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; [email protected] (C.R.); [email protected] (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; [email protected] 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; [email protected] 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; [email protected] * Correspondence: [email protected]; 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]. Figure1 summarizes the applications of chiral analysis in forensic chemistry.
Molecules 2018, 23, 262; doi:10.3390/molecules23020262 www.mdpi.com/journal/molecules Molecules 2018, 23, × FOR PEER REVIEW 2 of 47
Substances of forensic interest include pharmaceuticals and various classes of illicit drugs misused to improve sports performance or due to their psychotropic effects. The consumption of these substances can cause toxicological effects and/ or increased risk of death [10–13]. These substances 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 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.
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 (Figure1). 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) (Tables1 and2). 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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Structures of the Chiralof thechiralChiral chiralChiral chiral Compounds illicit illicitCompounds Compounds illicit drugs drugs drugs and and and pharmaceuticals pharmaceuticals.pharmaceuticals Molecules 2018, 23, × FOR PEER REVIEWStimulants Stimulants Stimulants and and andtheir their their metabolites metabolites metabolites 4 of 47 StimulantsStimulantsStimulants and and andtheir their their metabolites metabolites metabolites NH ChiralChiral Compounds CompoundsNHC H NH2 N2H2 StimulantsStimulantsChiralChiralChiral and Compounds and their Compounds Compounds theirN metabolitesHCH 3metabolitesNH C3H3 NH NHN2H2 Chiral CompoundsNHCNHHNCHHC3H3 NNHHCCHH NH2 NNHH2 2 NHCHN3HNCHHC3H3 O OO NHCH3 3 3 NH2 NTableH2 1. Structures of the chiral illicitNHCHN3H drugsCH3 and pharmaceuticals NHCNHNHCCHH3 StimulantsStimulantsStimulants and and andtheir their their metabolites metabolites metabolites O OO NHCNHNH3HCCHH33 StimulantsStimulantsStimulantsStimulants and and and andtheir their their their metabolites metabolites metabolites metabolites O O CHNCHCHCHN3HH3CH3 CH CHC3H3 CH CHC3H3 OO 3 3 3 3 O CHCCHH CH CNHCH3H23NH2 CH CNHCH3HC3HN3HCH3 O CH3CCHH3 3 CH3 CNCHHH3 3NNHH2 2 CH3 CNHCH3HC3HNNH HCCHH3 3 O O 33 NH22 NH2 2 Chiral CompoundsNHCHN33HCH3 3 OO CH3CH3 CH3 CH3 CH3 CH3 O O NHCNHNH3HCCHH3 O OO NHCNHNHH3C CHH33 Stimulants and their metabolites 3,4O -3,4MethylenedioxyO3,4O-Methylenedioxy-Methylenedioxy-3 - 3- AmphetamineAmphetamineAmphetamineC (AM)H C (AM)H (AM) NH 2 MetamphetamineMetamphetamineMetamphetamineCH C(MA)H (MA) (MA)N HCH3 3,4-3,4Methylenedioxy3,4-Methylenedioxy-MethylenedioxyCH3CCHH3 - - - CH3 CHC3H3 CH3 CHC3H3 3,4O -O3,4Methylenedioxy3,4O-Methylenedioxy-MethylenedioxyCH3CCHH33- -- CH33 CCHH3 3 CH33 CCHH3 3 methamphetamineOmethamphetamineOO 3 (MDMA)3 (MDMA) AmphetamineAmphetamineAmphetamine (AM) (AM) (AM) MetamphetamineMetamphetamineMetamphetamine (MA) (MA) (MA) methamphetamine3,4-3,4MethylenedioxyO-Methylenedioxy (MDMA)N-H C-H 3 AmphetamineAmphetamine (AM) (AM) MetamphetamineMetamphetamine (MA) (MA) methamphetaminemethamphetaminemethamphetamine (MDMA) (MDMA) (MDMA) methamphetaminemethamphetamine (MDMA) (MDMA) NH NNHH2 NNHH NH NCHHNH O OO CH2 3 2 O OO NH CHC3H CCHH3 3 3,4-3,4Methylenedioxy-Methylenedioxy3- - NHNNHH 3 3,4-Methylenedioxy-methamphetamine3,4-3,4Methylenedioxy3,4O-Methylenedioxy-MethylenedioxyNH - -- AmphetamineOAmphetamineOO (AM)NH (AM)2N NHH2 2 MetamphetamineO MetamphetamineOO NH NN HN(MA)HHC (MA)H C HC3H NH NCNHH3 CHC3H3 AmphetamineOAmphetamineAmphetamineOO (AM)NH (AM) (AM)N H22 MetamphetamineO MetamphetamineOMetamphetamineO (MA)C (MA)H (MA)3 CCHH3 3 O O 2 2 O O CHNCHCHNHH CH CH methamphetaminemethamphetamineNH (MDMA)NCH (MDMA)H3CHC3H3 CH CCHH3 3 O OO 3 3 3 3 3 methamphetaminemethamphetamineHmethamphetamineO HOHO (MDMA) (MDMA)C (MDMA)H (MDMA)3 CCHH3 3 O OO 3 CHCH 3,4-Methylenedioxy-CH3 CH3 CHCCHH OO CH3CCH3H3 HO O AmphetamineOO CH3CCH (AM)H333 O O MetamphetamineO CH CH3 3 (MA) HO HHOO O OO CHNCHH2N3 NHH2 O O N3H N3H HmethamphetamineO HO NH NHNH (MDMA) O OO N3H2NNHH22 O OO NH NNHHCH3C CHH3 3 NH NNHH O 3,43,4OO-Methylenedioxy-Methylenedioxy2 2 MethylenedioxyMethylenedioxyO OO -N-N-ethylamphetamine-ethylamphetamineCH3 CCHH3 3 p-pHydroxymethamphetamine-HydroxymethamphetamineCH 3,4-Methylenedioxy Methylenedioxy-N-ethylamphetamine p-HydroxymethamphetamineCH3CHC3H3 3,4O-MethylenedioxyNH 2 MethylenedioxyO -N-ethylamphetamineNH CH p-HydroxymethamphetamineCNH3 CCHH3 3 3,4-3,4Methylenedioxy3,4-Methylenedioxy-Methylenedioxy MethylenedioxyMethylenedioxyMethylenedioxy-N-Cethylamphetamine-HN-N3-Cethylamphetamine-CHethylamphetamineH3 3 p-Hydroxymethamphetaminepp-Hydroxymethamphetamine-Hydroxymethamphetamine amphetamineamphetamineCH 3C(MDA) C(MDA)HH3 O O (MDEA)C(MDEA)H3CCHH3 3 HO HO (pOHMA)(pOHMA) CH amphetamine3,4O-3,4MethylenedioxyOO-Methylenedioxy C(MDA)H3CCHH3 3 MethylenedioxyMethylenedioxyO OO (MDEA)-N--ethylamphetamineN3-ethylamphetamine 3 p-Hydroxymethamphetaminep-HHydroxymethamphetamineO HHO(pOHMA)O 3 amphetamineOamphetamineamphetamineOO (MDA) (MDA) (MDA) (MDEA)(MDEA)(MDEA)CH (pOHMA)(pOHMA)(pOHMA) amphetamineamphetamineamphetamine (MDA) (MDA) CH(MDA)3 O (MDEA)(MDEA)(MDEA) 3 HO(pOHMA)(pOHMA)(pOHMA) amphetamineamphetamineO (MDA) (MDA) (MDEA)(MDEA) (pOHMA)(pOHMA) N 3,4-3,4Methylenedioxy-Methylenedioxy MethylenedioxyMethylenedioxy-N--ethylamphetamineN-ethylamphetamine p-Hydroxymethamphetaminep-HydroxymethamphetamineN N 3,4-Methylenedioxy3,4-3,4Methylenedioxy3,4-Methylenedioxy-Methylenedioxy amphetamine MethylenedioxyMethylenedioxyMethylenedioxy-Methylenedioxy-N--ethylamphetamineN-N-ethylamphetamine-ethylamphetamine-ethylamphetamine p-Hydroxymethamphetaminepp-Hydroxymethamphetamine--HydroxymethamphetamineHydroxymethamphetamineN N N 3,4-3,4Methylenedioxy3,4-Methylenedioxy3,4-Methylenedioxy-MethylenedioxyNH NNHH MethylenedioxyMethylenedioxyMethylenedioxy-Methylenedioxy-N-ethylamphetamine-N-N-ethylamphetamine-ethylamphetamine-ethylamphetamine p-Hydroxymethamphetaminepp-Hydroxymethamphetamine-Hydroxymethamphetamine-HydroxymethamphetamineN N N HO HHOO NH NHN2H2 amphetamineamphetamine (MDA)NH (MDA)NCHNHH CCHH3 3 (MDEA)(MDEA) (pOHMA)(pOHMA)N N 2 HamphetamineOHamphetamineO(MDA) NH (MDA) N(MDA)NHH3 (MDEA)(MDEA)N N (pOHMA)(pOHMA) NHNH HamphetamineO amphetamineHamphetamineO (MDA) (MDA) (MDA) (MDEA)(MDEA)(MDEA)N (pOHMA)(pOHMA)(pOHMA)O O O NH 2 NHN2H2 HamphetamineO NH NC(MDA)HH3CCCHHH3 3 (MDEA)H (pOHMA)2 2 2 HO HO CH 3 3 3 HN NHN O NH NH2 CH3 CH3C3H CH N NN O O 2 3 3 H HH N OON N O OO CH CHC3H O OO HNOONHH O N O N N O CH3CCHH3 3 H H N N O OO CHN3CHHN3NHH O OO OO HO HHOO NH NNHH NH2 NHN2H2 HOOHHOO CH CCHH NH NHN2H 2 HO HHOO 3C CHH3 3 O OO O NH22 NH2 2 CH3CCHH3 3 N NN O O O 3 3 N NN O O 4-Hydroxy-3-methoxyCH H H H O OO 4-Hydroxy4-Hydroxy-3-CmethoxyH-33C-CHmethoxyH3 3 H HH O4-4HydroxyO-OHydroxyC-H3-3-3Cmethoxy-Hmethoxy3 3 O OO OO 4-OHydroxy44-OHydroxy-OHydroxy-3-methoxy-3-3-methoxy-methoxy O OO OO 4-methamphetamineHydroxy4-methamphetamineHydroxymethamphetamine-3-methoxy-3-methoxy MethylphenidateMethylphenidateMethylphenidateO O(MPH) (MPH) (MPH) AminorexAminorexAminorex methamphetaminemethamphetaminemethamphetamine MethylphenidateMethylphenidateMethylphenidate (MPH) (MPH) (MPH) AminorexAminorexAminorex methamphetamine4-Hydroxy-3-methoxymethamphetamine(HMMA)(HMMA)(HMMA) MethylphenidateMethylphenidate (MPH) (MPH) AminorexAminorex 44-Hydroxy-3-methoxy-Hydroxy4-4Hydroxy-Hydroxy(HMMA)(HMMA)(HMMA)-3-methoxy-3-- 3methoxy-methoxy Methylphenidate (MPH) Aminorex 4methamphetamine-Hydroxy44-Hydroxy-Hydroxy(HMMA)(HMMA)(HMMA)-3-methoxy-3- 3-methoxy-methoxy (HMMA) Methylphenidate (MPH) Aminorex methamphetamine(HMMA)(HMMA) (HMMA) methamphetaminemethamphetaminemethamphetamine MethylphenidateMethylphenidateMethylphenidateDrugDrugDrug precursors precursors precursors (MPH) (MPH) (MPH) AminorexAminorexAminorex DrugDrugDrug precursors precursors precursors (HMMA)(HMMA)(HMMA) DrugDrugDrug precursors precursors precursors (HMMA)O(HMMA)(HMMA)H OOHH OH OOHH OHOOHH OOHH OOCHH CCHCHH3 OH OOCHH CCHCHH OH OH 3 3 DrugDrugO precursorsH O precursorsH 3 3 3 CH3CCHH3 3 DrugDrugDrug precursors precursorsC precursorsH3CCHH3 3 3CH33 3CCHH33 CNHNHNH3H 3 NHCNHNHNH3CHH3 OHNOHH CH CCHCHH3 OHNOHOHNHCNHHHCCHCHH3 3 OHNOHOHNHNHH3 3 OHNOHHNNHH3 3 OH OOHHCHCCHH3 CHCCHH3 NH NCHH3C CHH3 3 NHCNHCH3HC3CHC CH3HH3 3 CHC3HCCHC H3H PseudoephedrineCHC3HCCHC H3H3 EphedrineCH3CC3H H3 33 PseudoephedrinePseudoephedrinePseudoephedrine3 3 3 3 EphedrineEphedrineEphedrine 3 3 PseudoephedrinePseudoephedrinePseudoephedrineNH NH EphedrineEphedrineEphedrineNH NNH H PseudoephedrinePseudoephedrinePseudoephedrineNH NNHH Opioids, morphineEphedrineEphedrine EphedrinederivativesNH NC HH3C H 3 and their metabolites PseudoephedrineNH NCHH CH EphedrineCH3C CHH3 3 PseudoephedrinePseudoephedrineCH3C C HH3 3 EphedrineEphedrine 3 33 O 3 3Opioids,3 Opioids,Opioids, morphine morphine morphine derivatives derivatives derivatives and and andtheir their their metabolites metabolites metabolites CH Opioids,Opioids,Opioids, morphine morphine morphine derivatives derivatives derivativesN and and andtheir their their metabolites metabolites metabolites 3 PseudoephedrinePseudoephedrine Opioids, Opioids,Opioids,Opioids, morphine morphine morphine morphine Ephedrinederivatives derivatives Ephedrinederivatives derivatives and and and andtheir their their their metabolites metabolites metabolites PseudoephedrinePseudoephedrinePseudoephedrineO Opioids, Opioids, morphine morphine Ephedrinederivatives EphedrinederivativesEphedrine and and their their metabolites metabolites O O O H C CCHH 3 N O COHO3 3 3 CH N NN HO OOCHCH 3 O COH3CCHH3 3 CH O OO Opioids, morphine derivativesN NN and their metabolites 3 3CH 3 Opioids,Opioids,Opioids, morphine morphine morphine derivatives Nderivatives derivativesNN and and andtheir their their metabolites metabolites metabolites CH3CNH3 3 H C H3HC3C N O Opioids,Opioids, morphine morphine derivatives NderivativesN and and their their metabolites metabolites 3 N N O OO HHOO H C H3HC C CH CHC3H HO CH H3C HHNC3C N N O O3 3 O 3 3N3 CNHN HO HOHO O OO CCHH H3C HC3CH3 NC3H3 CH3 CHC3H3 HO HHOO O COHOCCNHHN3 3 3 N CH3 CHC3H3 CNH3CCHH3 3 CH CHC3H N N HO HO CH3CCHHC3 3HC3H3 CH3 CHCH3 CH3 CH3 2-Ethylidine-1,5-dimethyl-3,3-N NN N CNHN3CCHH3 3 3 3 N N CNH CNNCHH3 CH3 CMethadoneH3 O O Tramadol3CH 3(T)C3H3 O OO N N H3C H3HNCC N diphenylpyrrolidine (EDDP) CH3CHC3H3 H3C HH3NC3C NN 3CCHH3 3 3 3N NN CH CH HO HHOO CH OH 3 CHC3H3 HO HHOO CH3 3 CH33 CCHH3 3 2-Ethylidine-1,5-Odimethyl-3,3- OH CH3 CHC3H 2-Ethylidine2-Ethylidine-1,5-dimethyl-1,5-dimethyl-3,3--3,3- CH3CCHH3 CH3 CCHH3 3 CH N CNHN3CCHH3 3 MethadoneCH3MethadoneCMethadoneH3 3 2-Ethylidine-1,5-dimethyl-3,3-diphenylpyrrolidine2-Ethylidine22-2-Ethylidine-Ethylidine-1,5---1,51,5-dimethyl1,5--dimethyl-dimethyl3 -3,3---3,33,3-3,3--- TramadolTramadolTramadolN (T)N N(T)3 (T) 3 3 Methadone diphenylpyrrolidinediphenylpyrrolidine (EDDP) (EDDP) TramadolCH C (T)H MethadoneMethadoneMethadone 2diphenylpyrrolidine-Ethylidine2-Ethylidine-1,5--1,5dimethyl-dimethyl (EDDP) - 3,3 - -3,3- TramadolTramadolTramadolC H(T)3C CH(T) (T)H3 3 MethadoneMethadone diphenylpyrrolidinediphenylpyrrolidinediphenylpyrrolidine(EDDP) (EDDP) (EDDP) (EDDP) TramadolTramadolC H(T)3CC H(T)H3 3 HO diphenylpyrrolidinediphenylpyrrolidinediphenylpyrrolidineHO (EDDP) (EDDP) (EDDP) HO OH OOHH diphenylpyrrolidinediphenylpyrrolidineOO (EDDP) (EDDP) OOHH CH 2-Ethylidine2-Ethylidine-1,5O--1,5dimethyl-dimethylCH-33,3--3,3- OH CH3 OH OOHHN 3 2-Ethylidine22-Ethylidine-Ethylidine-1,5--1,5-dimethylC1,5H-dimethylC-dimethylHCN3H3 -3,3--3,3-3,3- - N MethadoneMethadoneOH OOHH 2-Ethylidine22-Ethylidine-Ethylidine-1,5O-1,5dimethyl-1,5OO3O-dimethyl-dimethylH -3,3--3,3-3,3-- TramadolTramadolOH O O(T)HOHHH (T) MethadoneMethadoneMethadoneOH OH CH3CHC3H3 TramadolTramadolTramadol (T) (T) (T) CH3 diphenylpyrrolidinediphenylpyrrolidinediphenylpyrrolidineO O3 C CH(EDDP)H3 3 (EDDP) (EDDP) OH OH diphenylpyrrolidinediphenylpyrrolidinediphenylpyrrolidineCH3C H(EDDP)3 (EDDP) (EDDP) HO HHOO HO HOHO HO HOHO O-DesmethyltramadolOH OH N,O-Ddidesmethyltramadol HO HOHO OH OOHCHH CCHH3 3 HO HOHO O OOCNH3 CHC3H3 HO HOHO OH OOCHH3 CHC3H3 HO HHOO OHNOHNN3 N-DesmethyltramadolHHOO O CNHOOCHN (NDT) HO HHOO OHNOOHNHN HO HO CHCH HO HO CHH3CCHHCH3HC3HC3H3 HO HO CHHHCHC3H (ODT)N CNHN 3CCHH3 3 CNH3CNCHCHN3H3C3CHH3 3 (N,O-DDM-T)HN CNHN3 CCHH3 3 CNH CNNCHH 3 3 N CHNHN CH N NN N 3CNH33C 3H3 HN NHH3 3 HN CHNHH3CH3 CH3CHC3H3 H H H H HO HO 3CCHH3 3 AntidepressantsHO andHO their metabolites/ BenzodiazepineHO HO HO HHOO CH3CH3 HO HHOO HO HHOO O-DesmethyltramadolOO-Desmethyltramadol-DesmethyltramadolCH3CH3 CH3CHC3H N,ON-NDdidesmethyltramadol,O,O-Ddidesmethyltramadol-DdidesmethyltramadolCH3CHC3H N CNH CCHH3 3 N CNHN3CCHH3 3 N CNHN3 CCHH3 3 O-DesmethyltramadolOO-Desmethyltramadol-DesmethyltramadolF N CNHN3CH3 3 N-DesmethyltramadolNN-Desmethyltramadol-DesmethyltramadolF NH HNN3 (NDT)3 (NDT) (NDT) N,ON-N,DdidesmethyltramadolO,O-Ddidesmethyltramadol-Ddidesmethyltramadol-DdidesmethyltramadolNHN HNN3 3 O-DesmethyltramadolOFO-Desmethyltramadol-Desmethyltramadol H HH N,ON-NDdidesmethyltramadol,O,O-Ddidesmethyltramadol-DdidesmethyltramadolH HH O-DesmethyltramadolO-DesmethyltramadolO-Desmethyltramadol(ODT)(ODT)(ODT)C H3C CH H3 (ODT) N-DesmethyltramadolNN--DesmethyltramadolDesmethyltramadol-DesmethyltramadolF (NDT) (NDT) (NDT) N,ON-Ddidesmethyltramadol,O(-N,ODdidesmethyltramadol(N,O(-N,OHDDMO-DDM-DDM-T) - T)-T) CH3CCHH3 3 N-DesmethyltramadolNN-Desmethyltramadol-Desmethyltramadol (NDT) (NDT) (NDT) (N,O-DDM-T) F (ODT)(ODT)(ODT)(ODT) 3 3 N-DesmethyltramadolN-Desmethyltramadol (NDT) (NDT) (N,O((N,O(-N,ODDM--DDM-DDM-T) --T)-T) (ODT)(ODT) F (N,O(N,O-DDM-DDM-T) -T) O-DesmethyltramadolO-DesmethyltramadolO AntidepressantsN AntidepressantsCHAntidepressants 3 and and andtheir their their metabolites/ metabolites/ metabolites/ Benzodiazepine Benzodiazepine BenzodiazepineN,ON-Ddidesmethyltramadol,O -Ddidesmethyltramadol O-DesmethyltramadolOO-Desmethyltramadol-Desmethyltramadol Antidepressants and their metabolites/BenzodiazepineO NH2 N,,ON-NDdidesmethyltramadol,O,O-Ddidesmethyltramadol-Ddidesmethyltramadol AntidepressantsH AntidepressantsAntidepressants N-DesmethyltramadolNN --andDesmethyltramadol-Desmethyltramadol and andtheir their their metabolites/ metabolites/ metabolites/ (NDT) (NDT) (NDT) Benzodiazepine Benzodiazepine Benzodiazepine O (ODT)(ODT)(ODT)(ODT) AntidepressantsAntidepressants and and their their metabolites/ metabolites/ Benzodiazepine Benzodiazepine( N,O ((N,O(-N,ODDM--DDM-DDM-T) --T)-T) F FFluoxetineF (FLX) NorfluoxetineF (NFLX) VenlafaxineN