Analiza droga u urinu GC/MS tehnikom: stečena iskustva i primjena

This paper describes quantitative methods for determination of urinary drugs/metabolites. The analysis included indicators of opiate (morphine, codeine, 6-monoacetylmorphine) and methadone (methadone) consumption, indicator of marihuana/ hashish consumption (11-nor-9-tetrahydrocannabinol- 9-carboxylic acid), indicators of cocaine consumption (cocaine, benzoylecgonine, and ecgonine methyl ester) and of amphetamine consumption (amphetamine, methamphetamine, 3,4-methylenedioxyamphetamine, 3,4- methylenedioxymethamphetamine, and 3,4- methylenedioxethylamphetamine). The methods included solid-phase extraction of urine, concentration of eluent, derivatisation, and quantitative analysis by gas chromatography/ mass spectrometry (GC/MS) on a capillary column in the electron impact and selected ion monitoring (SIM) mode. Sensitivity, reproducibility, and accuracy were determined for all analytes (limit of detection between 3 and 12 ng/ml, precision 92%). The accuracy was checked through analysis of standard reference materials and participation in an international quality assessment programme. The methods were used in the analysis of spot urine samples of 60 subjects suspected of drug abuse. Negative findings indicated several disadvantages of urine as a biological sample.Za provjeru uzimanja droga razvijen je niz brzih, relativno jeftinih testova za analizu droga u urinu, kojima se razlikuju negativni od vjerojatno pozitivnih nalaza. Preporučeni analitički protokol zahtijeva potvrdu svih pozitivnih nalaza specifičnom i osjetljivom metodom. S tim ciljem opisane su kvantitativne metode za simultano određivanje pojedinih droga i njihovih metabolita u urinu: 1) morfina, kodeina i 6-acetilmorfina (6-MAM) pokazatelja uzimanja morfina, kodeina i heroina; 2) 11-nor-tetrahidrokanabinol-9-karboksilne kiseline (THCCOOH) pokazatelja uzimanja marihuane i hašiša; 3) kokaina, benzoilekgonina i metilekgonina pokazatelja uzimanja kokaina i 4) amfetamina, metamfetamina, 3,4-metilendioksiamfetamina (MDA), 3,4-metilendioksimetamfetamina (MDMA, Ecstasy) i 3,4-metilendioksietilamfetamina (MDEA) pokazatelja uzimanja tih istih spojeva, tehnikom plinske kromatografije sa spektrometrijom masa (GC/MS). Metode uključuju ekstrakciju urina na kolonama punjenim sorbensom, koncentriranje eluata, derivatizaciju te kvantitativnu analizu GC/MS sustavom s kapilarnom kolonom uz ionizaciju elektronskim snopom i detekciju karakterističnih iona. Osjetljivost, preciznost i točnost postupka određene su za sve analite. Točnost određivanja provjerena je analizom standardnih referentnih uzoraka. Sudjelovanjem u međunarodnom programu provjere kvalitete analiza droga u urinu potvrđena je točnost razvijenih metoda za sve analite. Opisane metode primijenjene su pri identifikaciji zloporabe droga u 60 osoba za koje se sumnjalo da uzimaju droge. U 35 uzoraka urina nađena je THCCOOH, amfetamini u dva, metadon u četiri, morfin/ kodein/6-MAM u 11 i kokain i metaboliti u tri uzorka urina. U četiri uzorka urina određeno je više droga/metabolita. Negativni nalazi droga u 10 uzoraka urina upućuju na nedostatke urina kao biološkog uzorka: može biti zamijenjen drugim urinom, izmijenjen dodacima raznih sredstava i razrijeđen vodom. Kako se većina droga/metabolita izlučuje urinom 2–3 dana nakon uzimanja droge (s izuzetkom THCCOOH), kritično je vrijeme uzimanja urina za analizu

Our experience in biological monitoring of exposure to some organic solvents

Prikazana su iskustva u primjeni biološkog monitoringa profesionalne izloženosti benzenu, toluenu, ksilenu, stirenu, trikloretilenu i tetrakloretilenu. Istaknuta je važnost pravilnog odabira pokazatelja, medija i vremena uzorkovanja. Uspoređena su vlastita istraživanja s postojećim biološkim graničnim vrijednostima te je naglašena potreba za njihovom realnom primjenom u svrhu zaštite zdravlja radnika.The paper describes our experience in the application of biological monitoring of occupational exposure to benzene, toluene, xylene, styrene, trichloroethylene and tatrachloroethylene. The importance of adequate selection of indicator, matrix and sampling time is pointed out. Own results are compared with the existing biological exposure indices. The need for actual and strict application of biological exposure indices is emphasized tram the point of view of workers\u27 healh protection

