3
HO HO O
HO O CH O O 3
H3C HCH 3NC 3 CH
NCH3 3 CH33 CH
CH CHCH
3 3
CH HO HO
HO NOOO3CHCHN33CH
NCH
CH
CH
CH 33
33 N NN
2-Ethylidine-1,5-dimethyl-3,3- HO HO CH CH
CH N33CH
3 3CH
CH33 CH
CH CH
3 3
CH
CH
Methadone
33
CH3 CH3 N N NN N 33CH
N
Tramadol
CH
N
NN
CHCH
3CH
(T)
CH
3CH 3 3
33
CH3 CH 3 O O
O O N 3CH
N 3 3 3 3
H3C H
H C H
H33C HH
3H
3
NC
C
C
N3CC N
N N
O
O OO diphenylpyrrolidine (EDDP) HO HO
CH33CH
CH CH CH
CH 3 3
33
NN HO
2-Ethylidine-1,5-dimethyl-3,3-
3N 3
CH3CH3
CH33Methadone
OHCH CH3 CH
CH33 CH CH
CH
CH
3 3
3 2-Ethylidine-1,5-dimethyl-3,3-
O HO HOHO
HO
N OHCH 3CH
Methadone 2-Ethylidine-1,5-dimethyl-3,3- Tramadol
Tramadol (T)
33(T)
33
Methadone 2-Ethylidine-1,5-dimethyl-3,3- Tramadol N(T)N
CH
CH3 CH NN
CH CH 3
CH3
CH CH
2-Ethylidine-1,5-dimethyl-3,3- NN CH
CH CH
CH
CH
3 3 3333
3
Methadone
33
Methadone 2-Ethylidine-1,5-dimethyl-3,3-diphenylpyrrolidine
diphenylpyrrolidine
diphenylpyrrolidine
diphenylpyrrolidine (EDDP) (EDDP)
(EDDP) Tramadol
Tramadol (T) (T)
Methadone
Methadone Methadone
Methadone 2-Ethylidine-1,5-dimethyl-3,3-
2-Ethylidine-1,5-dimethyl-3,3- Tramadol
Tramadol
CH(T)
Tramadol 3CH (T)(T)
MethadoneMethadone diphenylpyrrolidine
diphenylpyrrolidine
diphenylpyrrolidine
(EDDP)(EDDP)
diphenylpyrrolidine
diphenylpyrrolidine (EDDP)
(EDDP)
(EDDP)
(EDDP) Tramadol
Tramadol CH 3CHCH
3 (T)
(T) CHCH 3
3
33 3
HO OH OH OH diphenylpyrrolidine
diphenylpyrrolidine
HO (EDDP) (EDDP) HO
CH3 2-Ethylidine-1,5-dimethyl-3,3-
O OO
N CH3 OH OH OH CH
Methadone OH OH
Methadone N Tramadol
Tramadol (T)
OHH(T)
N
OH CH
CH 3
Methadone Tramadol (T)
OH CH
Methadone OH
OH
Tramadol
OH OH
(T)
O O 3
Methadone Tramadol (T)
O H 3
Methadone OH OH O
diphenylpyrrolidine
O3O
diphenylpyrrolidine
diphenylpyrrolidine
CHO33CH (EDDP)
CH 3 3 (EDDP)
(EDDP) TramadolOH
(T)
OHOH
CH3 diphenylpyrrolidine
diphenylpyrrolidine
O CH
diphenylpyrrolidine
CH3 CH
CH 3 3 (EDDP)
(EDDP)
CH
3 (EDDP) OH OH
HO HO HO
O-Desmethyltramadol HO HOHO HO HO HO
N,O-Ddidesmethyltramadol
HO HO OH OH OH
HO HO HO OH
HO OH
OH
OHCHNN3CH CH33 HO HO
HO HO O NO
N-Desmethyltramadol
HO
HO CH
O NN3 CH CH
3 3 (NDT) HO HO
HO HOHO OHNOH
HO CH
OH NN3 CH CH 33
N HO HO O CH CHO
OO CH CH OH OH
OHOH
HO HO (ODT)
N CH
CH
N 3CHN
NN
CH N 3
3
CH
CH CH
CH3 3
33
H
NN 3
3
CH
HCH
CH
3CH
CH
N
CH
NN
H
N33
3 CH
3
3
3
3
CH
3 3
CH3 3
HO HO (N,O-DDM-T)
H
N CH
N
CHNHH
NNN3CH
3 CHCH
CH 3 3
33
CH CH CH
3 3 HH
N N H
CH H3 CH 3 H
N N
H CHH H CH 3
N CH NCH 3 3 HH HH 3
HO HO
CH
CH 3CH
3 CH 3 3
3 3 Antidepressants and
HO HO their
H metabolites/
H Benzodiazepine
HO
H H
HO HO HO
O-Desmethyltramadol CH3CH3
HO
O-Desmethyltramadol N,O-Ddidesmethyltramadol
HO HO
O-Desmethyltramadol
HO HO HO
CH CH
HO HOHO
HO CH 3 CH N,O-Ddidesmethyltramadol
HO
HOHO
CH 3 CH
O-Desmethyltramadol
O-Desmethyltramadol
O-Desmethyltramadol
F N
O-Desmethyltramadol
O-Desmethyltramadol N CH N
N N333CH
NN CH CH
CH3
3
33 3 N-Desmethyltramadol
N-Desmethyltramadol
N-Desmethyltramadol
F
N CH
N
N
N
NNN CH
33 CH
CH
CH3
3 3(NDT)
(NDT)
3 3
(NDT) N,O-Ddidesmethyltramadol
N
N
N NCH N
NNN CH
33 CH
CH
CH 33 3
33
F (ODT)
O-Desmethyltramadol (ODT)
(ODT)
O-Desmethyltramadol
O-Desmethyltramadol CH3CH (ODT) N-Desmethyltramadol
H
N-Desmethyltramadol
N-Desmethyltramadol
N-Desmethyltramadol
F
H
N-Desmethyltramadol
N-Desmethyltramadol
H
H
H H
H
(NDT) (NDT)
(NDT)
(NDT)
(NDT)
(NDT) (N,O-DDM-T)
(N,O-DDM-T)
(N,O-DDM-T)
HO
H
H H
N,O-Ddidesmethyltramadol H
HHH
F (ODT)
(ODT) (ODT)
CH33CH
(ODT)
(ODT) CHCH
CH 3
3333
N-Desmethyltramadol
N-Desmethyltramadol (NDT) (NDT) (N,O-DDM-T)
(N,O-DDM-T)
(N,O-DDM-T)
(N,O-DDM-T)
(N,O-DDM-T)
(ODT)
(ODT) F (N,O-DDM-T)
(N,O-DDM-T)
O-Desmethyltramadol
O-Desmethyltramadol
O-Desmethyltramadol
O-Desmethyltramadol
O-Desmethyltramadol
O-Desmethyltramadol O Antidepressants
Antidepressants
Antidepressants
CH
N Antidepressants
3 andand
and
and their
theirtheir
their metabolites/
O metabolites/
metabolites/
metabolites/BenzodiazepineBenzodiazepine
Benzodiazepine
NH2 Benzodiazepine
H Antidepressants andand
Antidepressants
Antidepressants
Antidepressants
Antidepressants and
and
and their
N-Desmethyltramadol
their metabolites/
N-Desmethyltramadol
N-Desmethyltramadol
their metabolites/
N-Desmethyltramadol
their
their
N-Desmethyltramadol (NDT)
metabolites/
metabolites/
metabolites/
N-Desmethyltramadol (NDT) (NDT)
(NDT)Benzodiazepine
Benzodiazepine
Benzodiazepine
(NDT)
(NDT) Benzodiazepine
Benzodiazepine
(ODT)
(ODT) N-Desmethyltramadol
Antidepressants
(ODT) Antidepressants andand their
their (NDT)
N-Desmethyltramadol (NDT)
metabolites/
metabolites/ (N,O-DDM-T)
Benzodiazepine
Benzodiazepine (N,O-DDM-T)
(N,O-DDM-T) O
(ODT)
(ODT)
(ODT) (N,O-DDM-T)
(N,O-DDM-T)
(N,O-DDM-T)
F FFluoxetine
F (FLX) Norfluoxetine (NFLX)
F FF
Venlafaxine
N NN (VNF)
F F FF FF F
F
F
F FFF F
F
F
N
Antidepressants
Antidepressants
Fand
Antidepressants
Antidepressants
Antidepressants
Fand
F F their
FFand
FFandand
FFF
their
F metabolites/
their
their
their
Hmetabolites/
metabolites/
Nmetabolites/
metabolites/ Benzodiazepine
Benzodiazepine
Benzodiazepine
Benzodiazepine
Benzodiazepine HO HON N
HO
N
N NN
NNH
N
FF FF F FF FFF F HO HO
HO HO
HO
HO
HO O F HO O HO
F FF F F FF HO HO
F CHN3NCH CH33 F FF
F F F F F O OO N CH
HH 3 CH
CH
F
F FF F O O
O NH2 NH NH
22
N NN
F F FF OO OOO HNN CHN
NN 3N CH3 3
CH 33 F F OO OO NH22NH
O NH NH N N NN O OO
F F F H CH
H H CH F F F 2 2
NH
NH
F F FF O O HN N HH 3 3 F FF
F F FF O O NH2 NH2
22 HO HO
HO O O O
H H HO HO
HO
HO O OO
FFluoxetine (FLX) F Norfluoxetine (NFLX) Venlafaxine
F F
Fluoxetine
F Fluoxetine
Fluoxetine
F
FF (FLX)
(FLX) (FLX)OH Norfluoxetine
Norfluoxetine
F Norfluoxetine
F F (NFLX)
(NFLX)
(NFLX)O Venlafaxine
Venlafaxine
Venlafaxine O(VNF)
(VNF) (VNF)
O OH
(VNF)
F
Fluoxetine
Fluoxetine O (FLX)
Fluoxetine
(FLX)
Fluoxetine
Fluoxetine
O
O O (FLX)
N(FLX)
(FLX)
O-Desmethylvenlafaxine
N CH
CHN CH
3 CH
3N
CH
CH
3 CH
33 3(O- Norfluoxetine
FF
Norfluoxetine
Norfluoxetine
Norfluoxetine (NFLX)
O O(NFLX)
(NFLX)
O (NFLX)
NH Venlafaxine
Venlafaxine
Venlafaxine
Venlafaxine
Venlafaxine (VNF)
(VNF)(VNF)
(VNF)
(VNF)
N,O-Didesmethylvenlafaxine
Fluoxetine (FLX) Norfluoxetine (NFLX) Venlafaxine (VNF)
NH
Fluoxetine O
OO(FLX)
H
H H
N
NN
H
HH H
33
Norfluoxetine O O
N-Desmethylvenlafaxine O(NFLX)
O NH
NH222(N-DES-VNF)
NH
NH22 2
NH22 Venlafaxine (VNF)
O
O OO O
DES-VNF) (N,O-DES-VNF) OO
Fluoxetine (FLX)
Fluoxetine
Fluoxetine
Fluoxetine
Fluoxetine
Fluoxetine (FLX)
(FLX)
(FLX)
(FLX)
(FLX) Norfluoxetine (NFLX)
Norfluoxetine
Norfluoxetine
Norfluoxetine
(NFLX)
(NFLX)
(NFLX) Venlafaxine (VNF)
Venlafaxine
Venlafaxine
Venlafaxine
Venlafaxine (VNF)
Venlafaxine (VNF)
(VNF)
(VNF)
(VNF)
Molecules 2018, 23, 262 5 of 47
Table 1. Cont.
Molecules
Molecules
2018,
Molecules 23, ×23,
2018,
2018, FOR
23,××FOR
PEER
FORPEER
REVIEW
PEERREVIEW
REVIEW 5 of548
5ofof48
48
Chiral Compounds
H HH H HH
N NN N NN N NN
HO HO
HO HO HO
HO HO HO
HO
Molecules
MoleculesMolecules
2018,
Molecules
Molecules
Molecules 2018,
2018,
Molecules 2018,
2018,
23,2018,
×23,
23,FOR
2018, 23,
23,
××23, ××××FOR
PEER
FOR
FOR
23, FOR
FOR
PEER
PEER
FOR PEER
PEER
REVIEW
PEER REVIEW
REVIEW
REVIEW
REVIEW
REVIEW
PEER REVIEW 6 of6651 666ofof
ofof51
651 of 51
of5151
51
Molecules 2018,
Molecules 23, ×23,
2018, FOR× FORPEER REVIEW
PEER REVIEW 6 of651of 51
Molecules
Molecules 2018,
Molecules 2018, 23,× ×FOR
23, ×23,
2018, FOR FORPEER
PEERPEER REVIEW
REVIEW
REVIEW 6 of 6516ofof5151
OH OH
OH O OO OH OH
OH
Table Table
1.
Table Cont.
Table
Table 1.1.
Cont. Cont.
1.Cont.
1.1.Cont.
Table Cont.
O-Desmethylvenlafaxine Table 1. Cont.
Table
N-Desmethylvenlafaxine1. Cont.
N-Desmethylvenlafaxine
N-Desmethylvenlafaxine (N-DES-
(N-DES-
(N-DES- N,O-Didesmethylvenlafaxine
O-Desmethylvenlafaxine Table Cont.
1.1.Cont.
N-Desmethylvenlafaxine
Table 1. Cont.
Table N,O-Didesmethylvenlafaxine
(O-DES-VNF)
(O-DES-VNF)
(O-DES-VNF) VNF)VNF)
VNF)
(N-DES-VNF) (N,O-DES-VNF)
(N,O-DES-VNF)
(N,O-DES-VNF)
(N,O-DES-VNF)
NN NNN
N N
FF FF FFF HN HN
HN HN
HN
HNHN
HN F FF FF
F
FF
N N F F HH CC OO OO HN HN
H3HC33C H H
H33C CO
C333O O
OO OO O
OO F F
N NN F F F HN
HN HNHN
H3C H3C O O O O FF F F
HH33CC H3CH3CO O O OOO O OOO
O ONH
O NH NH
NHNH
OO NHNH
O O NH NH
O
O O O NH
NH NHNH
OO OOO
O O O OO OOO
O O
O O O O
O OO C CC CCC
C C O
O O O
CC CCC
C C
N NN CNNNN
N C
NN NNN
CN N C
N CN
C C C
N CN CC
N
N N N
N NN
Citalopram
Citalopram
Citalopram
Citalopram
Citalopram Reboxetine
Reboxetine
Reboxetine
Reboxetine
Reboxetine -citalopram
D-citalopram
-citalopram
-citalopram
-citalopram
DD DD
D D
D -citalopram
Citalopram
Reboxetine D-citalopram
D-citalopram
Citalopram
Citalopram
Citalopram
Reboxetine
Reboxetine
Reboxetine -citalopram
D-citalopram
-citalopram
D
DD-citalopram
OO OO
O O
O
NN NNN
N N O O
N N O OO
N NN
NN NN
NNN OH
OH OH
OH
OHOH
OH
N N OH OH
N NN NOH OHOH
NN N N
N NN
N
NN ClCl ClCl
Cl Cl
Cl NN NNN
N
N N
NN N N
N NN N Cl Cl
N NNN N Cl Cl Cl N NN
N
Mirtazapine
Mirtazapine
Mirtazapine
Mirtazapine
Mirtazapine Temazepam
Temazepam
Temazepam
Temazepam
Temazepam
Mirtazapine
Temazepam
Mirtazapine
Mirtazapine
Mirtazapine
Temazepam
Temazepam
Temazepam
Dissociative
Dissociative
Dissociative anaesthetic
anaesthetic
Dissociative
Dissociative and
anaesthetic
anaesthetic
anaesthetic
Dissociative and
and
anaesthetic and
itsand its
metabolitemetabolite
itsmetabolite
its
and its
metabolite
itsmetabolite
metabolite
Dissociative
anaesthetic and
and its
andits metabolite
its metabolite
metabolite
OO OOO
O O Dissociative
Dissociative
anaesthetic O and
anaesthetic
OO OOO
O and
its
and itsmetabolite
its metabolite
metabolite
O OClCl ClCl
Cl Cl
Cl O OClCl ClCl
Cl Cl
Cl
O OO Cl Cl O OO Cl Cl
Cl Cl Cl Cl Cl Cl
NH NH
NH
NH NH
NH
NH
NH NH
NH22 NHNH
NH
NH2 NH 22 222
NH
NH NH NH2 NH2
HC33C
H3H C H HHC33CC
H33H
NH C 3C NHNH NH22 NHNH
2 2
H3C H3C
Ketamine Norketamine
H33C H3H C3C
Ketamine
Ketamine
Ketamine
Norketamine
Norketamine
Norketamine
Ketamine
Ketamine
Norketamine
Norketamine
Ketamine
Ketamine
Norketamine
Norketamine
β-Blockers
β-Blockers and
β-Blockers
β-Blockers
β-Blockers and
β-Blockers
andand anti-arrhythmic
anti-arrhythmic
and and
anti-arrhythmic
and anti-arrhythmic
anti-arrhythmic
anti-arrhythmic
anti-arrhythmic
β-Blockers and anti-arrhythmic
β-Blockers and anti-arrhythmic
β-Blockers
β-Blockers
β-Blockers and
andand
β-Blockers anti-arrhythmic
anti-arrhythmic
anti-arrhythmic OH OHOH OHOH
OH
OHOH
OHH OHH
OHOH
OH OH
OH
OHOH
OH OOH H HH HH
H
OHOH
OH OH
OHOH
OHOH O OO O ON
O OH
O OH N
N OH NN NN
N
OH OH H H
OH OH H HH O O N N
O OO O OOOH
OH NN
OHOH
N N NN HH H
HNH H
H H H
OO NN O OOOH
OH OHN N O
O O O N
N N N
OO O
OO OHN N N
NN
O O N N H H
O O N N
O
O O O N
N N N HH H H
O
O O O NN N N
O OO O OO O OO
OO OO O
OO
O O O OO O O
OO O
OO
O
O O O O
O O O
O O
Metoprolol
Metoprolol
Metoprolol
Metoprolol
Metoprolol (MET)
(MET)
Metoprolol
(MET)(MET)
(MET)
(MET) Propranolol
Propranolol
Propranolol
Propranolol (PHO)
Propranolol
Propranolol (PHO)
(PHO)
(PHO)
(PHO)
(PHO)
O
O
Bisoprolol
Bisoprolol (BSP)
Bisoprolol
Bisoprolol
Bisoprolol (BSP)
Bisoprolol
O O
Bisoprolol (BSP)
(BSP)
(BSP)
(BSP)
(BSP)
Metoprolol
Metoprolol(MET)
(MET) Propranolol (PHO)
Propranolol (PHO) Bisoprolol (BSP)
Bisoprolol (BSP)
Metoprolol
Metoprolol
H
HH
N
HH(MET)
Metoprolol HH
N
H
N
(MET)
(MET) Propranolol
Propranolol (PHO)
Propranolol (PHO)
(PHO) Bisoprolol
Bisoprolol (BSP)
Bisoprolol (BSP)
(BSP)
NN N NN
H H
N N
H
H H H
N
N N N
O
OO OOO
O O
OHOHOH OH
OH
OH HOH
O O OH
OH
OH OH
OH
OH HH
H
O
O O O
OH OH OH HH
OH
HO
HO
HO HO
HO HO
HOOH OH
OH
O OO OO
OOH
O N OH
NH
OH
N HN
NN
N
HO
HO
HO HO
HO
HOHO
HO HO HO OH
OH OH
OHOH
HOH H
OH
HOH HHH O O H
N NH
H
HO HO NH
NHNH NH
NH
NHNH
NH
HO
HO HO
OO OOH
OO
O
OOH H NN
NOH N
NOH
H
N
N
O OO N NN
HO
HO HOHO H
NH NH O O N
H NH
H
NH
NH NHNH O OO N NN
HNHN HN
HN
HN HN
HN
HN HN
O
OO OOO
O O HN HNHN
O O
O
O O O
O
OO OOO
O O
O O
O
O O O
Carvedilol
ZAIZHELI
Carvedilol
Carvedilol
Carvedilol
Carvedilol
Carvedilol Nadolol
Nadolol
Nadolol
Nadolol
Nadolol
Nadolol Pindolol
Pindolol
Pindolol
Pindolol
Pindolol
Pindolol
Pindolol
Carvedilol
Carvedilol Nadolol
Nadolol Pindolol
Pindolol
Carvedilol
Carvedilol
Carvedilol Nadolol
Nadolol
Nadolol Pindolol
Pindolol
Pindolol
OHOH
OH OH
OH
O OO OO
OO
O OO
OO OO
O H OH
OH
OH
OH
HH
OH HH
HH
H
O O N NN N NN
N O O O NN N NNN OO OO
O O OO O OO
O
O OO
O OO O N OH
OOH H
N
NOH NN
N
HN
O OO N N O OO N N O O O O O O H
N NH
H
O OO OO
OO
O
N
N NN N NN
NN
N
N N NN O OO O OO O OO N NN
O O N N
O OO
OO OO NN NNNN OO O
H N
O 2 HH
OO
O H2N
2NHH
H
H22N
2N
22
NN
O OO N NN O O N
H H
HN HH
N H
H O O 2NH2N
O OO H HN
N O OO H 2NHH 2N 2N
H H H
Verapamil
Verapamil
Verapamil
Verapamil
Verapamil
Verapamil Norverapamil
Norverapamil
Norverapamil
Norverapamil
Norverapamil
Norverapamil Atenolol
Atenolol
Atenolol
Atenolol
Atenolol
Atenolol
Atenolol
Verapamil
Atenolol
Verapamil
Verapamil
Norverapamil
Atenolol
Atenolol
Molecules 2018, 23, 262 6 of 47
Molecules
Molecules 2018, 23, ×23,
2018, FOR× FOR PEER
PEER REVIEW
REVIEW 8 of851
of 51
Molecules 2018, 23, × FOR PEER REVIEW 8 of 51

