P54

SUPRAMOLECULAR RECEPTORS IN SOLID PHASE FOR ANIONIC

RADIONUCLIDES SEPERATION

R. Biesuz, L. Bertuzzi, G. Alberti, G. Bergamaschi, A. Miljkovic, V. Amendola,

Dipartimento Chimica, Università di Pavia, via Taramelli 12 – 27100 Pavia

Methods for the separation and concentration of anion species in solution are quite
uncommon, with respect to metal ions, despite the importance of anions in many
fields (e.g. environment, industry, biology and medicine).[1] and, in particular,
selective molecular receptors for hazardous and radioactive anions are of great
awareness. Our interest was towards the pertechnetate anion (i.e. stable form of
technetium), especially noteworthy among radioactive pollutants, and perrhenate
and pertechnetate, produced in the 188W/188Re or 99Mo/99mTc generators and used
in diagnostic and therapy.
The large size and low charge density of both pertechnetate and perrhenate make
the selective recognition and separation a great challenge.
In a previous work, we selected an azacryptand, that in the hexaprotonated form,
is the most suitable for Re(Tc)O4− encapsulation, together with the silica 63 µm as
solid phase, particularly for the best performance with respect to other solid phase
in column operation. [2-3] See figure 1.

Fig 1 the silica modified with (3-

iodopropyl) trimethoxysilane, after
rea tion with the azacrytpand
receptor.

From the these findings, we oriented the present investigation to set up the
operative conditions in an automated system for the production of pure Rhenium
(Technetium) in a suitable medium with high activity high Re(Tc)/Mo ratio, as
required for medical applications in the field.

[1] V. Amendola, et al Coord. Chem. Rev., 2006, 250, 11, 1451-1457.

[2] G. Alberti, V. Amendola, G. Bergamschi, R.Colleoni, C.Milanese, R.Biesuz
Dalton Trans, 42 (2013) 6227-6234
[3] Tesi laurea magistrale in Chimica di Ana Miljkovic, Uni-PV a.a . 2013-14.

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 P55

FLUORESCENT MESOPOROUS SILICA MATERIALS

DISCRIMINATING Ag(I) AND Hg(II)

R. Colleoni1, E. Climent2, K. Rurack2, R. Biesuz1.

1
Dipartimento di Chimica, Università degli Studi di Pavia, Corso Strada Nuova 65
– 27100 Pavia.
2
1.9 Division - Chemical and Optical Sensing, BAM - Federal Institute for
Materials Research and Testing, Richard-Willstätter-Str. 11 – 12489 Berlin.

A new hybrid sensing material

consisting of a fluoroionophore
embedded in mesoporous silica that can
discriminate between Ag(I) and
Hg(II) was prepared. Benzothiazole-
based molecule (1), already studied in
solution [1], is not soluble in water.
Simple steric adsorption into the
pores of suitably functionalized SBA-15 microparticles however is possible.
Various functionalization strategies for the silica were tested (two different silanes
grafted only onto the outer particle surface or onto the inner pore walls and outer
surface) leading to four different materials. After steric loading the materials were
washed with water and dried. The material functionalized with
propyltriethoxysilane on the inner and outer surface (PrpAA) proved to be the
best performer, possibly because of the non-polar environment which allows for
good immobilization of the dye and strong binding of the ions.
For the analyses, PrpAA, suspended in water, shows fluorescence enhancement
in presence of Ag(I) salts, while it undergoes quenching in presence of Hg(II).
Many other metals did not trigger any appreciable fluorescence variations. This
different behaviour towards metal ions is due to the molecular structure: this
fluoroionophore has a D1-A-D2 arrangement (D = Donor, A = Acceptor) with D2
as the selective receptor and D1 as additional donor, allowing for tuning of the
fluorescence properties [2].
Ag(I) enhances fluorescence emission since, after complexation, it can convert D2
into an acceptor yielding a D1-A-A2 pattern, creating a highly emissive CT
species. Hg(II) quenches the fluorescence due to the heavy atom effect. All the
other metals that we checked did not give any variations due to low affinity
towards the receptor moiety. Both enhancement and quenching can be used for
quantification analyses and LODs around 10-7 M were obtained after preliminary
tests.

