![\"\"](\"https:\/\/www.vtei.cz\/wp-content\/uploads\/2020\/05\/Pospichalova-1.jpg\")
<\/a>\nObr. 1. Zapojen\u00ed se \u0161esticetn\u00fdm ventilem, eluce proti sm\u011bru SPE
\nFig. 1. Conection of six-way valve, elution against the direction of SPE<\/h6>\n
Alkoholick\u00e9 n\u00e1poje jsou jednou z\u00a0nejpopul\u00e1rn\u011bj\u0161\u00edch l\u00e1tek ve spole\u010dnosti s\u00a0obsahem psychoaktivn\u00ed slou\u010deniny ethanol. Ethanol je v\u00a0j\u00e1trech oxidov\u00e1n p\u0159edev\u0161\u00edm na acetaldehyd (90\u201395 %). Mal\u00e1 \u010d\u00e1st (< 0,1 %) p\u0159ech\u00e1z\u00ed na metabolity ethylsulf\u00e1t (EtS) a\u00a0ethylglukuronid [13]. Ethylglukuronid nen\u00ed vhodn\u00fd jako biomarker v\u00a0odpadn\u00edch vod\u00e1ch, proto\u017ee je v\u00a0nich velmi nestabiln\u00ed [14]. Ethylsulf\u00e1t je specifick\u00fd biomarker pro monitorov\u00e1n\u00ed spot\u0159eby alkoholu. Jedn\u00e1 se o\u00a0metabolit ethanolu, kter\u00fd indikuje ned\u00e1vnou konzumaci alkoholu s\u00a0dobou detekce do 48\u00a0hodin u\u00a0zdrav\u00fdch jedinc\u016f a\u00a0je stabiln\u00ed v\u00a0odpadn\u00ed vod\u011b [14].<\/p>\n
Ke stanoven\u00ed nikotinu a\u00a0jeho metabolit\u016f se nejv\u00edce pou\u017e\u00edvaj\u00ed p\u0159\u00edstroje LC-MS\/MS [9, 11, 12, 15\u201320], v\u00fdjime\u010dn\u011b LC-QTOF [21] nebo hybridn\u00ed MS Q-Exactive s\u00a0vysok\u00fdm rozli\u0161en\u00edm [22\u201325]. Detekce a\u00a0kvantifikace se prov\u00e1d\u00ed s\u00a0hmotnostn\u00edm detektorem typu trojit\u00e9ho kvadrupolu s\u00a0ionizac\u00ed v\u00a0pozitivn\u00edm m\u00f3du ESI+. Odpadn\u00ed vody jako slo\u017eit\u00e1 matrice obsahuj\u00ed velk\u00e9 mno\u017estv\u00ed koeluuj\u00edc\u00edch slou\u010denin, kter\u00e9 mohou ve velk\u00e9 m\u00ed\u0159e ovliv\u0148ovat ionizaci sledovan\u00fdch slou\u010denin. Proto se p\u0159ed extrakc\u00ed p\u0159id\u00e1vaj\u00ed ke vzorku odpadn\u00ed vody deuterovan\u00e9 izomery stanovovan\u00fdch analyt\u016f a\u00a0kvantifikace se prov\u00e1d\u00ed metodou intern\u00edho standardu [9\u201312, 15\u201328]. Ke zv\u00fd\u0161en\u00ed citlivosti stanoven\u00ed a\u00a0\u010di\u0161t\u011bn\u00ed se pou\u017e\u00edv\u00e1 prekoncentrace vzorku extrakc\u00ed na pevn\u00e9 f\u00e1zi (SPE) v\u00a0zapojen\u00ed on-line [25\u201327] nebo off-line [9, 11, 15\u201324, 28]. Pou\u017e\u00edvaj\u00ed se kolonky Oasis HLB [11, 14, 19\u201321, 26\u201328], Oasis MCX [15], StrataX [12]. Pro on-line SPE se pou\u017e\u00edv\u00e1 zejm\u00e9na kolonka Hypersil Gold [23\u201325]. Stanoven\u00ed nikotinu a\u00a0jeho metabolit\u016f se prov\u00e1d\u00ed v\u011bt\u0161inou s\u00a0kolonami s\u00a0reverzn\u00ed f\u00e1z\u00ed C18 [9\u201312, 15\u201328] a\u00a0st\u0159edn\u011b pol\u00e1rn\u00ed mobiln\u00ed f\u00e1z\u00ed, kter\u00e1 obsahuje organickou f\u00e1zi (nej\u010dast\u011bji acetonitril nebo methanol) a\u00a0vodnou f\u00e1zi (voda s\u00a0p\u0159\u00eddavkem mraven\u010danu nebo octanu amonn\u00e9ho v\u00a0prom\u011bnliv\u00e9m pom\u011bru podle nastaven\u00e9ho programu gradientu).<\/p>\n
Tabulka 1. P\u0159ehled analyt\u016f, jejich z\u00e1kladn\u00ed charakteristiky a\u00a0rozd\u011blen\u00ed do metod stanoven\u00ed
\nTable 1. Overview of analytes, theirs characteristics and methods of detection<\/h5>\n![\"\"](\"data:image\/svg+xml;base64,PHN2ZyB3aWR0aD0iMSIgaGVpZ2h0PSIxIiB4bWxucz0iaHR0cDovL3d3dy53My5vcmcvMjAwMC9zdmciPjwvc3ZnPg==\")
<\/a>\nKe stanoven\u00ed ethylsulf\u00e1tu v\u00a0odpadn\u00ed vod\u011b se pou\u017e\u00edv\u00e1 metoda kapalinov\u00e9 chromatografie s\u00a0hmotnostn\u00edm detektorem s\u00a0trojit\u00fdm kvadrupolem a\u00a0elektrosprejem jako iontov\u00fdm zdrojem v\u00a0negativn\u00edm m\u00f3du (ESI-) [13, 29\u201335]. EtS je pol\u00e1rn\u00ed slou\u010denina a\u00a0na tradi\u010dn\u00edch kolon\u00e1ch C18, C8 s\u00a0reverzn\u00ed f\u00e1z\u00ed je m\u00e1lo zadr\u017eov\u00e1n. Proto se roz\u0161\u00ed\u0159ilo jeho stanoven\u00ed pomoc\u00ed iontov\u011b p\u00e1rov\u00e9 chromatografie. Pou\u017e\u00edvaj\u00ed se iontov\u011b p\u00e1rov\u00e1 \u010dinidla: 5 mM dibutylamonium acet\u00e1t [24], 7 mM dihexylamonium acet\u00e1t [12, 25]. Pou\u017e\u00edvan\u00e9 mobiln\u00ed f\u00e1ze jsou voda a\u00a0methanol, iontov\u011b p\u00e1rov\u00e9 \u010dinidlo se p\u0159id\u00e1v\u00e1 do jedn\u00e9 nebo obou f\u00e1z\u00ed. \u00dasp\u011b\u0161n\u011b byla ke stanoven\u00ed EtS pou\u017eita kolona Synergi Polar-RP (polar endcaped phenylpropyl reverzn\u00ed f\u00e1ze) [31, 32]. K\u00a0eluci se pou\u017e\u00edv\u00e1 voda s\u00a00,1 % kyselinou mraven\u010d\u00ed a\u00a0acetonitril. Ke zlep\u0161en\u00ed ionizace se za kolonu p\u0159id\u00e1v\u00e1 acetonitril. V\u00a0posledn\u00ed dob\u011b se vyr\u00e1b\u011bj\u00ed inertn\u00ed kolony s\u00a0nav\u00e1zan\u00fdm trifunk\u010dn\u00edm alkylov\u00fdm ligandem C18, kter\u00fd podporuje retenci pol\u00e1rn\u00edch slou\u010denin a\u00a0eliminuje pot\u0159ebu p\u0159\u00eddavku iontov\u00fdch p\u00e1r\u016f [27, 33, 34]. Jsou to nap\u0159. kolony: Atlantis T3 nebo Chrompack Inertsil ODS-3 (Varian). Pou\u017e\u00edvaj\u00ed se s\u00a0mobiln\u00edmi f\u00e1zemi voda s\u00a00,1 % kyselinou mraven\u010d\u00ed a\u00a0acetonitrilem a\u00a0s\u00a0post column p\u0159\u00eddavkem acetonitrilu ke zv\u00fd\u0161en\u00ed ionizace.<\/p>\n<\/a>\nObr. 2. Uk\u00e1zka kalibra\u010dn\u00ed k\u0159ivky pro metamfetamin (MAMP)
\nFig. 2. Example of calibration curve (methamphetamine)<\/h6>\n<\/a>\nObr. 3. Uk\u00e1zka kalibra\u010dn\u00ed k\u0159ivky pro kotinin (COT)
\nFig. 3. Example of calibration curve (cotinine)<\/h6>\n<\/a>\nObr. 4. Uk\u00e1zka chromatogramu extrahovan\u00fdch iont\u016f v\u00a0re\u017eimu ESI+ (amfetamin, benzoylecgonin, buprenorphin, EDDP, efedrin, ext\u00e1ze, fentanyl, heroin, kokaethylen, kokain, LSD, metadon, metamfetamin, morfin, nor-diazepam, nor-fentanyl, tramadol)
\nFig. 4. Example of extracted ion chromatograms in ESI+ (amphetamine, benzoylecgonine, buprenorphine, EDDP, ephedrine, MDMA, fentanyl, heroin, cocaethylene, cocaine, LSD, methadone, methamphetamine, morphine, nor-diazepam, nor-fentanyl, tramadol)<\/h6>\n<\/a>\nObr. 5. Uk\u00e1zka chromatogramu extrahovan\u00fdch iont\u016f v\u00a0re\u017eimu ESI+ (ethylcathinon, mefedron, 4-methylethcathinon, metylon, nor-mefedron, pentedron, tramadol, trans<\/em>-3-hydroxykotinin, \u03b1-pyrrolidinovalerofenon)
\nFig. 5. Example of extracted ion chromatograms in ESI+ (ethylcathinone, mephedrone, 4-methylethcathinone, metylone, nor-mephedrone, pentedrone, tramadol, trans<\/em>-3-hydroxykotinin, \u03b1-pyrrolidinovalerofenone)<\/h6>\n<\/a>\nObr. 6. Uk\u00e1zka chromatogramu extrahovan\u00fdch iont\u016f v\u00a0re\u017eimu ESI+ (nikotin, kotinin a\u00a0trans<\/em>-3-hydroxykotinin)
\nFig. 6. Example of extracted ion chromatograms in ESI+ (nicotine, cotinine and trans<\/em>-3-hydroxycotinine)<\/h6>\nStanoven\u00ed ethylsulf\u00e1tu se d\u00e1 prov\u00e1d\u011bt i\u00a0s\u00a0HILIC kolonami, nap\u0159. SeQuant ZIC-HILIC (Merck) a\u00a0mobiln\u00edmi f\u00e1zemi 5 mM octan amonn\u00fd (pH 6,8) a\u00a0acetonitril [33].<\/p>\n