Table
Table
Table 1. Cont.
1.1.Cont.
Cont.
Molecules
Molecules 2018,
Molecules 23, ×23,
2018,
2018, FOR
23,× ×FOR PEER
PEER
FOR REVIEW
REVIEW
PEER REVIEW 8 of8518ofof5151
Molecules
Molecules 23, × 23,
2018,2018, FOR × FOR
PEERPEER
REVIEW
REVIEW
Table 1. Cont. 7 of 51
7 of 51
Molecules
Molecules 2018,
23, ×23,
2018, FOR FORPEER
PEERPEER REVIEW
REVIEW
REVIEWAnticoagulants 8 of8518ofof5151
Anticoagulants
Chiral Compounds
Molecules
Molecules 2018,
Molecules 2018, 23,
23, ×23,
2018, FOR××FOR
FOR
PEERPEER REVIEW
REVIEW
PEER REVIEW 8ofof51
8 of 851 51
Table
Table
Table 1.1.Cont.
1. Cont. Cont.
Anticoagulants
OH OH
OH OH
Molecules 2018, 23, × FOR PEER REVIEW
H H TableTable
1.
Table 1.1.
Cont. Cont.
Cont.
H H
8 of 51
O O N N
O O OH
N N
O
O O
O O Table
Table 1. 1.
Anticoagulants
Cont. Cont.
Table 1. Cont.
Anticoagulants
Anticoagulants
OH O
S S
Anticoagulants
N N Anticoagulants
Anticoagulants
1. Cont.
H H TableAnticoagulants
OH O
Anticoagulants
Anticoagulants
Alprenolol
Alprenolol
Alprenolol
OH OHOH OO Sotalol
Sotalol
Sotalol
Anticoagulants
O
O O
O O
OH OHOH O O O Anticoagulants
Warfarin
Warfarin
OOH OH OH
O (WFN)
(WFN)
O OO
Warfarin (WFN)
O O
OH O O O OO
O O OO O O
Non-steroidal
Non-steroidal anti-inflamatory
anti-inflamatory drugs
drugs
Warfarin
Warfarin (WFN)
(WFN)
Warfarin (WFN)
O OO O OO
Non-steroidal anti-inflamatory drugs
Warfarin
Warfarin (WFN)
Warfarin (WFN)
(WFN)
Warfarin
Warfarin (WFN)(WFN) Non-steroidal OHOH drugs
anti-inflamatory
Warfarin OHOH Non-steroidal
(WFN) anti-inflamatory
Non-steroidal drugs
anti-inflamatory drugs OHOH
O O
Warfarin (WFN) Non-steroidal

Non-steroidal
HO anti-inflamatory
anti-inflamatory
Non-steroidal O drugs
anti-inflamatory
O OH drugs
drugs HO HO
Warfarin (WFN)O OH O HO
Non-steroidal
Non-steroidal anti-inflamatory
anti-inflamatory drugs
drugs
O
OOH
Non-steroidal
O
Non-steroidal
anti-inflamatory
anti-inflamatory
drugs
drugs
HO O O OHOHOH HO
O OHOH OH O OHOH OH
Non-steroidal anti-inflamatoryOH drugs
OHOH
Ibuprofen OHOH
Ibuprofen OH O
HOHOHO CarboxyibuprofenOH OH
Carboxyibuprofen
O OO HO HO 2-Hydroxyibuprofen
2-Hydroxyibuprofen
HO OHOH OH
O OHOO OH OH OO OH
O OH
OH HOHOHOOO O OO HOHO HO OH
Ibuprofen O OO OCarboxyibuprofen O OO HO2-Hydroxyibuprofen
HO O O OO
HOHOHO O O OH HO
O O
OHO OH O O O O OHOO
O
Ibuprofen
Ibuprofen OH
Ibuprofen OOCarboxyibuprofen
O Carboxyibuprofen
Carboxyibuprofen OH 2-Hydroxyibuprofen
2-Hydroxyibuprofen
2-Hydroxyibuprofen
HO O OH HO
Ibuprofen
Ibuprofen
Ibuprofen
O OH OCarboxyibuprofen
Carboxyibuprofen
Carboxyibuprofen N N
O2-Hydroxyibuprofen
2-Hydroxyibuprofen
2-Hydroxyibuprofen
O
Ibuprofen O O Carboxyibuprofen OHOH
2-Hydroxyibuprofen
O O Ibuprofen
Ibuprofen
Ibuprofen Carboxyibuprofen
O Carboxyibuprofen
Carboxyibuprofen
O OH
O 2-Hydroxyibuprofen
2-Hydroxyibuprofen
2-Hydroxyibuprofen
O OO NO OO O
O OHOH
OH O
OH
O Ibuprofen OH O OO
Carboxyibuprofen O OHOH
OH O OO
2-Hydroxyibuprofen
OHOH N NON O
OO OH
NaproxenO OH OH O OOKetoprofen OH
OHOH O O
Indoprofen OHOH
O
O O Naproxen Ketoprofen OO OH
O OH Indoprofen
N NN OH
O OO OH O N NN
O OO
OH OH
OH
O OO Naproxen O Ketoprofen O OO Indoprofen O O
OHOO
O OH
O OH
OH
OCl OO
Cl
OHOO
O
Naproxen Ketoprofen Indoprofen
N Indoprofen OOHOH
O
Naproxen
Naproxen O O Ketoprofen
Ketoprofen OHOH Indoprofen O
O Cl NaproxenO Ketoprofen IndoprofenOH
Naproxen OH OH
Naproxen
Naproxen Ketoprofen
Ketoprofen
Ketoprofen
O Indoprofen
Indoprofen
Indoprofen
OOOH
O
Cl ClCl Naproxen
Naproxen
Naproxen
N
O Ketoprofen
Ketoprofen
Ketoprofen O OH
O Indoprofen
Indoprofen
Indoprofen
N
H
H
OH O O F F O OHOH OH
Cl Cl Cl
Naproxen
N
O OO
Ketoprofen O OHOH OH Indoprofen OH
OH OH
Cl ClCl H
O OHO OH
O OH
OHOH F O OO OH
Carprofen Fenoprofen Flurbiprofen
OOH
Carprofen Fenoprofen O OH OO OH FlurbiprofenO OOOH OH
N NN O OH
O OH OOH
O OO OH
Cl H HH
O OO F FF
Carprofen OH OH Fenoprofen Flurbiprofen OOH OO
Bronchodilators
N NN OH
H HH O OO
BronchodilatorsOHOO
O F FF
O
N NN
Carprofen
Carprofen
Carprofen
CarprofenOH
H HH OO
OFenoprofen
Fenoprofen
Fenoprofen
Fenoprofen
Flurbiprofen
Flurbiprofen
FFO
Flurbiprofen
Flurbiprofen
F
Bronchodilators
OHOH H
Carprofen
NCarprofen
Carprofen O Fenoprofen
O
Fenoprofen
Fenoprofen
Bronchodilators Flurbiprofen
Flurbiprofen
Flurbiprofen
H H F Flurbiprofen
Carprofen
Carprofen
Carprofen N Fenoprofen
Bronchodilators
Fenoprofen
Bronchodilators
Bronchodilators
Fenoprofen Flurbiprofen
Flurbiprofen
HOHO OH N
H Bronchodilators
Bronchodilators
Bronchodilators
Carprofen OHOH
HO HOHO N Fenoprofen
Bronchodilators
Bronchodilators Flurbiprofen
OH H
H H Bronchodilators
HO OHOH OH NN
N H
HO HO
HO Salbutamol H H Bronchodilators
Salbutamol OH (SBT)
(SBT)
OHOHN N N
HOHOHO H HH
HOHOHO
Salbutamol (SBT) N NN
HOHOSalbutamol
HO OH(SBT)
HOHOHO H Antibiotics
Antibiotics
Antibiotics
Salbutamol
HO HOSalbutamol
HO
HO (SBT)
N(SBT)
(SBT)
Salbutamol
Antibiotics O O
Salbutamol
Salbutamol
OH(SBT)
Salbutamol (SBT)
(SBT)
O O
OHOH OH
Salbutamol (SBT)
F
HO Salbutamol (SBT)
Salbutamol (SBT)
F
Antibiotics
Antibiotics
Antibiotics OH
Cl Cl
O O OH
OH OH
O N HN HN F Antibiotics
Antibiotics
Antibiotics
OSalbutamol (SBT) ClOH Cl Cl Antibiotics
Antibiotics
N N
O O O OHO O
N
Antibiotics
N O
N
OH OHOH OH
OH
O O O O
O N HN N
N F F F
O
O O O OO O O
OH OHOH OH OH
ClClClCl N N OH OHOH
O OH OH
O
OHOH OHOH Antibiotics
F F F O OO O OO
OO HN
HN HN
Chloranfenicol ClClCl
Cl Cl N O
Ofloxacin OH OHOH
O N NNChloranfenicol FFNOfloxacin
Chloranfenicol Cl Ofloxacin
F
N N N N N
OH
HN HNHNCl Cl OH OH
ClCl ClCl
O O
O NOOOO O
N
N OH OH O O O N N NN N NO O ON N N
O O Chloranfenicol
NOOON
NO
HN HNHN
O OO Cl ClCl N
F
N NN
Ofloxacin
NN O O ON N N OH
Cl
O O O O OO
O N Chloranfenicol
Chloranfenicol
Chloranfenicol
HN N NN Ofloxacin
Ofloxacin
Ofloxacin
O OO
Cl N N
O Chloranfenicol
Chloranfenicol
Chloranfenicol
O Ofloxacin
Ofloxacin
Ofloxacin
Chloranfenicol Ofloxacin
N O
Chloranfenicol
Ofloxacin
Molecules 2018, 23, 262 7 of 47
Table 1. Cont.
Chiral Compounds
Molecules 2018,23,
Molecules2018, 23,× ×FOR
FORPEER Antiepileptic and their metabolites
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REVIEW 1010ofof5151
Molecules
FORPEER
PEER
Antiepileptic and their metabolites
REVIEW 10ofof5151
Molecules 2018,2018,
23, ×23,
FOR PEER REVIEW
REVIEW 10 of10
51
OH
Molecules
Molecules 2018,
2018, 23, ×23,
FOR× FOR PEER
PEER REVIEW
REVIEW Table1.1.Cont.
Table Cont. 10 of1051of 51
Molecules 2018, 23, × FOR PEER REVIEW Table
Table
Table 1. Cont. Cont.
1.1.Cont. 10 of 51
Proton
Protonpump
pumpinhibitors
inhibitors
Molecules 2018, 23, × FOR PEER 2018,
Molecules REVIEW
23, × FOR PEER REVIEW Table
Table 1. Cont.
1. Cont. 10 of 51 10 of 51
N Proton
Proton
Table 1.Proton
Cont. pump pump
pump inhibitors
inhibitors
inhibitors
Molecules 2018, 23, × FOR PEER REVIEW H H
H H
N N O O
7 of 48
HProton
Proton pump pump inhibitors
inhibitors
N N O O
H H
Table 1. Cont.
H H
O S SO O1. Cont.
H
Table
O N N
NH O O H H N N N S S N N
N N ZAIZGHELI
2 O O O
N N N N N
H HH
N NN N SN
O S SO O S S N N N
N SN S S Table 1. Cont.
N N N
10,11-Dihydro-10-hydroxycarbamazepine
10,11-Dihydro-10-
H H
S
N N S S N NN H H NN N N
N
ZAIZGHELINProton pump H inhibitors

O O O O
N N N
Molecules 2018, 23, × FOR PEER REVIEW
H H O O H
NF F 7 of 48
Ohydroxycarbamazepine
NN NN
ProtonProton pump inhibitors O O O
S pump N inhibitors
N O O N S S N N
OO H S N
N O O
F F FF S N
SO S N N F N N O O O
N N H HH
S N H H O OHO N N O NN
F O
O
N N N F F F FO
O O H
N
H H Table 1. Cont.
N ON N O O
O
N NO O N O F FF FF S N SS N
N O O
S N S O O O
O O N
Omeprazol N Lanzoprazol Rabeprazole
S N O NN
Omeprazol Lanzoprazol Rabeprazole
S N S N
Proton pump inhibitors
N N F F
Omeprazol
Omeprazol
Omeprazol HLanzoprazol
Lanzoprazol
Lanzoprazol Rabeprazole
Rabeprazole
Rabeprazole
N N N F F H
O O O O N O O
O
N O F
H O O
O
O
N OH H
N N
S N
O O S N O
Omeprazol
Omeprazol Lanzoprazol
Lanzoprazol Rabeprazole
Rabeprazole
F F F
HS HH N O O N
N NN O S SO O
Lanzop
N
F F F F O O
O
Omeprazol Rabeprazole
N
O N NS S S
razol Lanzoprazol
O O
F F H H O
N N Omeprazol Rabeprazole
Omeprazol
NN Lanzoprazol
Lanzop Rabeprazole
Omeprazol S S HN Lanzoprazol Rabeprazole
N O O
F OF O O O
Omeprazol Rabeprazole
F F F
O O
F H
N
O O
N NO S N
razol O
F O O H O OO
F F
Omeprazol
S N O Lanzoprazol Rabeprazole
FPantoprazole
N
F
HPantoprazole
H O
N
N N S
Pantoprazole
Pantoprazole
O O
Pantoprazole
N
F F O O O
F
S S N N N
O
Antineoplastic
Antineoplasticagents
agents and
andtheir
theirmetabolite
metabolite
F O O
N N
O O
PantoprazolePantoprazole
Pantoprazole O
Antineoplastic agents and their
O
metabolite
Antineoplastic
Antineoplastic agents
agents andand their
their metabolite
metabolite
F F
O
Pantoprazole Antineoplastic agents and their metabolite
Pantoprazole O O O O
Pantoprazole
Pantoprazole Antineoplastic
Antineoplastic agents
agents andand their
their metabolite
metabolite
Antineoplastic agents and their metabolite
Pantoprazole Antineoplastic agents and their metabolite
N O Antineoplastic
N O agents and their metabolite
HNHN OO
ClCl NAntineoplastic
O HN HN agents
HN POP and
OO their metabolite
HN O
NN POP OO
N
Antineoplastic agents and their metabolite
ClHN
HN P HNP P P P
Cl ClCl P P P HN
HN HN HN
HNN HNHN
N OOO
O HN HN
HN O
OO O
N O HN O
Cl Cl PO P OO O PO P OO O
Cl P P
HN
HN N O HN
HN HN
HN HN
Cl N O P O O HN P O
O O
Cl ClCl
O
PO HNHN ClClO
P
NHN
HNP ClCl
Cl Cl HN Cl Cl
Cl O Cl PCl O
3‐N‐
Ifosfamide
Ifosfamide O
HN 3-N-Dechloroethylifosfamide
3-N-Dechloroethylifosfamide
HN O
Ifosfamide
Cl Cl Ifosfamide
Ifosfamide
IfosfamideCl O Dechloroethylifo
Cl Cl O

Cl
sfamide
Ifosfamide Cl Antifungals
Antifungals
3‐N‐
IfosfamideCl
Ifosfamide 3-N-Dechloroethylifosfamide
3-N-DechloroethylifosfamideCl Antifungals
Antifungals
Antifungals
Ifosfamide N
Antifungals
Dechloroethylifosfamid
Cl
Cl N N
Antifungals N N N
Cl e
3-N- NN
N
Ifosfamide
N N N Antifungals
Antifungals N
N N N
Dechlor
HO
N N Cl Antifungals N N N NN
HO
N HO
3-N-
Cl Cl N N
Ifosfamide oethylif N NN
N N N N N N
Cl Cl Cl N N N N N N NHO HOHO
N
O O
O
Dechlor
Antifungals
osfamid O O
N
O N N
N
NHO N N
NN
Cl Ifosfamide oethylif
Cl N N N N
e
HO HO
Cl O Cl O O
Cl
O O
O Cl
Cl N
N
Cl
osfamid
Cl Cl N Cl Cl N N Cl Cl
Cl Cl Cl Cl Cl N
Cl Cl Cl N N
Cl Cl
O Econazole Cl Cl
O Miconazole N Tebuconazole Cl Cl Cl
e
Cl Cl
Cl Cl Cl Cl Cl O Cl Cl
Cl Cl Cl Cl Cl
Antifungals
O O O N
N N
Econazole
Econazole Miconazole
Miconazole Tebuconazole
Tebuconazole
O HO
Cl
Cl Cl Cl Cl
Cl Cl
EconazoleEconazole Miconazole
Miconazole
N N TebuconazoleN
Tebuconazole
Econazole
Econazole Miconazole
N Miconazole Tebuconazole
Tebuconazole
Cl Cl Cl Cl
N Cl Cl
Econazole Cl Cl
Miconazole
Cl Cl Cl Cl Cl Cl
Tebuconazole
N
O
CH
Antifungals 3
O N
N
N HN
N
O
Cl
Econazole
Econazole
N
N Cl
Miconazole
Miconazole
N N
N
HO
N
Tebuconazole
Tebuconazole
N
N NN
N N
Cl
Cl Cl Cl Cl Cl Cl Cl
N CH3 N HO
N N N N NN
O
N NN N H H H N
N O Cl N N NN
HO N
Econazole Miconazole Tebuconazole
Cl O N N N N H HH
N N N
Cl Cl Cl Cl Cl Cl O N N N NN NN N
N HOHO
Cl
O O O
Cl Cl N N
Cl Cl Cl O Cl OOClO
Cl Cl Cl Cl H H N
HO HOHO Cl Cl Cl
O O
O O NO N N NN
N
O Cl Cl Cl Cl
Micona
Cl O O O Cl N Cl
Cl Cl Cl N
Cl Cl
Econazole Tebuconazole
N N
Ketoconazole HO HO Hexaconazole Penconazole N
zole
O O O Cl Cl Cl Cl H
NO N
NO O O
N Cl Cl
N N N Cl Cl Cl
Micona
Cl N Cl ClN
Econazole Tebuconazole Cl Cl Cl
zole
Cl N OCl N O N
Ketoconazole
N N O Hexaconazole
HO N Penconazole Cl Cl Cl
Ketoconazole Hexaconazole H
Cl Penconazole
Ketoconazole Hexaconazole Penconazole
O Cl Cl Cl Cl
Ketoconazole Hexaconazole Penconazole
O N
N N O O
Ketoconazole
Ketoconazole
Ketoconazole O Hexaconazole
Hexaconazole
Hexaconazole Penconazole
Penconazole
Penconazole Cl Cl
O
N
Cl
N
H Cl Cl

N
Ketoconazole
Ketoconazole
NO
Cl N
Hexaconazole
Hexaconazole
O Penconazole
Penconazole
Cl
N Cl
N
Cl Ketoconazole
N Hexaconazole Penconazole
Cl
N
Triadimefon Imazalil
Molecules 2018, 23, 262 8 of 47
Table 1. Cont.
Molecules 2018,
Molecules 23, ×23,
2018, FOR PEER
× FOR REVIEW
PEER REVIEW 11 of1151of 51
Chiral Compounds
N N
O O N N
H H
O O O O
N N Cl Cl
Cl Cl N N
Molecules 2018,
Molecules 23, ×23,
2018,
Molecules 2018, FOR PEER
23,× ×FOR REVIEW
FORPEER
PEERREVIEW
REVIEW 12 of1251
12ofof5151
N N
Table
Table
TableCont.
Cl1. Cl 1.1.Cont.
Cont.
Triadimefon Table 1. Cont.
Imazalil
Triadimefon
Triadimefon Imazalil
Imazalil
Anti-helminthic
Anti-helminthic
Anti-helminthic
Anti-helminthic
Anti‐helminthic
O
O OO
N S
N
N N N NN S SS
N N N NN
N
N N N N
NN N SS S
N NN
N S
O
O OO