NN (VNF) F F F F F F FFF F F F F F H HON NNH F F F AntidepressantsAntidepressantsAntidepressants andF and andtheirFF their their metabolites/ Nmetabolites/ metabolites/ Benzodiazepine Benzodiazepine Benzodiazepine HO HNO NN F FF F N AntidepressantsAntidepressantsAntidepressantsF andF andF andtheirF their their metabolites/ metabolites/ metabolites/ Benzodiazepine Benzodiazepine Benzodiazepine H ON N FF F F FF FF F HO HHOO F F F HO CHCH F FF HO HO HHOO FF O OO N CHN3N 3 3 F FF O NH F FF CHCH O O NH2 N2H 2 F F O OO HN CNHH3NCCH3H 3 F FF F N N F FF O OO N NN 3 3 F FF O OO NH2NHN2H2 N NN O OO F F F H CHHH3 CH3 F F FF O OO NH2 NNHH2 2 F FF O O HN NHH F FF O O NH NH HO HHOO O OO H H F F 2 2 HO HHOO O OO F F F O H F F O O O O H F FluoxetineFFFluoxetineFluoxetine (FLX) (FLX) (FLX) NorfluoxetineF Norfluoxetine NorfluoxetineNorfluoxetineFF (NFLX) (NFLX) (NFLX) VenlafaxineVenlafaxineVenlafaxine (VNF) (VNF) (VNF) FluoxetineO O (FLX)N CNH3 CHC3H3 Norfluoxetine (NFLX) Venlafaxine (VNF) O-DesmethylvenlafaxineFluoxetineFluoxetineFluoxetineO O (FLX)O (FLX)N (FLX)C HN3N3 CC HH3 3(O- NorfluoxetineNorfluoxetineNorfluoxetineO O (NFLX) (NFLX) N(NFLX)H2 N H2 NVenlafaxine,O-DidesmethylvenlafaxineVenlafaxineVenlafaxine (VNF) (VNF) (VNF) O OO NH NN O OO NH2 NNHH2 2 FluoxetineFluoxetine (FLX) (FLX)H HH N-DesmethylvenlafaxineNorfluoxetineNorfluoxetineO O (NFLX) (NFLX)NH 2( N H-DES-VNF)2 2 VenlafaxineVenlafaxine (VNF)O (VNF) O DES-VNF)H H (N,O-DES-VNF)O OO FluoxetineFluoxetineFluoxetine (FLX) (FLX) (FLX) NorfluoxetineNorfluoxetineNorfluoxetine (NFLX) (NFLX) (NFLX) VenlafaxineVenlafaxineVenlafaxine (VNF) (VNF) (VNF)
Molecules 2018, 23, 262 5 of 47
Table 1. Cont. MoleculesMoleculesMolecules 2018 2018 2018, 23,, ×,23 23FOR, ,× × FOR FOR PEE PEE RPEE REVIEWRR REVIEW REVIEW 5 of5 485 of of 48 48 Chiral Compounds
H HH H HH N NN N NN N NN HO HHOO HO HHOO HO HHOO MoleculesMoleculesMoleculesMoleculesMoleculesMolecules 2018 2018 2018, 23 2018,, 2018 ,232018× 23 FOR, ,×,, ×, 23, FOR 23 23FOR ,,PEER ,×,× × × FOR FOR PEE FOR FORPEE REVIEW R PEEPEE R PEE PEEREVIEW REVIEWRRR REVIEW REVIEWREVIEW 6 of6 6 51of of 516 516 6 of of of 51 5151 MoleculesMolecules 2018 2018, 23,, ×23 FOR, × FOR PEE PEER REVIEWR REVIEW 6 of 651 of 51 MoleculesMoleculesMolecules 2018 2018, , 2018 23,,, ××23, FOR23FOR, ×, ×FOR PEEFORPEE PEER PEE REVIEWRR REVIEW REVIEW 6 of 651 6of of 51 51 OH OOHH O OO OH OOHH TableTableTable 1Table.TableTable Cont. 1 1. .Cont. Cont. 1 1 .1. .Cont.. Cont. Cont. TableTable 1. Cont. 1. Cont. O-Desmethylvenlafaxine-DesmethylvenlafaxineOO-Desmethylvenlafaxine-Desmethylvenlafaxine N-DesmethylvenlafaxineNN-Desmethylvenlafaxine-DesmethylvenlafaxineN-DesmethylvenlafaxineTableTableTable 1. Cont. 1 .1 Cont.. Cont.( N- (DES N(N -DES-DES- -- N,ONN-NDidesmethylvenlafaxine,O,,OO-Didesmethylvenlafaxine--DidesmethylvenlafaxineDidesmethylvenlafaxine (O-DES-VNF)-(DESO(O-DES-DES-VNF)-VNF)-VNF) (NVNF)-DES-VNF)VNF)VNF) (N,O(N,O((N,O-N,ODES-DES--DES-VNF)DES-VNF)-VNF)-VNF)
NN NN NN N F F FF HHNN F F F HN HNHN HHNNHN F F F FFFF N N F F HHCC OO OO HN HN H C H C33 O O O O F F N N N H3C3 H33HC3OC O O O O O N F F F HN HNHN F F H3C H3C O O O O F F OO NNHH H33C H3CH3CO O OOO OO OO O NOHNH NNHHNH O O NH NH O O O NH NHNH O OO OO OOOO OO OO O O O O O O O O C C C CCCC O O O CC CCCC O N N N CNNNN C N CNNN C N N N C C C C N C N C N CN C N N N N N N CitalopramCitalopramCitalopramCitalopramCitalopram ReboxetineReboxetineReboxReboxetineReboxetineReboxetineetine D-citalopramDD-citalopram-citalopramDDD--citalopram--citalopramcitalopram CitalopramCitalopram ReboxetineReboxetine D-citalopramD-citalopram CitalopramCitalopramCitalopram ReboxetineReboxetine Reboxetine D-citalopramD D-citalopramD-citalopram-citalopram OO OOOO NN NNNN O O N N O O O NN N N N NN NN OOHH OOOHHOHH N N OH OH N NN N NNOH OHOH NN ClCll CClClClll NN NNN NN NNNN NNN N N Cl Cl N N N N N N N N NNN NN Cll ClCl
MirtazapineMirtazapineMirtazapineMirtazapineMirtazapine TemazepamTemazepamTemazepamTemazepamTemazepam MirtazapineMirtazapine TemazepamTemazepam MirtazapineMirtazapineMirtazapine TemazepamTemazepam Temazepam DissociativeDissociativeDissociativeDissociativeDissociativeDissociative anaesthetic anaesthetic anaesthetic anaesthetic anaestheticanaesthetic and and and its and its andandmetaboliteits metabolite itsmetabolite itsits metabolite metabolitemetabolite DissociativeDissociativeDissociative anaesthetic anaesthetic anaesthetic and and and its its metaboliteits metabolite metabolite OO OOOO DissociativeDissociativeDissociative anaesthetic anaesthetic anaestheticOO OOOO and and andits metaboliteits its metabolite metabolite Cl CCl l Cl CCl l O OClCl CCllCl O OClCl CCllCl O O OCl Cl O O OCl Cl Cll Cl Cl Cll Cl Cl
NH NNHH NHNH NNHHNH NH2 NHNNHH22 NH2 2 NHN22H2 NH NH NH NH H C HH33CC 2 2 H3HC3C HHN33HCC33C NHNH NH NH22 NHN2H2 H3C H3C H C H C H33C H3C3 KetamineKetamineKetKetamineamineKetamineKetamine NorketamineNorketamineNorketamineNorketamineNorketamine KetamineKetamine Norketamine Norketamine KetamineKetamineKetamine NorketamineNorketamineNorketamine β-Blockersββ-Blockers-βBlockersβ-Blockersββ--Blockers--BlockersBlockers and and and anti and and antiand andanti- anti-arrhythmicarrhythmic anti- antiarrhythmic-antiarrhythmic--arrhythmic--arrhythmicarrhythmic β-Blockersβ-Blockers and and anti anti-arrhythmic-arrhythmic OH β-Blockersββ-Blockers-Blockers and and andanti anti -antiarrhythmic-arrhythmic-arrhythmic OH OHOH OOHHOH OH OH OH OOHH H H H HH OHOH OOHHOH OH O OO N H HNHN OHOH OOHHOH O O OO OOH ONHONH NN N OH OH H H OH OH O O N N O OO N NN H HH H H H OO OO OOH OHNOHN NN N O H H HHNH H O O OO OOH OHNONH NN N O O O N N N O O N N H H O O N N O O O N N N H H H O O O N N N
O OO OOO O OO O O O O OO O O O O OO O O O O OO O O O O O O O O O O O O MetoprololMetoprololMetoprololMetoprololMetoprolol (MET) (MET) (MET) (MET)(MET) PropranololPropranololPropranolol PropranololPropranolol (PHO) (PHO) (PHO) (PHO)(PHO) BisoprololBisoprololBisoprololBisoprololBisoprololBisoprolol (BSP) (BSP) (BSP) (BSP) (BSP)(BSP) MetoprololMetoprolol (MET) (MET) PropranololPropranolol (PHO) (PHO) BisoprololBisoprolol (BSP) (BSP) MetoprololMetoprololMetoprololH H HH(MET)HH (MET) (MET) PropranololPropranololPropranolol (PHO) (PHO) (PHO) BisoprololBisoprololBisoprolol (BSP) (BSP) (BSP) N N NNNN H H N N H H H N N N
OO OOOO O O OH OH OOHH OOHOHH OOHHOOHH O O O OHOHHH HHH HO HHOHOOOH OH O O N N HO HHOOOH OHOH OO OOOH ONOHHNH NHN HOHO HHOHOHOO HO HO OOHH OOHOHH O HN HH HH HHH O N HO HO NH NNHH HO HOHO OH HH O N NHNH NNHHNH O OOO ONH NNN O O N N HO HOHO O OOOH NOHHOH HNN NH NH O O HN HNH NH NHNH O OO N NN HNHHNNHHHNNN HN HN O OO OO OO O HN HHNN O O O O O
OO OOOO O O O O O O
CarvedilolZCarvedilolAIZHELICarvedilolCarvedilolCarvedilol NadololNadololNadolol NadololNadolol PindololPindololPindololPindololPindololPindolol CarvedilolCarvedilol NadololNadolol PindololPindolol CarvedilolCarvedilolCarvedilol NadololNadololNadolol PindololPindololPindolol OHOOHH OOOHHH O OO OOO OO OOO OH HH HHH N NN O O OO N OH NN O O NN NN O O NN NNN OO OOO O OO OOO OO OOOH ONOHNH NHN O OO N N O OO N N O O OO O O HN HNH N NN O O O O N NN O O OOO N N NNNN N NN O OO O OO O N O OO N NN O OOO NNN H N HH2NN O O N N OO OO NN NN OO OOO 2 HH2N2NHH22N2N O OO N NN O O HNH HNHH O O H 2NH2N O OO NH NHN O OO H2NHH2N2N H HH O VerapamilVerapamilVerapamilVerapamilVerapamil NorverapamilNorverapamilNorverapamil NorverapamilNorverapamil AtenololAtenololAtenololAtenololAtenololAtenolol VerapamilVerapamil NorverapamilNorverapamil AtenololAtenolol VerapamilVerapamilVerapamil NorverapamilNorverapamilNorverapamil AtenololAtenololAtenolol