[1] K. Rurack, A. Koval'chuck, J. L. Bricks, J. L. Slominskii; J. Am. Chem. Soc.,

123 (2001) 6205-6206.
[2] K. Rurack, W. Rettig, U. Resch-Genger; Chem. Comm., (2000), 407-408

242
 P56

MULTICLASS DETERMINATION OF PESTICIDES IN WHEAT FLOUR

BY MEPS FOLLOWED BY HPLC-MS/MS

F. Di Ottavio1, F. Della Pelle1, C. Montesano2, M.C. Simenoni1, D. Compagnone1,

R. Curini2, M. Sergi1, R. Scarpone3, G. Scortichini4
1
Facoltà di Bioscienze e Tecnologie Agroalimentari e Ambientali, Università di
Teramo – 64023 Mosciano S.A. (TE)
2
Dipartimento di Chimica, Università La Sapienza di Roma – 00185 Roma
3
Istituto Zooprofilattico dell’Abruzzo e del Molise, 64100 Teramo.
3
Istituto Zooprofilattico dell’Umbria e delle Marche, 06126 Perugia.

The continuous increasing of the population, combined with the steady diminution
of the areas designated to the cultivation, has placed the attention on the
importance of crop yields, therefore pesticides are used to prevent, destroy, repel,
regular or control pests [1]. The wheat is exposed to phytosanitary treatment
during planting, growing, harvesting and storage; to avoid serious effects on
humans and the ecosystem, for these reasons the European Union by Regulations
provided the restrictions on the use and applicability of these substances by laying
down strict guidelines, imposing Maximum Residue Limit (MRL) values of
pesticides in food and feed.
The aim of this work was the development of a sensitive, accurate method for
multiclass analysis of pesticide and fungicide residues in wheat flours; the
attention was focused on 16 pesticides with different physico-chemical
characteristics and different mechanism of action: acetylcholinesterase inhibitors
like organophosphorus, carbamates and neonicotinoids, and inhibitors of
ergosterol like imidazoles and triazoles..
The presented method involves a Micro Extraction by Packed Sorbent (MEPS) [2]
followed by High Performance Liquid Chromatography coupled to Tandem Mass
Spectrometry (HPLC-MS/MS).
The chromatographic separation was conducted using a core-shell column.
For the identification and quantification of the analytes an HPLC-MS/MS
equipped with a source TurboIonSpray operating in positive ionization (PI) was
used for all analytes. The total run time is 10 minutes.
The quantitative analysis was conducted in Multi-Reaction-Monitoring (MRM),
selecting two precursor ion/ion transitions for each analyte.
The acquisition window was divided in four periods to allow the acquisition of all
optimized MRM transitions with an appropriate dwell time, in order to improve
sensitivity and maintain the quality of the chromatographic peaks, by acquiring a
suitable number of points for the shape definition.

[1] http://www.epa.gov/agriculture/tpes.html
[2] M. Moein, A. Abdel-Rehim, M. Abdel-Rehim, Trends in Analytical
Chemistry, 67 (2015) 33-44

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 P57

DIRECT INJECTION - HPLC ANALYSIS FOR THE DETERMINATION

OF FURANIC COMPOUNDS IN OIL AS MARKERS OF SOLID
INSULATION DEGRADATION IN POWER TRANSFORMERS

R.M. De Carlo1, M.C. Bruzzoniti1, L. Rivoira1, C. Sarzanini1, S. Kapila3, V.