Phenyltetrahydroimidazo
Praziquantel Tetramisole Phenyltetrahydroimidazo thiazole
thiazole (PTHIT,
Praziquantel Tetramisole Phenyltetrahydroimidazo
(PTHIT, levamisole/dexamisole)
levamisole/dexamisole)
Praziquantel
Praziquantel
Praziquantel Tetramisole
Tetramisole
Tetramisole thiazole (PTHIT,
thiazole
thiazole(PTHIT,
(PTHIT,
Antihistaminic
Antihistaminic levamisole/dexamisole)
O
OH Antihistaminic
Antihistaminic
Antihistaminic
N

OH
OH
N N N
OH OHOH

Fexofenadine
Fexofenadine
OH OHOH
3. Chiral Analyses in Biological Samples
Fexofenadine
Fexofenadine
Fexofenadine
This study reviewed 58 articles that have been published between 1996 and 2017 based on
This study
ScienceDirect reviewed
and ISI Web of 58Knowledge
articles that have been
databases. The published
investigated between
compounds 1996 and 2017 based on
included
3. Chiral
ScienceDirect
synthetic Analyses
3.3.Chiral
Chiraland
psychoactive ISIin
Analyses
Analyses WebBiological
drugs of Samples
ininBiological
Biological
Knowledge Samples
Samples
(stimulants), databases.
synthetic The investigated
opioids, compounds
β‐blockers, included synthetic
antidepressants,
psychoactivebronchodilators
anticoagulants, drugs (stimulants), and syntheticanesthetics
dissociative opioids, (Table
β-blockers,
1 and antidepressants,
2). Figure 2 shows anticoagulants,
the on
This study
This
This study
studyreviewed
reviewed 58 58
reviewed articles
58articlesthatthat
articles have
that been
have
have published
been
been published
publishedbetween
between
between1996 andand
1996
1996 2017
and based
2017
2017 based
basedonon
bronchodilators
ScienceDirect
and
ScienceDirect
dissociative
and ISI Web
Web
anesthetics
of Knowledge
Knowledge
(Tables 1
databases.and
databases.
2).
TheThe
databases.
Figure 2
relative number of studies of each class of chiral drug investigated and the analytical methods used
ScienceDirect andand ISI ISI
Web of of
Knowledge investigated
The
shows
investigated
investigated
the
compounds relative
compounds
compounds
number of
included
included
included
for analysis of these compounds in different biological matrices.
studies of eachpsychoactive
synthetic class of chiral drug
drugs investigated
(stimulants), and the analytical
synthetic opioids, methods
β-blockers,usedantidepressants,
for analysis of these
synthetic
synthetic psychoactive
psychoactive drugs
drugs (stimulants),
(stimulants), synthetic
synthetic opioids,
opioids, β-blockers,
β-blockers, antidepressants,
antidepressants,
anticoagulants,
anticoagulants,
compounds anticoagulants,bronchodilators
in different biologicaland
bronchodilators
bronchodilators anddissociative
and
matrices. anesthetics
dissociative
dissociativeanesthetics(Table
anesthetics (Table1 and
(Table 1 1and2). 2).
and Figure
2).Figure 2 shows
Figure thethe
2 2shows
shows the
relative
The number
relative
relativenumber
use of of studies
numberofofstudies
racemates of each class
studiesofofeach
eachclass
typically of chiral drug
classofofchiral
results investigated
chiraldrug
in druginvestigated and
investigatedand
stereoselective the
andtheanalytical
theanalytical methods
analyticalmethods
pharmacological used
methodsused
used
activity and
for for
analysis of these
foranalysis
analysis
pharmacokinetic compounds
ofaffecting
ofthese
these compounds
compounds in different
bioavailability, biological
inindifferent
different matrices.
biological
biological
metabolism matrices.
matrices.
and excretion that may contribute to the
toxicity, increase risk of death or serious adverse effects [11,56,57]. Though there is a tendency
for manufacturing pharmaceuticals as single enantiomers, many pharmaceuticals are still supplied as
racemates [57]. Concerning illicit drugs, these compounds are also available as racemates or single
enantiomers depending on the manufacturing procedure [5,6,57]. Regarding illicit administrations,
consumption of pure enantiomer (eutomers), in some cases, may cause overdose or even might
lead to lethal cases [10,12]. Thus, significance of chiral analysis has increased since it is possible to
determine whether the drug of concern is derived from a controlled or illicit substance [58–60]. In fact,
some controlled substances are commercialized in the enantiomeric pure form due to their advantages
in therapeutic activities [5,6,57]. On the other hand, illicit production of these drugs leads to either
racemic or single enantiomers depending on the manufacturing procedure, i.e, racemic or enantiomer
Figure 2. Relative number of each class of chiral drug referred in the reviewed enantioselective
published
pure studies
precursors. and the
Thus, in analytical
forensic methods
chemistry, used for separation
evaluation of the
of the EFchiral
maydrugs in the
aid in biological
discrimination of the
fluids.
consumption of legal and illegal substances, give information about method of synthesis used or profile
Figure 2. Relative
Figure
Figure number
2.2.Relative
Relative of each
number
number class
ofofeach
each of chiral
class
class drug
ofofchiral
chiral referred
drug
drug in the
referred
referred reviewed
ininthe enantioselective
thereviewed
reviewedenantioselective
enantioselective
The use of racemates typically results in stereoselective pharmacological activity and
published
publishedstudies
published andand
studies
studies the analytical
andthe methods
theanalytical
analytical used
methods
methods for for
used
used separation of the
forseparation
separation chiral
ofofthe drugs
thechiral
chiral in biological
drugs
drugs ininbiological
biological
pharmacokinetic affecting bioavailability, metabolism and excretion that may contribute to the
fluids.
fluids.
fluids.
toxicity, increase risk of death or serious adverse effects [11,56,57]. Though there is a tendency for
manufacturing pharmaceuticals as single enantiomers, many pharmaceuticals are still supplied as
TheThe useuseof of
racemates typically results in instereoselective pharmacological activity andand
The use of racemates
racemates typically
typically results
results in stereoselective
stereoselective pharmacological
pharmacological activity
activity and
racemates [57]. Concerning illicit drugs, these compounds are also available as racemates or single
pharmacokinetic affecting bioavailability, metabolism andandexcretion thatthat
may contribute to the
pharmacokinetic
pharmacokinetic affecting
affecting bioavailability,
bioavailability, metabolism
metabolism and excretion
excretion thatmay
may contribute
contribute totothe
the
Fexofenadine

This
Molecules study
2018, 23, 262reviewed58 articles that have been published between 1996 and 2017 based 9 ofon
47
ScienceDirect and ISI Web of Knowledge databases. The investigated compounds included
synthetic psychoactive drugs (stimulants), synthetic opioids, β-blockers, antidepressants,
among different bronchodilators
anticoagulants, seizures linking among them, consumers
and dissociative and traffickers.
anesthetics Chiral
(Table 1 and analysis2 can
2). Figure alsothe
shows be
applied in doping control, as an example, the use of preparations containing dextromethorphan
relative number of studies of each class of chiral drug investigated and the analytical methods used by
athletes is allowed,
for analysis of thesewhereas
compoundsthe use of levorphanol
in different is expressly
biological matrices.banned by the International Olympic
Committee [61].
Figure
Figure 2. Relative number
2. Relative number ofof each
each class
class of
of chiral
chiral drug
drug referred
referred in
in the
the reviewed
reviewed enantioselective
enantioselective
published studies and the analytical methods used for separation of the chiral drugs in biological fluids.
published studies and the analytical methods used for separation of the chiral drugs in biological
fluids.
Though various works have been published concerning chiral separation of the different classes of
pharmaceuticals
The use ofandracemates
illicit drugstypically
of forensic interestinin biological
results matrices
stereoselective most works do not
pharmacological determine
activity and
the enantiomeric affecting
pharmacokinetic composition. Data aboutmetabolism
bioavailability, the enantiomeric composition
and excretion of parent
that may compounds
contribute to the
and metabolites
toxicity, increaseisrisk
of high importance
of death or seriousforadverse
accurate data [11,56,57].
effects interpretation and there
Though for further analysis for
is a tendency of
results [62]. Also,
manufacturing metabolites of as
pharmaceuticals achiral compounds
single enantiomers, canmany
also be chiral and should
pharmaceuticals are be considered
still supplied inas
biological
racematessamples.
[57]. Concerning illicit drugs, these compounds are also available as racemates or single
enantiomers depending on the manufacturing procedure [5,6,57]. Regarding illicit administrations,
3.1. Synthetic Psychoactive Drugs
consumption of pure enantiomer (eutomers), in some cases, may cause overdose or even might lead
to lethal
This cases
class [10,12].
of chiralThus,
drugssignificance
was the most of chiral analysis
studied has1 increased
(Tables and 2 andsince it is2).possible
Figure Among to
synthetic psychoactive drugs are the amphetamine-like drugs, a group of structurally related
compounds with vast potential for abuse, addiction and toxicity [63]. Among most studied
compounds are AM, methamphetamine (MA), 3,4-methylenedioxymethylamphetamine (MDMA),
3,4-methylenedioxy-ethylamphetamine (MDEA) and methylphenidate (MPH) (Tables 1 and 2).
Enantiomers of these drugs have been discriminated in plasma, urine, blood and hair. Analytical
methods used for the separation of enantiomers of these drugs included LC-MS, GC-FID and GC-MS
and CE. Amphetamine-like drugs can be used for the treatment of some disorders such as selegiline
(L-deprenyl, SG) used for treatment of Parkinson’s disease, Adderall or Elvanse for treatment of
attention-deficit hyperactivity disorder or famprofazone, a nonsteroidal anti-inflammatory agent,
used for pain control [8,59]. Consumption of SG produces S-MA and its metabolite S-AM while
famprofazone produces both R-MA and S-MA and their metabolites in human body [7]. Adderall
contains both R-AM and S-AM. On the other hand, these substances are often misused for recreation
purposes and even by healthy individuals to enhance work or school performance (e.g., MPH) or
doping in sport practice [64,65]. Thus, illicit preparations have been an alternative route of access these
substances by consumers and abusers. Illicit production of AM may use 1-phenyl-2-propanone and
other reagents such as formic acid, ammonium formate or formamide are used, which is designated as
the Leuckart method, yielding a racemic product [17].
Manufacture of MDMA and related drugs can use safrole, isosafrole, piperonal or
3,4-methylenedioxyphenyl-2-propanone (PMK). Many illicit drug syntheses start with PMK and
use either the Leuckart route or various reductive aminations, producing a racemic MDMA [17].
Therefore, studies based on determining the ratios of R- and S-isomers of the parent compounds
and metabolites are important for distinctive medical drug administration or illegal abuse of
Molecules 2018, 23, 262 10 of 47
amphetamine like drugs [8]. Also, chiral information is useful and essential to identify the precursor,
the synthetic pathway, and intrinsic characteristics of the seized samples [5,6]. In this context, analysis
of the enantiomers of AM and metabolites have revealed to be very useful, since it can provide
information about the origins of the drug consumed (legal or illicit) [58,60].
In the majority of chiral amphetamine-like drugs, the S-enantiomers exhibit greater potency than
the R-enantiomers [60,66–70].
Nishida et al. described a LC-MS method for the determination of the enantiomers of MA,
AM, SG and its metabolite, desmethylselegiline (DMSG), in hair samples [59]. In this study, authors
showed differences in the enantiomer ratio of MA and AM and between MA abuse consumers and SG
consumers [59].
Molecules 2018, 23, ×Besides,
FOR PEERitREVIEW
was also shown that the existence of DMSG in SG users that is not normally 10 of 47
found in urine demonstrating that the method can be useful for distinguish therapeutic users of SG and
MA
usersabusers.
of SG and Fujii MA
et al.abusers.
described a GC-MS
Fujii et al. method based
described on the method
a GC-MS formation of diastereoisomers
based on the formation usingof
CDR TPC for the separation of the enantiomers of MA, AM, MDMA and MDA
diastereoisomers using CDR TPC for the separation of the enantiomers of MA, AM, MDMA and in urine samples [8].
This
MDAmethod
in urine cansamples
be used[8].
for This
discrimination
method can between legal
be used forand illegal consumption
discrimination betweenoflegal
theseand
drugs [8].
illegal
Rasmussen
consumption et al.
of developed
these drugs for [8].
the first time a GC-MS
Rasmussen et al. enantioselective
developed for method
the firstfortime
the separation
a GC-MS
of AM, MA, MDA,
enantioselective MDMA
method forand
the MDEA in whole
separation blood
of AM, MA,based
MDA,inMDMA
the formation of diastereomers
and MDEA in whole blood[67]
(Figure
based in3).the formation of diastereomers [67] (Figure 3).
Figure 3. Chromatogram
Chromatogramrepresenting
representingthe
theenantioseparation
enantioseparationof of
R/S-AM,
R/S-AM, R/S-MA,
R/S-MA,R/S-MDA, R/S-
R/S-MDA,
MDMA and and
R/S-MDMA R/S-MDEA
R/S-MDEAas R-MTPCl derivatives
as R-MTPCl in whole
derivatives blood
in whole concentrations
blood at (A)
concentrations 2µg/g
at (A) andand
2 µg/g (B)
at 0.002
(B) µg/g,
at 0.002 respectively.
µg/g, Reproduction
respectively. with
Reproduction permission
with of Elsevier
permission (Figure
of Elsevier 1 from
(Figure Rasmussen
1 from Rasmussenet
et
al.al. [67]).
[67]).
The method was validated and applied to four whole blood samples from forensic cases
including a suspected case of driving under the influence of drugs. In all cases amphetamines were
ingested as racemates with stereoselective metabolism since the R/S ratio for most enantiomers
were >1 showing that the R-enantiomer is metabolised faster than the S-enantiomer. Hädener et al.
described a two dimensional LC-MS/MS method for quantification of AM enantiomers in human
urine [16]. The study was applied to 67 urine samples from suspected AM abusers, subjects treated
Molecules 2018, 23, 262 11 of 47
The method was validated and applied to four whole blood samples from forensic cases including
a suspected case of driving under the influence of drugs. In all cases amphetamines were ingested as
racemates with stereoselective metabolism since the R/S ratio for most enantiomers were >1 showing
that the R-enantiomer is metabolised faster than the S-enantiomer. Hädener et al. described a two
dimensional LC-MS/MS method for quantification of AM enantiomers in human urine [16]. The study
was applied to 67 urine samples from suspected AM abusers, subjects treated with S-AM prodrug and
suspected MA abusers. In each 40 samples obtained from suspected AM abusers both enantiomers were
present and mean R/S ration was 1.25 indicating a predominance of the R-enantiomer. The excepted
value R/S would be 1 but due to stereoselective metabolism of AM in which S-AM is metabolized
faster than R-AM resulting in higher concentrations of R-AM. In the consumers of the prodrug it
was found only S-AM and R-AM in one sample at <LOQ probably due to imputities of the drug
manufacture itself [16]. Considering individuals suspected of MA consumer, in 80% of the samples,
both enantiomers were found although with a predominance of S-AM. R/S ration ranged from 0.01 to
0.47. Five samples from MA abusers contained only S-AM. This result was explained by the faster
metabolization of S-enantiomer of MA. Thus, more S-MA is converted to S-AM than R-MA to R-AM.
Binz et al. developed an analytical method for chiral analysis of AM in hair [7]. In this study,
analysis of hair samples from nine Elvanse patients revealed only S-AM in eight cases. One subject
showed both enantiomers indicating a (side-) consumption of street AM. The analysis of the 16 AM
abusers samples showed only racemic AM. Furthermore, it could be shown in a controlled study
that S-AM can be detected after administration of even very low doses of lisdexamfetamine and
dexamphetamine, which can be of interest in forensic toxicology and especially in drug-facilitated
crime [7].
MPH, another CNS stimulant, is used in the treatment of attention deficit hyperactivity disorder
(ADHD) and narcolepsy. Abuse of this substance has been reported [71]. This drug is commercialized
in the racemic mixture though only the D-threo-form is responsible for the desired therapheutic
effect. MPH is enantioselectively metabolized, preferring S-MPH over R-MPH to ritanilic acid (RA)
that is pharmacologically inactive. Individual variations on MPH metabolization classified some
individuals as poor metabolizers. Thomsen et al. developed an LC-MS/MS enantioselective method
for determination of MPH and RA in femoral blood applied to forensic cases [65] in order to evaluate
poor metabolizers by estimating R/S ratio of MPH. Postmortem blood samples from autopsy cases and
antemortem blood samples from mixture of traffic, violence and sexual assault cases were analyzed.
Apart from one case, R-MPH showed the highest concentration in the postmortem cases, a similar
pattern to the found in living organisms. Concentration of RA was higher in all cases than MPH with
equal distribution of R and S enantiomers. In antemortem individuals the same pattern was observed
with higher levels of R-MPH and equal quantities of R and S forms of RA.
These reports demonstrate that knowledge about the enantioselective behavior and measure of
the enantiomeric ratio of these types of drugs can provide useful and valuable data in the forensic
field giving information about consumption of licit and illicit amphetamine like drugs and other
psychoactive drugs and essential to aid in the correct interpretation of the use of these substances.
3.2. Synthetic Opioids

The second most studied classe of chiral compounds are the synthetic opioids (Tables 1 and 2).
Among reported compounds are tramadol, methadone and methorphan [72,73]. Opioid abuse,
addiction, and overdose are considered of a serious public health [54]. In the European Monitoring
Centre of Drug and Drug Addiction report of 2017, opioids were the third most consumed class
of drugs of abuse in Europe and the first with more fatal cases [74]. Concerning tramadol, it is a
synthetic opioid that acts as agonist by selective activity at the µ-opioid receptors commercialized
as a racemate of the more active 1R,2R-enantiomer ((+)-tramadol) and the less active 1S,2S-tramadol
((−)-tramadol), with both enantiomers acting through different mechanisms, but in a synergistic
manner [75]. However, there are differences in their binding properties leading to considerable
Molecules 2018, 23, 262 12 of 47
differences in pharmacological activities [75]. Tramadol is metabolized to O-desmethyltramadol (ODT)