Molecules 2018, 23, 262 6 of 47 MoleculesMolecules 2018 2018, 23,, ×23 FOR, × FOR PEE PEER REVIEWR REVIEW 8 of8 51 of 51
Molecules 2018, 23, × FOR PEER REVIEW 8 of 51 TableTableTable 1 1.. Cont. 1Cont.. Cont. MoleculesMoleculesMolecules 2018 2018 ,2018 23,, ×23, 23FOR, ×, ×FOR FORPEE PEER PEE REVIEWRR REVIEW REVIEW 8 of8 51 8of of 51 51 MoleculesMolecules 2018 ,2018 23, ×, 23FOR, × FORPEE RPEE REVIEWR REVIEW 7 of 517 of 51 MoleculesMoleculesMolecules 2018 2018, 201823,, ×23, FOR23, ×, ×FOR PEEFOR PEER PEE REVIEWR RREVIEW REVIEW Table 1. Cont. 8 of 851 8of of 51 51 ChiralAnticoagulantsAnticoagulants Compounds MoleculesMolecules 2018 ,2018 23, ×, 23FOR, × FORPEER PEE REVIEWR REVIEW 8 of 518 of 51 Molecules 2018, 23, × FOR PEER REVIEW TableTableTable 1. Cont. 1 .1 Cont.. Cont. 8 of 51 AnticoagulantsOH O H OH OH TableTableTable 1. Cont. 1 .1 Cont.. Cont. H H Molecules 2018, 23, × FOR PEER REVIEWH H O O N N 8 of 51 O O N N O O OH O O O AnticoagulantsTableAnticoagulantsAnticoagulantsTableTable 1. Cont. 1 1. .Cont. Cont. OH O S S N N AnticoagulantsAnticoagulantsAnticoagulants OH O H HTable 1. Cont. AnticoagulantsAnticoagulantsAnticoagulants Alprenolol Sotalol AlprenololAlprenololOH OHOH O O O SotalolSotalol O O O O Anticoagulants OH OHOH O O O Anticoagulants OOH OOH O O WarfarinWarfarinO H(WFN) (WFN) O
O O O O WarfarinOH (WFN)O O O Non-steroidal anti-inflamatory drugs O O O O O O Non-steroidal anti -inflamatory drugs WarfarinWarfarinWarfarinO O(WFN)O (WFN) (WFN)O O O Non-steroidal anti-inflamatory drugs WarfarinWarfarinWarfarin (WFN) (WFN) (WFN) OH O WarfarinO (WFN) OH WarfarinWarfarin (WFN) (WFN)O HO H NonNonNon-steroidal-steroidal-steroidal anti anti - antiinflamatory- inflamatory- inflamatory drugs drugs drugs OHOH Non-steroidal anti-inflamatoryOOH drugs Warfarin (WFN) NonHNonO-HsteroidalO-steroidal anti anti-inflamatory-inflamatoryO drugs drugs HOHO Warfarin (WFN)O OOH NonNon-steroidal-steroidal anti -antiinflamatory-inflamatory drugs drugs OOOH Non-steroidalNon-steroidalO anti-inflamatory anti-inflamatory drugs drugs HO O O OHO OHH HO O OHOOHH O OHOOHH Non-steroidal anti-inflamatoryO drugsHOOHH O O IbuprofenIbuprofen OHOOHH HOHHOOCarboxyibuprofenCarboxyibuprofen O O HO2H-HOHydroxyibuprofenO2-HydroxyibuprofenO HO OHH O OO OHOOHH O OO OHO OHH O OO HO OHO OHH HOHOHOOOO HOHO Ibuprofen O OO Carboxyibuprofen O 2-HydroxyibuprofenO OO HOHHOO OOHO HOHHOO O O O OO O O O O O OHO O H O OHO IbuprofenIbuprofenIbuprofen OH O CarboxyibuprofenOOCarboxyibuprofenCarboxyibuprofen 2-Hydroxyibuprofen2-2Hydroxyibuprofen-Hydroxyibuprofen HO O OHOH HO IbuprofenIbuprofenIbuprofenO CarboxyibuprofenOCarboxyibuprofenCarboxyibuprofen 2-Hydroxyibuprofen2O-2NHydroxyibuprofen-HydroxyibuprofenN O OOH OH IbuprofenIbuprofenO O CarboxyibuprofenCarboxyibuprofen 2-Hydroxyibuprofen2-HydroxyibuprofenOH O O IbuprofenIbuprofen CarboxyibuprofenCarboxyibuprofenO O OO H 2-Hydroxyibuprofen2O-HydroxyibuprofenO OH O O N O O O O OH OH OH O O OO OHOOHH O OO Ibuprofen OHOOHH Carboxyibuprofen O 2-HydroxyibuprofenN NN O O O OO O NaproxenO OO OH OKetoprofen OHOOHH Indoprofen OHOOHH O Naproxen OHOH Ketoprofen O NIndoprofenNN O O O OOH OH O O OO O H N N O OOHO OHH O OO Naproxen O Ketoprofen O OO OIndoprofenN Cl OOHOO O OOOH OOHH OCl OO OOHOO N OOHOOOH NaproxenNaproxenNaproxen O KetoprofenKetoprofenKetoprofen OH IndoprofenIndoprofenIndoprofen Cl NaproxenO O KetoprofenOH IndoprofenOH O NaproxenNaproxenNaproxen OH KetoprofenKetoprofenKetoprofen O IndoprofenIndoprofenIndoprofen O OH OOOH O OOH Cl ClCl NaproxenNaproxenNaproxenN KetoprofenKetoprofenKetoprofen O IndoprofenIndoprofenIndoprofen N H O H OH O F O OHOOHH Cl Cl Cl F O O O OHOOHH NaproxenN Ketoprofen O Indoprofen OHOOHH Cl ClCl H O O OHOOOHOH OH F O OO Carprofen Fenoprofen OHOH FlurbiprofenO H OOHH CarprofenN Fenoprofen O OO Flurbiprofen N N OOH OOHOH OHOOHH O OO H H H O O Cl O O F FF CarprofenN N N OH OOHH Fenoprofen O O Flurbiprofen OHOO H H H O OO O BronchodilatorsBronchodilatorsO HO F FF N NN O O CarprofenH HH O OO Fenoprofen FlurbiprofenF F CarprofenCarprofenCarprofen O H FenoprofenFenoprofenFenoprofen FlurbiprofenFlurbiprofenFlurbiprofenF O OH Bronchodilators NCarprofenCarprofenCarprofenOH FenoprofenBronchodilatorsFenoprofenFenoprofen O FlurbiprofenFlurbiprofenFlurbiprofen H H H O F CarprofenCarprofenCarprofen N N BronchodilatorsFenoprofenBronchodilatorsFenoprofenBronchodilatorsFenoprofen FlurbiprofenFlurbiprofenFlurbiprofen HOHO OH H BronchodilatorsBronchodilatorsBronchodilators N HO HCarprofenO OHOOHH FenoprofenBronchodilatorsBronchodilators Flurbiprofen HO H HH Bronchodilators HO OHOOHNH NN HOHOHO H HH Bronchodilators SalbutamolSalbutamol O(SBT)H (SBT)OOH NH NN HOHHOO H HH HO N N HOHHOSalbutamolSalbutamolOHO (SBT) (SBT) N OH Antibiotics HOHHOO H AntibioticsAntibiotics HO HOSalbutamolHSalbutamolOSalbutamol (SBT) (SBT)N (SBT) HO O O AntibioticsO O SalbutamolSalbutamolSalbutamolOH (SBT)OHO (SBT)H (SBT)O H F F HO Salbutamol (SBT) OH SalbutamolSalbutamol (SBT) (SBT) C l AntibioticsAntibiotics Antibiotics O O OH OH OH Cl O HN HN F AntibioticsAntibioticsAntibiotics O N Cl N NO OHO SalbutamolN (SBT) Cl N NO O O O OH OOHOHCHlOOHH AntibioticsAntibioticsAntibiotics O O N F O O O O O HN O N F F O O O O O N OH OOHH OCOHl H N N OH OHO H Cl Cl O O Cl F F F O O O O O OH OOOHH OH Antibiotics HNOHHHNNOH N O OH OHOH O OOChloranfenicolCl C lCl F N F OfloxacinN N NNChloranfenicolC l CCl l N F NOfloxacinN N Chloranfenicol Ofloxacin OH OHOH HN HNHNCl CCl l O O O OOOOO O OO N N N N O N N N N NN OH OH Cl ClC l N N O O HN HHNN F O OChloranfenicolO OfloxacinN N ON ONNO O OOCl CCl l N N NN N O O ON N OH Cl O O O O N O O O N N O O O ChloranfenicolChloranfenicolChloranfenicolHN OfloxacinOfloxacinOfloxacin N Cl N N O ChloranfenicolChloranfenicolChloranfenicolO OfloxacinOfloxacinOfloxacin N O ChloranfenicolChloranfenicolChloranfenicol OfloxacinOfloxacinOfloxacin
Chloranfenicol Ofloxacin
Molecules 2018, 23, 262 7 of 47
Table 1. Cont. Molecules 2018, 23, × FOR PEER REVIEW 9 of 51
Chiral Compounds MoleculesMolecules 2018 2018, 23, 23, ×, ×FOR FOR PEE PEER RREVIEW REVIEWAntiepileptic and their metabolites 1010 of of 51 51 Antiepileptic and their metabolites MoleculesMoleculesMolecules 2018 2018, 232018, ,× 23 ,FOR 23, ×, ×FOR PEE FOR RPEE PEEREVIEWRR REVIEW REVIEW 10 of10 5110 of of 51 51 OH MoleculesMolecules 2018 2018, 23,, ×23 FOR, × FOR PEE PEER REVIEWR REVIEW TableTable 1 .1 Cont.. Cont. 10 of10 51 of 51 Molecules 2018, 23, × FOR PEER REVIEW TableTableTable 1. Cont. 1 .1 Cont.. Cont. 10 of 51 ProtonProton pump pump inhibitors inhibitors Molecules 2018, 23, × FORMolecules PEER 2018REVIEW, 23, × FOR PEER REVIEW TableTable 1. Cont. 1. Cont. 10 of 51 10 of 51 N Table 1Proton. Cont.ProtonProton pump pump pump inhibitors inhibitors inhibitors H H Molecules 2018, 23, × FOR PEER REVIEW H N 7 of 48 H N O O N N O H H O H H H N S N O NH2N O O ProtonHProtonH pumpH pump inhibitors inhibitors N N N S N N N S N O O O H H H Table 1. Cont.N TableO SO O1. Cont.N N N N O S SO O N N ZAIZGHELI N SN S S N N N N SN S S N N N Table 1. Cont. H H 10,11-Dihydro-10-hydroxycarbamazepineN S S S N N N H H NN N N O N O O 10,11O -DihydroH H -10- NN N N O O H Molecules 2018, 23, × FOR PEER REVIEW ZAIZGHELIProton pumpH inhibitorsNF F 7 of 48 NN N N O O