Metodika<\/h2>\n
Postupn\u011b bylo v\u00a0z\u00e1vislosti na chemick\u00fdch vlastnostech a\u00a0mo\u017enostech stanoven\u00ed jednotliv\u00fdch l\u00e1tek zavedeno stanoven\u00ed t\u0159\u00ed des\u00edtek l\u00e1tek metodou kapalinov\u00e9 chromatografie s\u00a0hmotnostn\u00ed detekc\u00ed za podm\u00ednek ionizace elektrosprejem v\u00a0pozitivn\u00edm a\u00a0negativn\u00edm m\u00f3du. P\u0159ehled stanovovan\u00fdch analyt\u016f v\u010detn\u011b jejich z\u00e1kladn\u00ed charakteristiky a\u00a0rozd\u011blen\u00ed do postup\u016f stanoven\u00ed je uveden v\u00a0tabulce 1<\/em>.<\/p>\nPou\u017eit\u00e1 technika: kapalinov\u00fd chromatograf Agilent 1200 RR s\u00a0bin\u00e1rn\u00edm \u010derpadlem, hmotnostn\u00ed detektor Applied Biosystem 4000 Q Trap s\u00a0trojit\u00fdm kvadrup\u00f3lem, autosampler Aspec GX-271 (Gilson). Stanoven\u00ed v\u0161ech l\u00e1tek bylo prov\u00e1d\u011bno s\u00a0chromatografickou kololnou Synergi Hydro-RP80A (150 \u00d7 2,0 mm, 4 \u00b5m).<\/p>\n
Multirezidu\u00e1ln\u00ed stanoven\u00ed t\u00e9to skupiny analyt\u016f se nedalo prov\u00e9st vzhledem k\u00a0jejich odli\u0161n\u00fdm chemick\u00fdm vlastnostem a\u00a0koncentra\u010dn\u00edm hladin\u00e1m jejich v\u00fdskytu. Z\u00a0tohoto d\u016fvodu bylo stanoven\u00ed rozd\u011bleno do \u010dty\u0159 separ\u00e1tn\u00edch postup\u016f.<\/p>\n
Anal\u00fdzy byly prov\u00e1d\u011bny v\u00a0odpadn\u00ed vod\u011b odeb\u00edran\u00e9 v\u00a0n\u00e1toku na \u010dist\u00edrnu komun\u00e1ln\u00edch odpadn\u00edch vod. Tato voda je velmi komplexn\u00ed matrice a\u00a0jako nezbytn\u00e9 se jev\u00ed pou\u017eit\u00ed vnit\u0159n\u00edch standard\u016f. Matrice siln\u011b ovliv\u0148uje ionizaci. Bylo pozorov\u00e1no potla\u010den\u00ed ionizace vlivem l\u00e1tek p\u0159\u00edtomn\u00fdch ve vzorku. Proto bylo pracov\u00e1no s\u00a0deuterovan\u00fdmi standardy v\u0161ech stanovovan\u00fdch l\u00e1tek.<\/p>\n
Pro anal\u00fdzy se zpracov\u00e1vaj\u00ed 24hodinov\u00e9 kompozitn\u00ed vzorky odeb\u00edran\u00e9 na n\u00e1toku na \u010dist\u00edrnu komun\u00e1ln\u00edch odpadn\u00edch vod, kter\u00e9 se po odb\u011bru uchov\u00e1vaj\u00ed v\u00a0chladu a\u00a0temnu. Pokud nelze vzorek analyzovat do 72 hodin od odb\u011bru, vzorek se zmraz\u00ed a\u00a0do vlastn\u00ed anal\u00fdzy se skladuje p\u0159i teplot\u011b -20 \u00b1 4 \u00b0C.<\/p>\n
P\u0159ed vlastn\u00edm stanoven\u00edm se vzorky odst\u0159ed\u00ed a\u00a0ze vzorku se odstran\u00ed pevn\u00e9 \u010d\u00e1stice filtrac\u00ed p\u0159es jednor\u00e1zov\u00e9 membr\u00e1nov\u00e9 filtry o\u00a0porozit\u011b 0,45 \u00b5m. Po p\u0159\u00eddavku sm\u011bsn\u00e9ho roztoku vnit\u0159n\u00edch standard\u016f je vzorek p\u0159ipraven k\u00a0LC-MS\/MS anal\u00fdze. P\u0159ed chromatogrfickou anal\u00fdzou se vzorky zakoncentruj\u00ed z\u00a0objemu 1\u20135\u00a0ml on-line SPE (drogy, nikotin a\u00a0jeho metabolity). K\u00a0on-line prekoncentraci vzork\u016f byl pou\u017eit autosampler Aspec GX-271(Gilson) a\u00a0kolonka Hypersil Gold (20\u00a0\u00d7 2,1 mm, 12 \u00b5m), na kterou bylo 1\u20135 ml vzorku vlo\u017eeno pr\u016ftokem 0,5 ml\/min. Potom byla kolonka je\u0161t\u011b promyta 1 ml UHQ vody, aby se dokon\u010dil transfer vzorku a\u00a0odstranily n\u011bkter\u00e9 interference. Eluce analyt\u016f na chromatografickou kolonu byla prov\u00e1d\u011bna mobiln\u00ed f\u00e1z\u00ed jej\u00edm pr\u016ftokem koncentra\u010dn\u00ed kolonkou v\u00a0protism\u011bru v\u00a0zapojen\u00ed podle obr.\u00a01<\/em>. Pouze anal\u00fdza ethylsulf\u00e1tu se vzhledem k\u00a0jeho vysok\u00fdm koncentrac\u00edm (des\u00edtky\u00a0\u00b5g\/l) prov\u00e1d\u00ed bez prekoncentrace s\u00a0n\u00e1st\u0159ikem 5 \u00b5l vzorku na kolonu.<\/p>\nV\u0161echny pou\u017e\u00edvan\u00e9 chemik\u00e1lie jsou \u010distoty p. a. nebo vy\u0161\u0161\u00ed. Pro kalibraci jsou pou\u017e\u00edv\u00e1ny certifikovan\u00e9 referen\u010dn\u00ed chemik\u00e1lie, nap\u0159. fy Merc, Lipomed, Chromservis. Jako vnit\u0159n\u00ed standardy jsou pou\u017e\u00edv\u00e1ny deuterovan\u00e9 l\u00e1tky, tak\u00e9 certifikovan\u00e9. Podm\u00ednky kapalinov\u00e9 chromatografie pro jednotliv\u00e9 postupy shrnuje tabulka 2<\/em>.<\/p>\nTabulka 2. Podm\u00ednky kapalinov\u00e9 chromatografie
\nTable 2. Liquide chromatography settings<\/h5>\n<\/a>\nIdentifikace a\u00a0kvantifikace se ve v\u0161ech p\u0159\u00edpadech prov\u00e1d\u00ed za pomoci vyhodnocovac\u00edho syst\u00e9mu Analyst\u00ae Software. Jednotliv\u00e9 l\u00e1tky se identifikuj\u00ed porovn\u00e1n\u00edm reten\u010dn\u00edch \u010das\u016f s\u00a0reten\u010dn\u00edmi \u010dasy standard\u016f a\u00a0shodnosti pom\u011bru intenzit dvou sledovan\u00fdch p\u0159echod\u016f dan\u00e9 l\u00e1tky ve vzorku a\u00a0v\u00a0kalibra\u010dn\u00edm roztoku. Kvantifikace se prov\u00e1d\u00ed na z\u00e1klad\u011b hodnot vypo\u010dten\u00fdch pomoc\u00ed kalibra\u010dn\u00edch k\u0159ivek metodou kalibrace na vnit\u0159n\u00ed standard (deuterovan\u00fd standard). Ve stanovovan\u00e9m rozsahu byly v\u0161echny kalibra\u010dn\u00ed k\u0159ivky line\u00e1rn\u00ed (korela\u010dn\u00ed koeficient 0,99 a\u00a0lep\u0161\u00ed).<\/p>\n
Pro uk\u00e1zku jsou na obr. 2\u20138<\/em> uvedeny kalibra\u010dn\u00ed k\u0159ivky a\u00a0chromatogramy extrahovan\u00fdch iont\u016f standard\u016f. Je zobrazen prekurzorov\u00fd, produktov\u00fd ion a\u00a0vnit\u0159n\u00ed standard.<\/p>\nMeze stanovitelnosti (LOQ) dosa\u017een\u00e9 popsan\u00fdmi postupy v\u00a0matrici odpadn\u00ed voda jsou uvedeny v\u00a0tabulce 3<\/em>.<\/p>\nTabulka 3. Meze stanovitelnosti (LOQ) v\u00a0matrici odpadn\u00ed voda
\nTable 3. Limits of quantification for wastewater<\/h6>\n<\/a>\nPodrobn\u011bji jsou pou\u017eit\u00e9 analytick\u00e9 metody pro stanoven\u00ed nikotinu a\u00a0jeho metabolit\u016f a\u00a0ethylsulf\u00e1tu pops\u00e1ny ve sborn\u00edku z\u00a0konference Hydroanalytika 2019 [36].<\/p>\n<\/a>\nObr. 7. Uk\u00e1zka chromatogramu extrahovan\u00fdch iont\u016f v\u00a0re\u017eimu ESI- (nor-THC)
\nFig. 7. Example of extracted ion chromatograms in ESI- (nor-THC)<\/h6>\n<\/a>\nObr. 8. Uk\u00e1zka chromatogramu extrahovan\u00fdch iont\u016f v\u00a0re\u017eimu ESI- (ethylsulf\u00e1t)
\nFig. 8. Example of extracted ion chromatograms in ESI- (ethylsulfate)<\/h6>\n
Pro skupinu syntetick\u00fdch kathinon\u016f byly hled\u00e1ny mo\u017enosti sn\u00ed\u017een\u00ed mez\u00ed stanovitelnosti, proto\u017ee nebyl zaznamen\u00e1n pozitivn\u00ed n\u00e1lez t\u011bchto l\u00e1tek (s\u00a0v\u00fdjimkou dvou hodnot mefedronu t\u011bsn\u011b na \u00farovni LOQ, obdob\u00ed hodnocen\u00ed 2018). Zkou\u0161en\u00e1 metodika vych\u00e1zela z\u00a0d\u0159\u00edv\u011bj\u0161\u00ed metodiky off-line SPE vyvinut\u00e9 v\u00a0projektu Program IGA 2013\u00a0\u2013 Projekt \u201eNov\u00e9 drogy\u00a0\u2013 anal\u00fdza trhu epidemiologie u\u017e\u00edv\u00e1n\u00ed a\u00a0identifikace mo\u017enosti pro sni\u017eov\u00e1n\u00ed \u0161kod\u201c [37]. Ani p\u0159i zpracov\u00e1n\u00ed 500\u00a0ml vzorku nebylo dosa\u017eeno lep\u0161\u00edch mez\u00ed stanovitelnosti ne\u017e p\u0159i on-line SPE metod\u011b [38].<\/p>\n