and N-desmethyltramadol (NDT). O-Demethylation of tramadol is carried out by CYP2D6; enzyme
expressed polymorphically [76]. Polymorphisms play an important role in inter-individual drug
response [77]. The metabolite ODT is pharmacologically active, has longer half-life and is more potent
than parent compounds. Tramadol has been used for nonmedical purposes due to it euphoric and
mood enhancing effects [78,79]. Tramadol abusers develop physiological dependence which can
cause negative effects such as convulsion and seizures [80]. Besides, genetic polymorphisms can
influence biological properties including toxicity. A LC-MS/MS method for separation of tramadol,
and its principal metabolites, ODT and NDT for pharmacokinetic applications in plasma samples was
reported [81]. Authors showed that plasma binding was not enantioselective, nevertheless kinetic
disposition of tramadol and its NDT metabolite was enantioselective, with plasma accumulation of
(+)-tramadol and (+)-NDT, whereas the pharmacokinetics of ODT was not enantioselective in patients
with neuropathic pain phenotyped as extensive metabolizers of CYP2D6. Thus, enantioselective
methods for both tramadol and its metabolites are essential for an accurate evaluation of their biological
properties and toxicity.
Methadone is a synthetic opioid frequently used for treatment of opiate dependent persons,
pharmacologically similar to morphine, but lacks the euphoric effects [82,83]. Methadone can be
fatal by itself or by interaction with other drugs such as depressors of the CNS. Methadone is a
substrate for CYP2B6 and CYP2C19, which are all stereoselective [83]. Plus, CYP P450 isoenzymes
are known to have individual variability (polymorphisms) that leads to poor metabolizers, rapid
metabolizers and ultra-rapid metabolizers [83]. This chiral drug, when administred as racemate gives
rise to enantiomeric metabolites S-(−)-2-ethylidine-1,5-dimethyl-3,3-diphenylpyrrolidine (EDDP) and
the R-(+)-enantiomer. The R-(−)-enantiomer of methadone, is the one with higher affinity for the
µ-opioid receptor having a higher analgesic potency (over fifty times more) [82,83]. On the other
hand, the S-enantiomer is responsible for the poor cardiac tolerance [84]. In order to investigate
the enantiomeric ratios of methadone and EDDP in postmortem samples, Jantos et al. applied a
LC-MS/MS method in femoral blood, urine, bile, brain, lungs, kidneys and muscle tissue samples [82].
The study was based in sixteen samples, from eleven man and five women with ages ranging between
23 to 43 years old. Concentrations of R-methadone and R-EDDP were found in all body fluids and
tissues, while S-enantiomers were only found in thirteen of the cases. The R/S ratios ranged from
0.58 to 4.19 for methadone, and from 0.38 to 1.38 for EDDP [82]. Because methadone does not suffer
racemization in the human body, the enantiomeric ratios found in postmortem samples, can reflect if
the substance consumed was either racemic or not; also can identify if the cause of death is related to
toxic exposure [82]. Moody et al. developed an enantioselective method by LC-MS/MS for methadone
and EDDP in human plasma, urine and liver microsomes [85]. This study demonstrated differences in
the pharmacokinetic between enantiomers of methadone and its main metabolite EDDP and suggest
greater production of and lesser clearance of S-EDDP.
The antitussive dextromethorphan (allowed drug) and the narcotic analgesic levomethorphan
(banned drug, not commercially available) are the R- and R- isomers of 3-methoxy-N-methylmorphinan.
Aumatell and Wells developed a CE chiral method for separation of methorphan (racemate of
dextromethorphan and levomethorphan) [86]. Distinction of these compounds is not only of interest in
forensic science (such as the elucidation of the cause of death after intake of levomethorphan), but also
for the treatment of intoxicated patients. Also, the use of preparations containing dextromethorphan by
athletes is allowed, whereas the use of levorphanol is expressly banned by the International Olympic
Committee [61].
3.3. Antidepressants
Although antidepressants are considered non-addictive, many people abuse of these drugs [87,88].
Users can become physically dependent and non-compliance may arise as a result, with fatal
consequences in some cases [89]. Among studied antidepressants are fluoxetine (FLX), citalopram,
Molecules 2018, 23, 262 13 of 47
reboxetine, venlafaxine (VNF) and its metabolites (Tables 1 and 2). FLX is an example of antidepressants
administrated as racemate, that both enantiomers have the same biological active. It acts by selective
inhibition of the serotonin reuptake pump, increasing the extracellular catecholamines, such as
serotonine, dopamine and norepinephrine. In the human body, it is metabolized to norfluoxetine
(NFLX). FLX enantiomers are approximately equipotent in blocking the 5-HT reuptake, while the
enantiomers of NFLX show marked differences in pharmacological activity. The enantiomer S-NFLX
shows approximately 20 times more potency than the R-enantiomer as 5-HT reuptake inhibitor, both
in vitro and in vivo [90–92]. Shen et al. considered the enantioseparation of FLX in human plasma
but its metabolite was not considered [93]. Nevertheless, Unceta et al. developed an LC-FD method
for simultaneous separation of FLX and NFLX enantiomers, in order to investigate potential sources
of variability, in rats receiving chronic treatments, on concentrations of FLX and NFLX and their
enantiomers [91]. The plasma levels of R-NFLX were considerably increased in comparison to the
S-enantiomer. In plasma FLX R/S ratios were of 1.02 compared to 1.05 in cerebral cortex, which was in
contrast with NFLX R/S ratios, that were 1.81 in plasma and 1.5 in cerebral cortex [91].
Citalopram, used in the treatment of depression, commercialized as racemate and its enantiomer
S-(+)-citalopram (escitalopram, marketed in the enantiomerically pure form) is 100 times more potent
as a serotonin reuptake inhibitor as compared to R-(−)-citalopram.
VNF is a phenylethylamine derivative that affects brain neurotransmission by blocking
presynaptic reuptake of serotonin and noradrenaline [94], and administrated in the treatment of
psychiatric disorders [95]. VNF undergoes extensive first-pass metabolism by CYP P450 enzymes
into its major active metabolite O-desmethylvenlafaxine (OD-VNF), and two minor metabolites,
N-desmethylvenlafaxine (ND-VNF) and N,O-didesmethylvenlafaxine (N,O-DD-VNF). OD-VNF
inhibits the reuptake of serotonin and noradrenaline in similar potency to that of VNF [96].
Stereoselective metabolism has been observed both in vitro and in vivo, where CYPD2D6 displays and
appreciable stereoselectivity towards the R-enantiomer [96].
Reboxetine is used as a selective noradrenaline reuptake inhibitor for the treatment of major
depressive disorders, commercialized as racemate (S,S- and R,R-reboxetine). Ohman et al. developed
an enantioseletive method for analysis of reboxetine in serum in patients with chronic medication [97].
Authors found that the median S,S/R,R ratio in steady state was 0.5 and ranged from 0.22 to 0.88.
It was also shown that women have an approximately 30% higher S,S/R,R ratio than men. The S,S/R,R
ratios of reboxetine were not found to correlate with reboxetine concentrations. Authors also found
a correlation between selective noradrenaline reuptake inhibitor activity that is higher in women than
in men and that may alter the enantiomeric ratio.
3.4. β-Blockers
β-Blockers, also known as β-adrenergic blocking agents, are a class of chiral drugs that are used
for the management of cardiac arrhythmias. Usually one enantiomer presents higher potency than the
other. For instances, S-(−)-propranolol (PHO) is 100 times more than R-(+)-PHO. Most of β-blockers
(except timolol: S-isomer) are marketed as racemates, such as acebutolol, atenolol (ATE), alprenolol,
betaxolol, carvediol, metoprolol (MET), labetalol, pindolol and sotalol [4]. In addition to therapeutic
properties, these compounds exhibit calming neurological effects decreasing anxiety, nervousness and
stabilizing motor performance. Thus, these compounds are included in prohibited list according to the
World Anti Doping Agency (WADA) regulation because of the improved psychomotor performance
that may be beneficial in sports requiring precision and accuracy such as shooting archery among
others [61]. Among β-blockers only PHO, MET, carvediol, verapamil and its metabolite enantiomers
were discriminated in plasma and urine (Tables 1 and 2) [98–101]. Analytical methods used for
the separation of enantiomers of these drugs included LC-MS and GC-MS. PHO is administered
as racemate to treat hypertension and normalize tachycardia response; however, the S-enantiomer
shows greater cardiosympatholytic activity [102]. Siluk et al. developed an analytical method for
separation of R,S-PHO in human plasma for determination of pharmacokinetic difference among
Molecules 2018, 23, 262 14 of 47
the two enantiomers and even drugs interaction [98]. In this study, authors suggest that R-PHO
is eliminated faster than S-PHO. Concerning MET, Kim et al. developed an analytical method for
enantioseparation of its enantiomers in urine. This method can be applied in pharmacological and
pharmacokinetic studies of both enantiomers in biological samples [100]. Beyond carvediol and
verapamil, there are not studies concerning the enantioseparation of other used β-blockers in biological
samples. Methods of enantioseparation for these substances are important to evaluate pharmacological
and pharmacokinetic differences among enantiomers and possible toxicity due to interaction with
other administered drugs.
3.5. Anticoagulants
Warfarin (WFN) is one of the most commonly prescribed cardiovascular medication anticoagulant
drugs used to manage thromboembolic disease. WFN is administered as an oral medication consisting
of a racemate though the S-enantiomer has higher activity than the R-enantiomer. Several factors
increase the risk of over-anticoagulation such as genetic polymorphisms as well as others factors,
including age, sex, and histories of smoking and alcohol consumption and diets rich in vitamin
K [103]. Genetic factors and drug interactions mostly account for the risk of over-anticoagulation [103].
Knowledge about enantioselective pharmacodynamic and pharmacokinetic is not only important to
assure efficiency and safety but also because genetic polymorphisms may have an important impact in
biological properties including toxicity. Separation of WFN enantiomers was achieved using different
analytical methods: SFC-MS/MS, LC-MS/MS and Micellar electrokinetic chromatography MEKC-MS
in plasma (Tables 1 and 2) [104–106]. Knowledge about pharmacokinetic of enantiomers of WFN and
its metabolites may add in the development of enantiopure commercialized forms of WFN that may
be safer and for studies of possible toxicological and interaction of WFN with other pharmaceuticals
concomitant administered. Beyond the use of WFN as anticoagulant, this compound was used as a
poison, and is still marketed as a pesticide against rats and mices.
3.6. Dissociative Anaesthetics

Ketamine (K) began to be a widespread drug of abuse in many countries and primarily available
through illicit means. At sub anaesthetic doses this drug provides hallucinogenic effects [107,108].
Because of these desire effects K is often used in recreational purposes and particularly dangerous with
regards to traffic and workplace safety. In fact, K can be bought in the internet from suspected veterinary
distributers and clinics [109]. Chiral discrimination of K and its main metabolite norketamine (NK)
was done in in plasma and hair [110,111]. S-(+)-K is an anesthetic and analgesic but R-(−)-ketamine
is associated to hallucinations and agitation. K is a dissociative anaesthetic that induces loss of
consciousness, amnesia, immobility, and in a lesser extent analgesia [110]. It is used in paediatric
emergency retrieval and in veterinary surgery, because of its reduced tendency to give respiratory
depression [110]. Its main advantage is to induce profound analgesia and amnesia, while maintaining
the cardiopulmonary functions and the protective airway reflexes stable [110]. K undergoes extensive
first-pass metabolism to produce various free and glucurinated hydroxylated derivatives [110].
However, its main metabolic pathway occurs through N-demethylation to NK which appears to
have 20–30% activity of its parent drug [110]. Although is used as a racemate, the S-enantiomer
showed to have four times higher affinity for the phencyclidine site of the NMDA receptor, as well as
a greater potency when compare to the R-form and the racemate [110].
3.7. Bronchodilators
Among bronchodilators are β-adrenoreceptor agonists (or β2-agonists) that are drugs commonly
used for the treatment of asthma and other pulmonary disorders. They have bronchodilator and
anabolic activities. Because of these properties, these compounds may be used by athletes to enhance
performance and as a safer alternative to anabolic steroids though the use by asthmatic athletes
is not forbidden. In this sense, like β-blockers, the β-adrenergic compounds are scheduled in the
Molecules 2018, 23, 262 15 of 47
Prohibit List of the WADA [61]. Only one report was found that describes the enantioseparation of a
bronchodilator, the salbutamol (SBT). This compound is commercialized as racemate, however the
R-enantiomer of SBT binds to β2-adrenergic receptors with greater affinity than the S-enantiomer,
which does not act through β-adrenergic receptor activation. S-SBT has adverse effects associated,
such like augmentation of bronchospasm and pro-inflammatory activities. Studies have reported
that the S-enantiomer can potentiate the effects of spasmogens in airway of smooth muscle from
both guinea pigs and humans, with a number of clinical studies also reporting worsening of airways
hyper-responsiveness in animals and in subjects with asthma [112]. The initial step in metabolism of
both enantiomers is sulfate conjugation, a stereoselective process that occurs in human airway epithelial
cells, as also in other cells and tissues [113]. The greater rate of sulfate conjugation of R-SBT might
lead to lower plasma levels of R- than S-enantiomer in human subjects, which can be responsible for
increasing the adverse effects related with the latter [112,113]. Since bronchodilator pharmacodynamic
is enantioselective the development of enantioselective methods for bronchodilators is essential for
stereo-pharmacokinetics and enantioselective safety studies. Data from pharmacokinetic studies can
contribute to the development of enantiopure broncodilators therapeutic drugs that can be safer and
used in the control of broncodilators abuse.
3.8. Anti-Helmintic
Levamisole and dexamisol [(phenyltetrahydroimidazothiazole (PTHIT)] were widely used as
an anti-worm medication for both humans and animals, but they are no longer approved for use in
the United States or Canada due to their toxicity. In South America, illicit cocaine laboratories
have been known to add PTHIT to cocaine preparations to extend their effects [10]. Casale et
al. developed an analytical method by GC-FID for the determination of PTHIT enantiomers. i.e.,
levamisole and dexamisole in illicit cocaine seizures and in urine cocaine abusers [10]. Beyond the
possibility of linking origin of cocaine seizures, the addiction of PTHIT have been shown to cause
agranulocytosis/neutropenia in cocaine abusers expanding the adverse effects of the consumption of
this drug. Approximately 78% of cocaine samples contained PTHIT in an average concentration of
23%. Enantiomeric compositions of dexamisole/levamisole were different among samples. 66% of the
samples contained levamisole, 19% the racemate and 15% the higher levels of levamisole. Samples
containing only dexamisole were not detected. The higher content in levamisole may be due to
traffickers adding tetramisole and levamisole to the cocaine or illegitimate preparation of levamisole.
Considering urine samples the majority of urine extracts contained levamisole (46%), and levamisole
enhanced enrichment (20%) and dexamisole-enhanced enrichment (26%). Levamisole has high toxicity
affecting all organ systems, with agranulocytosis, manifested primarily as acute, profound neutropenia
and have been found in concaine consumers leading to serious clinical complications. Thus, this
methodology allowed clinical toxicologists and forensic chemists to establish specific enantiomer
composition of PTHIT in cocaine samples and in the urine specimens of cocaine abusers [10]
Molecules 2018, 23, 262 16 of 47
Table 2. Chromatographic analytical methods described for the analysis of chiral illicit drugs and pharmaceuticals in biological matrices.
Matrix Concentration Range/ER

Drug Method Stationary Phase LOD LOQ Reference
Application (When Mencioned)
0.049 ng/mL (R)
SGE-BPX5 (15 m × 0.25 mm, 0.25 µm film thickness) 50 fg 0.006–50 ng/mL [30]
0.195 ng/mL (S)
Plasma GC/NICI-MS 5–250 µg/L;

HP-5MS (30 m × 0.25 mm, 0.25 µm film thickness) n.r. 5 µg/L ER (R/S): 0.97–1.66, with a [114]
mean value of 1.15
GC/EI-MS HP-5MS (30 m × 0.25 mm, 0.25 µm film thickness) n.r. 5 ng/g plasma 5–400 ng/g plasma [31]
GC/EI-MS HP-5MS (30 m × 0.25 mm, 0.25 µm film thickness) n.r. 0.004 µg/g 0.004–3 µg/g [67]
Blood
0.003–60 ng/mg (R)
0.8 pg/mg (R) 2.7 pg/mg (R)
GC/NICI-MS HP-5MS (30 m × 0.25 mm, 0.25 µm film thickness) 0.002–60 ng/mg (S) [49]
0.7 pg/mg (S) 2.4 pg/mg (S)
ER (R/S): 0.03–0.95
HP5-MS (30 m × 0.25 mm, 0.25 µm film thickness) n.r. 2.5 ng/sample 2.5–100 ng/sample [31]
GC/EI-MS
Hair 5% phenyl-methylsilicone capillary column (17 m × 0.2
0.1 ng/mg 0.2 ng/mg 0.2–20 ng/mg [69]
mm, 0.33 µm film thickness)
AM LC/MS/MS n.r. 20 pg/mg 50 pg/mg 50–20000 pg/mg [7]
HPLC/ESI-MS Chiral DRUG (150 mm × 2 mm) 0.05 ng/mg n.r. 0.2–40 ng/mg [59]
DB-5MS (20 m × 0.18 mm × 0.18 mm) 10 ng/mL n.r. 25–10000 ng/mL [115]
GC/EI-MS HP-5MS (20 m × 0.25 mm, 0.25 µm film thickness) n.r. 10 µg/L 10–500 µg/L [42]
45–1000 (l; d)
HP-5MS (30 m × 0.2 mm, 0.33 µm film thickness) 40 ng/mL 45 ng/mL [70]
45–2000 (d,l)
Urine 5% phenyl polysiloxane (15 m × 0.2 mm, 0.2 µm df) n.r. n.r. 25–10000 ng/mL [116]
1.1 ng/mL (R) 3.7 ng/mL (R)
HP-5MS (20 m × 0.25 mm, 0.25 µm film thickness) 5–500 µg/L [117]
GC/MS 1.3 ng/mL (S) 4.3 ng/mL (S)
DB-5 (10 m × 0.1 mm, 0.4 µm film thickness) 0.5 ng/mL n.r. 20–1000 ng/mL [8]
0.02 µg/mL n.r. 0.05–10 µg/mL [33]
CE/ESI-MS Uncoated fused silica capillary (50 µm, 100 cm)
0.03 µg/mL n.r. 0.2–10 µg/mL (S) [118]
PVA chemically modified diol capillary column
CE n.r. n.r. n.r. [36]
(40 cm × 50 µm)
Lux AMP (150 × 3 mm, 5 µm) n.r. 0.05 mg/L 0.05–25 mg/L [16]
LC/MS-MS
Chirobiotic V2 (250 × 2.1 mm, 5 µm) 0.02 mg/L 0.05 mg/L 0.05–50.00 mg/L [60]
Adsorbosphere HS, C18 (150 × 4.6 mm, 5µm); C18
HPLC-UV n.r. n.r. 0.1–100 mg/L [119]
precolumn (7.5 × 4.6 mm)
Molecules 2018, 23, 262 17 of 47
Table 2. Cont.

5–250 µg/L
Plasma GC/NICI-MS HP-5MS (30 m × 0.25 mm, 0.25 µm film thickness) n.r. 5 µg/L ER (R/S): 1.02–1.63, with a [114]
mean value of 1.33
Blood GC/EI-MS HP-5MS (30 m × 0.25 mm, 0.25 µm film thickness) n.r. 0.004 µg/g 0.004–3 µg/g [67]
0.007–60 ng/mg (R)
2.1 pg/mg (R) 6.9 pg/mg (R)
GC/NICI-MS HP-5MS (30 m × 0.25 mm, 0.25 µm film thickness) 0.005–60 ng/mg (S) [49]
1.5 pg/mg (S) 5.0 pg/mg (S)
ER (R/S): 0.01–0.82
Hair
5% phenyl-methylsilicone capillary column (17 m × 0.2
GC/MS 0.1 ng/mg 0.2 ng/mg 0.2–20 ng/mg [69]
mm i.d., 0.33 µm film thickness)
HPLC/ESI-MS Chiral DRUG (150 mm × 2 mm) 0.01 ng/mg n.r. 0.04–40 ng/mg [59]
DB-5MS (20 m × 0.18 mm i.d. × 0.18 mm) 10 ng/mL n.r. 25–10000 ng/mL [115]
GC/EI-MS 45–1000 (l; d)
MA HP-5MS (30 m × 0.2 mm, 0.33 µm film thickness) 40 ng/mL 45 ng/mL [70]
45–2000 (d,l)
HP-5MS (20 m × 0.25 mm, 0.25 µm film thickness) n.r. 10 µg/L 10–500 µg/L [42]
5% phenyl polysiloxane (15 m × 0.25 mm, 0.2 µm df) n.r. n.r. 25–10000 ng/mL [116]
HP-1 (12 m × 0.25 mm, 0.25 µm film thickness) n.r. 10 ng/mL 10–2000 ng/mL [120]
GC/MS 2.0 ng/mL (R) 6.8 ng/mL (R)
HP-5MS (20 m × 0.25 mm, 0.25 µm film thickness) 5–500 µg/L [117]
1.6 ng/mL (S) 5.2 ng/mL (S)
Urine
DB-5 (10 m × 0.1 mm, 0.4 µm film thickness) 3 ng/mL n.r. 20–1000 ng/mL [8]
0.02 µg/mL n.r. 0.05–10 µg/mL [33]
CE/ESI-MS Uncoated fused silica capillary (50 µm, 100 cm)
0.03 µg/mL n.r. 0.2–10 µg/mL (S) [118]
PVA chemically modified diol capillary column (40 cm
CE n.r. n.r. n.r. [36]
× 50 µm)
LC/MS/MS Chirobiotic V2 (250 × 2.1 mm, 5 µm) 0.02 mg/L 0.05 mg/L 0.05–50.00 mg/L [60]
Adsorbosphere HS, C18 (150 × 4.6 mm, 5µm); C18
HPLC-UV n.r. n.r. 0.1–100 mg/L [119]
precolumn (7.5 × 4.6 mm)
ODS-1 (150 mm × 4.6 mm) with precolumn (20 × 4.0
Plasma HPLC-DAD n.r. 7 ng/mL n.r. [121]
mm)
GC/EI-MS HP-5MS (30 m × 0.25 mm, 0.25 µm film thickness) n.r. 0.004 µg/g 0.004–3 µg/g [67]
MDMA Blood Kinetex C18 column (100 × 2.1 mm, 2.6 µm film
LC/MS/MS n.r. 0.0025 µg/L 0.0025–1.25 µg/L (R, S) [122]
thickness)
1.7 pg/mg (R) 5.6 pg/mg (R) 0.006–60 ng/mg (R)
Hair GC/NICI-MS HP-5MS (30 m × 0.25 mm, 0.25 µm film thickness) [49]
1.5 pg/mg (S) 5.1 pg/mg (S) 0.005–60 ng/mg (S)
Molecules 2018, 23, 262 18 of 47
Table 2. Cont.