N O S S N N O O OhydroxycarbamazepineO O H ProtonProtonS pumpS N inhibitorspumpN inhibitors N O O OF F F S N S S N N F F N N O O O N N H H O O O N H S N H H H NF O NN O N N N N F F F F O O O O H H H Table 1N. Cont. O N O O N N N S N S N N O O O F F F F F S N O O O S N S O O O S N S N O O N ONN S N F F OmeprazolOmeprazol Proton pumpN Lanzoprazol inhibitorsLanzoprazolN N RabeprazoleRabeprazole N N N F F H O O O O H N O O OmeprazolOmeprazolOmeprazol LanzoprazolLanzoprazolNLanzoprazol ORabeprazoleRabeprazoleRabeprazole H O F O OO N OH H S N O N N O O S N O F F F HS H H N O O N OmeprazolOmeprazolN N N O S SO O LanzoprazolLanzoprazolN RabeprazoleRabeprazole F F F F O N Lanzop OO O N NS S S O OOmeprazol Rabeprazole F F H H razol O NN N N O O F OF OOmeprazolO Omeprazol Lanzoprazol LanzoprazolO Rabeprazole F F F Omeprazol H LanzopLanzoprazol Rabeprazole Omeprazol S S N O O Rabeprazole F H O O razol N N N O S N O F O O F F H O O O N O OmeprazolS N N Lanzoprazol Rabeprazole F O FPantoprazoleHPantoprazoleH N NN O O S F F O F PantoprazolePantoprazolePantoprazole O O S S N N N O F O O N N O O AntineoplasticAntineoplastic agents agents and and their their metabolite metabolite F F O O PantoprazolePantoprazole Pantoprazole AntineoplasticO AntineoplasticAntineoplastic agents agents agents and and theirand their their metabolite metabolite metabolite Pantoprazole Antineoplastic agents and their metabolite Pantoprazole O O O O AntineoplasticAntineoplastic agents agents and and their their metabolite metabolite PantoprazolePantoprazole Antineoplastic agents and their metabolite Pantoprazole AntineoplasticNN AntineoplasticOO agents andagents their andHNH metaboliteN their metaboliteOO N O HN O ClCl N NN POP OO AntineoplasticHN H HNagentsN POP andOO their metabolite Cl P P Cl ClCl AntineoplasticHNHPN PP agents and their HmetaboliteNHPN PP HN HN HNN HNHN OO O HHNN HHNN O O N O HN OO Cl Cl P P O P P O Cl OP OO OP OO HN NHN O HN HHNN HN HN HN P O Cl N OP O O O O Cl HN ClCOPl HNHN Cl PO N O Cl Cl CHlN ClCl Cl Cl P O CHlN CPCl l O 3‐N‐ IfosfamideIfosfamideHN O 3-3N-N-Dechloroethylifosfamide-DechloroethylifosfamideHN O Cl Cl Ifosfamide Cl Cl Dechloroethylifo IfosfamideIfosfamideIfosfamide ClO 3-N-3Dechloroethylifosfamide-3N-N-Dechloroethylifosfamide-DechloroethylifosfamideCl O sfamide IfosfamideCl 3-N-DechloroethylifosfamideAntifungals3Antifungals‐N‐ IfosfamideIfosfamide 3-N3-Dechloroethylifosfamide-N-DechloroethylifosfamideCl Antifungals Ifosfamide Cl DechloroethylifosfamidAntifungalsAntifungalsAntifungals N AntifungalsCl Cl N N N N N Cle N N N N 3-N- N N N N AntifungalsAntifungalsN N HO N N Cl N N N N N Ifosfamide 3Antifungals-N-DechloroethylifosfamideDechlo r HOHO Cl Cl N N N N N N N N N N N 3-N- N HO NHOHO Cl Cl CIfosfamidel N N oethylifN N N N O O N Dechlor N N NN N N O O osfamidAntifungalsO O NHO N N Cl N N N N N HO HO Cl IfosfamideCl O Cl O O oethylifO O ClO Cl Cl e Cl N Cl Cl N N Cl N Cl N Cl Cl Cl Cl osfamidCl Cl Cl Cl Cl Cl N N O Econazole O Miconazole N Tebuconazole Cl Cl Cl Cl Cl Cl Cl Cl Cl O O Cl Cl Cl Cl OCl CCll O Cl Cl Cl Cl Cl N N e N Antifungals O HO Cl Cl Econazole Cl Miconazole TebuconazoleC l Econazole Cl MiconazoleCl Cl N TebuconazoleN Cl Cl EconazoleCl Cl MiconazoleN Tebuconazole Cl Cl Cl Cl Cl Cl Cl Cl Cl Cl N EconazoleEconazoleEconazoleEconazole N MiconazoleMiconazoleCHMiconazole3Miconazole TebuconazoleTebuconazoleTebuconazoleTebuconazole N N H O Antifungals O N N N O N N N N N N N EconazoleEconazole MiconazoleMiconazoleN TebuconazoleTebuconazoleN N Cl N Cl N N HO N Cl Cl Cl Cl Cl Cl Cl Cl N CH3 N HON N N N N N N N N O N N H H H N O Cl N N HO N N N Cl O N N N N H H H Econazole Miconazole N Tebuconazole O N N N N Cl Cl Cl Cl Cl Cl N N N N N N N HOHO Cl O O Cl Cl N N Cl Cl Cl O Cl COOl Cl O Cl Cl Cl H H O HO HOHO Cl Cl Cl N O ONO O O O N NN N Cl ClCl Cl Cl O O O Cl N Cl N Cl Cl N Cl NCl N Cl Micona HO HO N EconazoleO O O Ketoconazole Cl Cl HexaconazoleTebuconazole Penconazole N O O Cl Cl H NO NO zole N Cl Cl N N N Cl ClCl Cl N ClClN Micona Cl Cl Cl EconazoleN O N ON Tebuconazole Cl N NCl Ketoconazole O HexaconazolezoleHO N PenconazoleC l ClCl O H KetoconazoleO Cl Cl HexaconazoleCl CNl Penconazole KetoconazoleKetoconazole HexaconazoleHexaconazole ClPenconazolePenconazole N N O O Cl Cl KetoconazoleKetoconazoleKetoconazoleO HexaconazoleHexaconazoleHexaconazole PenconazolePenconazolePenconazole N O N H N Cl Cl Cl KetoconazoleKetoconazoleO HexaconazoleHexaconazole PenconazolePenconazole N Cl N O Cl N N Cl Cl KetoconazoleN Hexaconazole Penconazole Cl
N Triadimefon Imazalil
Cl Triadimefon Imazalil
Molecules 2018, 23, 262 8 of 47
Table 1. Cont. MoleculesMolecules 2018 2018, 23,, ×23 FOR, × FOR PEE PEER REVIEWR REVIEW 11 of11 51 of 51 Chiral Compounds
N N
O O N N H H O O O O
N N Cl Cl Cl Cl N N MoleculesMoleculesMolecules 2018 2018, 201823,, ×23, FOR23, ×, ×FOR PEEFOR PEER PEE REVIEWRR REVIEW REVIEW 12 of12 5112 of of 51 51 N N Molecules 2018, 23, × FOR PEER REVIEW 8 of 47 TableTableTableC l1. CCont. l1 .1 Cont.. Cont. TriadimefonTable 1. Cont. Imazalil TriadimefonTriadimefon AntiImazalilAntiAnti-helminthicImazalil-helminthic-helminthic Anti‐helminthicAnti-helminthic O O O O
N S N N N N N S S S N N N N N N N N N N N N N N N S N N N S S S O O O O
Phenyltetrahydroimidazo Phenyltetrahydroimidazo thiazole PraziquantelPraziquantel Tetramisole Tetramisole thiazolePhenyltetrahydroimidazoPhenyltetrahydroimidazoPhenyltetrahydroimidazo (PTHIT, (PTHIT, levamisole/dexamisole) PraziquantelPraziquantelPraziquantel TetramisoleTetramisoleTetramisole levamisole/dexamisole)thiazolethiazolethiazole (PTHIT, (PTHIT, (PTHIT, AntihistaminicAntihistaminic levamisole/dexamisole)levamisole/dexamisole)levamisole/dexamisole)
O
OH AntihistaminicAntihistaminicAntihistaminic
N OH
OH N N N OH OHOH
FexofenadineOFexofenadineH OHOH
3. Chiral Analyses in Biological Samples 3. Chiral Analyses in Biological Samples FexofenadineFexofenadineFexofenadine This study reviewed 58 articles that have been published between 1996 and 2017 based on ScienceDirectThis study and ISI reviewed Web of 58 Knowledge articles that databases. have been The published investigated between compounds 1996 andincluded 2017 based on syntheticScienceDirect3. Chiral3. 3. Chiralpsychoactive Chiral Analyses and Analyses Analyses ISI in Webdrugs Biologicalin in Biological of Biological Knowledge(stimulants), Samples Samples Samples databases.synthetic Theopioids, investigated β‐blockers, compounds antidepressants, included synthetic anticoagulants,psychoactive bronchodilators drugs (stimulants), and dissociative synthetic anesthetics opioids, (Tableβ-blockers, 1 and antidepressants,2). Figure 2 shows anticoagulants,the ThisThisThis study study study reviewed reviewed reviewed 58 58articles 58 articles articles that that thathave have have been been been published published published between between between 1996 1996 1996 and and and2017 2017 2017 based based based on on on relative number of studies of each class of chiral drug investigated and the analytical methods used bronchodilatorsScienceDirectScienceDirectScienceDirect andand and and dissociativeISI ISIWebISI WebWeb of anestheticsofKnowledgeof KnowledgeKnowledge (Tablesdatabases. databases.databases.1 and The2 ). The The Figureinvestigated investigatedinvestigated2 shows compounds thecompoundscompounds relative included number includedincluded of for analysis of these compounds in different biological matrices. studiessyntheticsyntheticsynthetic of each psychoactive class psychoactive psychoactive of chiral drugs drug drugs drugs (stimulants),investigated (stimulants), (stimulants), and synthetic synthetic syntheticthe analytical opioids, opioids, opioids, methods β -blockers, β β-blockers,-blockers, used antidepressants, for antidepressants,antidepressants, analysis of these compoundsanticoagulants,anticoagulants,anticoagulants, in different bronchodilators bronchodilators bronchodilators biological and matrices. and anddissociative di dissociativessociative anesthetics anesthetics anesthetics (Table (Table (Table 1 and 1 1and and2). 