Tumiatti2, R. Maina2
1
Department of Chemistry, University of Turin, Via Giuria 5, 10125 Torino
2
Sea Marconi Technologies, Via Ungheria 20, 10093 Collegno (Torino)
3
Department of Chemistry, Missouri University of Science and Technology, 142
Schrenk Hall, 400 W. 11th St., Rolla, MO 65409 (USA)

The presence of 2-furaldheyde (2-FAL) and related furanic compounds in

insulating mineral oils is correlated to thermal degradation and mechanical
properties of the Kraft paper used as solid insulation in large electrical equipment
(e.g. power transformers). Besides 2-FAL, the main compounds formed by paper
degradation are 5-(hydroxymethyl)-2-furaldehyde (5-HMF), 2-furfuryl alcohol (2-
FOL), 2-acetylfuran (2-ACF), 5-methyl-2-furfuraldehyde (5-MEF). Nevertheless,
2-FAL is usually the molecule which is found at higher concentrations in the oil,
since it is the compound of final degradation of 5-HMF, 2-FOL, 2-ACF and 5-
MEF.
The presence of the above-mentioned compounds in the oil is therefore an
indication of the health status of solid insulation in power transformers, and their
detection represents an important tool for planning maintenance procedure
throughout the whole lifetime of the electrical equipment. In order to perform
accurate and reliable surveillance of the degradation state of paper, simple, robust
and fast analytical methods are required. As regards the determination of furanic
compounds in mineral oil, the International Standard IEC 61198 details the
analytical methods to be used, which are basically based on a L/L or SPE
extraction, followed by HPLC-UV.
The aim of this work is to study the feasibility of the determination of 2-FAL, 5-
HMF, 2-FOL, 2-ACF and 5-MEF by direct injection HPLC-UV analysis, without
any sample pretreatment.
The method has been developed on a core-shell C18 column, evaluating the effect
of sample volume injected, in order to avoid column saturation due to the direct
injection of oil. Eluent composition as well as gradient programming have been
tuned in order to ensure fast analysis time, and the resolution between 2-FAL and
2-FOL peaks for which the column exhibits similar selectivity. A proper washing
procedure has been developed in order to remove the organic compounds which
characterize the typical oil profile.
Precision of the method has been calculated by analyzing oil samples from in-
service transformers. Accuracy was tested inside three round-robin tests (2012-
2014).

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 P58

FORENSIC INVESTIGATION ON TEXTILES: CAPABILITIES OF

RAMAN SPECTROSCOPY

F. Bianchi1, V. Trolla1, N. Riboni1, G. Avantaggiato2, G. Iacobellis2, G. Furlan2,

M. Careri1
1
Dipartimento di Chimica, Università di Parma, Parco Area delle Scienze, 17/A –
43124 Parma
2
Reparto Carabinieri Investigazioni Scientifiche di Parma, Via Parco Ducale 3 –
43125 Parma

Raman spectroscopy is a widely utilized technique in the field of cultural heritage

for dating and assessing authenticity of artifacts like paintings, frescoes,
manuscripts and scrolls. More recently, attention has been paid to the
determination of contaminants in drugs and food [1]. Interesting results have been
achieved also in the field of forensic sciences for the analysis of varnishes,
narcotics, explosives and textiles [2]. Being a rapid and non-destructive technique
that does not require sample preparation, Raman microscopy has been
successfully exploited for both the identification of different types of fibers like
nylon, cotton and polyamide [3].
Aging is another important factor to be considered when forensic comparison
among fibers is carried out. Fibers collected on the crime scene can undergo
physical, photochemical, thermal, chemical and mechanical changes, thus making
their comparison more difficult.
In this study we demonstrate the capabilities of Raman spectroscopy in
distinguishing dyed textiles after aging, in a non-destructive way. The proper
optimization of the experimental conditions in terms of choice of laser
wavelength, power, signal acquisition time followed by chemometric processing
like principal component analysis or discriminant analysis allowed to correctly
classify not only textiles produced using similar dyes, but also the samples aged
under different conditions. Finally, good values of cross validation demonstrated
the feasibility of the proposed technique for forensic science investigations.