Z\u00e1v\u011br<\/h2>\n
Pro \u0159e\u0161en\u00ed projektu \u201e\u010cist\u00e1 voda\u00a0\u2013 zdrav\u00e9 m\u011bsto: Komun\u00e1ln\u00ed odpadn\u00ed voda jako diagnostick\u00e9 m\u00e9dium hlavn\u00edho m\u011bsta Prahy\u201c, ve\u00a0kter\u00e9m jsou monitorov\u00e1ny vedle nez\u00e1konn\u00fdch drog a\u00a0jejich metabolit\u016f (v\u010detn\u011b syntetick\u00fdch kathinon\u016f) tak\u00e9 n\u011bkter\u00e1 l\u00e9\u010diva a\u00a0dal\u0161\u00ed leg\u00e1ln\u00ed drogy, jako jsou ethanol a\u00a0nikotin, resp. jejich metabolity, byly vypracov\u00e1ny a\u00a0zvalidov\u00e1ny postupy pro stanoven\u00ed t\u011bchto l\u00e1tek. V\u00a0z\u00e1vislosti na chemick\u00fdch vlastnostech a\u00a0mo\u017enostech stanoven\u00ed jednotliv\u00fdch l\u00e1tek byly o\u00a0tyto l\u00e1tky roz\u0161\u00ed\u0159eny ji\u017e d\u0159\u00edve pou\u017e\u00edvan\u00e9 metody kapalinov\u00e9 chromatografie s\u00a0hmotnostn\u00ed detekc\u00ed za podm\u00ednek ionizace elektrosprejem v\u00a0pozitivn\u00edm m\u00f3du a\u00a0v\u00a0negativn\u00edm m\u00f3du. Metody byly akreditov\u00e1ny. Celkem jsou uveden\u00fdmi metodami stanovov\u00e1ny t\u0159i des\u00edtky l\u00e1tek. Metody rozvinut\u00e9 ve V\u00daV TGM, v. v. i., spl\u0148uj\u00ed po\u017eadavky na citlivost pro anal\u00fdzu v\u011bt\u0161iny zneu\u017e\u00edvan\u00fdch drog a\u00a0jejich metabolit\u016f i\u00a0dal\u0161\u00edch l\u00e1tek sledovan\u00fdch v\u00a0r\u00e1mci Projektu.<\/p>\n","protected":false},"excerpt":{"rendered":"
The wastewater-based epidemiology approach to the analysis of illicit drugs was conceptualized in 2001 and for the first time was applied in 2005 for cocaine estimation in Italy. Further, other illicit drugs as heroine, cannabis and amphetamine have been estimated.<\/p>\n","protected":false},"author":8,"featured_media":7943,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":{"_acf_changed":false,"footnotes":""},"categories":[90,89],"tags":[1105,1890,663,1107,1889,213],"coauthors":[1855,327,599,1856,1857],"class_list":["post-7990","post","type-post","status-publish","format-standard","has-post-thumbnail","hentry","category-waste-management","category-water-technology-water-supply-waste-water-treatment","tag-alcohol","tag-ethylsulfate","tag-illicit-drugs","tag-nicotine","tag-nicotine-metabolites","tag-wastewater"],"acf":[],"_links":{"self":[{"href":"https:\/\/www.vtei.cz\/en\/wp-json\/wp\/v2\/posts\/7990","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/www.vtei.cz\/en\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/www.vtei.cz\/en\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/www.vtei.cz\/en\/wp-json\/wp\/v2\/users\/8"}],"replies":[{"embeddable":true,"href":"https:\/\/www.vtei.cz\/en\/wp-json\/wp\/v2\/comments?post=7990"}],"version-history":[{"count":4,"href":"https:\/\/www.vtei.cz\/en\/wp-json\/wp\/v2\/posts\/7990\/revisions"}],"predecessor-version":[{"id":30580,"href":"https:\/\/www.vtei.cz\/en\/wp-json\/wp\/v2\/posts\/7990\/revisions\/30580"}],"wp:featuredmedia":[{"embeddable":true,"href":"https:\/\/www.vtei.cz\/en\/wp-json\/wp\/v2\/media\/7943"}],"wp:attachment":[{"href":"https:\/\/www.vtei.cz\/en\/wp-json\/wp\/v2\/media?parent=7990"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/www.vtei.cz\/en\/wp-json\/wp\/v2\/categories?post=7990"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/www.vtei.cz\/en\/wp-json\/wp\/v2\/tags?post=7990"},{"taxonomy":"author","embeddable":true,"href":"https:\/\/www.vtei.cz\/en\/wp-json\/wp\/v2\/coauthors?post=7990"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}
<\/a>\nObr. 2. Uk\u00e1zka kalibra\u010dn\u00ed k\u0159ivky pro metamfetamin (MAMP)
\nFig. 2. Example of calibration curve (methamphetamine)<\/h6>\n<\/a>\nObr. 3. Uk\u00e1zka kalibra\u010dn\u00ed k\u0159ivky pro kotinin (COT)
\nFig. 3. Example of calibration curve (cotinine)<\/h6>\n<\/a>\nObr. 4. Uk\u00e1zka chromatogramu extrahovan\u00fdch iont\u016f v\u00a0re\u017eimu ESI+ (amfetamin, benzoylecgonin, buprenorphin, EDDP, efedrin, ext\u00e1ze, fentanyl, heroin, kokaethylen, kokain, LSD, metadon, metamfetamin, morfin, nor-diazepam, nor-fentanyl, tramadol)
\nFig. 4. Example of extracted ion chromatograms in ESI+ (amphetamine, benzoylecgonine, buprenorphine, EDDP, ephedrine, MDMA, fentanyl, heroin, cocaethylene, cocaine, LSD, methadone, methamphetamine, morphine, nor-diazepam, nor-fentanyl, tramadol)<\/h6>\n<\/a>\nObr. 5. Uk\u00e1zka chromatogramu extrahovan\u00fdch iont\u016f v\u00a0re\u017eimu ESI+ (ethylcathinon, mefedron, 4-methylethcathinon, metylon, nor-mefedron, pentedron, tramadol, trans<\/em>-3-hydroxykotinin, \u03b1-pyrrolidinovalerofenon)
\nFig. 5. Example of extracted ion chromatograms in ESI+ (ethylcathinone, mephedrone, 4-methylethcathinone, metylone, nor-mephedrone, pentedrone, tramadol, trans<\/em>-3-hydroxykotinin, \u03b1-pyrrolidinovalerofenone)<\/h6>\n<\/a>\nObr. 6. Uk\u00e1zka chromatogramu extrahovan\u00fdch iont\u016f v\u00a0re\u017eimu ESI+ (nikotin, kotinin a\u00a0trans<\/em>-3-hydroxykotinin)
\nFig. 6. Example of extracted ion chromatograms in ESI+ (nicotine, cotinine and trans<\/em>-3-hydroxycotinine)<\/h6>\nStanoven\u00ed ethylsulf\u00e1tu se d\u00e1 prov\u00e1d\u011bt i\u00a0s\u00a0HILIC kolonami, nap\u0159. SeQuant ZIC-HILIC (Merck) a\u00a0mobiln\u00edmi f\u00e1zemi 5 mM octan amonn\u00fd (pH 6,8) a\u00a0acetonitril [33].<\/p>\n
Metodika<\/h2>\n
Postupn\u011b bylo v\u00a0z\u00e1vislosti na chemick\u00fdch vlastnostech a\u00a0mo\u017enostech stanoven\u00ed jednotliv\u00fdch l\u00e1tek zavedeno stanoven\u00ed t\u0159\u00ed des\u00edtek l\u00e1tek metodou kapalinov\u00e9 chromatografie s\u00a0hmotnostn\u00ed detekc\u00ed za podm\u00ednek ionizace elektrosprejem v\u00a0pozitivn\u00edm a\u00a0negativn\u00edm m\u00f3du. P\u0159ehled stanovovan\u00fdch analyt\u016f v\u010detn\u011b jejich z\u00e1kladn\u00ed charakteristiky a\u00a0rozd\u011blen\u00ed do postup\u016f stanoven\u00ed je uveden v\u00a0tabulce 1<\/em>.<\/p>\nPou\u017eit\u00e1 technika: kapalinov\u00fd chromatograf Agilent 1200 RR s\u00a0bin\u00e1rn\u00edm \u010derpadlem, hmotnostn\u00ed detektor Applied Biosystem 4000 Q Trap s\u00a0trojit\u00fdm kvadrup\u00f3lem, autosampler Aspec GX-271 (Gilson). Stanoven\u00ed v\u0161ech l\u00e1tek bylo prov\u00e1d\u011bno s\u00a0chromatografickou kololnou Synergi Hydro-RP80A (150 \u00d7 2,0 mm, 4 \u00b5m).<\/p>\n
Multirezidu\u00e1ln\u00ed stanoven\u00ed t\u00e9to skupiny analyt\u016f se nedalo prov\u00e9st vzhledem k\u00a0jejich odli\u0161n\u00fdm chemick\u00fdm vlastnostem a\u00a0koncentra\u010dn\u00edm hladin\u00e1m jejich v\u00fdskytu. Z\u00a0tohoto d\u016fvodu bylo stanoven\u00ed rozd\u011bleno do \u010dty\u0159 separ\u00e1tn\u00edch postup\u016f.<\/p>\n