Identification of drug abuse

Zlouporaba droga opasna je za zdravlje i život, kako na radnome mjestu tako i u široj zajednici. U razvijenim zemljama naglašava se sve više potreba za provjerom uzimanja droga pojedinaca; uobičajena je provjera prilikom prijave za različita radna mjesta kao i periodička kontrola u određenim zanimanjima. Budući da su posljedica u slučaju svakog pozitivnog nalaza teške i dalekosežne, postoji potreba za razvojem ujednačenih i međunarodno priznatih metoda za identifikaciju zlouporabe droga. Opisan je odabir bioloških uzoraka za analizu najčešće rabljenih droga, glavne karakteristike tih droga te preporučene analitlčke tehnike za njihovo određivanje. Istaknuta je važnost osiguranja kvalitete laboratorija u kojem se analiziraju droge u biološkim uzorcima.Drug abuse poses health and safety hazards, both in the workplace and in the wider community. There is increasing pressure to use urine drug tests to determine the prevalence of drug abuse, to deter illicit use of drugs, and to identify drug abusers for rehabilitation. Drug-abuse testing programs have been implemented for employees, job applicants, policemen, firemen, enlisted personnel of the army, athletes, and workers in occupations that are considered critical to public safety and health, such as those in nuclear power plants and the transportation industry. Since the consequences ot a positive test can be quite severe, there is a need to develop uniform and internationally recognised methods for identifying drug abuse The paper discusses selection ot biological specimens for drug-abuse testing, main characteristics of drugs of abuse and recommended analytical techniques tor their determination. The impoitance of a quality assurance program for drug-testing laboratories is emphasised

Hair - Biological Sample for the Analysis of Drugs of Abuse

Opisane su glavne značajke kose kao biološkog uzorka, mehanizmi ulaza droga u kosu, njihova zadržavanja i nestajanja. Budući da se većina metoda za određivanje droga u kosi koristi vezanim sustavom plinski kromatografspektrometar masa (GC/MS), samo su te metode i prikazane. U pravilu svaka se metoda sastoji od pranja kose, usitnjavanja, ekstrakcije, pročišćavanja i derivatizacije. Dok je analitički postupak obrade kose gotovo do potankosti razrađen, u interpretaciji rezultata droga u kosi još je mnogo teškoća. Kosa nije jednolika sastava, ulaz droge u kosu nije isključivo preko korijena, postoje razlike u rastu kose, na koncentraciju droga u kosi utječu kozmetičko tretiranje kose, čistoća uzete droge. Sve su to čimbenici koji se moraju uzeti u obzir pri interpretaciji nalaza. Nalaz služi kao dokaz da je ispitanik kronično ili ponavljano uzimao drogu i nije moguće sa sigurnošću ustanoviti kolika je to količina droge bila kao ni vrijeme uzimanja.Hair testing for drugs of abuse has the advantage of prolonged detection over blood, saliva, or urine analysis and is a useful diagnostic complement to them. This paper describes the main characteristics of hair as a biological sample and the mechanisms of incorporation, retention, and loss of drugs of abuse. The overview is confined to the most common procedures for determination of drugs of abuse, that is, those which involve gas chromatography – mass spectrometry (GC-MS). Each procedure basically includes the same steps: hair washing (decontamination), cutting, extraction/digestion, clean-up, and derivatisation. Although the analysis of hair has been worked out in detail, interpretation of findings of drugs of abuse in hair is complex. The content of hair is not uniform all along; a drug does not necessarily enter through the root; there are differences in hair growth; cosmetic treatment, purity of the taken drug, and the hair colour may affect the concentration of drugs of abuse in hair. The interpretation of findings must take all that into account. Positive findings of drugs of abuse in hair reflect chronic/recurring consumption but do not positively establish the quantity or time of consumption

Određivanje benzena u urinu plinskokromatografskom tehnikom analize para iznad otopine