2 Figure).2). Figure Figure 2 shows 2 2shows shows the the the relativeTherelativerelativeuse number number number of of racemates studiesof of studies studies of eachtypicallyof of each each class class class of results chiralof of chiral chiral drug in drug drug stereoselectiveinvestigated investigated investigated and and andthe pharmacological the analyticalthe analytical analytical methods methods methods activity used used used and pharmacokineticfor foranalysisfor analysis analysis of theseof affectingof these these compounds compounds compounds bioavailability, in differentin in different different biological metabolism biological biological matrices. matrices. andmatrices. 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 racemicFigure or2. singleRelative enantiomers number of each depending class of chiral on thedrug manufacturing referred in the reviewed procedure, enantioselective i.e, racemic or enantiomer purepublished precursors. studies Thus, and the in analytical forensic methods chemistry, used evaluation for separation of theof the EF chiral may drugs aid in in thebiological discrimination of the consumptionfluids. of legal and illegal substances, give information about method of synthesis used or profile
FigureFigureFigure 2. Relative2 .2 .Relative Relative number number number of eachof of each eachclass class classof chiralof of chiral chiral drug drug drug referred referred referred in thein in the reviewedthe reviewed reviewed enantioselective enantioselective enantioselective The use of racemates typically results in stereoselective pharmacological activity and publishedpublishedpublished studies studies studies and and theand theanalytical the analytical analytical methods methods methods used used used for forseparation for separation separation of theof of thechiral the chiral chiral drugs drugs drugs in biologicalin in biological 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 Theuse use useof ofracematesof racematesracemates typically typicallytypically results resultsresults in instereoselectivein stereoselectivestereoselective pha pharmacologicalpharmacologicalrmacological activity activityactivity and and and racemates [57]. Concerning illicit drugs, these compounds are also available as racemates or single pharmacokineticpharmacokineticpharmacokinetic affecting affecting affecting bioavailability, bioavailability, bioavailability, metabolism metabolism metabolism and and andexcretion excretion excretion that that thatmay may may contribute contribute contribute to theto to the the enantiomers depending on the manufacturing procedure [5,6,57]. Regarding illicit administrations, toxicity,toxicity,toxicity, increase increase increase risk risk riskof deathof of death death or seriousor or serious serious adverse adverse adverse effects effects effects [11,56,57]. [11,56,57]. [11,56,57]. Though Though Though there there there is ais tendencyis a atendency tendency for for for consumption of pure enantiomer (eutomers), in some cases, may cause overdose or even might lead
Molecules 2018, 23, × FOR PEER REVIEW 8 of 47
Table 1. Cont.
Anti-helminthic O
N S N N
N N N S O
Phenyltetrahydroimidazo Praziquantel Tetramisole thiazole (PTHIT, levamisole/dexamisole) Antihistaminic
N OH
OH
Fexofenadine
3. Chiral Analyses in Biological Samples
MoleculesThis2018 study, 23, 262 reviewed 58 articles that have been published between 1996 and 2017 based9 of on 47 ScienceDirect and ISI Web of Knowledge databases. The investigated compounds included synthetic psychoactive drugs (stimulants), synthetic opioids, β-blockers, antidepressants, amonganticoagulants, different bronchodilators seizures linking amongand dissociative them, consumers anesthetics and traffickers.(Table 1 and Chiral 2). Figure analysis 2 canshows also the be appliedrelative innumber doping of control,studies asof aneach example, class of thechiral use drug of preparations investigated containing and the analytical dextromethorphan methods used by athletesfor analysis is allowed, of these whereas compounds the use in different of levorphanol biological is expressly matrices. banned by the International Olympic Committee [61].
Figure 2.2. Relative number of each class of chiral drugdrug referred inin thethe reviewedreviewed enantioselectiveenantioselective 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 pharmaceuticalsThe use of and racemates illicit drugs typically of forensic results interest in in biologicalstereoselective matrices pharmacological most works do notactivity determine and thepharmacokinetic enantiomeric affecting composition. bioavailability, Data about metabolism the enantiomeric and excretion composition that may of parent contribute compounds to the andtoxicity, metabolites increase isrisk of highof death importance or serious for adverse accurate effects data [11,56,57]. interpretation Though and there for further is a tendency analysis for of resultsmanufacturing [62]. Also, pharmaceuticals metabolites of as achiral single compounds enantiomers, can many also bepharmaceuticals chiral and should are bestill considered supplied inas biologicalracemates samples.[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 lethalThis cases class [10,12]. of chiral Thus, drugs significance was the most of chiral studied analysis (Tables has1 andincreased2 and Figuresince it2 ).is possible 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) (Tables1 and2). 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 consumersMolecules 2018 [,59 23]., × Besides,FOR PEER it REVIEW was also shown that the existence of DMSG in SG users that is not normally10 of 47 found in urine demonstrating that the method can be useful for distinguish therapeutic users of SG and MAusers abusers. of SG and Fujii MA et al. abusers. described Fu ajii GC-MS et al. methoddescribed based a GC-MS on the meth formationod based of diastereoisomers on the formation using of CDRdiastereoisomers TPC for the separationusing CDR of TPC the enantiomersfor the separation of MA, of AM, the MDMAenantiomers and MDAof MA, in AM, urine MDMA samples and [8]. ThisMDA method in urine can samples be used [8]. for This discrimination method can between be used legal for and discrimination illegal consumption between of legal these and drugs illegal [8]. Rasmussenconsumption et al.of developedthese drugs for [8]. the firstRasmussen time a GC-MS et al. enantioselectivedeveloped for methodthe first for time the separationa GC-MS ofenantioselective AM, MA, MDA, method MDMA for and the MDEAseparation in whole of AM, blood MA, based MDA, in MDMA the formation and MDEA of diastereomers in whole blood [67] (Figurebased in3). the formation of diastereomers [67] (Figure 3).