[1] Y. Li, J. Church, J. Food Drug Anal. 22 (2014) 29-48

[2] J. Palus, M. Kunicki, Forensic Sci. Int. 158 (2006) 164-172
[3] M.M.L. Yu, P. M. L. Sandercock, J. Forensic Sci. 57 (2012) 70-74

245
 P59

ANALYSIS OF DRUGS OF ABUSE: SYNTHETIC CANNABINOIDS AND

ALL AROUNDERS

D. Merli, S. Protti, M. Pesavento, S. Tinivella, L. Cucca, A. Profumo

Dipartimento di Chimica, Università di Pavia, Via Taramelli, 12 – 27100 Pavia

The determination of drugs of abuse is an important challenge, and the continuous

discovery and commercialization of thousands of designed drugs made the effort
very arduous. The two main difficulties from the analytical point of view lie in the
doses administered (e.g. LSD, in the order of 100 µg), and in the introduction on
the market of continuously new substances. In particular, in recent years we
assisted to the introduction of the so-called “synthetic cannabinoids”, belonging to
different chemical classes, often misregulated and freely available from internet
(where they are sold as “salt bath”, “spice” or “herbal incense”). Most of them
belong to the class of indoles, and the most representative drugs are JWH-018 and
the up to date non-regulated adamantly derivative AB-001.
In the present communication, we propose user friendly methods for the on-field
qualitative characterization and semi-quantitative determination of some synthetic
cannabinoids, based on colorimetric detection of the substances and the analysis
of the color intensity (on the RGB scale) with a free i-phone App. This will be a
screening test before a deeper analytical characterization of the seized drug.

246
 P60

LUMICYANO: EVALUATION OF A NEW FLUORESCENT

CYANOACRYLATE IN FINGERMARKS DETECTION

R. Risoluti1, S. Materazzi1, V. Filetti1, G. Iuliano2, L. Niola2, L. Ripani2

1
Dipartimento di Chimica, “Sapienza” Università di Roma, p.le A.Moro 5 –
00185 Roma
2
Reparto Investigazioni Scientifiche RIS – viale Tor di Quinto 119 – 00191 Roma

Fingermarks play a key role in crime scene investigations because their friction
ridge pattern can be used for identification purposes. In most cases, fingermarks
are invisible and influenced by substrates or environmental factors. Latent
fingermarks developed by traditional cyanoacrylate fuming process, often lack
contrast with the substrates; therefore further enhancements are required, such as
dye staining or powder dusting. This second step, that is part of the conventional
detection, aims at improving contrast and at increasing the legibility of details. [1]
To avoid the second step, several commercially available formulations of
cyanoacrylate have been recently manufactured and marketed as‘one-step’ fuming
reagents for latent fingerprints revelation. In particular Lumicyano is found to be
very promising, combining the cyanoacrylate fuming and the dyeing procedures
into a one-step process offering the potential to save time and effort in the
detection of latent fingermarks. In this work, a detailed comparative examination
between conventional, two-step process (cyanoacrylate fuming followed by
staining with Basic Yellow 40) and Lumicyano is proposed for fingermarks
detection, in cooperation with the Forensic Science Department (Carabinieri-RIS)
of Rome. The study has been conducted on fresh as well as on aged fingermarks
(up to 3 months) and applied to non-porous surfaces without changing the fuming
chamber settings of forensic laboratories. The fluorescence has been observed
either under UV (scenescope, 315-340 nm) or visible (crimescope, 450-550 nm)
light irradiation, in order to ensure a good compatibility with the lightning
material available within most police forces. The possibility of further DNA
detection after marks relevation, has been also investigated.
The results indicate that good ridges clarity and excellent contrast are observed
with one step process, concluding that Lumicyano detects fingermarks with equal
or better sensitivity and ridge details than currently used cyanoacrylate, extending
the acquisition time till three month. Moreover, this study has shown that further
enhancement with BY40 after Lumicyano can still be carried out if needed,
allowing the identification of marks not revealed in a singol step, for best
performing results.