Anal\u00fdzy byly prov\u00e1d\u011bny v\u00a0odpadn\u00ed vod\u011b odeb\u00edran\u00e9 v\u00a0n\u00e1toku na \u010dist\u00edrnu komun\u00e1ln\u00edch odpadn\u00edch vod. Tato voda je velmi komplexn\u00ed matrice a\u00a0jako nezbytn\u00e9 se jev\u00ed pou\u017eit\u00ed vnit\u0159n\u00edch standard\u016f. Matrice siln\u011b ovliv\u0148uje ionizaci. Bylo pozorov\u00e1no potla\u010den\u00ed ionizace vlivem l\u00e1tek p\u0159\u00edtomn\u00fdch ve vzorku. Proto bylo pracov\u00e1no s\u00a0deuterovan\u00fdmi standardy v\u0161ech stanovovan\u00fdch l\u00e1tek.<\/p>\n
Pro anal\u00fdzy se zpracov\u00e1vaj\u00ed 24hodinov\u00e9 kompozitn\u00ed vzorky odeb\u00edran\u00e9 na n\u00e1toku na \u010dist\u00edrnu komun\u00e1ln\u00edch odpadn\u00edch vod, kter\u00e9 se po odb\u011bru uchov\u00e1vaj\u00ed v\u00a0chladu a\u00a0temnu. Pokud nelze vzorek analyzovat do 72 hodin od odb\u011bru, vzorek se zmraz\u00ed a\u00a0do vlastn\u00ed anal\u00fdzy se skladuje p\u0159i teplot\u011b -20 \u00b1 4 \u00b0C.<\/p>\n
P\u0159ed vlastn\u00edm stanoven\u00edm se vzorky odst\u0159ed\u00ed a\u00a0ze vzorku se odstran\u00ed pevn\u00e9 \u010d\u00e1stice filtrac\u00ed p\u0159es jednor\u00e1zov\u00e9 membr\u00e1nov\u00e9 filtry o\u00a0porozit\u011b 0,45 \u00b5m. Po p\u0159\u00eddavku sm\u011bsn\u00e9ho roztoku vnit\u0159n\u00edch standard\u016f je vzorek p\u0159ipraven k\u00a0LC-MS\/MS anal\u00fdze. P\u0159ed chromatogrfickou anal\u00fdzou se vzorky zakoncentruj\u00ed z\u00a0objemu 1\u20135\u00a0ml on-line SPE (drogy, nikotin a\u00a0jeho metabolity). K\u00a0on-line prekoncentraci vzork\u016f byl pou\u017eit autosampler Aspec GX-271(Gilson) a\u00a0kolonka Hypersil Gold (20\u00a0\u00d7 2,1 mm, 12 \u00b5m), na kterou bylo 1\u20135 ml vzorku vlo\u017eeno pr\u016ftokem 0,5 ml\/min. Potom byla kolonka je\u0161t\u011b promyta 1 ml UHQ vody, aby se dokon\u010dil transfer vzorku a\u00a0odstranily n\u011bkter\u00e9 interference. Eluce analyt\u016f na chromatografickou kolonu byla prov\u00e1d\u011bna mobiln\u00ed f\u00e1z\u00ed jej\u00edm pr\u016ftokem koncentra\u010dn\u00ed kolonkou v\u00a0protism\u011bru v\u00a0zapojen\u00ed podle obr.\u00a01<\/em>. Pouze anal\u00fdza ethylsulf\u00e1tu se vzhledem k\u00a0jeho vysok\u00fdm koncentrac\u00edm (des\u00edtky\u00a0\u00b5g\/l) prov\u00e1d\u00ed bez prekoncentrace s\u00a0n\u00e1st\u0159ikem 5 \u00b5l vzorku na kolonu.<\/p>\nV\u0161echny pou\u017e\u00edvan\u00e9 chemik\u00e1lie jsou \u010distoty p. a. nebo vy\u0161\u0161\u00ed. Pro kalibraci jsou pou\u017e\u00edv\u00e1ny certifikovan\u00e9 referen\u010dn\u00ed chemik\u00e1lie, nap\u0159. fy Merc, Lipomed, Chromservis. Jako vnit\u0159n\u00ed standardy jsou pou\u017e\u00edv\u00e1ny deuterovan\u00e9 l\u00e1tky, tak\u00e9 certifikovan\u00e9. Podm\u00ednky kapalinov\u00e9 chromatografie pro jednotliv\u00e9 postupy shrnuje tabulka 2<\/em>.<\/p>\nTabulka 2. Podm\u00ednky kapalinov\u00e9 chromatografie
\nTable 2. Liquide chromatography settings<\/h5>\n<\/a>\nIdentifikace a\u00a0kvantifikace se ve v\u0161ech p\u0159\u00edpadech prov\u00e1d\u00ed za pomoci vyhodnocovac\u00edho syst\u00e9mu Analyst\u00ae Software. Jednotliv\u00e9 l\u00e1tky se identifikuj\u00ed porovn\u00e1n\u00edm reten\u010dn\u00edch \u010das\u016f s\u00a0reten\u010dn\u00edmi \u010dasy standard\u016f a\u00a0shodnosti pom\u011bru intenzit dvou sledovan\u00fdch p\u0159echod\u016f dan\u00e9 l\u00e1tky ve vzorku a\u00a0v\u00a0kalibra\u010dn\u00edm roztoku. Kvantifikace se prov\u00e1d\u00ed na z\u00e1klad\u011b hodnot vypo\u010dten\u00fdch pomoc\u00ed kalibra\u010dn\u00edch k\u0159ivek metodou kalibrace na vnit\u0159n\u00ed standard (deuterovan\u00fd standard). Ve stanovovan\u00e9m rozsahu byly v\u0161echny kalibra\u010dn\u00ed k\u0159ivky line\u00e1rn\u00ed (korela\u010dn\u00ed koeficient 0,99 a\u00a0lep\u0161\u00ed).<\/p>\n
Pro uk\u00e1zku jsou na obr. 2\u20138<\/em> uvedeny kalibra\u010dn\u00ed k\u0159ivky a\u00a0chromatogramy extrahovan\u00fdch iont\u016f standard\u016f. Je zobrazen prekurzorov\u00fd, produktov\u00fd ion a\u00a0vnit\u0159n\u00ed standard.<\/p>\nMeze stanovitelnosti (LOQ) dosa\u017een\u00e9 popsan\u00fdmi postupy v\u00a0matrici odpadn\u00ed voda jsou uvedeny v\u00a0tabulce 3<\/em>.<\/p>\nTabulka 3. Meze stanovitelnosti (LOQ) v\u00a0matrici odpadn\u00ed voda
\nTable 3. Limits of quantification for wastewater<\/h6>\n<\/a>\nPodrobn\u011bji jsou pou\u017eit\u00e9 analytick\u00e9 metody pro stanoven\u00ed nikotinu a\u00a0jeho metabolit\u016f a\u00a0ethylsulf\u00e1tu pops\u00e1ny ve sborn\u00edku z\u00a0konference Hydroanalytika 2019 [36].<\/p>\n<\/a>\nObr. 7. Uk\u00e1zka chromatogramu extrahovan\u00fdch iont\u016f v\u00a0re\u017eimu ESI- (nor-THC)
\nFig. 7. Example of extracted ion chromatograms in ESI- (nor-THC)<\/h6>\n<\/a>\nObr. 8. Uk\u00e1zka chromatogramu extrahovan\u00fdch iont\u016f v\u00a0re\u017eimu ESI- (ethylsulf\u00e1t)
\nFig. 8. Example of extracted ion chromatograms in ESI- (ethylsulfate)<\/h6>\n
Pro skupinu syntetick\u00fdch kathinon\u016f byly hled\u00e1ny mo\u017enosti sn\u00ed\u017een\u00ed mez\u00ed stanovitelnosti, proto\u017ee nebyl zaznamen\u00e1n pozitivn\u00ed n\u00e1lez t\u011bchto l\u00e1tek (s\u00a0v\u00fdjimkou dvou hodnot mefedronu t\u011bsn\u011b na \u00farovni LOQ, obdob\u00ed hodnocen\u00ed 2018). Zkou\u0161en\u00e1 metodika vych\u00e1zela z\u00a0d\u0159\u00edv\u011bj\u0161\u00ed metodiky off-line SPE vyvinut\u00e9 v\u00a0projektu Program IGA 2013\u00a0\u2013 Projekt \u201eNov\u00e9 drogy\u00a0\u2013 anal\u00fdza trhu epidemiologie u\u017e\u00edv\u00e1n\u00ed a\u00a0identifikace mo\u017enosti pro sni\u017eov\u00e1n\u00ed \u0161kod\u201c [37]. Ani p\u0159i zpracov\u00e1n\u00ed 500\u00a0ml vzorku nebylo dosa\u017eeno lep\u0161\u00edch mez\u00ed stanovitelnosti ne\u017e p\u0159i on-line SPE metod\u011b [38].<\/p>\n
Z\u00e1v\u011br<\/h2>\n
Pro \u0159e\u0161en\u00ed projektu \u201e\u010cist\u00e1 voda\u00a0\u2013 zdrav\u00e9 m\u011bsto: Komun\u00e1ln\u00ed odpadn\u00ed voda jako diagnostick\u00e9 m\u00e9dium hlavn\u00edho m\u011bsta Prahy\u201c, ve\u00a0kter\u00e9m jsou monitorov\u00e1ny vedle nez\u00e1konn\u00fdch drog a\u00a0jejich metabolit\u016f (v\u010detn\u011b syntetick\u00fdch kathinon\u016f) tak\u00e9 n\u011bkter\u00e1 l\u00e9\u010diva a\u00a0dal\u0161\u00ed leg\u00e1ln\u00ed drogy, jako jsou ethanol a\u00a0nikotin, resp. jejich metabolity, byly vypracov\u00e1ny a\u00a0zvalidov\u00e1ny postupy pro stanoven\u00ed t\u011bchto l\u00e1tek. V\u00a0z\u00e1vislosti na chemick\u00fdch vlastnostech a\u00a0mo\u017enostech stanoven\u00ed jednotliv\u00fdch l\u00e1tek byly o\u00a0tyto l\u00e1tky roz\u0161\u00ed\u0159eny ji\u017e d\u0159\u00edve pou\u017e\u00edvan\u00e9 metody kapalinov\u00e9 chromatografie s\u00a0hmotnostn\u00ed detekc\u00ed za podm\u00ednek ionizace elektrosprejem v\u00a0pozitivn\u00edm m\u00f3du a\u00a0v\u00a0negativn\u00edm m\u00f3du. Metody byly akreditov\u00e1ny. Celkem jsou uveden\u00fdmi metodami stanovov\u00e1ny t\u0159i des\u00edtky l\u00e1tek. Metody rozvinut\u00e9 ve V\u00daV TGM, v. v. i., spl\u0148uj\u00ed po\u017eadavky na citlivost pro anal\u00fdzu v\u011bt\u0161iny zneu\u017e\u00edvan\u00fdch drog a\u00a0jejich metabolit\u016f i\u00a0dal\u0161\u00edch l\u00e1tek sledovan\u00fdch v\u00a0r\u00e1mci Projektu.<\/p>\n","protected":false},"excerpt":{"rendered":"