This paper describes the application of static headspace gas chromatography in determining benzene in urine. The method was analytically validated for sensitivity (DL=42 ng/l), repeatability (RSD=3% and 4%), and accuracy (71%), and was applied in measuring urine benzene in nonsmokers (N=14) and smokers (N=18). All urine samples had measurable benzene concentrations. The method proved sensitive enough to establish a significant statistical difference (P<0.000614) in urine benzene concentrations between smokers (mean=760; range =181–1869 ng/l) and nonsmokers (mean=214; range=61–515 ng/l).Opisana je metoda za određivanje benzena u urinu plinskokromatografskom tehnikom analize para iznad otopine (“headspace”). Metoda je analitički vrednovana s obzirom na osjetljivost (granica detekcije = 42 ng/l), ponovljivost (RSD = 3 i 4%) i točnost (71%). Primijenjena je u analizi benzena u urinu nepušača (N=14) i pušača (N=18). Benzen je bio mjerljiv u svim uzorcima urina. Dobivena je statistički značajna razlika (p<0.000614) između koncentracija benzena u urinu pušača (x=760; raspon=181–1869 ng/l) i nepušača (x=214; raspon=61–515 ng/l). Benzen ne dolazi u urin normalnim fiziološkim procesima; u nepušača uglavnom potječe iz ispušnih plinova automobila i izgaranja benzina, a u pušača je glavni izvor izloženosti benzenu dim cigarete

Hair Testing for Drugs of Abuse

Hair testing for drugs of abuse is a developing technology, which offers the possibility of longer detection times than is commonly obtained with urine analysis. It is the main method for evaluation of an individual’s drugs of abuse history. In many countries hair analysis is routinely used to detect drug abuse in forensic cases, occupational and traffic medicine and clinical toxicology. Hair analysis in pregnant women, neonates and infants is a useful tool for the detection of drug exposure in utero. In Croatia hair testing for drugs of abuse is performed at the Institute for Medical Research and Occupational Health. Three-year experience in drugs of abuse analysis in hair is described. In 331 hair samples (270 from adolescents and 61 from adults) opiates and metabolites, cocaine, methadone, and amphetamines were analyzed by gas chromatography/mass spectrometry. Most prevalent drugs of abuse in adolescents were amphetamines, and in adults heroin. From the examples cited and samples analyzed it is evident that hair testing is emerging as a reliable biological marker for cumulative account of individual exposure to drugs of abuse

Procjena zloporabe droga analizom kose: dvogodišnje iskustvo

This article gives a brief account of a two–year experience with gas chromatography/mass spectrometry methods developed for the analysis of opiates (morphine, codeine, heroin and 6–acetylmorphine), cocaine, methadone, and amphetamines (amphetamine; methamphetamine; 3,4–methylenedioxyamphetamine – MDA, and 3,4–methylenedioxymethamphetamine – MDMA, Ecstasy) in hair . The methods developed were reproducible (RSD=5.0–16.1%), accurate (85.1–100.6%) and sensitive (LD=0.05–0.30 ng/mg). They were applied in the analysis of 221 hair samples obtained from young subjects aged 15–25 years, who were suspected of drug abuse. Seventy–seven hair samples were found positive for drugs of abuse. Forty–two subjects were found to have consumed amphetamine, of whom 40 were found MDMA (Ecstasy). Heroin consumption, based on positive 6–acetylmorphine, was found in 26 subjects. Cocaine alone was present in three subjects, two were found cocaine and heroin, two cocaine and methadone, and one cocaine and MDMA. These results could indicate the trend in drug abuse among young people in Croatia.Razvijene su GC/MS metode za analizu opijata (morfin, kodein, heroin i 6–acetilmorfin), kokaina, metadona i amfetamina (amfetamin; metamfetamin; 3,4–metilendioksiamfetamin –MDA i 3,4–metilendioksimetamfetamin – MDMA) u kosi. Osjetljivost, preciznost i točnost metode određene su za sve analite. Opisane metode primijenjene su pri identifikaciji zloporabe droga u kosi 221 mlade osobe (15–25 god.) za koje se sumnjalo da uzimaju drogu. U 77 osoba dokazano je uzimanje droga, dok su u uzorcima kose 144–ju ispitanika rezultati negativni. U 54,5% (N=42) osoba za koje je utvrđeno da uzimaju droge nađeni su amfetamini, pretežno MDMA (N=40), što upućuje na uzimanje tableta “Ecstasy”. Konzumiranje heroina dokazano je u 33,8% (N=26) osoba na temelju prisutnosti 6–acetilmorfina, jedinog specifičnog metabolita heroina. Uzimanje kokaina dokazano je u osam ispitanika. Razrađenim metodama moguće je procijeniti učestalost i vrstu droga koju uzima mlada populacija u Hrvatskoj