GC/NICI-MS n.r. (chiral derivatization S-HFBPrCl)
n.r. n.r. n.r. [68]
LC/HRMS Chirex3012 (250 × 4.6 mm, 5 µm film thickness)
HP-5MS (20 m × 0.25 mm, 0.25 µm film thickness) n.r. 10 µg/L 10–500 µg/L [42]
Urine GC/EI-MS
GC/MS HP-5MS (20 m × 0.25 mm, 0.25 µm film thickness) 1.7 ng/mL 5.7–5.8 ng/mL 5–500 µg/L [117]
HPLC-DAD ODS-1 (150 × 4.6 mm) with precolumn (20 × 4.0 mm) n.r. 7 ng/mL n.r. [121]
Plasma HPLC-DAD ODS-1 (150 × 4.6 mm) with precolumn (20 × 4.0 mm) n.r. 5 ng/mL n.r. [121]
Kinetex C18 column (100 × 2.1 mm, 2.6 µm film
Blood LC/MS/MS n.r. 0.0025 µg/L 0.0025–0.25 µg/L (R, S) [122]
thickness)
GC/NICI-MS HP-5MS (30 m × 0.25 mm, 0.25 µm film thickness) [49]
Hair
MDA 5% phenyl-methylsilicone capillary column (17 m × 0.2
n.r. n.r. n.r. [68]
LC/HRMS Chirex3012 (250 × 4.6 mm, 5 µm film thickness)
Urine HP-5MS (20 m × 0.25 mm, 0.25 µm film thickness) n.r. 2 µg/L 2–100 µg/L [42]
GC/EI-MS
HPLC-DAD ODS-1 (150 × 4.6 mm) with precolumn (20 × 4.0 mm) n.r. 5 ng/mL n.r. [121]
Whole blood GC/EI-MS HP-5MS (30 m × 0.25 mm, 0.25 µm film thickness) n.r. 0.004 µg/g 0.004–3 µg/g [67]
GC/NICI-MS HP-5MS (30 m × 0.25 mm, 0.25 µm film thickness) [49]
MDEA Hair
mm i.d., 0.33 µm film thickness)
Urine GC/EI-MS 5% phenyl polysiloxane (15 m × 0.25 mm, 0.2 µm df) n.r. n.r. 25–5000 ng/mL [116]
Kinetex C18 column (100 × 2.1 mm, 2.6 µm film
Blood LC/MS/MS n.r. 0.0025 µg/L 0.0025–0.25 µg/L (R, S) [122]
thickness)
n.r. n.r. n.r. [68]
pOHMA LC/HRMS Chirex3012 (250 × 4.6 mm, 5 µm film thickness)
0.02 µg/mL n.r. 0.05–10 µg/mL [33]
Urine CE/ESI-MS Uncoated fused silica capillary (50 µm, 100 cm)
0.05 µg/mL n.r. 0.2–10 µg/mL (S) [118]
PVA chemically modified diol capillary column (40 cm
CE n.r. n.r. n.r. [36]
× 50 µm)
Molecules 2018, 23, 262 19 of 47
Table 2. Cont.

HMMA Blood LC/MS/MS Kinetex C18 column (100 × 2.1 mm, 2.6 µm film thickness) n.r. 0.0025 µg/L 0.0025–0.25 µg/L (R, S) [122]
LC/EI-MS/MS Chiral-AGP (50 × 2.0 mm, 5 µm) n.r. 2.5 ng/mL 0–500 ng/mL [85]
Plasma Chiral-AGP (100 × 4.0 mm, 5 µm); Chiral-AGP guard
LC/MS 0.02 ng/mL 1 ng/mL 1–300 ng/mL [83]
column (10 × 2.0 mm, 5 µm)
Chiral-AGP (100 × 4.0 mm, 5 µm); Chiral-AGP guard
LC/ESI-MS/MS n.r. n.r. n.r. [12]
column (10 × 2.0 mm, 5 µm)
Methadone Blood
α-1-acid glycoprotein (100 × 4.0 mm, 5 µm) and a α-1-acid
LC/MS n.r. 0.02 mg/L 0.05–2.1 mg/L [123]
glycoprotein guard column (10 × 2.0 mm, 5 µm)
Blood 0.65 ng/L (R) 2.40 ng/L (R)
LC/MS/MS Chiral-AGP column (150 × 3 mm, 5 µm) 50–1000 ng/L [82]
Tissues 0.49 ng/L (S) 1.82 ng/L (S)
0–500 ng/mL
Urine LC/EI-MS/MS Chiral-AGP (50 × 2.0 mm, 5 µm) n.r. 2.5 ng/mL [85]
ER (R/S): 1.42–2.96
Liver LC/EI-MS/MS Chiral-AGP (50 × 2.0 mm, 5 µm) n.r. 2.5 ng/mL 0–500 ng/mL [85]
LC/MS 0.01 ng/mL 0.1 ng/mL 1–25 ng/mL [83]
column (10 × 2.0 mm, 5 µm)
Plasma
LC/EI-MS/MS Chiral-AGP (50 × 2.0 mm, 5 µm) n.r. 2.5 ng/mL 0–500 ng/mL [85]
Blood LC/ESI-MS/MS n.r. n.r. n.r. [12]
EDDP column (10 × 2.0 mm, 5 µm)
Blood 0.77 ng/L (R) 2.82 ng/L (R)
LC/MS/MS Chiral-AGP column (150 × 3 mm, 5 µm) 50–1000 ng/L [82]
Tissues 0.76 ng/L (S) 2.79 ng/L (S)
0–500 ng/mL
Urine LC/EI-MS/MS Chiral-AGP (50 × 2.0 mm, 5 µm) n.r. 2.5 ng/mL [85]
ER (R/S): 0.76–0.89
Liver LC/EI-MS/MS Chiral-AGP (50 × 2.0 mm, 5 µm) n.r. 2.5 ng/mL 0–500 ng/mL [85]
0.2 ng/mL (total) 0.2–600 ng/mL (total)
LC/ESI-MS/MS Chiralpak AD (250 × 4.6 mm, 10 µm) n.r. [81]
0.5 ng/mL (unbound) 0.5–250 ng/mL (unbound)
Lux Cellulose-2 (150 × 4.6 mm, 3 µm); Lux Cellulose-2
0.15 ng/mL 1 ng/mL 1–800 ng/mL [75]
LC/APCI-MS/MS guard column (4 × 3 mm)
Chiralpak AD (250 × 4.6 mm, 10 µm) 1 ng/mL 3 ng/mL 25–1000 ng/mL [34]
Plasma
Chiralcel OD-R (250 × 4.6 mm, 10 µm); LiChrospher 0.18 ng/mL (+)
T HPLC-DAD 1 ng/mL 1–500 ng/mL [124]
60-RP-selected B (250 × 4 mm, 5 µm) 0.16 ng/mL (-)
Chiralpak AD (250 × 4.6 mm, 10 µm) 1 nM 5 nM 0.01–1.55 µM [125]
Chiralpak AD (250 × 4.6 mm); LichroCART 4-4
HPLC-FD n.r. 2.5 ng/mL 2.5–250 ng/mL [126]
LiChrospher 100 Diol (5 µm) precolumn
n.r. 2 ng/mL 2–200 ng/mL [127]
column (10 × 4.0 mm, 5 µm)
GC/EI-MS Rt-βDEXcst (30m × 0.25 mm, 0.25 µm film thickness) 0.01 µg/mL 0.1 µg/mL 0.1–20 µg/mL [128]
Urine
HPLC-FD Chiralpak AD (250 × 4.6 mm, 10 µm) 2 nM 25 nM 0.1–3.0 µM [125]
Molecules 2018, 23, 262 20 of 47
Table 2. Cont.

Lux Cellulose-2 (150 × 4.6 mm, 3 µm); Lux Cellulose-2 guard 0.20 ng/mL (+)
1 ng/mL 1–400 ng/mL [75]
LC/APCI-MS/MS column (4 × 3 mm) 0.30 ng/mL (-)
Chiralpak AD (250 × 4.6 mm, 10 µm) 1.3 ng/mL 4 ng/mL 25–1000 ng/mL [34]
Plasma
ODT Chiralcel OD-R (250 × 4.6 mm, 10 µm); LiChrospher 0.08 ng/mL (+)
HPLC-DAD 0.5 ng/mL 0.5–100 ng/mL [124]
Chiralpak AD (250 × 4.6 mm, 10 µm) 1 nM 5 nM 0.01–1.55 µM [125]
Chiralpak AD (250 × 4.6 mm); LichroCART 4-4 LiChrospher
100 Diol (5 µm) precolumn
Chiral-AGP (150 × 4.0 mm × 5 µm); Chiral-AGP guard
n.r. 2.5 ng/mL 2.5–100 ng/mL [127]
column (10 × 4.0 mm × 5 µm)
GC/EI-MS Rt-βDEXcst (30 m × 0.25 mm, 0.25 µm film thickness) 0.03 µg/mL 0.1 µg/mL 0.1–20 µg/mL [128]
Urine
HPLC-FD Chiralpak AD (250 × 4.6 mm, 10 µm) 2 nM 25 nM 0.1–3.0 µM [125]
Chiralcel OD-R (250 × 4.6 mm, 10 µm); LiChrospher 0.15 ng/mL (+)
NDT Plasma HPLC-DAD 0.5 ng/mL 0.5–250 ng/mL [124]
Chiral-AGP (150 × 4.0 mm, 5 µm); Chiral-AGP guard column
(10 × 4.0 mm, 5 µm)
Chiralpak AD (250 × 4.6 mm); LichroCART 4-4 LiChrospher
N,O-DDM-T Plasma HPLC-FD n.r. 2.5 ng/mL 2.5–250 ng/mL [126]
100 Diol (5 µm) precolumn
Chiralcel OD-R (250 × 4.6 mm × 10 µm); LiChrospher 100
Citalopram Plasma LC/ESI-MS/MS n.r. 0.1 ng/mL 0.1–20 ng/mL [129]
RP-8 precolumn (4 × 4.0 mm × 5 µm)
FLX Plasma LC/APCI-MS/MS Chirobiotic V (250 × 4.6 mm, 5 µm) n.r. 2 ng/mL 2–1000 ng/mL [93]
Plasma LC/MS 0.25 ng/mL 1 ng/mL 1–125 ng/mL [111]
(10 × 2.0 mm, 5 µm)
K
Hair CE-UV-DAD Uncoated fused-silica capillary (450 × 50 mm) 0.08 ng/mg 0.25 ng/mg 0.5–8.0 ng/mg [110]
NK Plasma LC/MS 0.25 ng/mL 1 ng/mL 1–125 ng/mL [111]
(10 × 2.0 mm, 5 µm)
Hair CE-UV-DAD Uncoated fused-silica capillary (450 × 50 mm) 0.08 ng/mg 0.25 ng/mg 0.5–8.0 ng/mg [110]
n.r. 0.006 ng/mL 0.006–12.5 ng/mL [130]
Plasma GC/NICI-MS BPX5 fused silica (15 m × 0.25 mm)
n.r. 0.072 ng/mL 0.072–18.25 ng/mL [131]
MPH Chiral AGP (100 × 4.0 mm, 5 µm); guard column (10 × 2.0
Blood LC/MS/MS n.r. n.r. 0.2–500 ng/g [65]
mm, 5 µm)
Urine GC/EI-MS DB-5 (30 m × 0.32 mm, 0.25 µm film thickness) n.r. 10 ng/mL 0–10000 ng/mL [132]
Molecules 2018, 23, 262 21 of 47
Table 2. Cont.

50–500 nmol/L
Reboxetine Serum LC/MS Chiral-AGP (2 × 100 mm, 5 µm) <1 nmol/L n.r. ER (S/R): 0.22–0.88, with a [97]
mean value of 0.5
1–1000 nM
LC/ESI-MS/MS Chirobiotic V (250 × 2.1 mm, 5 µm) n.r. 0.5 nM [95]
ER (S/R): 1.01–4.33
Plasma
5.2 ng/mL (R)
VNF HPLC/ESI-MS Chirobiotic V (250 × 4.6 mm, 5 µm) 1 ng/mL 5–400 ng/mL [133]
5 ng/mL (S)
10–4000 nM
Whole blood LC/ESI-MS/MS Chirobiotic V (250 × 2.1 mm, 5 µm) n.r. 0.5 nM [95]
ER (S/R): 0.59–1.11
1–1000 nM
LC/ESI-MS/MS Chirobiotic V (250 × 2.1 mm, 5 µm) n.r. 0.5 nM [95]
ER (S/R): 0.70–12.3
Plasma
3.5 ng/mL (R)
OD-VNF HPLC/ESI-MS Chirobiotic V (250 × 4.6 mm, 5 µm) 1.5 ng/mL 4–280 ng/mL [133]
4.3 ng/mL (S)
10–4000 nM
ER (S/R): 0.59–1.11
0.5–500 nM
Plasma LC/ESI-MS/MS Chirobiotic V (250 × 2.1 mm, 5 µm) n.r. 0.25 nM [95]
ND-VNF ER (S/R): 1.24–2.91
5–2000 nM
ER (S/R): 0.46–1.53
0.5–500 nM
Plasma LC/ESI-MS/MS Chirobiotic V (250 × 2.1 mm, 5 µm) n.r. 0.25 nM [95]
ER (S/R): 0.42–1.18
N,O-DD-VNF
5–2000 nM
ER (S/R): 0.90–1.99
0.1–4 ng/mL
MET Urine GC/EI-MS HP-5MS (30 m × 0.25 mm, 0.25 µm film thickness) 0.5 ng/mL n.r. [100]
ER (R/S): 0.83
Chirobiotic V (250 × 4.6 mm, 5 µm); Chirobiotic V
PHO Plasma LC/APCI-MS 0.03 ng/mL 0.25 ng/mL 0.25–200 ng/mL [98]
guard column (20 × 4.0 mm, 5 µm)
Chiralpak AD (250 × 4.6 mm); Chiralpak AD-H guard
SFC/APCI-MS/MS n.r. 13.6 ng/mL 13.6–2500 ng/mL [104]
column (10 × 4.0 mm)
Plasma Chirobiotic V (250 × 4.6 mm, 5 µm); Cyclobond I guard 5–1500 ng/mL
WFN LC/ESI-MS/MS 1.5 ng/mL 5 ng/mL [105]
column (20 × 4.0 mm, 5 µm) ER (S/R): 0.47±0.14
0.1 µg/mL (instr. 0.25–5 µg/mL
MEKC/ESI-MS Fused silica capillaries (120 cm, 375 µm o.d., 50 µm) n.r. [106]
limit) ER (R/S): 2.24–6.20
Chirobiotic T (250 × 4.6 mm, 10 µm) n.r. 0.2 ng/mL 0.2–500 ng/mL [99]
Carvedilol Plasma LC/ESI-MS/MS
Ace 3 C18 (50 × 2.0 mm, 3 µm) n.r. 0.2 ng/mL 0.2–200 ng/mL [134]
Verapamil Plasma CE-UV Fused-silica capillaries n.r. n.r. 2.5–250 ng/mL [101]
Molecules 2018, 23, 262 22 of 47
Table 2. Cont.

Norverapamil Plasma CE-UV Fused-silica capillaries n.r. n.r. 2.5–250 ng/mL [101]
SBT Urine NACE/ESI-MS Fused silica capillaries (48.5 cm, 375 µm o.d., 50 µm) 8–14 ng/mL 18–20 ng/mL 15–150 ng/mL [135]
PTHIT Urine GC-FID Rt-β-DEXsm (30 m × 0.25 mm, 0.25 µm film thickness n.r. n.r. n.r. [10]
AM: amphetamine; CE: capillary electrophoresis; CI: chemical ionization; DAD: diode array detection; EDDP: 2-ethylidene-l,5-dimethyl-3,3-diphenylpyrrolidine; EI: electron
impact; ER: enantiomeric ratio; ESI: electrospray ionization; FD: fluorescence detector; FID: flame ionization detector; FLX: fluoxetin; GC: gas chromatography; HMMA:
4-hydroxy-3-methoxymethamphetamine; HPLC: high performance liquid chromatography; K: ketamine; LC: liquid chromatography; LOD: limit of detection; LOQ: limit of quantification;
MA: methamphetamine; MDA: 3,4-Methylenedioxyamphetamine; MDMA: 3,4 Methylenedioxymethamphetamine; MDEA: N-methyl-diethanolamine; MEKC: micellar electrokinetic
chromatography; MET: metoprolol; MPH: methylphenidate; MS: mass spectrometry; MS/MS: tandem mass spectrometry; NACE: nonaqueous capillary electrophoresis; NDT:
N-desmetil-tramadol; ND-VNF: N-desmethylvenlafaxine; NICI: negative ion chemical ionization; N,O-DD-VNF: N,O-didesmethylvenlafaxine; NFLX: norfluoxetin; NK: norketamine;
ODT: O-desmetil-tramadol; OD-VNF: O-desmetil-venlafaxine; PHO: propranolol; PTHIT: phenyltetrahydroimidazothiazole; SBT: salbutamol; T: tramadol; UV: ultraviolet detector; VNF:
venlafaxine; WFN: warfarin. n.r.: not referred.
Molecules 2018, 23, 262 23 of 47
4. Chiral Analyses in the Aquatic Environment

4. Chiral Analyses in the Aquatic Environment
This study reviewed 33 articles that have been published between 2005 and 2017 based in
This study reviewed 33 articles that have been published between 2005 and 2017 based in
ScienceDirect and ISI web of Knowledge databases (Table 3 and Figure 4). The target compounds
ScienceDirect and ISI web of Knowledge databases (Table 3 and Figure 4). The target compounds
included antidepressants,
included β-blockers, nonsteroidal
antidepressants, β-blockers, nonsteroidal anti-inflamatory
anti-inflamatory drugsdrugs (NSAIDs),
(NSAIDs), synthetic
synthetic
psychoactive drugs, antibiotics, synthetic opioids, antiepileptics, antihistaminic; bronchodilators,
psychoactive drugs, antibiotics, synthetic opioids, antiepileptics, antihistaminic; bronchodilators,
antineoplasticagents
antineoplastic agentsand
and proton
proton pump
pump inhibitors
inhibitors (Table
(Table 1). Figure
1). Figure 4 shows
4 shows the relative
the relative number
number of
of studies
studies
of of each
each class class of
of chiral chiral
drug drug investigated
investigated and the analytical
and the analytical methods usedmethods used for
for analysis of analysis of these
these compounds
compounds
in in different environmental
different environmental matrices. matrices.
Figure 4. Relative number of each class of chiral drug referred in the reviewed
Figure 4. Relative number of each class of chiral drug referred in the reviewed enantioselective published
enantioselective published studies and the analytical methods for separation of the chiral drugs
studies and the analytical methods for separation of the chiral drugs in environmental samples.
in environmental samples.
The entrance of pharmaceuticals and illicit drugs into the aquatic environment may occur through
The from
effluents entrance of pharmaceuticals
WWTPs which are unable andtoillicit drugs
totally removeinto these
the aquatic environment
micro-pollutants or may
directoccur throughof
discharged
effluentsWWTPs
sewage. from WWTPs which
biological are unablecan
treatments, to totally
alter theremove
EF of these micro-pollutants
the enantiomers present or direct
in the discharged
influent, as
microbiota
of sewage. action
WWTPs generally
biologicalis stereoselective
treatments, can[136,137].
alter the Disposed drugs will usually
EF of the enantiomers present beinfound in their
the influent,
parent form, either
as microbiota as racemate
action generallyorissingle enantiomers.
stereoselective On the Disposed
[136,137]. other hand,drugs
excretedwilldrugs
usually willbebefound
normallyin
found as metabolites,
their parent frequently
form, either chiral,orofsingle
as racemate the parent compound
enantiomers. On[138].
the other hand, excreted drugs will be
The possible
normally found asadverse effectsfrequently
metabolites, of the enantiomers
chiral, of theon parent
aquaticcompound
and human[138]. life lead to the studies of
occurrence
The possible adverse effects of the enantiomers on aquatic and humandrugs
of chiral drugs in environmental matrices [20,136,137,139,140]. Illicit can also
life lead be studies
to the found in
environmental
of occurrence of samples, and environmental
chiral drugs in environmental data are important
matrices resources for a forensic
[20,136,137,139,140]. approach.
Illicit drugs This
can also
includes the usage of environmental data in order to: (1) verify patterns of
be found in environmental samples, and environmental data are important resources for a forensic illicit and prescribed drugs
usage in local communities (2) application of drugs as chemical markers of faecal water contamination
approach. This includes the usage of environmental data in order to: (1) verify patterns of illicit and
with (human) sewage and (3) verify the source of drugs (legal or illicit) [20,136,137,139,140].
prescribed drugs usage in local communities (2) application of drugs as chemical markers of faecal
The estimation of the consumption of substances of abuse and illicit drugs can be measured by the
water contamination with (human) sewage and (3) verify the source of drugs (legal or illicit) [20,136,
concentrations of these compounds in wastewater. Drugs are consumed and metabolised in human body
137,139,140].
and excreted as parent compounds or as metabolites, and finally reach WWTPs through the sewage [139].
SinceThe
the estimation of the consumption
metabolic patterns of most availableof substances
drugs areofunderstood,
abuse and illicit drugs canthat
it is assumed be the
measured
amountbyof
the concentrations of these compounds in wastewater. Drugs are consumed
drug or its metabolite quantified in raw sewage will correspond with the consumed dose—sewage and metabolised in human
body and excreted
epidemiology [139]. as
Theparent compounds
application of theorchiral
as metabolites,
discriminationand finally
has also reach
been WWTPs
used for through the
distinction
sewage [139]. Since the metabolic patterns of most available drugs are understood,
between legal and illicit use of drugs, verification of the method of synthesis of illicit drugs, identificationit is assumed that
the amount
that of drugresults
drug residue or its metabolite
either fromquantified
consumptionin raw of sewage will correspond
illicit drugs or metabolism with of theother
consumed
drugs,
dose—sewage
verification epidemiology
of route [139]. Theverification
of administration, applicationofofpotency
the chiralof discrimination has also been
abused drugs, monitoring of used for
changing
distinction between legal and illicit use of drugs, verification of the method of synthesis of illicit drugs,
identification that drug residue results either from consumption of illicit drugs or metabolism of other
Molecules 2018, 23, 262 24 of 47
drugs, verification of route of administration, verification of potency of abused drugs, monitoring of