[1] G.Groeneveld, S.Kuijer, M.DePuit. Science and Justice 54 (2014) 42–48

247
 P61

PRESSURIZED LIQUID EXTRACTION FOR THE DETERMINATION

OF CANNABINOIDS AND METABOLITES IN HAIR: DETECTION OF
CUT-OFF VALUES BY HPLC-HRMS/MS

M. Sergi1, M.C. Simeoni1, G. Vannutelli2, C. Montesano2, A. Gregori3, L. Ripani3,

D. Compagnone1, R. Curini2
1
Facoltà di Bioscienze e Tecnologie Agroalimentari e Ambientali, Università di
Teramo, Via C. Lerici, 1 – 64023 Mosciano S.A. (TE)
2
Dipartimento di Chimica, Università La Sapienza di Roma, P.le A.Moro – 00185
Roma
3
Department of Scientific Investigation (RIS), Carabinieri, Via di Tor di Quinto
151 - 00191 Rome

Hair analysis has become a routine procedure in most forensic laboratories since it
presents clear advantages: wider time window, non-invasive sampling and good
stability of the analytes over time [1]. Cannabinoids analysis in hair is still not
straightforward: major challenges arise from low concentration of ∆9-
tetrahydrocannabinol (THC) and even lower for the main metabolite 11-nor-9-
carboxy-THC (THC-COOH). Furthermore the determination of THC-COOH has
shown to be crucial to distinguish among passive drug exposure and active
consumption since this molecule is an exclusive product of metabolism and can be
considered as marker of drug abuse [2].
The aim of the present work was to develop a sensitive and accurate method for
the determination of cannabinoids in hair.
The extraction of analytes from hair (50 mg) is based on an automated pressurized
liquid extraction (PLE), followed by SPE: this procedure allows both the
reduction of matrix effect and the enrichment of the analytes which is particularly
useful for the detection of THC-COOH. The analysis is carried out by HPLC-
HRMS/MS with an Orbitrap system. Chromatographic run obtained with a fused-
core column provided a good separation of the analytes in less than 4 min.
The whole procedure has been validated according to SWGTOX guidelines. The
presented PLE-SPE procedure provides an efficient extraction/sample clean-up
with few simple steps and with a minimum use of organic solvents.
To the best of our knowledge, this is the first LC-MS/MS based method that
allows the detection of THC-COOH in hair at values lower than the cut-off.

[1] F. Pragst,M.A. Balikova, State of the art in hair analysis for detection of drug
and alcohol abuse, Clin. Chim. Acta. 370 (2006) 17-49
[2] S. Dulaurent, J.M. Gaulier, L. Imbert, A. Morla,G. Lachatre, Simultaneous
determination of THC, cannabidiol, cannabinol and THC-COOH acid in hair
using LC-MS/MS, Forensic Sci. Int. 236 (2014) 151-6.

248
 P62

DETERMINATION OF ANTICOAGULANT RODENTICIDES AND Α-

CHLORALOSE IN HUMAN HAIR BY ULTRA-HIGH PERFORMANCE
LIQUID CHROMATOGRAPHY–TANDEM MASS SPECTROMETRY
AND APPLICATION TO A REAL CASE

A. Salomone1, M. Leporati1, G. Golè2, E. Gerace1, M. Vincenti1,3

1
Centro Regionale Antidoping e di Tossicologia “A. Bertinaria”, regione Gonzole
10/1, 10043 Orbassano (TO), Italy
2
Medicina Legale ASL TO2, Via Foligno 14, 10149 Torino, Italy
3
Dipartimento di Chimica, Università degli Studi di Torino, via P. Giuria 7, 10125
Torino, Italy