Figure 3. ChromatogramChromatogram representing representing the the enantioseparation enantioseparation of ofR/SR-AM,/S-AM, R/SR-MA,/S-MA, R/S-MDA,R/S-MDA, R/S- RMDMA/S-MDMA and andR/S-MDEAR/S-MDEA as R-MTPCl as R-MTPCl derivatives derivatives in whole in whole blood blood concentrations concentrations at ( atA) ( A2µg/g) 2 µg/g and and (B) (atB )0.002 at 0.002 µg/g,µg/g, respectively. respectively. Reproduction Reproduction with with perm permissionission of Elsevier of Elsevier (Fig (Figureure 1 from 1 from Rasmussen Rasmussen et etal. 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 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 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 3.2. Synthetic Opioids The second most studied classe of chiral compounds are the synthetic opioids (Tables1 and2). 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 (Tables1 and2). 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 (Tables1 and2)[ 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 (Tables1 and2)[ 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 5% phenyl-methylsilicone capillary column (17 m × 0.2 Hair 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. Matrix Concentration Range/ER Drug Method Stationary Phase LOD LOQ Reference Application (When Mencioned) 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] 45–1000 (l; d) GC/EI-MS HP-5MS (30 m × 0.2 mm, 0.33 µm film thickness) 40 ng/mL 45 ng/mL [70] MA 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. Matrix Concentration Range/ER Drug Method Stationary Phase LOD LOQ Reference Application (When Mencioned) 5% phenyl-methylsilicone capillary column (17 m × 0.2 GC/MS 0.2 ng/mg 0.5 ng/mg 0.5–20 ng/mg [69] mm, 0.33 µm film thickness) 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 5% phenyl polysiloxane (15 m × 0.25 mm, 0.2 µm df) n.r. n.r. 25–10000 ng/mL [116] 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) 1.6 pg/mg (R) 5.3 pg/mg (R) 0.005–60 ng/mg (R) GC/NICI-MS HP-5MS (30 m × 0.25 mm, 0.25 µm film thickness) [49] 1.3 pg/mg (S) 4.3 pg/mg (S) 0.004–60 ng/mg (S) Hair MDA 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, 0.33 µm film thickness) 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. 2 µg/L 2–100 µg/L [42] Urine GC/EI-MS 5% phenyl polysiloxane (15 m × 0.25 mm, 0.2 µm df) n.r. n.r. 25–10000 ng/mL [116] 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] 2.7 pg/mg (R) 8.9 pg/mg (R) 0.009–60 ng/mg (R) GC/NICI-MS HP-5MS (30 m × 0.25 mm, 0.25 µm film thickness) [49] 2.3 pg/mg (S) 7.7 pg/mg (S) 0.008–60 ng/mg (S) MDEA Hair 5% phenyl-methylsilicone capillary column (17 m × 0.2 GC/MS 0.2 ng/mg 0.5 ng/mg 0.5–20 ng/mg [69] 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) GC/NICI-MS n.r. (chiral derivatization S-HFBPrCl) n.r. n.r. n.r. [68] × µ pOHMA LC/HRMS Chirex3012 (250 4.6 mm, 5 m 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. Matrix Concentration Range/ER Drug Method Stationary Phase LOD LOQ Reference Application (When Mencioned) 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] Chiral-AGP (100 × 4.0 mm, 5 µm); Chiral-AGP guard 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] Chiral-AGP (100 × 4.0 mm, 5 µm); Chiral-AGP guard 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 (+) HPLC-DAD 1 ng/mL 1–500 ng/mL [124] T 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 n.r. 2.5 ng/mL 2.5–250 ng/mL [126] HPLC-FD LiChrospher 100 Diol (5 µm) precolumn Chiral-AGP (150 × 4.0 mm, 5 µm); Chiral-AGP guard 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. Matrix Concentration Range/ER Drug Method Stationary Phase LOD LOQ Reference Application (When Mencioned) 0.1 ng/mL (total) 0.1–300 ng/mL (total) LC/ESI-MS/MS Chiralpak AD (250 × 4.6 mm, 10 µm) n.r. [81] 0.25 ng/mL (unbound) 0.25–125 ng/mL (unbound) 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 Chiralcel OD-R (250 × 4.6 mm, 10 µm); LiChrospher 0.08 ng/mL (+) ODT HPLC-DAD 0.5 ng/mL 0.5–100 ng/mL [124] 60-RP-selected B (250 × 4 mm, 5 µm) 0.06 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 LiChrospher n.r. 2.5 ng/mL 2.5–250 ng/mL [126] HPLC-FD 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] 0.1 ng/mL (total) 0.1–300 ng/mL (total) LC/ESI-MS/MS Chiralpak AD (250 × 4.6 mm, 10 µm) n.r. [81] 0.25 ng/mL (unbound) 0.25–125 ng/mL (unbound) Chiralcel OD-R (250 × 4.6 mm, 10 µm); LiChrospher 0.15 ng/mL (+) Plasma HPLC-DAD 0.5 ng/mL 0.5–250 ng/mL [124] NDT 60-RP-selected B (250 × 4 mm, 5 µm) 0.16 ng/mL (-) Chiral-AGP (150 × 4.0 mm, 5 µm); Chiral-AGP guard column HPLC-FD n.r. 2.5 ng/mL 2.5–75 ng/mL [127] (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] Chiral-AGP (100 × 4.0 mm, 5 µm); Chiral-AGP guard column 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] Chiral-AGP (100 × 4.0 mm, 5 µm); Chiral-AGP guard column 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. Matrix Concentration Range/ER Drug Method Stationary Phase LOD LOQ Reference Application (When Mencioned) 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) HPLC/ESI-MS Chirobiotic V (250 × 4.6 mm, 5 µm) 1 ng/mL 5–400 ng/mL [133] VNF 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 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 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 Whole blood LC/ESI-MS/MS Chirobiotic V (250 × 2.1 mm, 5 µm) n.r. 0.25 nM [95] 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 Whole blood LC/ESI-MS/MS Chirobiotic V (250 × 2.1 mm, 5 µm) n.r. 0.25 nM [95] 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) Chirobiotic V (250 × 4.6 mm, 5 µm); Cyclobond I guard 5–1500 ng/mL Plasma LC/ESI-MS/MS 1.5 ng/mL 5 ng/mL [105] WFN 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. Matrix Concentration Range/ER Drug Method Stationary Phase LOD LOQ Reference Application (When Mencioned) 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 Molecules 2018, 23, × FOR PEER REVIEW 22 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 (Table3 and Figure4). The target compounds ScienceDirect and ISI web of Knowledge databases (Table 3 and Figure 4). The target compounds β included antidepressants,antidepressants, β-blockers, nonsteroidalnonsteroidal anti-inflamatoryanti-inflamatory drugsdrugs (NSAIDs),(NSAIDs), syntheticsynthetic psychoactive drugs, antibiotics,antibiotics, syntheticsynthetic opioids,opioids, antiepileptics,antiepileptics, antihistaminic;antihistaminic; bronchodilators,bronchodilators, antineoplastic agents agents and and proton proton pump pump inhibitors inhibitors (Table (Table 1). Figure1). Figure 4 shows4 shows the relative the relative number number of studies of ofstudies each class of each of chiral class ofdrug chiral investigated drug investigated and the analytical and the analyticalmethods used methods for analysis used for of analysisthese compounds of these incompounds different environmental in 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 effluentsThe from entrance WWTPs of pharmaceuticals which are unable and to illicit totally drugs remo intove these the aquatic micro-pollutants environment or maydirect occur discharged through of sewage.effluents WWTPs from WWTPs biological which treatments are unable, can to totallyalter the remove EF of thesethe enantiomers micro-pollutants present or directin the dischargedinfluent, as microbiotaof sewage. action WWTPs generally biological is stereoselective treatments, can [136,137]. alter the Disposed EF of the enantiomersdrugs will usually present be in found the influent, in their parentas microbiota form, either action as racemate generally or is single stereoselective enantiomers. [136 On,137 the]. Disposedother hand, drugs excreted will drugs usually will be be found normally in foundtheir parent as metabolites, form, either frequently as racemate chiral, or of single the parent enantiomers. compound On [138]. the other hand, excreted drugs will be normallyThe possible found as adverse metabolites, effects frequently of the enantiomers chiral, of theon parentaquaticcompound and human [138 life]. lead to the studies of occurrenceThe possible of chiral adverse drugs in effects environmental of the enantiomers matrices [20,136,137,139,140]. on aquatic and human Illicit drugs life leadcan also to the be studiesfound in environmentalof occurrence ofsamples, chiral drugsand environmental in environmental data are matrices important [20,136 resources,137,139 for,140 a]. forensic Illicit drugs approach. can alsoThis includesbe found the in environmentalusage of environmental samples, data and in environmental order to: (1) verify dataare patterns important of illicit resources and prescribed for a forensic 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 estimationmetabolic patterns of the consumption of most available of substances drugs are of understood, abuse and illicitit is assumed drugs can that be the measured amount by of drugthe concentrations or its metabolite of these quantified compounds in raw in wastewater.sewage will Drugs correspond are consumed with the and consumed metabolised dose—sewage in human epidemiologybody and excreted [139]. asThe parent application compounds of the or chiral as metabolites, discrimination and finallyhas also reach been WWTPs used for through distinction the betweensewage [legal139]. and Since illicit the use metabolic of drugs, patterns verification of most of th availablee method drugs of synthesis are understood, of illicit drugs, it is assumed identification that thatthe amountdrug residue of drug results or its metaboliteeither from quantified consumption in raw of sewageillicit drugs will correspondor metabolism with of the other consumed drugs, verificationdose—sewage of route epidemiology of administrati [139].on, The verification application of of potency the chiral of discriminationabused drugs, monitori has alsong been of usedchanging for 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, Molecules 2018, 23, × FOR PEER REVIEW 24 of 47 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 weeklyHashim, pattern N.of increased H. & Khan, daily S. J.loads, studied during the EFofweekends ibuprofen, [148]. naproxen and ketoprofen in wastewater samples,Hashim, taken N. H. from & Khan, a WWTPs S. J. instudied Sydney the (Australia)EFof ibuprofen, with naproxen tertiary treatment and ketoprofen over seven in wastewater separate samples,sampling taken events, from duringa WWTPs June in Sydney and August (Australia) 2010 wi [147th tertiary]. For ibuprofen,treatment over EF seven ranged separate from 0.49sampling and events,0.62; 0.66 during and June 0.86 and for August naproxen 2010 and [147]. 