The wastewater-based epidemiology approach to the analysis of illicit drugs was conceptualized in 2001 and for the first time was applied in 2005 for cocaine estimation in Italy. Further, other illicit drugs as heroine, cannabis and amphetamine have been estimated.<\/p>\n","protected":false},"author":8,"featured_media":7943,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":{"_acf_changed":false,"footnotes":""},"categories":[90,89],"tags":[1105,1890,663,1107,1889,213],"coauthors":[1855,327,599,1856,1857],"class_list":["post-7990","post","type-post","status-publish","format-standard","has-post-thumbnail","hentry","category-waste-management","category-water-technology-water-supply-waste-water-treatment","tag-alcohol","tag-ethylsulfate","tag-illicit-drugs","tag-nicotine","tag-nicotine-metabolites","tag-wastewater"],"acf":[],"_links":{"self":[{"href":"https:\/\/www.vtei.cz\/en\/wp-json\/wp\/v2\/posts\/7990","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/www.vtei.cz\/en\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/www.vtei.cz\/en\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/www.vtei.cz\/en\/wp-json\/wp\/v2\/users\/8"}],"replies":[{"embeddable":true,"href":"https:\/\/www.vtei.cz\/en\/wp-json\/wp\/v2\/comments?post=7990"}],"version-history":[{"count":4,"href":"https:\/\/www.vtei.cz\/en\/wp-json\/wp\/v2\/posts\/7990\/revisions"}],"predecessor-version":[{"id":30580,"href":"https:\/\/www.vtei.cz\/en\/wp-json\/wp\/v2\/posts\/7990\/revisions\/30580"}],"wp:featuredmedia":[{"embeddable":true,"href":"https:\/\/www.vtei.cz\/en\/wp-json\/wp\/v2\/media\/7943"}],"wp:attachment":[{"href":"https:\/\/www.vtei.cz\/en\/wp-json\/wp\/v2\/media?parent=7990"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/www.vtei.cz\/en\/wp-json\/wp\/v2\/categories?post=7990"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/www.vtei.cz\/en\/wp-json\/wp\/v2\/tags?post=7990"},{"taxonomy":"author","embeddable":true,"href":"https:\/\/www.vtei.cz\/en\/wp-json\/wp\/v2\/coauthors?post=7990"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}
\nFig. 3. Example of calibration curve (cotinine)<\/h6>\n
<\/a>\nObr. 4. Uk\u00e1zka chromatogramu extrahovan\u00fdch iont\u016f v\u00a0re\u017eimu ESI+ (amfetamin, benzoylecgonin, buprenorphin, EDDP, efedrin, ext\u00e1ze, fentanyl, heroin, kokaethylen, kokain, LSD, metadon, metamfetamin, morfin, nor-diazepam, nor-fentanyl, tramadol)
\nFig. 4. Example of extracted ion chromatograms in ESI+ (amphetamine, benzoylecgonine, buprenorphine, EDDP, ephedrine, MDMA, fentanyl, heroin, cocaethylene, cocaine, LSD, methadone, methamphetamine, morphine, nor-diazepam, nor-fentanyl, tramadol)<\/h6>\n<\/a>\nObr. 5. Uk\u00e1zka chromatogramu extrahovan\u00fdch iont\u016f v\u00a0re\u017eimu ESI+ (ethylcathinon, mefedron, 4-methylethcathinon, metylon, nor-mefedron, pentedron, tramadol, trans<\/em>-3-hydroxykotinin, \u03b1-pyrrolidinovalerofenon)
\nFig. 5. Example of extracted ion chromatograms in ESI+ (ethylcathinone, mephedrone, 4-methylethcathinone, metylone, nor-mephedrone, pentedrone, tramadol, trans<\/em>-3-hydroxykotinin, \u03b1-pyrrolidinovalerofenone)<\/h6>\n<\/a>\nObr. 6. Uk\u00e1zka chromatogramu extrahovan\u00fdch iont\u016f v\u00a0re\u017eimu ESI+ (nikotin, kotinin a\u00a0trans<\/em>-3-hydroxykotinin)
\nFig. 6. Example of extracted ion chromatograms in ESI+ (nicotine, cotinine and trans<\/em>-3-hydroxycotinine)<\/h6>\nStanoven\u00ed ethylsulf\u00e1tu se d\u00e1 prov\u00e1d\u011bt i\u00a0s\u00a0HILIC kolonami, nap\u0159. SeQuant ZIC-HILIC (Merck) a\u00a0mobiln\u00edmi f\u00e1zemi 5 mM octan amonn\u00fd (pH 6,8) a\u00a0acetonitril [33].<\/p>\n
Metodika<\/h2>\n
Postupn\u011b bylo v\u00a0z\u00e1vislosti na chemick\u00fdch vlastnostech a\u00a0mo\u017enostech stanoven\u00ed jednotliv\u00fdch l\u00e1tek zavedeno stanoven\u00ed t\u0159\u00ed des\u00edtek l\u00e1tek metodou kapalinov\u00e9 chromatografie s\u00a0hmotnostn\u00ed detekc\u00ed za podm\u00ednek ionizace elektrosprejem v\u00a0pozitivn\u00edm a\u00a0negativn\u00edm m\u00f3du. P\u0159ehled stanovovan\u00fdch analyt\u016f v\u010detn\u011b jejich z\u00e1kladn\u00ed charakteristiky a\u00a0rozd\u011blen\u00ed do postup\u016f stanoven\u00ed je uveden v\u00a0tabulce 1<\/em>.<\/p>\nPou\u017eit\u00e1 technika: kapalinov\u00fd chromatograf Agilent 1200 RR s\u00a0bin\u00e1rn\u00edm \u010derpadlem, hmotnostn\u00ed detektor Applied Biosystem 4000 Q Trap s\u00a0trojit\u00fdm kvadrup\u00f3lem, autosampler Aspec GX-271 (Gilson). Stanoven\u00ed v\u0161ech l\u00e1tek bylo prov\u00e1d\u011bno s\u00a0chromatografickou kololnou Synergi Hydro-RP80A (150 \u00d7 2,0 mm, 4 \u00b5m).<\/p>\n
Multirezidu\u00e1ln\u00ed stanoven\u00ed t\u00e9to skupiny analyt\u016f se nedalo prov\u00e9st vzhledem k\u00a0jejich odli\u0161n\u00fdm chemick\u00fdm vlastnostem a\u00a0koncentra\u010dn\u00edm hladin\u00e1m jejich v\u00fdskytu. Z\u00a0tohoto d\u016fvodu bylo stanoven\u00ed rozd\u011bleno do \u010dty\u0159 separ\u00e1tn\u00edch postup\u016f.<\/p>\n
Anal\u00fdzy byly prov\u00e1d\u011bny v\u00a0odpadn\u00ed vod\u011b odeb\u00edran\u00e9 v\u00a0n\u00e1toku na \u010dist\u00edrnu komun\u00e1ln\u00edch odpadn\u00edch vod. Tato voda je velmi komplexn\u00ed matrice a\u00a0jako nezbytn\u00e9 se jev\u00ed pou\u017eit\u00ed vnit\u0159n\u00edch standard\u016f. Matrice siln\u011b ovliv\u0148uje ionizaci. Bylo pozorov\u00e1no potla\u010den\u00ed ionizace vlivem l\u00e1tek p\u0159\u00edtomn\u00fdch ve vzorku. Proto bylo pracov\u00e1no s\u00a0deuterovan\u00fdmi standardy v\u0161ech stanovovan\u00fdch l\u00e1tek.<\/p>\n
Pro anal\u00fdzy se zpracov\u00e1vaj\u00ed 24hodinov\u00e9 kompozitn\u00ed vzorky odeb\u00edran\u00e9 na n\u00e1toku na \u010dist\u00edrnu komun\u00e1ln\u00edch odpadn\u00edch vod, kter\u00e9 se po odb\u011bru uchov\u00e1vaj\u00ed v\u00a0chladu a\u00a0temnu. Pokud nelze vzorek analyzovat do 72 hodin od odb\u011bru, vzorek se zmraz\u00ed a\u00a0do vlastn\u00ed anal\u00fdzy se skladuje p\u0159i teplot\u011b -20 \u00b1 4 \u00b0C.<\/p>\n