changing patterns of drugs abused, and differentiation between consumption and disposal of unused
drugs [17,141].
Concerning biodegradation, ecotoxicity and environmental fate, the recognition of
enantioselectivity is essential to provide a more realistic risk assessment of chiral compounds.
The fate of chiral drugs in the environment can be studied by monitoring their EF during biological
processes [20,38,136]. Degradation of these compounds relies on both abiotic and biotic processes [142].
Biodegradation in WWTP is expected to be stereoselective, which changes the EF of a given molecule
in the sample, consequently bringing different removals/degradation rates [142]. Over the past five
years, the amount of research published in chiral environmental analysis has been increasing on a
high rate. Knowledge on how chiral micropollutant, such as pharmaceuticals and illicit drugs, behave
in the environment, especially in water samples, either wastewater or superficial water, has been
providing valuable information, both for risk assessment and WWTPs efficiency [139,142]. The EF of
certain pharmaceuticals, such as PHO, alprenolol, VNF and climbazole [136,143,144] in surface waters,
can reveal the efficiency of different WWTPs [136,139,140,143–146]. Additional these compounds
have been pointed as indicators to differentiate between treated and untreated water. Analysis of
wastewater samples are mostly done by comparing the EF of the influent and effluent of the target
analytes, which gives an overview of the efficiency and of the WWTPs [9,147,148].
According to Kasprzyk-Hordern et al., since WWTPs are fed by fresh sewage, a long-term
monitoring programme of drugs might reveal their usage patterns in local communities and their
changes over longer periods of time [139]. This is the main route that chiral drugs enter the
environment, and these can be found either in a modified form (metabolites) and/or with alterations in
their enantiomeric EF due to human metabolism [138]. In the first attempt to apply chirality to sewage
epidemiology, Kasprzyk-Hordern et al., collected wastewater samples over a period of 8 months,
from seven WWTPs in London, during five sampling campaigns and, quantified the levels of AM, MA,
MDMA, MDA, ephedrine and pseudoephedrine enantiomers [149]. The samples were enriched with
R-AM, S-MA, S-MDA, 1R,2S-(−)-ephedrine and 1S,2S-(+)-pseudoephedrine. However, the authors
could not reach any conclusion according to the use of illicit drugs, since AM and MA enantiomers can
also result from the metabolism of chiral pharmaceuticals. On the other hand, when comes to MDMA
and MDA, the enantiomeric profiling proved to be invaluable in making distinction between MDA
abuse and its formation due to metabolism of MDMA, suggesting that this profiling could also help
with making a distinction between actual consumption and direct disposal [17].
Vasquez-Roig et al., in a two week study of three WWTPs located in the city of Valencia (Spain)
and surroundings, described the enantiomeric profile of some chiral drugs [148]. Although for
some of the target analytes it was not possible to study their enantiomeric fate, since these were
present in very low concentrations, which was the case of MDMA and AM, they were able to
observe enantiomeric enrichment ofATE, where the S-enantiomer was in higher abundance in raw
wastewater, meanwhile during the wastewater treatment, enrichment of both R- or S-enantiomer were
observed [148]. This difference in enantiomeric enrichment seemed to be related with the technology
used by the treatment plant. Although all of three used activated sludge, one of the plants had
also biological nitrogen removal, which the authors believe that different bacteria were involved
in this process (in aerobic conditions), that could favour the degradation of R-ATE, leading to an
enrichment of S-ATE [148]. They also found enrichment at similar levels of 1R,2S-(−)-ephedrine and
1S,2S-(+)-pseudoephedrine in raw wastewater. In terms of elimination rates these ranged from 29 to
100% and showed to be compound and enantiomer dependent. AM and MA were not detected in
effluents, however stereoselective degradation was observed for MDMA, where the S-enantiomer was
more readily degraded than the R-MDMA. Atenolol was found to be poorly removed, thus S-atenolol
removal efficiency was higher than R-atenolol [148]. VNF concentrations increased in two of the
WWTPs after sewage treatment, which in according to the authors, was due to biotic effects, such like,
elimination of glucuronide metabolites, back-reversion of the demethylated metabolite, or desorption
Molecules 2018, 23, 262 25 of 47
from particulate matter [148]. The application of wastewater enantiomeric profiling revealed usage
patterns of chiral drugs in the region, where the consumption of AM showed an irregular pattern
throughout the two-week sampling campaign, while MA showed a slight increase in daily loads,
throughout the weekend in one of the WWTPs. MDMA showed a clear weekly pattern of increased
daily loads, during weekends [148].
slight increase in daily loads, throughout the weekend in one of the WWTPs. MDMA showed a clear
weekly Hashim,
pattern N. H. & Khan,
of increased S. J.loads,
daily studied the EFof
during ibuprofen,
weekends [148]. naproxen and ketoprofen in wastewater
samples, taken from a WWTPs in Sydney
Hashim, N. H. & Khan, S. J. studied the EFof ibuprofen, (Australia) with naproxen
tertiary treatment over seven
and ketoprofen separate
in wastewater
sampling
samples, events,
taken from during
a WWTPs June and August
in Sydney 2010with
(Australia) [147]. For ibuprofen,
tertiary treatment overEF seven
ranged from 0.49
separate and
sampling
0.62; 0.66 and 0.86 for naproxen and 0.54 and 0.66 for ketoprofen [147]. Also
events, during June and August 2010 [147]. For ibuprofen, EF ranged from 0.49 and 0.62; 0.66 and 0.86 for Barreiro et al in 2010,
found forand
naproxen the0.54
firstand
time0.66
the for
occurrence
ketoprofen of (+)-omeprazole
[147]. Also Barreiroandet(−al)-omeprazole,
in 2010, found while simultaneously
for the first time the
developing
occurrence of a(+)-omeprazole
column switching, liquid chromatography
and (-)-omeprazole, method fordeveloping
while simultaneously the chiral aseparation of these
column switching,
drugs,chromatography
liquid in wastewater and estuarine
method for the samples [150]. Ribeiro
chiral separation et al studied,
of these drugs, inthe EF of FLX,and
wastewater NFLX, VNF,
estuarine
samples [150]. Ribeiro
SBT, alprenolol, MET, et PHOal studied,
and bisoprololthe EF (BSPof FLX,
= fromNFLX, VNF,effluent
the final SBT, alprenolol, MET, PHO
of the secondary and
clarifier
bisoprolol (BSP= from
of three WWTPs the final
located effluent
in the North of of
thePortugal,
secondaryFigure
clarifier of three
5 [39]. WWTPs located
Regarding in the North
antidepressants, onlyof
Portugal,
R-FLX was Figure
detected5 [39]. Regarding
in two antidepressants,
of the WWTPs, indicating only R-FLX
a faster was detected
degradation in S-enantiomer
of the two of the WWTPs, during
indicating
the biologicala faster degradation
degradation [39]. ofVNF the enantiomers
S-enantiomerwere during
foundthebetween
biological degradation
40.4 and 129 ng/L [39].inVNF
the
enantiomers were found between 40.4 and 129 ng/L in the three WWTPs
three WWTPs studied, with similar EF, which varied between 0.54 and 0.55, proving that VNF studied, with similar EF, found
which
varied
was not between
racemic 0.54 andConcerning
[39]. 0.55, proving thethat VNF found
β-blockers was not racemic
enantiomers, BSP and [39].
PHO Concerning
were found theinβ-blockers
all three
enantiomers,BSP and PHO were found in all three WWTPs, while MET
WWTPs, while MET was only found in two, however all of them under the medium quantification was only found in two, however
all of them under the medium quantification level (MQL) [39].
level (MQL) [39].
Figure
Figure5.5.Chromatogram
Chromatogramand
andmass
massspectra
spectraofofWWTP
WWTP effluent
effluent sample
sample showing
showing the enantiomers of FLX,
the enantiomers FLX,
VNF,
VNF,BSP,
BSP,MET
METand
andPHO.
PHO.Reproduction with
Reproduction withpermission of Elsevier
permission (Figure
of Elsevier 2 from
(Figure Ribeiro
2 from et al.et[39]).
Ribeiro al. [39]).
Molecules 2018, 23, 262 26 of 47
Table 3. Analytical methods of separation of several chiral illicit drugs and pharmaceuticals in different environmental matrices.
Matrix
Drugs Method Stationary Phase LOD/MDL LOQ/MQL Concentration Range/EF Reference
Application
0.5–1000 ng/L; EF:
AM: 5.1 ng/L
0.52–0.84 (mean 0.64)
AM
MA Chiral-CBH (100 × 2 mm, MA: 0.6 ng/L 0.05–1000 ng/L; EF ≥ 0.5
MDMA Wastewater UPLC/ESI-MS/MS 5 µm); Chiral-CBH guard n.r. [17]
MDMA: 0.7 ng/L 0.1–1000 ng/L; EF = 0.68
MDA column (10 × 2 mm)
Ephedrine MDA: 4.2 ng/L 0.1–1000 ng/L; EF > 0.5
0.5–1000 ng/L; EF:
Ephedrine: 5.6 ng/L
0.81–0.96 (mean 0.91)
0.025–250 µg/L;
AM (R/S): 0.38/0.39 ng/L (IW); 0.28/0.41 AM (R/S): 1.28/1.32 ng/L (IW); 0.94/1.36
EF: 0.5 (IW); 0.6 (EW); 0.3
ng/L (EW); 4.92/5.15 ng/L (DS) ng/L (EW); 16.56/17.28 ng/L (DS)
(DS)
0.025–250 µg/L
MA (R/S): 0.12/0.13 ng/L (IW); 0.09/0.08 MA (R/S): 0.38/0.41 ng/L (IW); 0.28/0.27
EF: 0.6 (IW); 0.5 (EW); 0.5
ng/L (EW); 0.73/0.77 ng/L (DS) ng/L (EW); 3.24/2.45 ng/L (DS)
(DS)
0.025–250 µg/L
MDMA: 0.05 ng/L (IW); 0.04 ng/L (EW); MDMA (R/S): 0.17/0.18 ng/L (IW);
AM, MA, MDMA, Chiral-CBH (100 × 2 mm, EF: 0.7 (IW); 0.9 (EW); 0.4
1.43/1.79 ng/L (R/S) (DS) 0.13/0.14 ng/L (EW); 4.75/5.96 ng/L (DS)
MDA, Ephedrine, 5 µm); Chiral-CBH guard (DS)
Pseudoephedrine, column (10 × 2 mm) 0.025–250 µg/L
MDA (R/S): 0.33/0.36 ng/L (IW); 0.21/0.25 MDA (R/S): 1.13/1.19 ng/L (IW); 0.72/0.83
Norephedrine, Influent EF: 0.6 (IW); 0.5 (EW); 0.3
LC/ESI-MS/MS ng/L (EW); 2.21/4.14 ng/L (DS) ng/L (EW); 7.46/13.74 ng/L (DS)
Atenolol, wastewater (DS)
Alprenolol, PHO, (IW); effluent
Ephedrine (1R,2S/1S,2R): 0.23/0.16 ng/L Ephedrine (1R,2S/1S,2R): 0.01/0.02 ng/L 0.025–250 µg/L
MET, T, SBT, wastewater
(IW); 0.14/0.07 ng/L (EW) (IW); 0.48/0.25 ng/L (EW) EF: 0 (IW, EW)
Sotalol, FLX, (EW); digested
Mirtazapine, sludge (DS) Pseudoephedrine (1R,2R/1S,2S): 0.26/0.1 Pseudoephedrine (1R,2R/1S,2S): 0.01/0.03 [62]
0.025–250 µg/L
VNF, OD-VNF, ng/L (IW); 0.15/0.11 ng/L (EW); ng/L (IW); 0.52/0.36 ng/L (EW);
EF: 1 (IW); 0.2 (EW)
Citalopram, 15.54/44.53 ng/L (DS) 51.62/148.5 ng/L (DS)
D-citalopram 0.025–250 µg/L
Norephedrine (E1/E2): 0.33/0.15 ng/L (IW); Norephedrine (E1/E2): 0.01/0.02 ng/L (IW);
EF: 0 (IW); 0.3 (EW); 0.1
0.19/0.21 ng/L (EW); 9.54/13.05 ng/L (DS) 0.63/0.71 ng/L (EW); 31.52/43.38 ng/L (DS)
(DS)
Atenolol (R/S): 95.81/58 ng/L (IW);
Atenolol (R/S): 28.74/17.40 ng/L (IW); 0.025–250 µg/L
102.68/109.08 ng/L (EW); 25.15/23.70 ng/L
30.80/32.73 ng/L (EW); 7.55/7.12 ng/L (DS) EF: 0.5 (IW, EW); 0.4 (DS)
(DS)
Chirobiotic V Alprenolol (R/S): 0.14/0.07 ng/L (IW); Alprenolol (R/S): 0.47/0.24 ng/L (IW); 0.025–250 µg/L
(250 × 2.1 mm, 5 µm) 0.06/0.03 ng/L (EW); 0.14 ng/L (DS) 0.21/0.09 ng/L (EW); 0.48/0.46 ng/L (DS) EF: 0.5 (IW, EW); 0.7 (DS)
PHO (R/S): 0.08/0.09 ng/L (IW); 0.06/0.05 PHO (R/S): 0.26/0.3 ng/L (IW); 0.20/0.17 0.025–250 µg/L
ng/L (EW); 0.07/0.06 ng/L (DS) ng/L (EW); 0.23/0.20 ng/L (DS) EF: 0.4 (IW, EW); 0.5 (DS)
Molecules 2018, 23, 262 27 of 47
Table 3. Cont.
Matrix
Application
MET (R/S): 0.08/0.06 ng/L (IW); 0.05/0.04 MET (R/S): 0.27/0.18 ng/L (IW); 0.15/0.12 0.025–250 µg/L
ng/L (EW); 0.05/0.07 ng/L (DS) ng/L (EW); 0.15/0.22 ng/L (DS) EF: 0.3 (IW, DS)
T (E1/E2): 0.09/0.43 ng/L (IW); 0.05/0.24 T (E1/E2): 0.29/1.43 ng/L (IW); 0.16/0.79 0.025–250 µg/L
ng/L (EW); 0.05/0.34 ng/L (DS) ng/L (EW); 0.18/1.13 ng/L (DS) EF: 0.7 (IW, EW, DS)
SBT (R/S): 2.22/2.20 ng/L (IW); 1.31/0.98 SBT (R/S): 7.41/7.32 ng/L (IW); 4.36/3.26 0.025–250 µg/L
ng/L (EW); 80.03/65.03 ng/L (DS) ng/L (EW); 265.10/225.10 ng/L (DS) EF: 0.5 (IW, EW)
Sotalol (E1/E2): 0.66/0.61 ng/L (IW); Sotalol (E1/E2): 2.20/2.05 ng/L (IW); 0.025–250 µg/L
0.53/0.46 ng/L (EW); 1.64/0.76 ng/L (DS) 1.76/1.53 ng/L (EW); 5.47/5.87 ng/L (DS) EF: 0.5 (IW, EW, DS)
FLX (R/S): 0.08/0.07 ng/L (IW); 0.05/0.04 FLX (R/S): 0.26/0.22 ng/L (IW); 0.17/0.14 0.025–250 µg/L
ng/L (EW); 0.09/0.07 ng/L (DS) ng/L (EW); 0.30/0.23 ng/L (DS) EF: 0.7 (IW, EW, DS)
0.025–250 µg/L
Mirtazapine (R/S): 0.40/1.17 ng/L (IW); Mirtazapine (R/S): 1.32/3.89 ng/L (IW);
EF: 0.3 (IW); 0.2 (EW); 0.5
0.40/0.86 ng/L (EW); 0.31/0.73 ng/L (DS) 1.32/2.86 ng/L (EW); 1.02/2.44 ng/L (DS)
(DS)
VNF (R/S): 0.03/0.04 ng/L (IW); 0.02/0.03 VNF (R/S): 0.11/0.12 ng/L (IW); 0.07/0.11 0.025–250 µg/L
ng/L (EW); 0.03 ng/L (DS) ng/L (EW); 0.08 ng/L (DS) EF: 0.5 (IW, EW, DS)
OD-VNF (R/S): 0.32/0.16 ng/L (IW); OD-VNF (R/S): 1.05/3.85 ng/L (IW); 0.025–250 µg/L
0.38/0.08 ng/L (EW); 0.75/1.02 ng/L (DS) 1.28/2.30 ng/L (EW); 2.51/3.41 ng/L (DS) EF: 0.5 (IW, EW, DS)
Citalopram (R/S): 0.31/0.24 ng/L (IW); Citalopram (R/S): 13.69/13.07 ng/L (IW); 0.025–250 µg/L
0.27/0.21 ng/L (EW); 0.21/0.09 ng/L (DS) 11.78/11.15 ng/L (EW); 9.09/4.69 ng/L (DS) EF: 0.6 (IW, DS); 0.7 (EW)
D-Citalopram (R/S): 0.50/0.36 ng/L (IW); D-Citalopram (R/S): 1.68/1.21 ng/L (IW); 0.025–250 µg/L
0.40/0.29 ng/L (EW); 0.42/0.36 ng/L (DS) 1.34/0.96 ng/L (EW); 1.99/1.22 ng/L (DS) EF: 1 (IW); 0.6 (DS)
AM (R/S): 0.85/0. 9 ng/L (IW); 0.9/0.85 AM (R/S): 4.35/4.4 ng/L (IW); 4.4/4.35 0.25–1900 ng/L
ng/L (EW) ng/L (EW) EF n.r.
MA: 2.8/2.95 ng/L (R/S) (IW); 3.35 ng/L 0.25–1900 ng/L
MA: 0.85/0. 9 ng/L (R/S) (IW); 1 ng/L (EW)
(EW) EF n.r.
0.25–1900 ng/L
MDMA: 0.9 ng/L (IW); 0.95/1 ng/L (E1/E2) MDMA: 2.4 ng/L (IW); 2.55–2.65 ng/L
EF: 0.53–0.72 (mean 0.63)
(EW) (E1/E2) (EW)
AM, MA, WWTP (IW); 0.71 (EW) [149]
MDMA, influent (IW); Chiral-CBH (100 × 2 mm,
UPLC/ESI-MS/MS 5 µm); Chiral-CBH guard MDA: 1.95/2 ng/L (E1/E2) (IW); 2 ng/L 0.25–1900 ng/L
MDA, MDEA, WWTP MDA: 9.7 ng/L (IW); 10.1 ng/L (EW)
column (10 × 2 mm) (EW) EF n.r.
Norephedrine, effluent (EW)
VNF MDEA: 2.25/2.2 ng/L (E1/E2) (IW); 2.4 0.25–1900 ng/L
MDEA: 0.55 ng/L (IW); 0.6 ng/L (EW)
ng/L (EW) EF n.r.
Norephedrine (E1/E2): 3.5/3.3 ng/L (IW); Norephedrine (E1/E2): 11.75/10.9 ng/L 0.25–1900 ng/L
1.4/1.35 ng/L (EW) (IW); 4.6/4.5 ng/L (EW) EF n.r.
0.25–1900 ng/L
VNF: 4.95/5.05 ng/L (E1/E2) (IW); 5.1 ng/L EF: 0.45–0.50 (mean 0.48)
VNF: 1.6 ng/L (IW); 1.65 ng/L (EW)
(EW) (IW); 0.37–0.48 (mean 0.43)
(EW)
Molecules 2018, 23, 262 28 of 47
Table 3. Cont.
Matrix
Application
AM: 1–500 ng/L
EF: 0.52–0.84 (mean 0.64) (IW); 0.57–1 (mean 0.78) (EW); 0.86
(RW before WWTP); 0.81 (RW after WWTP)
MA: 1–500 ng/L
EF: 0.22–0.53 (mean 0.34) (IW); 0.7–1 (mean 0.86) (EW)
MDMA: 1–500 ng/L
EF: 0.5–0.8 (mean 0.66) (IW); 0.64–0.91 (mean 0.75) (EW);
0.56–0.81 (mean 0.68) (RW before WWTP); 0.61–0.80 (mean
0.69) (RW after WWTP)
AM, MA, WWTP MDA: 1–500 ng/L