Oštećenje vida za boje u radnica izvrgnutih niskim razinama toluena

Colour vision was examined by the Lanthony-D-15 desaturated test in 41 women exposed to toluene and in 29 non-exposed referents. Toluene exposure was evaluated by methods ot environmental and biological monitoring. In the exposed group the median value of toluene in air was 35 ppm (range 11.2-49.9 ppm). Quantitative colour vision impairment was expressed as colour confusion index and colour confusion index corrected for alcohol intake. Qualitative impairment was expressed as normal, yellow-blue, red-green range or complex impairment. Statistical analysis showed the index values to be significantly correlated with age in both groups. In the exposed group they were significantly higher than in the non-exposed group. There was no significant difference in the prevalence of impairment in the blue-yellow range between the examined groups, although the prevalence of impairment in the exposed group was higher than in the non-exposed one. Results suggest that exposure to low toluene concentrations may induce colour vision impairment in women.Desaturacijskim testom Lanthony D-15 ispitivan je vid za boje u 41 radnice izvrgnute prosječnoj koncentraciji toluena u zraku od 35 ppm (raspon 11,2-49,9 ppm) i u 29 radnica kontrolne skupine. Kvalitativni vid za boje izražen je kao indeks pogreške i kao indeks pogreške korigiran za utjecaj alkohola, a kvalitativni vid kao normalan, s oštećenjern u plavo-žutom dijelu spektra ili kao složeno oštećenje. Rezultati su statistički analizirani t-testom i Mann-Whitneyevim U-testom. U obje su ispitivane skupine vrijednosti indeksa pogreške i indeksa pogreške korigiranog za utjecaj alkohola bile značajno povezane s dobi ispitanika. U skupini izloženoj toluenu njihove su vrijednosti bile statistički značajno više (P) nego u kontrolnoj skupini. Nije bilo statistički značajno učestalijeg oštećenja vida u žuto-plavom dijelu spektra. Rezultati upućuju na zaključak da u radnica izvrgnutih niskim koncentracijama toluena indeks pogreške značajno odstupa od očekivanoga, ali da kvalitativno oštećenje vida za boje nije učestalije nego u poredbenoj skupini

Usporedba dinamičke analize para iznad otopine i mikroekstrakcije analita na čvrstoj fazi za plinskokromatografsko određivanje BTEX-a u urinu

The aim of this study was to compare two extraction procedures: dynamic headspace-purge and trap (PT) and headspace solid-phase microextraction (HS-SPME) for gas chromatographic determination of benzene, toluene, ethylbenzene, and isomeric xylenes (BTEX) in urine with photoionization (PID) and mass spectrometric (MS) detection, respectively. Both methods showed linearity in the range of interest [(50-2000) ng L-1], good accuracy (80% to 100 %), and repeatability (RSD≤11 %). Detection limits were in the low ng L-1 level for both methods, although slightly greater sensitivity was found for the PT method. In comparison with PT, HS-SPME was simpler and required less time for analysis. Although the analytical features of both examined methods are appropriate for biomonitoring of environmental exposure to BTEX, only the HS-SPME-GC-MS method is recommended for routine analysis of BTEX in urine. The method was applied for the quantitative analysis of BTEX in urine samples collected from non-smokers (n=10) and smokers (n=10).Cilj ovog rada bio je usporediti dva postupka ekstrakcije za plinskokromatografsko određivanje benzena, toluena, etilbenzena i izomera ksilena u urinu. Uspoređene su dinamička analiza para iznad otopine (tzv. purge and trap) uz fotoionizacijski detektor i mikroekstrakcija analita na čvrstoj fazi uz detektor spektrometar masa. Rezultati upućuju na linearnost odziva detektora u ispitivanome koncentracijskom području [(50- 2000) ng L-1], zadovoljavajuću točnost (80 %-100 %) i ponovljivost (RSD ≤11 %). Postignute su niske granice detekcije za obje metode. Mikroekstrakcija analita na čvrstoj fazi uz detektor spektrometar masa pokazala se jednostavnijom i bržom za izvođenje pa se preporučuje za rutinsko određivanje BTEX-a u urinu. Metoda je primijenjena za analizu tih spojeva u uzorcima urina nepušača (n=10) i pušača (n=10)