P\u0159ed vlastn\u00edm stanoven\u00edm se vzorky odst\u0159ed\u00ed a\u00a0ze vzorku se odstran\u00ed pevn\u00e9 \u010d\u00e1stice filtrac\u00ed p\u0159es jednor\u00e1zov\u00e9 membr\u00e1nov\u00e9 filtry o\u00a0porozit\u011b 0,45 \u00b5m. Po p\u0159\u00eddavku sm\u011bsn\u00e9ho roztoku vnit\u0159n\u00edch standard\u016f je vzorek p\u0159ipraven k\u00a0LC-MS\/MS anal\u00fdze. P\u0159ed chromatogrfickou anal\u00fdzou se vzorky zakoncentruj\u00ed z\u00a0objemu 1\u20135\u00a0ml on-line SPE (drogy, nikotin a\u00a0jeho metabolity). K\u00a0on-line prekoncentraci vzork\u016f byl pou\u017eit autosampler Aspec GX-271(Gilson) a\u00a0kolonka Hypersil Gold (20\u00a0\u00d7 2,1 mm, 12 \u00b5m), na kterou bylo 1\u20135 ml vzorku vlo\u017eeno pr\u016ftokem 0,5 ml\/min. Potom byla kolonka je\u0161t\u011b promyta 1 ml UHQ vody, aby se dokon\u010dil transfer vzorku a\u00a0odstranily n\u011bkter\u00e9 interference. Eluce analyt\u016f na chromatografickou kolonu byla prov\u00e1d\u011bna mobiln\u00ed f\u00e1z\u00ed jej\u00edm pr\u016ftokem koncentra\u010dn\u00ed kolonkou v\u00a0protism\u011bru v\u00a0zapojen\u00ed podle obr.\u00a01<\/em>. Pouze anal\u00fdza ethylsulf\u00e1tu se vzhledem k\u00a0jeho vysok\u00fdm koncentrac\u00edm (des\u00edtky\u00a0\u00b5g\/l) prov\u00e1d\u00ed bez prekoncentrace s\u00a0n\u00e1st\u0159ikem 5 \u00b5l vzorku na kolonu.<\/p>\nV\u0161echny pou\u017e\u00edvan\u00e9 chemik\u00e1lie jsou \u010distoty p. a. nebo vy\u0161\u0161\u00ed. Pro kalibraci jsou pou\u017e\u00edv\u00e1ny certifikovan\u00e9 referen\u010dn\u00ed chemik\u00e1lie, nap\u0159. fy Merc, Lipomed, Chromservis. Jako vnit\u0159n\u00ed standardy jsou pou\u017e\u00edv\u00e1ny deuterovan\u00e9 l\u00e1tky, tak\u00e9 certifikovan\u00e9. Podm\u00ednky kapalinov\u00e9 chromatografie pro jednotliv\u00e9 postupy shrnuje tabulka 2<\/em>.<\/p>\nTabulka 2. Podm\u00ednky kapalinov\u00e9 chromatografie
\nTable 2. Liquide chromatography settings<\/h5>\n<\/a>\nIdentifikace a\u00a0kvantifikace se ve v\u0161ech p\u0159\u00edpadech prov\u00e1d\u00ed za pomoci vyhodnocovac\u00edho syst\u00e9mu Analyst\u00ae Software. Jednotliv\u00e9 l\u00e1tky se identifikuj\u00ed porovn\u00e1n\u00edm reten\u010dn\u00edch \u010das\u016f s\u00a0reten\u010dn\u00edmi \u010dasy standard\u016f a\u00a0shodnosti pom\u011bru intenzit dvou sledovan\u00fdch p\u0159echod\u016f dan\u00e9 l\u00e1tky ve vzorku a\u00a0v\u00a0kalibra\u010dn\u00edm roztoku. Kvantifikace se prov\u00e1d\u00ed na z\u00e1klad\u011b hodnot vypo\u010dten\u00fdch pomoc\u00ed kalibra\u010dn\u00edch k\u0159ivek metodou kalibrace na vnit\u0159n\u00ed standard (deuterovan\u00fd standard). Ve stanovovan\u00e9m rozsahu byly v\u0161echny kalibra\u010dn\u00ed k\u0159ivky line\u00e1rn\u00ed (korela\u010dn\u00ed koeficient 0,99 a\u00a0lep\u0161\u00ed).<\/p>\n
Pro uk\u00e1zku jsou na obr. 2\u20138<\/em> uvedeny kalibra\u010dn\u00ed k\u0159ivky a\u00a0chromatogramy extrahovan\u00fdch iont\u016f standard\u016f. Je zobrazen prekurzorov\u00fd, produktov\u00fd ion a\u00a0vnit\u0159n\u00ed standard.<\/p>\nMeze stanovitelnosti (LOQ) dosa\u017een\u00e9 popsan\u00fdmi postupy v\u00a0matrici odpadn\u00ed voda jsou uvedeny v\u00a0tabulce 3<\/em>.<\/p>\nTabulka 3. Meze stanovitelnosti (LOQ) v\u00a0matrici odpadn\u00ed voda
\nTable 3. Limits of quantification for wastewater<\/h6>\n<\/a>\nPodrobn\u011bji jsou pou\u017eit\u00e9 analytick\u00e9 metody pro stanoven\u00ed nikotinu a\u00a0jeho metabolit\u016f a\u00a0ethylsulf\u00e1tu pops\u00e1ny ve sborn\u00edku z\u00a0konference Hydroanalytika 2019 [36].<\/p>\n<\/a>\nObr. 7. Uk\u00e1zka chromatogramu extrahovan\u00fdch iont\u016f v\u00a0re\u017eimu ESI- (nor-THC)
\nFig. 7. Example of extracted ion chromatograms in ESI- (nor-THC)<\/h6>\n<\/a>\nObr. 8. Uk\u00e1zka chromatogramu extrahovan\u00fdch iont\u016f v\u00a0re\u017eimu ESI- (ethylsulf\u00e1t)
\nFig. 8. Example of extracted ion chromatograms in ESI- (ethylsulfate)<\/h6>\n
Pro skupinu syntetick\u00fdch kathinon\u016f byly hled\u00e1ny mo\u017enosti sn\u00ed\u017een\u00ed mez\u00ed stanovitelnosti, proto\u017ee nebyl zaznamen\u00e1n pozitivn\u00ed n\u00e1lez t\u011bchto l\u00e1tek (s\u00a0v\u00fdjimkou dvou hodnot mefedronu t\u011bsn\u011b na \u00farovni LOQ, obdob\u00ed hodnocen\u00ed 2018). Zkou\u0161en\u00e1 metodika vych\u00e1zela z\u00a0d\u0159\u00edv\u011bj\u0161\u00ed metodiky off-line SPE vyvinut\u00e9 v\u00a0projektu Program IGA 2013\u00a0\u2013 Projekt \u201eNov\u00e9 drogy\u00a0\u2013 anal\u00fdza trhu epidemiologie u\u017e\u00edv\u00e1n\u00ed a\u00a0identifikace mo\u017enosti pro sni\u017eov\u00e1n\u00ed \u0161kod\u201c [37]. Ani p\u0159i zpracov\u00e1n\u00ed 500\u00a0ml vzorku nebylo dosa\u017eeno lep\u0161\u00edch mez\u00ed stanovitelnosti ne\u017e p\u0159i on-line SPE metod\u011b [38].<\/p>\n
Z\u00e1v\u011br<\/h2>\n
Pro \u0159e\u0161en\u00ed projektu \u201e\u010cist\u00e1 voda\u00a0\u2013 zdrav\u00e9 m\u011bsto: Komun\u00e1ln\u00ed odpadn\u00ed voda jako diagnostick\u00e9 m\u00e9dium hlavn\u00edho m\u011bsta Prahy\u201c, ve\u00a0kter\u00e9m jsou monitorov\u00e1ny vedle nez\u00e1konn\u00fdch drog a\u00a0jejich metabolit\u016f (v\u010detn\u011b syntetick\u00fdch kathinon\u016f) tak\u00e9 n\u011bkter\u00e1 l\u00e9\u010diva a\u00a0dal\u0161\u00ed leg\u00e1ln\u00ed drogy, jako jsou ethanol a\u00a0nikotin, resp. jejich metabolity, byly vypracov\u00e1ny a\u00a0zvalidov\u00e1ny postupy pro stanoven\u00ed t\u011bchto l\u00e1tek. V\u00a0z\u00e1vislosti na chemick\u00fdch vlastnostech a\u00a0mo\u017enostech stanoven\u00ed jednotliv\u00fdch l\u00e1tek byly o\u00a0tyto l\u00e1tky roz\u0161\u00ed\u0159eny ji\u017e d\u0159\u00edve pou\u017e\u00edvan\u00e9 metody kapalinov\u00e9 chromatografie s\u00a0hmotnostn\u00ed detekc\u00ed za podm\u00ednek ionizace elektrosprejem v\u00a0pozitivn\u00edm m\u00f3du a\u00a0v\u00a0negativn\u00edm m\u00f3du. Metody byly akreditov\u00e1ny. Celkem jsou uveden\u00fdmi metodami stanovov\u00e1ny t\u0159i des\u00edtky l\u00e1tek. Metody rozvinut\u00e9 ve V\u00daV TGM, v. v. i., spl\u0148uj\u00ed po\u017eadavky na citlivost pro anal\u00fdzu v\u011bt\u0161iny zneu\u017e\u00edvan\u00fdch drog a\u00a0jejich metabolit\u016f i\u00a0dal\u0161\u00edch l\u00e1tek sledovan\u00fdch v\u00a0r\u00e1mci Projektu.<\/p>\n","protected":false},"excerpt":{"rendered":"
The wastewater-based epidemiology approach to the analysis of illicit drugs was conceptualized in 2001 and for the first time was applied in 2005 for cocaine estimation in Italy. Further, other illicit drugs as heroine, cannabis and amphetamine have been estimated.<\/p>\n","protected":false},"author":8,"featured_media":7943,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":{"_acf_changed":false,"footnotes":""},"categories":[90,89],"tags":[1105,1890,663,1107,1889,213],"coauthors":[1855,327,599,1856,1857],"class_list":["post-7990","post","type-post","status-publish","format-standard","has-post-thumbnail","hentry","category-waste-management","category-water-technology-water-supply-waste-water-treatment","tag-alcohol","tag-ethylsulfate","tag-illicit-drugs","tag-nicotine","tag-nicotine-metabolites","tag-wastewater"],"acf":[],"_links":{"self":[{"href":"https:\/\/www.vtei.cz\/en\/wp-json\/wp\/v2\/posts\/7990","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/www.vtei.cz\/en\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/www.vtei.cz\/en\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/www.vtei.cz\/en\/wp-json\/wp\/v2\/users\/8"}],"replies":[{"embeddable":true,"href":"https:\/\/www.vtei.cz\/en\/wp-json\/wp\/v2\/comments?post=7990"}],"version-history":[{"count":4,"href":"https:\/\/www.vtei.cz\/en\/wp-json\/wp\/v2\/posts\/7990\/revisions"}],"predecessor-version":[{"id":30580,"href":"https:\/\/www.vtei.cz\/en\/wp-json\/wp\/v2\/posts\/7990\/revisions\/30580"}],"wp:featuredmedia":[{"embeddable":true,"href":"https:\/\/www.vtei.cz\/en\/wp-json\/wp\/v2\/media\/7943"}],"wp:attachment":[{"href":"https:\/\/www.vtei.cz\/en\/wp-json\/wp\/v2\/media?parent=7990"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/www.vtei.cz\/en\/wp-json\/wp\/v2\/categories?post=7990"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/www.vtei.cz\/en\/wp-json\/wp\/v2\/tags?post=7990"},{"taxonomy":"author","embeddable":true,"href":"https:\/\/www.vtei.cz\/en\/wp-json\/wp\/v2\/coauthors?post=7990"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}