MDMA, influent (IW); Chiral-CBH (100 × 2 mm, EF: 0.26–0.47 (mean 0.34) (IW); 0.38–0.58 (mean 0.45) (EW);
MDA, WWTP effluent LC/ESI-MS/MS 5 µm); Chiral-CBH guard n.r. n.r. 0.58 (RW before WWTP); 0.56–0.57 (RW after WWTP) [9]
Ephedrine, (EW); river column (10 × 2 mm) Ephedrine: 1–500 ng/L
Atenolol, VNF water (RW) EF: 0.81–1 (mean 0.99) (IW); 0.22–1 (mean 0.92) (EW); 0.79–1
(mean 0.97) (RW before WWTP); 0.80–1 (mean 0.99) (RW after
WWTP)
DF: 0.02–0.66 (mean 0.26) (IW); 0.04–0.82 (mean 0.36) (EW);
0–1 (mean 0.6) (RW before WWTP); 0–1 (mean 0.46) (RW after
WWTP)
VNF: 1–500 ng/L
EF: 0.35–0.65 (mean 0.48) (IW); 0.46–0.69 (mean 0.52) (EW);
0.40–0.65 (mean 0.52) (RW before WWTP); 0.47–0.62 (mean
1–500 ng/L
Atenolol: 1.7 ng/L (IW, EF: 0.30–0.47 (mean 0.40) (IW); 0.40–0.61 (mean 0.46) (EW);
EW); 0.3 ng/L (RW) 0.38–0.56 (mean 0.46) (RW before WWTP); 0.39–0.50 (mean
AM (R/S): 12.4/11.5
AM: 4.6/4.4 ng/L (R/S) 0.5–500 ng/L
ng/L (SE); 5.0/4.8 ng/L
(SE); 1.8 ng/L (RW) EF n.r.
(RW)
MA (R/S): 11.9/14.2 MA (R/S): 47.6/47.3
0.25–500 ng/L
ng/L (SE); 4.6/5.5 ng/L ng/L (SE); 18.5/18.3
EF n.r.
Chirobiotic V (250 × 4.6 (RW) ng/L (RW)
AM, MA, MDMA, Sewage
MDA, Atenolol, effluent (SE); mm, 5 µm); Chirobiotic V MDMA (E1/E2): MDMA (E1/E2):
LC/QTOF-MS 5–500 ng/L [151]
PHO, MET, FLX, River water guard column (20 × 40 22.8/21.8 ng/L (SE); 85.7/81.9 ng/L (SE);
EF n.r.
VNF (RW) mm, 5 µm) 9.6/10.4 ng/L (RW) 35.8/39 ng/L (RW)
Atenolol (R/S): 5/5.3 Atenolol (R/S): 11.4/11
0.25–100/5–500 ng/L (R); 0.5–100/5–500 ng/L (S)
ng/L (SE); 2.1/2.2 ng/L ng/L (SE); 4.8/4.7 ng/L
EF: 0.55 (SE); 0.47 (RW)
(RW) (RW)
PHO (R/S): 3.4/2.6
PHO (R/S): 1.4/1 ng/L 0.25–100/5–500 ng/L
ng/L (SE); 1.4/1.2 ng/L
(SE); 0.6/0.4 ng/L (RW) EF: 0.43 (SE); 0.45 (RW)
(RW)
Molecules 2018, 23, 262 29 of 47
Table 3. Cont.
Matrix
Application
MET: 0.6/0.7 ng/L (E1/E2) (SE); 0.2 ng/L MET: 1.3 ng/L (SE); 0.3/0.4 ng/L 0.25–100/5–500 ng/L
(RW) (E1/E2) (RW) EF: 0.54 (SE)
FLX: 2.4/2.6 ng/L (R/S) (SE); 0.8 ng/L FLX (R/S): 7.6/6.5 ng/L (SE); 2.5/2 0.25–100/5–500 ng/L
(RW) ng/L (RW) EF n.r.
VNF (R/S): 4.8/3.9 ng/L (SE); 2.5/2.2 VNF (R/S): 15.1/14.4 ng/L (SE); 0.5–100/5–500 ng/L
ng/L (RW) 7.9/8.1 ng/L (RW) EF: 0.43 (SE); 0.58 (RW)
0.5–500 ng/L
AM (R/S): 4.8/5 ng/L (RW) AM (R/S): 9.7/10 ng/L (RW)
EF n.r.
Chiral-CBH (100 × 2 mm,
2.5–500 ng/L
5 µm); Chiral-CBH guard MA (R/S): 4.1/3.6 ng/L (RW) AM (R/S): 20.6/18.1 ng/L (RW)
EF n.r.
column (10 × 2 mm, 5 µm)
MDMA (R/S): 26.8/25.6 ng/L 12.5–500 ng/L
MDMA (R/S): 10.7/10.2 ng/L (RW)
(RW) EF n.r.
1.75–500 ng/L
MDA (R/S): 2.4/2.3 ng/L (RW) MDA (R/S): 9.6/9.1 ng/L (RW)
EF n.r.
Atenolol (R/S): 22.9/20.7 ng/L 0.5–500 ng/L
Atenolol (R/S): 2.3/2.1 ng/L (RW)
(RW) EF n.r.
5–500 ng/L
VNF (E1/E2): 10.3/9.6 ng/L (RW) VNF (E1/E2): 51.7/47.9 ng/L (RW)
EF n.r.
Atenolol: 1.8 ng/L (IW); 1.4 ng/L (EW) 6 ng/L (IW); 5 ng/L (EW)
PHO: 0.5 ng/L (IW, EW) 2 ng/L (IW, EW)
MET: 2.3 ng/L (IW); 0.6 ng/L (EW) 8 ng/L (IW); 2 ng/L (EW)
Atenolol, PHO, Influent 1–500 ng/mL
MET, SBT, wastewater Chirobiotic V (250 × 4.6 mm, SBT: 0.7 ng/L (IW); 0.6 ng/L (EW) 2 ng/L (IW, EW) EF n.r.
Sotalol, Nadolol, (IW); Effluent LC/ESI-MS/MS 5 µm) with a nitrile guard Sotalol: 7.5 ng/L (IW); 7.2 ng/L (EW) 25 ng/L (IW); 24 ng/L (EW) [152]
Pindolol, FLX, wastewater cartridge (10 × 3 mm)
Nadolol: 1.8 ng/L (IW); 1.7 ng/L (EW) 12 ng/L (IW); 3 ng/L (EW)
Citalopram (EW)
Pindolol: 0.4 ng/L (IW); 0.2 ng/L (EW) 1 ng/L (IW, EW)
FLX: 2.2 ng/L (IW); 0.6 ng/L (EW) 7 ng/L (IW); 2 ng/L (EW)
Citalopram: 2.4 ng/L (IW); 0.5 ng/L (EW) 8 ng/L (IW); 2 ng/L (EW)
25–1000 ng/mL
Atenolol: 110 ng/L IW); 12 ng/L (EW) n.r.
EF ≈ 0.5 (IW; EW)
WWTP Chirobiotic V (250 × 4.6 mm,
Atenolol, PHO, influent (IW); HPLC/ESI-MS/MS 5 µm) with a nitrile guard 25–1000 ng/mL [153]
PHO: 17 ng/L (IW); 4.4 ng/L (EW) n.r.
MET WWTP cartridge (10 × 3 mm) and an EF ≈ 0.5 (IW; EW)
effluent (EW) in-line filter 25–1000 ng/mL
MET: 42 ng/L (IW); 17 ng/L (EW) n.r.
EF: 0.5 (IW); 6=0.5 (EW)
Molecules 2018, 23, 262 30 of 47
Table 3. Cont.
Matrix
Application
Atenolol EF: 0.40–0.52 (mean
0.46)
MET EF: 0.39–0.52 (mean
Atenolol, MET, Chirobiotic V (250 × 4.6 mm, n.r. n.r. 0.46)
Sotalol, Effluent LC/ESI-MS/MS 5 µm) Sotalol EF: 0.34–0.41 (mean
Citalopram, wastewater 0.36) [142]
Temazepam
Citalopram EF: 0.44–0.62
(mean 0.58)
Chiralpak AD-RH (150 × 4.6 mm, Temazepam EF: 0.39–0.49
n.r. n.r.
5 µm) (mean 0.47)
Atenolol: 22 µg/L 70 µg/L
PHO: 3 µg/L 10 µg/L 12.5–100 µg/mL
Atenolol, PHO, Lux Cellulose-1 (250 × 4.6 mm,
River water LC-UV EF n.r. [154]
MET, Bisoprolol 5 µm) MET: 20 µg/L 40 µg/L
Bisoprolol: 3 µg/L 10 µg/L
Alprenolol (R/S): 8.08/4.52 ng/L Alprenolol (R/S): 18.5/13.7 ng/L
PHO (R/S): 1.97/0.65 ng/L PHO (R/S): 5.96/1.98 ng/L
20–400 ng/L
MET (R/S): 11.5/3.37 ng/L MET (R/S): 14.8/10.2 ng/L EF n.r.
Alprenolol, PHO,
MET, SBT, Chirobiotic V (150 × 2.1 mm, SBT (R/S): 5.07/6.29 ng/L SBT (R/S): 15.4/19.1 ng/L
WWTP effluent LC/ESI-MS/MS [39]
Bisoprolol, FLX, 5 µm) Bisoprolol (E1/E2): 2.78/4.54 ng/L Bisoprolol (E1/E2): 8.44/13.8 ng/L
NFLX, VNF
FLX (R/S): 8.41/3.74 ng/L FLX (R/S):19.5/11.3 ng/L
30–400 ng/L
NFLX (R/S): 0.97/5.27 ng/L NFLX (R/S): 2.95/16 ng/L
EF n.r.
20–400 ng/L
VNF (R/S): 9.82/1.71 ng/L VNF (R/S): 19.7/5.18 ng/L
EF: 0.54–0.55 (mean 0.55)
5–1000 µg/L
Alprenolol (R/S): 0.2/0.1 ng/L Alprenolol (R/S): 0.5/0.4 ng/L
EF n.r.
5–1000 µg/L
PHO (R/S): 0.6/0.5 ng/L PHO (R/S): 2.1/1.7 ng/L
Alprenolol, PHO, Chirobiotic V (250 × 4.6 mm, EF: 0.44–0.56 (mean 0.49)
MET, FLX, VNF, 5 µm); Chirobiotic V guard 5–1000 µg/L
Ibuprofen, Surface water LC/ESI-MS/MS column (20 × 4 mm, 5 µm) MET: 0.2 ng/L MET (R/S): 0.6/0.5 ng/L
EF: 0.48–0.64 (mean 0.55) [37]
Naproxen,
Flurbiprofen FLX: 0.1 ng/L 0.5 ng/L 5–1000 µg/L; EF: 0.5–0.63
5–1000 µg/L; EF: 0.46–0.51
VNF: 0.1 ng/L 0.5 ng/L
(mean 0.49)
Ibuprofen (R/S): 11/9.6 ng/L Ibuprofen (R/S): 37/32 ng/L
5–1000 µg/L
Chiralpak AD-RH (150 × 4.6 mm, Naproxen: 0.4 ng/L Naproxen (R/S): 1.4/1.2 ng/L EF n.r.
5 µm)
Flurbiprofen (R/S): 3.3/2.4 ng/L Flurbiprofen (R/S): 11/7.9 ng/L
Molecules 2018, 23, 262 31 of 47
Table 3. Cont.
Matrix Concentration
Drugs Method Stationary Phase LOD/MDL LOQ/MQL Reference
Application Range/EF
Influent
wastewater (IW);
Effluent MDN-5S (30 m × 0.25 mm, 0.25 µm film EF: 0.5 (IW); ≤0.42
PHO GC/ESI-MS/MS n.r. n.r. [155]
wastewater (EW); thickness) (EW); 0.42–0.53 (SW)
Surface water
(SW)
PHO River water LC-UV Lux-Cellulose 1 (250 × 4.6 mm, 5 µm) 0.4 µg/L 1.3 µg/L 0.125–50 µg/mL; EF n.r. [156]
Influent
wastewater (IW); 3.7/3.5 ng/L (IW); 1.9/1.5 ng/L 12.4/11.5 ng/L (IW); 6.5/5.1 ng/L EF: 0.48–0.52 (IW);
MET LC/ESI-MS/MS Chirobiotic V (250 × 4.6 mm, 5 µm) [40]
Effluent (EW) (R/S) (EW) (R/S) 0.5–0.7 (EW)
wastewater (EW)
Chiral-CBH (100 × 2 mm, 5 µm);
Treated
MET LC/MS/MS Chiral-CBH guard column and in-line 0.96/2.9 pM (E1/E2) 5.8/11.6 pM (E1/E2) EF: 0.51–0.55 [157]
wastewater
high-pressure filter (4 mm, 0.5 µm)
Effluent
MDN-5S (30 m × 0.25 mm, 0.25 µm film EF: 0.5 (EW); 0.31–0.44
MET wastewater (EW); GC/ESI-MS/MS n.r. n.r. [158]
thickness) (RW)
River water (RW)
Raw wastewater Chiral-AGP (100 × 2 mm, 5 µm); in-line FLX: 3 pM (RaW); 2/1 pM (R/S) 0–500 pM
FLX: 12.4 pM (RaW); 3 pM (TW)
FLX, NFLX (RaW); Treated LC/ESI-MS/MS high-pressure filter with a replaceable (TW) EF: 0.71 (RaW, TW) [159,160]
wastewater (TW) cap frit (4 mm, 5 µm); Chiral-AGP guard 0–500 pM
NFLX: 12.1/14.3 pM (E1/E2)
column (10 × 2 mm) NFLX: 2.4 pM (RaW); 2 pM (TW) EF: 0.69 (RaW); 0.68
(RaW); 4 pM (TW)
(TW)
FLX: 0.8–2 ng/mL 4 ng/mL 4–60 ng/mL; EF n.r.
FLX, NFLX WWTP effluent HPLC-FD Chirobiotic V (150 × 4.6 mm, 5 µm) [137]
NFLX: 0.8–2 ng/mL 2 ng/mL 2–30 ng/mL; EF n.r.
Chirobiotic V (250 × 2.1 mm, 5 µm);
VNF River water LC/ESI-MS/MS 6/4 ng/L (R/S) n.r. EF: 0.46–0.74 [144]
Chirobiotic guard column (10 × 2 mm)
Ibuprofen (R/S): 1319/1111 ng/L Ibuprofen (R/S): 5403/4551 ng/L
250–400 µg/L
(IW); 498/383 ng/L (EW); (IW); 2039/1570 ng/L (EW);
EF: 1 (IW)
263/114 ng/L (SW) 1076/466 ng/L (SW)
Carboxyibuprofen (E1/E2): Carboxyibuprofen (E1/E2): 32.7–300 µg/L (IW);
Ibuprofen, 71/63.6 ng/L (IW); 71.4/58.6 232/208 ng/L (IW); 233/191 ng/L 250–400 µg/L (EW, SW)
Carboxyibuprofen, Influent
ng/L (EW); 21.5/22.3 ng/L (SW) (EW); 70.2/72.7 ng/L (SW) EF: 0.83 (IW)
2-Hydroxyibuprofen, wastewater (IW);
Effluent LC/ESI-MS/MS Chirobiotic T (250 × 2.1 mm, 5 µm) 2-Hydroxyibuprofen (E1/E2): 2-Hydroxyibuprofen (E1/E2): 16.3–400 µg/L (E1);
Naproxen, [161]
Ketoprofen, wastewater (EW); 31.7/20.4 ng/L (IW); 28/30.4 104/66.4 ng/L (IW); 91.3/99.3 16.3–300 µg/L (E2)
Indoprofen, Surface water (SW) ng/L (EW); 10.9/10.4 ng/L (SW) ng/L (EW); 35.4/33.9 ng/L (SW) EF: 0.76 (IW)
Chloramphenicol, Naproxen (R/S): 11/7.53 ng/L Naproxen (R/S): 38.1/26.1 ng/L
Ifosfamide, 8.66–50 µg/L
(IW); 14.4/13.4 ng/L (EW); (IW); 49.9/46.5 ng/L (EW);
Praziquantel EF: 1 (IW)
7.5/6.83 ng/L (SW) 25.9/23.7 ng/L (SW)
Ketoprofen (R/S): 2.08/2.61 ng/L Ketoprofen (R/S): 6.85/8.59 ng/L
1.65–400 µg/L
(IW); 2.28/2.56 ng/L (EW); (IW); 7.51/8.44 ng/L (EW);
EF n.r.
1.60/1.32 ng/L (SW) 5.29/4.37 ng/L (SW)
Molecules 2018, 23, 262 32 of 47
Table 3. Cont.
Indoprofen (E1/E2): 2.23/3.44 ng/L Indoprofen (E1/E2): 7.59/11.7
1.70–100 µg/L
(IW); 2.20/2.59 ng/L (EW); 1.54/1.46 ng/L (IW); 7.47/8.81 ng/L (EW);
EF n.r.
ng/L (SW) 5.24/4.95 ng/L (SW)
Chloramphenicol (1R,2R/1S,2S): Chloramphenicol (1R,2R/1S,2S): 17–400 µg/L (1R,2R);
29.1/5.66 ng/L (IW); 26.1/4.84 ng/L 98.9/18.8 ng/L (IW); 88.6/16.1 3.33–800 µg/L (1S,2S)
(EW); 13.5/2.59 ng/L (SW) ng/L (EW); 45.8/8.61 ng/L (SW) EF n.r.
Ifosfamide (E1/E2): 0.24/0.28 ng/L Ifosfamide (E1/E2): 0.82/0.96 ng/L
0.17–50 µg/L
(IW); 0.23/0.22 ng/L (EW); 0.12/0.13 (IW); 0.78/0.74 ng/L (EW);
EF n.r.
ng/L (SW) 0.41/0.44 ng/L (SW)
Praziquantel (E1/E2): 3.02/3.11 ng/L Praziquantel (E1/E2): 10.1/10.4
1.67–400 µg/L
(IW); 2.78/2.82 ng/L (EW); 1.34/1.39 ng/L (IW); 9.26/9.40 ng/L (EW);
EF n.r.
ng/L (SW) 4.47/4.63 ng/L (SW)
Influent Ibuprofen EF: 0.73–0.90