Anticoagulant rodenticides are the largest group of pesticides used for control of
harmful rodents. They are classified into two main groups, depending on their
chemical structure: hydroxycoumarine and indandione rodenticides. Their
fundamental mode of action is represented by the inhibition of the vitamin K
epoxide reductase, which causes blood-clotting alteration, leading to extensive
hemorrhages as the ultimate cause of death.
In this study, we developed a UHPLC-MS/MS for the simultaneous determination
of 10 anticoagulant rodenticides (coumatetralyl, brodifacoum, bromadiolone,
difenacoum, flocoumafen, coumachlor, acenocoumarol, coumafuryl, dicoumarol,
warfarin), plus α-chloralose in human hair, with the scope of detecting potential
chronological trace of poisons exposure in clinical and forensic cases. The
developed method was applied to a real case of alleged poisoning.
The optimized UHPLC-MS/MS method allowed the simultaneous determination
of 10 anticoagulant rodenticides plus α-chloralose. The whole chromatographic
run, comprehensive of the time required for column re-equilibration, was
completed in 8.5 min. Retention times ranged between 1.39 min (coumafuryl) and
4.33 min (brodifacoum). In the real case, a segmental hair analyses was
performed. Difenacoum was detected in the first hair segment (0-3 cm) at the
concentration of 2.9 pg/mg, while α-chloralose was detected at the concentration
of 85 pg/mg. The two remaining, consecutive segments (3-6 and 6-9 cm) showed
traces of difenacoum (below the LOQ) and low but quantifiable levels of
chloralose (29 pg/mg and 6 pg/mg, respectively).
In conclusion an UPLC-MS/MS method for the simultaneous determination of 10
anticoagulant rodenticides and α-chloralose in human hair was developed and
validated. The method proved to be simple, accurate, rapid and highly sensitive,
allowing the simultaneous detection of all compounds. Finally, the method was
applied to a real case of difenacoum and α-chloralose poisoning and proved
sensitive enough to detect the occasional exposure to both analytes.

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 P63

THE NEVER ENDING STORY OF CANNABINOIDS IN HAIR

D. Di Corcia1, F. Seganti1, E. Gerace1, A. Salomone1, M. Vincenti1,2

1
Centro Regionale Antidoping e di Tossicologia “A. Bertinaria”, Regione
Gonzole 10/1, 10043 Orbassano (TO), Italy
2
Dipartimento di Chimica, Università degli Studi di Torino, via P. Giuria 7, 10125
Torino, Italy

In hair analysis, the distinction between active Cannabis consumption and

external contamination is a well-known problem. The SoHT recommends the use
of cut-offs for THC (50 pg/mg) and its metabolite THC-COOH (0.2 pg/mg) to
prove the active consumption. Nevertheless, THC-COOH is frequently not
detectable in hair, even if considerable THC concentrations were present and
highly sensitive analytical methods were applied. Alternatively, THCA-A has
been proposed as a specific marker to prove external contamination and the THC-
A/THC ratio as a discrimination factor.
In this study, we analyzed 78 hair samples (60 head and 18 pubic) previously
tested positive for THC, in order to (i) evaluate the frequency of THC-COOH
positive samples, in comparison to THC positive; (ii) evaluate the possible
correlation between THC and THC-COOH levels, and (iii) evaluate the reliability
of ratio THCA-A/THC as a valid marker to discriminate between active
consumption and external contamination.
A specific UHPLC method coupled to hybrid QqQ-LIT-MS3 was developed and
validated for the detection of THC, THC-COOH and THC-A. LODs for THC,
THC-COOH and THC-A were, respectively, 5.30, 0.07 and 0.60 pg/mg. Among
78 samples, 30 tested negative for THC-COOH or below LOQ. Among the 48
positive samples (true active consumers), THC-COOH levels were in the range
0.15-8.93 pg/mg (median: 1.40 pg/mg) while THC and THC-COOH
concentrations resulted uncorrelated (R2=0.2238). Among head hair samples, the
ratio THCA-A/THC was in the range 0.29-4.95. In these cases, a combined effect
of active use and external contamination was likely to account for these THC and
THCA-A levels. Among pubic hair, the ratio THCA-A/THC was in the range
0.48-1.55. For these cases, no external contamination is likely to occur and THC
and THCA-A levels should be attributed to active use only. Among 30 samples
negative for THC-COOH, the ratio THCA-A/THC was in the range 0.68-4.94
(median: 1.99).
Whenever THC-COOH is not detected, the ratio THCA-A/THC may be used to
discriminate active consumption from external contamination. A cut-off value of
1.6 is proposed. When THCA-A/THC ratio exceeds 1.6, external contamination
may be considered prevalent with respect to active use. Otherwise, absence of
THC-COOH combined with THCA-A/THC ratios below 1.6, and low THC
absolute levels do not provide conclusive evidences with regard to frequent
Cannabis consumers identification.

250