\nFig. 5. Example of extracted ion chromatograms in ESI+ (ethylcathinone, mephedrone, 4-methylethcathinone, metylone, nor-mephedrone, pentedrone, tramadol, trans<\/em>-3-hydroxykotinin, \u03b1-pyrrolidinovalerofenone)<\/h6>\n
<\/a>\nObr. 6. Uk\u00e1zka chromatogramu extrahovan\u00fdch iont\u016f v\u00a0re\u017eimu ESI+ (nikotin, kotinin a\u00a0trans<\/em>-3-hydroxykotinin)
\nFig. 6. Example of extracted ion chromatograms in ESI+ (nicotine, cotinine and trans<\/em>-3-hydroxycotinine)<\/h6>\nStanoven\u00ed ethylsulf\u00e1tu se d\u00e1 prov\u00e1d\u011bt i\u00a0s\u00a0HILIC kolonami, nap\u0159. SeQuant ZIC-HILIC (Merck) a\u00a0mobiln\u00edmi f\u00e1zemi 5 mM octan amonn\u00fd (pH 6,8) a\u00a0acetonitril [33].<\/p>\n
Metodika<\/h2>\n
Postupn\u011b bylo v\u00a0z\u00e1vislosti na chemick\u00fdch vlastnostech a\u00a0mo\u017enostech stanoven\u00ed jednotliv\u00fdch l\u00e1tek zavedeno stanoven\u00ed t\u0159\u00ed des\u00edtek l\u00e1tek metodou kapalinov\u00e9 chromatografie s\u00a0hmotnostn\u00ed detekc\u00ed za podm\u00ednek ionizace elektrosprejem v\u00a0pozitivn\u00edm a\u00a0negativn\u00edm m\u00f3du. P\u0159ehled stanovovan\u00fdch analyt\u016f v\u010detn\u011b jejich z\u00e1kladn\u00ed charakteristiky a\u00a0rozd\u011blen\u00ed do postup\u016f stanoven\u00ed je uveden v\u00a0tabulce 1<\/em>.<\/p>\nPou\u017eit\u00e1 technika: kapalinov\u00fd chromatograf Agilent 1200 RR s\u00a0bin\u00e1rn\u00edm \u010derpadlem, hmotnostn\u00ed detektor Applied Biosystem 4000 Q Trap s\u00a0trojit\u00fdm kvadrup\u00f3lem, autosampler Aspec GX-271 (Gilson). Stanoven\u00ed v\u0161ech l\u00e1tek bylo prov\u00e1d\u011bno s\u00a0chromatografickou kololnou Synergi Hydro-RP80A (150 \u00d7 2,0 mm, 4 \u00b5m).<\/p>\n
Multirezidu\u00e1ln\u00ed stanoven\u00ed t\u00e9to skupiny analyt\u016f se nedalo prov\u00e9st vzhledem k\u00a0jejich odli\u0161n\u00fdm chemick\u00fdm vlastnostem a\u00a0koncentra\u010dn\u00edm hladin\u00e1m jejich v\u00fdskytu. Z\u00a0tohoto d\u016fvodu bylo stanoven\u00ed rozd\u011bleno do \u010dty\u0159 separ\u00e1tn\u00edch postup\u016f.<\/p>\n
Anal\u00fdzy byly prov\u00e1d\u011bny v\u00a0odpadn\u00ed vod\u011b odeb\u00edran\u00e9 v\u00a0n\u00e1toku na \u010dist\u00edrnu komun\u00e1ln\u00edch odpadn\u00edch vod. Tato voda je velmi komplexn\u00ed matrice a\u00a0jako nezbytn\u00e9 se jev\u00ed pou\u017eit\u00ed vnit\u0159n\u00edch standard\u016f. Matrice siln\u011b ovliv\u0148uje ionizaci. Bylo pozorov\u00e1no potla\u010den\u00ed ionizace vlivem l\u00e1tek p\u0159\u00edtomn\u00fdch ve vzorku. Proto bylo pracov\u00e1no s\u00a0deuterovan\u00fdmi standardy v\u0161ech stanovovan\u00fdch l\u00e1tek.<\/p>\n
Pro anal\u00fdzy se zpracov\u00e1vaj\u00ed 24hodinov\u00e9 kompozitn\u00ed vzorky odeb\u00edran\u00e9 na n\u00e1toku na \u010dist\u00edrnu komun\u00e1ln\u00edch odpadn\u00edch vod, kter\u00e9 se po odb\u011bru uchov\u00e1vaj\u00ed v\u00a0chladu a\u00a0temnu. Pokud nelze vzorek analyzovat do 72 hodin od odb\u011bru, vzorek se zmraz\u00ed a\u00a0do vlastn\u00ed anal\u00fdzy se skladuje p\u0159i teplot\u011b -20 \u00b1 4 \u00b0C.<\/p>\n
P\u0159ed vlastn\u00edm stanoven\u00edm se vzorky odst\u0159ed\u00ed a\u00a0ze vzorku se odstran\u00ed pevn\u00e9 \u010d\u00e1stice filtrac\u00ed p\u0159es jednor\u00e1zov\u00e9 membr\u00e1nov\u00e9 filtry o\u00a0porozit\u011b 0,45 \u00b5m. Po p\u0159\u00eddavku sm\u011bsn\u00e9ho roztoku vnit\u0159n\u00edch standard\u016f je vzorek p\u0159ipraven k\u00a0LC-MS\/MS anal\u00fdze. P\u0159ed chromatogrfickou anal\u00fdzou se vzorky zakoncentruj\u00ed z\u00a0objemu 1\u20135\u00a0ml on-line SPE (drogy, nikotin a\u00a0jeho metabolity). K\u00a0on-line prekoncentraci vzork\u016f byl pou\u017eit autosampler Aspec GX-271(Gilson) a\u00a0kolonka Hypersil Gold (20\u00a0\u00d7 2,1 mm, 12 \u00b5m), na kterou bylo 1\u20135 ml vzorku vlo\u017eeno pr\u016ftokem 0,5 ml\/min. Potom byla kolonka je\u0161t\u011b promyta 1 ml UHQ vody, aby se dokon\u010dil transfer vzorku a\u00a0odstranily n\u011bkter\u00e9 interference. Eluce analyt\u016f na chromatografickou kolonu byla prov\u00e1d\u011bna mobiln\u00ed f\u00e1z\u00ed jej\u00edm pr\u016ftokem koncentra\u010dn\u00ed kolonkou v\u00a0protism\u011bru v\u00a0zapojen\u00ed podle obr.\u00a01<\/em>. Pouze anal\u00fdza ethylsulf\u00e1tu se vzhledem k\u00a0jeho vysok\u00fdm koncentrac\u00edm (des\u00edtky\u00a0\u00b5g\/l) prov\u00e1d\u00ed bez prekoncentrace s\u00a0n\u00e1st\u0159ikem 5 \u00b5l vzorku na kolonu.<\/p>\nV\u0161echny pou\u017e\u00edvan\u00e9 chemik\u00e1lie jsou \u010distoty p. a. nebo vy\u0161\u0161\u00ed. Pro kalibraci jsou pou\u017e\u00edv\u00e1ny certifikovan\u00e9 referen\u010dn\u00ed chemik\u00e1lie, nap\u0159. fy Merc, Lipomed, Chromservis. Jako vnit\u0159n\u00ed standardy jsou pou\u017e\u00edv\u00e1ny deuterovan\u00e9 l\u00e1tky, tak\u00e9 certifikovan\u00e9. Podm\u00ednky kapalinov\u00e9 chromatografie pro jednotliv\u00e9 postupy shrnuje tabulka 2<\/em>.<\/p>\nTabulka 2. Podm\u00ednky kapalinov\u00e9 chromatografie
\nTable 2. Liquide chromatography settings<\/h5>\n<\/a>\nIdentifikace a\u00a0kvantifikace se ve v\u0161ech p\u0159\u00edpadech prov\u00e1d\u00ed za pomoci vyhodnocovac\u00edho syst\u00e9mu Analyst\u00ae Software. Jednotliv\u00e9 l\u00e1tky se identifikuj\u00ed porovn\u00e1n\u00edm reten\u010dn\u00edch \u010das\u016f s\u00a0reten\u010dn\u00edmi \u010dasy standard\u016f a\u00a0shodnosti pom\u011bru intenzit dvou sledovan\u00fdch p\u0159echod\u016f dan\u00e9 l\u00e1tky ve vzorku a\u00a0v\u00a0kalibra\u010dn\u00edm roztoku. Kvantifikace se prov\u00e1d\u00ed na z\u00e1klad\u011b hodnot vypo\u010dten\u00fdch pomoc\u00ed kalibra\u010dn\u00edch k\u0159ivek metodou kalibrace na vnit\u0159n\u00ed standard (deuterovan\u00fd standard). Ve stanovovan\u00e9m rozsahu byly v\u0161echny kalibra\u010dn\u00ed k\u0159ivky line\u00e1rn\u00ed (korela\u010dn\u00ed koeficient 0,99 a\u00a0lep\u0161\u00ed).<\/p>\n
Pro uk\u00e1zku jsou na obr. 2\u20138<\/em> uvedeny kalibra\u010dn\u00ed k\u0159ivky a\u00a0chromatogramy extrahovan\u00fdch iont\u016f standard\u016f. Je zobrazen prekurzorov\u00fd, produktov\u00fd ion a\u00a0vnit\u0159n\u00ed standard.<\/p>\nMeze stanovitelnosti (LOQ) dosa\u017een\u00e9 popsan\u00fdmi postupy v\u00a0matrici odpadn\u00ed voda jsou uvedeny v\u00a0tabulce 3<\/em>.<\/p>\nTabulka 3. Meze stanovitelnosti (LOQ) v\u00a0matrici odpadn\u00ed voda