Astec Chiraldex (20 m × (IW); 0.60–0.76 (EW)
Ibuprofen, Naproxen wastewater (IW); GC/MS 0.1 µg/L n.r. [44]
0.25 mm, 0.12 µm film
Effluent Naproxen EF: 0.88–0.90
thickness)
wastewater (EW) (IW); 0.71–0.86 (EW)
Ibuprofen EF: 0.88–0.94
(IW); 0.38–0.40 (EW)
Influent
Ibuprofen, Naproxen, wastewater (IW); GC/EI-MS/MS HP5-MS (30 m × 0.25 mm, n.r. n.r. Naproxen EF: 0.99 (IW); [162]
Ketoprofen Effluent 0.25 µm film thickness) 0.86–0.94 (EW)
wastewater (EW) Ketoprofen EF: 0.56–0.60
(IW); 0.54–0.68 (EW)
Ibuprofen (R/S): 1410/1525 ng/L Ibuprofen (R/S): 4695/5080 ng/L 415–2000 µg/L
(IW); 1458/1452 ng/L (EW) (IW); 4854/4837 ng/L (EW) EF: 1 (IW)
2-Hydroxyibuprofen (E1/E2): 2-Hydroxyibuprofen (E1/E2): 163.5–2000 µg/L
409/415 ng/L (IW) 1360/1382 ng/L (IW) EF: 0.2 (IW)
Naproxen: 233/267 ng/L (R/S) (IW); Naproxen: 777/891 ng/L (R/S) 84.3–2000 µg/L
Ibuprofen,
539 ng/L (R) (EW) (IW); 1796 ng/L (R) (EW) EF: 1 (IW, EW)
2-Hydroxyibuprofen,
Naproxen, Indoprofen, 0.85–250 µg/L (E1);
Carprofen, Fenoprofen, Polysaccharide amylose 0.85–500 µg/L (E2)
(IW); 2.88/2.65 ng/L (EW) ng/L (IW); 9.60/8.84 ng/L (EW)
Flurbiprofen, Influent tris-(3,5-dimethylpheny EF n.r.
Chloramphenicol, wastewater (IW); lcarbamate) column
UHPSFC/ESI-MS/MS Carprofen (E1/E2): 378/287 ng/L Carprofen (E1/E2): 1259/956 ng/L 168–500 µg/L [163]
Aminorex, Tetramisole, Effluent (IW); 584/705 ng/L (EW) (IW); 1945/2347 ng/L (EW) EF n.r.
Omeprazole, Ifosfamide, wastewater (EW)
3-N-Dechloroethylifosfamide, Fenoprofen (E1/E2): 571/538 ng/L Fenoprofen (E1/E2): 1900/1793 171–4000 µg/L
Praziquantel, Imazalil, (IW); 499/489 ng/L (EW) ng/L (IW); 1660/1632 ng/L (EW) EF n.r.
Ofloxacin Flurbiprofen: 331 ng/L (IW); 252/378 Flurbiprofen (E1/E2): 838/1101 83.8–2000 µg/L
ng/L (E1/E2) (EW) ng/L (IW); 838/1259 ng/L (EW) EF n.r.
Chloramphenicol (1R,2R/1S,2S): Chloramphenicol (1R,2R/1S,2S): 16.9–500 µg/L (1R,2R);
45.6/43.5 ng/L (IW); 53.4/50.1 ng/L 152/145 ng/L (IW); 178/167 ng/L 16.7–500 µg/L (1S,2S)
(EW) (EW) EF n.r.
Molecules 2018, 23, 262 33 of 47
Table 3. Cont.
Matrix
Application
Aminorex (E1/E2): 1.82/2.57 ng/L Aminorex (E1/E2): 6.05/8.56 ng/L 0.83–500 µg/L
(IW); 2.16/3.02 ng/L (EW) (IW); 7.20/10 ng/L (EW) EF n.r.
Tetramisole (R/S): 2.54/2.94 ng/L Tetramisole (R/S): 8.46/9.79 ng/L 0.83–500 µg/L
(IW); 2.83/2.87 ng/L (EW) (IW); 9.43/9.54 ng/L (EW) EF n.r.
0.82–125 µg/L (E1); 0.82–250 µg/L
Omeprazole: 24.5 ng/L (IW, EW) 81.6 ng/L (IW, EW) (E2)
EF n.r.
Ifosfamide (E1/E2): 0.51/0.58 ng/L Ifosfamide (E1/E2): 1.70/1.93 ng/L 0.17–125 µg/L
(IW); 0.51/0.54 ng/L (EW) (IW); 1.69/1.78 ng/L (EW) EF n.r.
3-N-Dechloroethyl-ifosfamide 3-N-Dechloroethyl-ifosfamide 0.17–50 µg/L (E1); 0.83–125 µg/L
(E1/E2): 0.46/3.22 ng/L (IW); (E1/E2): 1.54/10.70 ng/L (IW); (E2)
1.35/8.62 ng/L (EW) 4.50/28.70 ng/L (EW) EF n.r.
Praziquantel (E1/E2): 2.66/2.47 ng/L Praziquantel (E1/E2): 8.86/8.23 0.83–50 µg/L
(IW); 2.64/2.77 ng/L (EW) ng/L (IW); 8.78/9.23 ng/L (EW) EF n.r.
1.69–500 µg/L (E1); 1.69–250 µg/L
(E2)
EF n.r.
Aminorex (E1/E2): 2.32/2.54 ng/L Aminorex (E1/E2): 7.74/8.44 ng/L 0.83–500 µg/L
(IW); 2.23/3.44 ng/L (EW) (IW); 7.59/11.70 ng/L (EW) EF: 0.4 (IW)
Tetramisole (R/S): 2.72/3.08 ng/L Tetramisole (R/S): 9.06/10.30 ng/L 0.83–250 µg/L (R); 0.83–500 µg/L (S)
(IW); 3.16/2.60 ng/L (EW) (IW); 10.50/8.65 ng/L (EW) EF: 0.6 (IW, EW)
1.63–500 µg/L
Omeprazole: 49 ng/L (IW, EW) 163 ng/L (IW, EW)
EF n.r.
Cellulose
tris-(3-chloro-4-methylpheny 3-N-Dechloroethyl-ifosfamide 3-N-Dechloroethyl-ifosfamide
0.83–125 µg/L
lcarbamate) column (E1/E2): 2.81/2.99 ng/L (IW); (E1/E2): 9.35/9.97 ng/L (IW);
EF: 0.4 (IW)
7.69/8.68 ng/L (EW) 25.60/28.90 ng/L (EW)
1.67–500 µg/L (E1); 1.67–250 µg/L
Praziquantel (E1/E2): 6.62/5.59 ng/L Praziquantel (E1/E2): 22/18.60
(E2)
EF n.r.
1.74–500 µg/L (E1); 1.74–250 µg/L
Imazalil (E1/E2): 5.12/5.16 ng/L Imazalil (E1/E2): 17/17.20 ng/L
(E2)
(IW); 7.09/6.45 ng/L (EW) (IW); 23.60/21.50 ng/L (EW)
EF: 0 (IW)
16.4–500 µg/L (E1); 16.4–250 µg/L
Ofloxacin (E1/E2): 98.20/63.10 ng/L Ofloxacin (E1/E2): 327/210 ng/L
(E2)
(IW); 65.20/79 ng/L (EW) (IW); 218/263 ng/L (EW)
EF: 0 (IW)
0.08–300 ng/L; EF: 0.49–0.62 (mean
Ibuprofen: 0.7 ng/L (S) n.r.
0.53)
Ibuprofen, Effluent HP5-MS (30 m × 0.25 mm,
Naproxen, GC/EI-MS/MS 0.08–300 ng/L; EF: 0.66–0.86 (mean [147]
wastewater 0.25 µm film thickness) Naproxen: 0.7 ng/L (S) n.r.
Ketoprofen 0.79)
Ketoprofen: 2.2 ng/L (S) n.r. 3–300 ng/L; EF: 0.54–0.66 (mean 0.60)
Molecules 2018, 23, 262 34 of 47
Table 3. Cont.
Matrix
Application
Influent HP5-MS (30 m × 0.25 Ibuprofen EF: 0.6–0.8 (IW); 0.5

wastewater (IW); mm, 0.25 µm film (EW)
Ibuprofen, GC/EI-MS/MS n.r. n.r.
Effluent thickness) [164]
Naproxen Naproxen EF: 1 (IW); 0.7–0.9
wastewater (EW) (EW)
0.4–4000 µg/L
Ibuprofen: 0.7 ng/L (IW); 0.5 ng/L (EW) n.r. EF: 0.79–0.86 (IW); 0.63–0.68
(EW)
Ibuprofen, Influent Sumichiral OA-2500 1.2–4000 µg/L
Naproxen, wastewater (IW); LC/MS/MS (250 × 4.6 mm, 5 µm); Naproxen: 1.2/1.1 ng/L (R/S) (IW); 1.1 [165]
n.r. EF: 0.98–0.99 (IW); 0.93–0.96
Ketoprofen Effluent Chirex 3005 guard ng/L (EW)
(EW)
wastewater (EW) column (30 × 4.6 mm,
5 µm) 1–4000 µg/L
Ketoprofen (R/S): 0.9/0.8 ng/L (IW); 0.8/0.7
n.r. EF: 0.54–0.68 (IW); 0.61–0.68
ng/L (EW)
(EW)
Ibuprofen (R/S): 16.45/23.15 ng/L (EW); Ibuprofen (R/S): 67.37/94.81 ng/L 41–492 µg/L
9.15/9.39 ng/L (SW) (EW); 37.47/38.46 ng/L (SW) EF: 0.65 (EW)
8.66–416 µg/L (R); 8.66–312
Naproxen (R/S): 3.45/4.16 ng/L (EW); Naproxen (R/S): 11.96/14.39 ng/L
µg/L (S)
2.45/3.39 ng/L (SW) (EW); 8.49/11.73 ng/L (SW)
Ibuprofen, EF: 0.92 (EW)
Naproxen, 0.83–297 µg/L (R); 0.83–396
Ketoprofen, Ketoprofen: 0.52 ng/L (EW); 0.26/0.27 ng/L Ketoprofen (R/S): 1.73/1.70 ng/L
µg/L (S)
Chloramphenicol, (R/S) (SW) (EW); 0.86/0.88 ng/L (SW)
EF n.r.
Aminorex,
Chloramphenicol (1R,2R/1S,2S): 3.40–612 µg/L (1R,2R); 3.33–400
Tetramisole, Chloramphenicol (1R,2R/1S,2S): 2.18/2.43
Effluent LC/ESI-MS/MS Chiral-AGP (100 × 2 7.39/8.09 ng/L (EW); 3.46/3.96 ng/L µg/L (1S,2S)
Ifosfamide, ng/L (EW); 1.02/1.19 ng/L (SW)
wastewater (EW); mm, 5 µm); Chiral-AGP (SW) EF n.r.
3-N-Dechloroethy [32]
Surface water (SW) guard column (10 × 2
lifosfamide, 0.17–100 µg/L
mm, 5 µm) Aminorex: 0.12 ng/L (EW); 0.06 ng/L (SW) 0.39 ng/L (EW); 0.20 ng/L (SW)
Fexofenadine, EF n.r.
10,11-Dihydro-10
1.65–396 µg/L (R); 1.65–297
-hydroxy- Tetramisole (R/S): 1.04/0.93 ng/L (EW); Tetramisole (R/S): 3.42/3.08 ng/L
µg/L (S)
carbamazepine, 0.48/0.47 ng/L (SW) (EW); 1.58/1.56 ng/L (SW)
EF: 0.50 (EW)
Praziquantel
Ifosfamide (E1/E2): 0.31/0.29 ng/L
Ifosfamide: 0.09/0.08 ng/L (E1/E2) (EW); 0.17–51 µg/L
(EW);
0.04 ng/L (SW) EF n.r.
0.14/0.15 ng/L (SW)
3-N-Dechloroethy-lifosfamide
3-N-Dechloroethy-lifosfamide (E1/E2): 0.08–40 µg/L
(E1/E2): 11.10/9.79 ng/L (EW);
3.33/2.94 ng/L (EW); 1.09 /1.14 ng/L (SW) EF n.r.
3.62/3.78 ng/L (SW)
136–306 µg/L (E1); 136–408
Fexofenadine (E1/E2): 56.02/58.10 ng/L Fexofenadine (E1/E2): 190.29/197.33
µg/L (E2)
(EW); 33/34.66 ng/L (SW) ng/L (EW); 112.10/117.73 ng/L (SW)
EF: 0.55 (EW)
Molecules 2018, 23, 262 35 of 47
Table 3. Cont.
10,11-Dihydro-10-hydroxy-carbama-zepine 10,11-Dihydro-10-hydroxy-carbama-zepine 1.67–100 µg/L (E1);
(E1/E2): 1.08/1.06 ng/L (EW); 0.53/0.54 (E1/E2): 3.58/3.53 ng/L (EW); 1.67–300 µg/L (E2)
ng/L (SW) 1.75/1.79 ng/L (SW) EF n.r.
8.33–400 µg/L (E1);
Praziquantel (E1/E2): 4.54/4.83 ng/L Praziquantel (E1/E2): 15.12/16.07
8.33–200 µg/L (E2)
(EW); 2.52/2.21 ng/L (SW) ng/L (EW); 8.38/7.37 ng/L (SW)
EF n.r.
Influent
wastewater (IW);
Chiralpak AD-RH (150 × 4.6 EF: 1 (IW); 0.88–0.91
Naproxen Effluent LC/ESI-MS/MS n.r. n.r. [41]
mm) (EW); 0.84–0.98 (RW)
wastewater (EW);
River water (RW)
Omeprazole: 2.03/2.29 ng/L (R/S) (IW); Omeprazole: 2.03/2.29 ng/L (R/S) 2–500 µg/L
0.74 ng/L (EW); 0.67/0.68 ng/L (R/S) (IW); 2.81 ng/L (EW); 2.55/2.59 ng/L EF: 0.70 (IW); 0.53 (EW);
(RW) (R/S) (RW) 0.54 (RW)
Lansoprazole: 0.96/1.02 ng/L (R/S) Lansoprazole (R/S): 4.34/4.63 ng/L 2–500 µg/L
Omeprazole, Influent (IW); 0.69/0.70 ng/L (R/S) (EW); 0.67 (IW); 3.13/3.20 ng/L (EW); 3.05/3.06 EF: 0.51 (IW); 0.52 (EW,
Lansoprazole, wastewater (IW); ng/L (RW) ng/L (RW) RW) [43]
Rabeprazole, Effluent LC/ESI-MS/MS Chiralpak IC (250 × 4.6 mm,
Pantoprazole wastewater (EW); 5 µm) Rabeprazole: 0.94/0.95 ng/L (R/S) (IW); Rabeprazole (R/S): 3.37/3.40 ng/L
2–500 µg/L
River water (RW) 0.71/0.73 ng/L (R/S) (EW); 0.78 ng/L (IW); 2.54/2.62 ng/L (EW); 2.81/2.78
EF: 0.52 (IW); 0.51 (RW)
(RW) ng/L (RW)
Pantoprazole (E1/E2): 0.96/0.94 ng/L Pantoprazole (E1/E2): 2.99/2.94 ng/L 2–500 µg/L
(IW); 0.93/1 ng/L (EW); 0.96/0.91 ng/L (IW); 2.90/3.12 ng/L (EW); 2.99/2.83 EF: 0.54 (IW); 0.51 (EW);
(RW) ng/L (RW) 0.53 (RW)
Econazole: 0.5 ng/L (RaW); 0.3 ng/L 0.5–250 ng/mL
(TW); 3 ng/g (Sd) EF: 0.50 (Sd)
0.5–250 ng/mL
Miconazole: 0.5 ng/L (RaW); 0.3 ng/L
Econazole, Raw wastewater AGP column (100 × 4 mm, 5 µm); n.r. EF: 0.5 (RaW); 0.47 (TW);
(TW); 3 ng/g (Sd)
Miconazole, (RaW); Treated LC/ESI-MS/MS AGP guard column (10 × 4 mm) 0.5 (Sd) [166]
Tebuconazole, wastewater (TW); Tebuconazole: 0.8 ng/L/0.9 ng/L
Ketoconazole Sludge (Sd) 0.5–250 ng/mL
(E1/E2) (RaW); 0.3 ng/L (TW); 4/5
EF n.r.
ng/g (E1/E2) (Sd)
HSA column (100 × 2 mm, 5 µm); Ketoconazole: 10 ng/L (RaW); 5 ng/L 5–250 ng/mL
n.r.
HSA guard column (10 × 2 mm) (TW); 29 ng/g (Sd) EF: 0.48 (RaW, TW, Sd)
Molecules 2018, 23, 262 36 of 47
Table 3. Cont.
0.5–250 ng/mL (Sd)
Econazole: 0.5 ng/L (RW); 3 ng/g (Sd)
EF: 0.52 (RW); 0.50 (Sd)
Econazole, AGP column (100 × 4 mm, 5 µm); n.r. 0.5–250 ng/mL (Sd)
Miconazole, River water (RW); LC/ESI-MS/MS AGP guard column (10 × 4 mm) Miconazole: 0.6 ng/L (RW); 3ng/g (Sd) EF: 0.49–0.54 (RW);
[167]
Tebuconazole, Sludge (Sd) 0.50–0.52 (Sd)
Ketoconazole Tebuconazole: 0.6 ng/L (RW) EF: 0.47–0.61 (RW)
HSA column (100 × 2 mm, 5 µm); Ketoconazole: 7 ng/L (RW); 29 ng/g 5–250 ng/mL (Sd)
n.r.
HSA guard column (10 × 2 mm) (Sd) EF: 0.48–0.49 (Sd)
Tebuconazole: 19.8 µg/L (−); 25.4 µg/L
60 µg/L (−); 76.2 µg/L (+)
(+)
Tebuconazole, 30–1500 µg/L
Chiralpak IC (250 × 4.6 mm, Hexaconazole: 9.1 µg/L (−); 8.6 µg/L
Hexaconazole, River water LC/ESI-MS/MS EF n.r.
5 µm) 27.7 µg/L (−); 25.8 µg/L (+) [168]
Penconazole, (+)
Triadimefon Penconazole: 29 µg/L (−); 27.6 µg/L (+) 88.1 µg/L (−); 83.8 µg/L (+)
Triadimefon: 8.5 µg/L 25.5 µg/L
AM: amphetamine; D-citalopram: desmethyl-citalopram; EF: enantiomeric fraction; ESI: electrospray ionization; FD: fluorescence detector; FLX: fluoxetin; GC: gas chromatography;
HPLC: high performance liquid chromatography; LC: liquid chromatography; LOD: limit of detection; LOQ: limit of quantification; MA: methamphetamine; MDA:
3,4-methylenedioxyamphetamine; MDL: method detection limit; MDMA: 3,4 methylenedioxymethamphetamine; MDEA: N-methyl-diethanolamine; MET: metoprolol; MS: mass
spectrometry; MS/MS: tandem mass spectrometry; MQL: method quantification limit; NFLX: norfluoxetin; PHO: propranolol; QTOF: quadrupole time of flight mass spectrometer;
OD-VNF: O-desmethylvenlafaxine; SBT: salbutamol; T: tramadol; UHPSFC: ultra high performance supercritical fluid chromatography; UPLC: ultra performance liquid chromatography;
UV: ultraviolet detector; VNF: venlafaxine. n.r.: not referred.
Molecules 2018, 23, 262 37 of 47
5. General Conclusions and Further Perspectives

The LC-MS/MS is the first choice for environmental and biological matrices analyses due to the
low quantification limits, the selectivity and unequivocal identification. Regarding environmental
analysis the direct method by LC using CSP are mostly described. However methods for a complex
mixture of chiral drugs are still scarce. Chiral analyses in biological matrices describe many indirect
methods by GC, but the trend is the direct method by LC.
Despite the importance of the chiral analysis in forensic chemistry, this type of data are not
yet currently in used in certificated laboratories for doping control, criminal offense, environmental
monitoring and chiral drug control in general. In this sense more research is needed regarding new
enantioselectivity methods with different CSP and demonstrations with practical applications to
establish the importance of the chiral analysis in forensic chemistry.
Acknowledgments: This work was partially supported through national funds provided by
FCT/MCTES-Foundation for Science and Technology from the Minister of Science, Technology
and Higher Education (PIDDAC) and European Regional Development Fund (ERDF) through the
COMPETE-Programa Operacional Factores de Competitividade (POFC) programme, under the Strategic Funding
UID/Multi/04423/2013, the project PTDC/MAR-BIO/4694/2014 (reference POCI-01-0145-FEDER-016790;
Project 3599-Promover a Producao Cientifica e Desenvolvimento Tecnologico e a Constituicao de Redes Tematicas
(3599-PPCDT)) in the framework of the programme PT2020 as well as by the project INNOVMAR-Innovation and
Sustainability in the Management and Exploitation of Marine Resources (reference NORTE-01-0145-FEDER-000035,
within Research Line NOVELMAR), supported by North Portugal Regional Operational Programme (NORTE
2020), under the PORTUGAL 2020 Partnership Agreement, through the European Regional Development Fund
(ERDF), and Chiral_Drugs_CESPU_2017.
Author Contributions: Maria Elizabeth Tiritan, Cláudia Ribeiro and Carlos Afonso conceived and designed
the work; Cláudia Ribeiro, Cristiana Santos, Valter Gonçalves and Ana Ramos analyzed and reunited the data;
Maria Elizabeth Tiritan, Cláudia Ribeiro and Cristiana Santos contributed in writing up the paper.
Conflicts of Interest: The authors declare no conflict of interest.
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molecules

Received: 30 December 2017; Accepted: 25 January 2018; Published: 28 January 2018

Keywords: chiral drugs; forensic chemistry; enantiomers; pharmaceuticals; illicit drugs

Molecules 2018, 23, 262; doi:10.3390/molecules23020262 www.mdpi.com/journal/molecules

Figure 1. Application of chiral drug analysis in forensic chemistry.

2. Chromatography in Chiral Analyses

Molecules 2018, 23, × FOR PEER REVIEW 8 of 51

Warfarin (WFN) Non-steroidal

ZAIZGHELINProton pump H inhibitors

3. Chiral Analyses in Biological Samples

3.2. Synthetic Opioids

differences in pharmacological activities [75]. Tramadol is metabolized to O-desmethyltramadol (ODT)

3.6. Dissociative Anaesthetics

Matrix Concentration Range/ER

Plasma GC/NICI-MS 5–250 µg/L;

Matrix Concentration Range/ER

Matrix Concentration Range/ER

Matrix Concentration Range/ER

Matrix Concentration Range/ER

Matrix Concentration Range/ER

Matrix Concentration Range/ER

4. Chiral Analyses in the Aquatic Environment

drugs, verification of route of administration, verification of potency of abused drugs, monitoring of

AM, MA, WWTP MDA: 1–500 ng/L

Influent Ibuprofen EF: 0.73–0.90

Influent HP5-MS (30 m × 0.25 Ibuprofen EF: 0.6–0.8 (IW); 0.5

5. General Conclusions and Further Perspectives

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