0.54 For and ibuprofe 0.66 forn, ketoprofenEF ranged from [147 0.49]. Also and Barreiro 0.62; 0.66 et and al in0.86 2010, for naproxenfound for and the 0.54 first and time 0.66 the for occurrence ketoprofen of (+)-omeprazole[147]. Also Barreiro and et (− al)-omeprazole, in 2010, found while for the simultaneously first time the occurrencedeveloping of a(+)-omeprazole column switching, and (-)-omeprazole, liquid chromatography while simultaneously method for developing the chiral a separation column switching, of these liquiddrugs, chromatography in wastewater andmethod estuarine for the samples chiral separa [150].tion Ribeiro of these et al studied,drugs, in the wastewater EF of FLX, and NFLX, estuarine VNF, samplesSBT, alprenolol, [150]. Ribeiro MET, et PHO al studied, and bisoprolol the EF (BSPof FLX, = from NFLX, the finalVNF, effluentSBT, alprenolol, of the secondary MET, PHO clarifier and bisoprololof three WWTPs(BSP= from located the final in theeffluent North of ofthe Portugal, secondary Figure clarifier5[ of39 three]. Regarding WWTPs located antidepressants, in the North only of Portugal,R-FLX was Figure detected 5 [39]. in twoRegarding of the WWTPs,antidepressants, indicating only a faster R-FLX degradation was detected of the in Stwo-enantiomer of the WWTPs, during indicatingthe biological a faster degradation degradation [39]. of VNF the enantiomersS-enantiomer were during found the betweenbiological 40.4 degradation and 129 ng/L [39].in VNF the enantiomersthree WWTPs were studied, found withbetween similar 40.4 EF,and which 129 ng/L varied in the between three WWTPs 0.54 and studied, 0.55, proving with similar that VNF EF, foundwhich variedwas not between racemic 0.54 [39 and]. Concerning 0.55, proving the thatβ-blockers VNF found enantiomers, was not racemic BSP and [39]. PHO Concerning were found the inβ-blockers all three enantiomers,BSPWWTPs, while METand PHO was were only found in in all two, three however WWTPs, all while of them MET under was theonly medium found in quantification two, however all of them under the medium quantification level (MQL) [39]. level (MQL) [39]. FigureFigure 5. 5. ChromatogramChromatogram and and mass mass spectra spectra of of WWTP WWTP effluent effluent sample sample showing the enantiomers of FLX,FLX, VNF,VNF, BSP, BSP, MET MET and and PHO. PHO. Reproduction Reproduction with with permission permission of Elsevier of Elsevier (Figure (Figure 2 from 2 from Ribeiro Ribeiro et al. et [39]). 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 [17] Wastewater UPLC/ESI-MS/MS 5 µm); Chiral-CBH guard n.r. 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); × EF: 0.7 (IW); 0.9 (EW); 0.4 AM, MA, MDMA, Chiral-CBH (100 2 mm, 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 ng/L (EW); 2.21/4.14 ng/L (DS) ng/L (EW); 7.46/13.74 ng/L (DS) Atenolol, wastewater LC/ESI-MS/MS (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 0.025–250 µg/L [62] 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 Drugs Method Stationary Phase LOD/MDL LOQ/MQL Concentration Range/EF Reference 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 MDA: 1.95/2 ng/L (E1/E2) (IW); 2 ng/L 0.25–1900 ng/L MDA, MDEA, 5 µm); Chiral-CBH guard MDA: 9.7 ng/L (IW); 10.1 ng/L (EW) WWTP (EW) EF n.r. Norephedrine, effluent (EW) column (10 × 2 mm) MDEA: 2.25/2.2 ng/L (E1/E2) (IW); 2.4 0.25–1900 ng/L VNF 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 Drugs Method Stationary Phase LOD/MDL LOQ/MQL Concentration Range/EF Reference 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 0.51) (RW after WWTP) 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 0.45) (RW after WWTP) 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. (RW) ng/L (RW) AM, MA, MDMA, Sewage Chirobiotic V (250 × 4.6 mm, 5 µm); Chirobiotic V MDMA (E1/E2): MDMA (E1/E2): MDA, Atenolol, effluent (SE); 5–500 ng/L LC/QTOF-MS guard column (20 × 40 22.8/21.8 ng/L (SE); 85.7/81.9 ng/L (SE); [151] PHO, MET, FLX, River water 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 Drugs Method Stationary Phase LOD/MDL LOQ/MQL Concentration Range/EF Reference 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, influent (IW); 5 µm) with a nitrile guard 25–1000 ng/mL Atenolol, PHO, HPLC/ESI-MS/MS PHO: 17 ng/L (IW); 4.4 ng/L (EW) n.r. [153] 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 Drugs Method Stationary Phase LOD/MDL LOQ/MQL Concentration Range/EF Reference 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 12.5–100 µg/mL Atenolol, PHO, Lux Cellulose-1 (250 × 4.6 mm, PHO: 3 µg/L 10 µg/L 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, µ Flurbiprofen FLX: 0.1 ng/L 0.5 ng/L 5–1000 g/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) FLX: 3 pM (RaW); 2/1 pM (R/S) 0–500 pM Raw wastewater Chiral-AGP (100 × 2 mm, 5 µm); in-line 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) Influent Carboxyibuprofen, 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 8.66–50 µg/L Ifosfamide, (IW); 14.4/13.4 ng/L (EW); (IW); 49.9/46.5 ng/L (EW); EF: 1 (IW) Praziquantel 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. Matrix Concentration Drugs Method Stationary Phase LOD/MDL LOQ/MQL Reference Application Range/EF 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); 0.1 µg/L GC/MS 0.25 mm, 0.12 µm film n.r. [44] 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); Indoprofen (E1/E2): 2.38/2.68 ng/L Indoprofen (E1/E2): 7.91/8.91 Carprofen, Fenoprofen, 0.85–500 µg/L (E2) Polysaccharide amylose (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 Carprofen (E1/E2): 378/287 ng/L Carprofen (E1/E2): 1259/956 ng/L 168–500 µg/L Aminorex, Tetramisole, UHPSFC/ESI-MS/MS [163] 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 Drugs Method Stationary Phase LOD/MDL LOQ/MQL Concentration Range/EF Reference 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 Indoprofen (E1/E2): 5.53/3.94 ng/L Indoprofen (E1/E2): 18.4/13.1 (E2) (IW); 6.82/5.52 ng/L (EW) ng/L (IW); 22.7/18.4 ng/L (EW) 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) (IW); 6.80/5.30 ng/L (EW) ng/L (IW); 22.60/17.60 ng/L (EW) 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 µ 0.08–300 ng/L; EF: 0.66–0.86 (mean [147] wastewater 0.25 m 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 Drugs Method Stationary Phase LOD/MDL LOQ/MQL Concentration Range/EF Reference Application Influent HP5-MS (30 m × 0.25 Ibuprofen EF: 0.6–0.8 (IW); 0.5 (EW) Ibuprofen, wastewater (IW); GC/EI-MS/MS mm, 0.25 µm film n.r. n.r. [164] Naproxen Effluent thickness) 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 × µ Naproxen: 1.2/1.1 ng/L (R/S) (IW); 1.1 Naproxen, wastewater (IW); LC/MS/MS (250 4.6 mm, 5 m); n.r. EF: 0.98–0.99 (IW); 0.93–0.96 [165] ng/L (EW) Ketoprofen Effluent Chirex 3005 guard (EW) wastewater (EW) column (30 × 4.6 mm, 1–4000 µg/L 5 µm) 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: 0.52 ng/L (EW); 0.26/0.27 ng/L Ketoprofen (R/S): 1.73/1.70 ng/L Ketoprofen, µg/L (S) (R/S) (SW) (EW); 0.86/0.88 ng/L (SW) Chloramphenicol, 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 Chiral-AGP (100 × 2 7.39/8.09 ng/L (EW); 3.46/3.96 ng/L µg/L (1S,2S) Ifosfamide, LC/ESI-MS/MS ng/L (EW); 1.02/1.19 ng/L (SW) wastewater (EW); mm, 5 µm); Chiral-AGP (SW) EF n.r. 3-N-Dechloroethy Surface water (SW) guard column (10 × 2 [32] 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. Matrix Concentration Drugs Method Stationary Phase LOD/MDL LOQ/MQL Reference Application Range/EF 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); [43] × ng/L (RW) ng/L (RW) RW) Rabeprazole, Effluent LC/ESI-MS/MS Chiralpak IC (250 4.6 mm, 5 µm) Rabeprazole: 0.94/0.95 ng/L (R/S) (IW); Rabeprazole (R/S): 3.37/3.40 ng/L Pantoprazole wastewater (EW); 2–500 µg/L 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 River water (RW) 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 n.r. EF: 0.5 (RaW); 0.47 (TW); Econazole, Raw wastewater AGP column (100 × 4 mm, 5 µm); (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 0.5–250 ng/mL Ketoconazole Sludge (Sd) (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. Matrix Concentration Drugs Method Stationary Phase LOD/MDL LOQ/MQL Reference Application Range/EF 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, AGP guard column (10 × 4 mm) Miconazole: 0.6 ng/L (RW); 3ng/g (Sd) EF: 0.49–0.54 (RW); River water (RW); LC/ESI-MS/MS [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, River water LC/ESI-MS/MS Hexaconazole: 9.1 µg/L (−); 8.6 µg/L EF n.r. 5 µm) 27.7 µg/L (−); 25.8 µg/L (+) Penconazole, (+) [168] 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. 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