\nTable 3. Limits of quantification for wastewater<\/h6>\n<\/a>\nPodrobn\u011bji jsou pou\u017eit\u00e9 analytick\u00e9 metody pro stanoven\u00ed nikotinu a\u00a0jeho metabolit\u016f a\u00a0ethylsulf\u00e1tu pops\u00e1ny ve sborn\u00edku z\u00a0konference Hydroanalytika 2019 [36].<\/p>\n<\/a>\nObr. 7. Uk\u00e1zka chromatogramu extrahovan\u00fdch iont\u016f v\u00a0re\u017eimu ESI- (nor-THC)
\nFig. 7. Example of extracted ion chromatograms in ESI- (nor-THC)<\/h6>\n<\/a>\nObr. 8. Uk\u00e1zka chromatogramu extrahovan\u00fdch iont\u016f v\u00a0re\u017eimu ESI- (ethylsulf\u00e1t)
\nFig. 8. Example of extracted ion chromatograms in ESI- (ethylsulfate)<\/h6>\n
Pro skupinu syntetick\u00fdch kathinon\u016f byly hled\u00e1ny mo\u017enosti sn\u00ed\u017een\u00ed mez\u00ed stanovitelnosti, proto\u017ee nebyl zaznamen\u00e1n pozitivn\u00ed n\u00e1lez t\u011bchto l\u00e1tek (s\u00a0v\u00fdjimkou dvou hodnot mefedronu t\u011bsn\u011b na \u00farovni LOQ, obdob\u00ed hodnocen\u00ed 2018). Zkou\u0161en\u00e1 metodika vych\u00e1zela z\u00a0d\u0159\u00edv\u011bj\u0161\u00ed metodiky off-line SPE vyvinut\u00e9 v\u00a0projektu Program IGA 2013\u00a0\u2013 Projekt \u201eNov\u00e9 drogy\u00a0\u2013 anal\u00fdza trhu epidemiologie u\u017e\u00edv\u00e1n\u00ed a\u00a0identifikace mo\u017enosti pro sni\u017eov\u00e1n\u00ed \u0161kod\u201c [37]. Ani p\u0159i zpracov\u00e1n\u00ed 500\u00a0ml vzorku nebylo dosa\u017eeno lep\u0161\u00edch mez\u00ed stanovitelnosti ne\u017e p\u0159i on-line SPE metod\u011b [38].<\/p>\n
Z\u00e1v\u011br<\/h2>\n
Pro \u0159e\u0161en\u00ed projektu \u201e\u010cist\u00e1 voda\u00a0\u2013 zdrav\u00e9 m\u011bsto: Komun\u00e1ln\u00ed odpadn\u00ed voda jako diagnostick\u00e9 m\u00e9dium hlavn\u00edho m\u011bsta Prahy\u201c, ve\u00a0kter\u00e9m jsou monitorov\u00e1ny vedle nez\u00e1konn\u00fdch drog a\u00a0jejich metabolit\u016f (v\u010detn\u011b syntetick\u00fdch kathinon\u016f) tak\u00e9 n\u011bkter\u00e1 l\u00e9\u010diva a\u00a0dal\u0161\u00ed leg\u00e1ln\u00ed drogy, jako jsou ethanol a\u00a0nikotin, resp. jejich metabolity, byly vypracov\u00e1ny a\u00a0zvalidov\u00e1ny postupy pro stanoven\u00ed t\u011bchto l\u00e1tek. V\u00a0z\u00e1vislosti na chemick\u00fdch vlastnostech a\u00a0mo\u017enostech stanoven\u00ed jednotliv\u00fdch l\u00e1tek byly o\u00a0tyto l\u00e1tky roz\u0161\u00ed\u0159eny ji\u017e d\u0159\u00edve pou\u017e\u00edvan\u00e9 metody kapalinov\u00e9 chromatografie s\u00a0hmotnostn\u00ed detekc\u00ed za podm\u00ednek ionizace elektrosprejem v\u00a0pozitivn\u00edm m\u00f3du a\u00a0v\u00a0negativn\u00edm m\u00f3du. Metody byly akreditov\u00e1ny. Celkem jsou uveden\u00fdmi metodami stanovov\u00e1ny t\u0159i des\u00edtky l\u00e1tek. Metody rozvinut\u00e9 ve V\u00daV TGM, v. v. i., spl\u0148uj\u00ed po\u017eadavky na citlivost pro anal\u00fdzu v\u011bt\u0161iny zneu\u017e\u00edvan\u00fdch drog a\u00a0jejich metabolit\u016f i\u00a0dal\u0161\u00edch l\u00e1tek sledovan\u00fdch v\u00a0r\u00e1mci Projektu.<\/p>\n","protected":false},"excerpt":{"rendered":"
The wastewater-based epidemiology approach to the analysis of illicit drugs was conceptualized in 2001 and for the first time was applied in 2005 for cocaine estimation in Italy. Further, other illicit drugs as heroine, cannabis and amphetamine have been estimated.<\/p>\n","protected":false},"author":8,"featured_media":7943,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":{"_acf_changed":false,"footnotes":""},"categories":[90,89],"tags":[1105,1890,663,1107,1889,213],"coauthors":[1855,327,599,1856,1857],"class_list":["post-7990","post","type-post","status-publish","format-standard","has-post-thumbnail","hentry","category-waste-management","category-water-technology-water-supply-waste-water-treatment","tag-alcohol","tag-ethylsulfate","tag-illicit-drugs","tag-nicotine","tag-nicotine-metabolites","tag-wastewater"],"acf":[],"_links":{"self":[{"href":"https:\/\/www.vtei.cz\/en\/wp-json\/wp\/v2\/posts\/7990","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/www.vtei.cz\/en\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/www.vtei.cz\/en\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/www.vtei.cz\/en\/wp-json\/wp\/v2\/users\/8"}],"replies":[{"embeddable":true,"href":"https:\/\/www.vtei.cz\/en\/wp-json\/wp\/v2\/comments?post=7990"}],"version-history":[{"count":4,"href":"https:\/\/www.vtei.cz\/en\/wp-json\/wp\/v2\/posts\/7990\/revisions"}],"predecessor-version":[{"id":30580,"href":"https:\/\/www.vtei.cz\/en\/wp-json\/wp\/v2\/posts\/7990\/revisions\/30580"}],"wp:featuredmedia":[{"embeddable":true,"href":"https:\/\/www.vtei.cz\/en\/wp-json\/wp\/v2\/media\/7943"}],"wp:attachment":[{"href":"https:\/\/www.vtei.cz\/en\/wp-json\/wp\/v2\/media?parent=7990"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/www.vtei.cz\/en\/wp-json\/wp\/v2\/categories?post=7990"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/www.vtei.cz\/en\/wp-json\/wp\/v2\/tags?post=7990"},{"taxonomy":"author","embeddable":true,"href":"https:\/\/www.vtei.cz\/en\/wp-json\/wp\/v2\/coauthors?post=7990"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}
Pou\u017eit\u00e1 technika: kapalinov\u00fd chromatograf Agilent 1200 RR s\u00a0bin\u00e1rn\u00edm \u010derpadlem, hmotnostn\u00ed detektor Applied Biosystem 4000 Q Trap s\u00a0trojit\u00fdm kvadrup\u00f3lem, autosampler Aspec GX-271 (Gilson). Stanoven\u00ed v\u0161ech l\u00e1tek bylo prov\u00e1d\u011bno s\u00a0chromatografickou kololnou Synergi Hydro-RP80A (150 \u00d7 2,0 mm, 4 \u00b5m).<\/p>\n
Multirezidu\u00e1ln\u00ed stanoven\u00ed t\u00e9to skupiny analyt\u016f se nedalo prov\u00e9st vzhledem k\u00a0jejich odli\u0161n\u00fdm chemick\u00fdm vlastnostem a\u00a0koncentra\u010dn\u00edm hladin\u00e1m jejich v\u00fdskytu. Z\u00a0tohoto d\u016fvodu bylo stanoven\u00ed rozd\u011bleno do \u010dty\u0159 separ\u00e1tn\u00edch postup\u016f.<\/p>\n
Anal\u00fdzy byly prov\u00e1d\u011bny v\u00a0odpadn\u00ed vod\u011b odeb\u00edran\u00e9 v\u00a0n\u00e1toku na \u010dist\u00edrnu komun\u00e1ln\u00edch odpadn\u00edch vod. Tato voda je velmi komplexn\u00ed matrice a\u00a0jako nezbytn\u00e9 se jev\u00ed pou\u017eit\u00ed vnit\u0159n\u00edch standard\u016f. Matrice siln\u011b ovliv\u0148uje ionizaci. Bylo pozorov\u00e1no potla\u010den\u00ed ionizace vlivem l\u00e1tek p\u0159\u00edtomn\u00fdch ve vzorku. Proto bylo pracov\u00e1no s\u00a0deuterovan\u00fdmi standardy v\u0161ech stanovovan\u00fdch l\u00e1tek.<\/p>\n
Pro anal\u00fdzy se zpracov\u00e1vaj\u00ed 24hodinov\u00e9 kompozitn\u00ed vzorky odeb\u00edran\u00e9 na n\u00e1toku na \u010dist\u00edrnu komun\u00e1ln\u00edch odpadn\u00edch vod, kter\u00e9 se po odb\u011bru uchov\u00e1vaj\u00ed v\u00a0chladu a\u00a0temnu. Pokud nelze vzorek analyzovat do 72 hodin od odb\u011bru, vzorek se zmraz\u00ed a\u00a0do vlastn\u00ed anal\u00fdzy se skladuje p\u0159i teplot\u011b -20 \u00b1 4 \u00b0C.<\/p>\n
P\u0159ed vlastn\u00edm stanoven\u00edm se vzorky odst\u0159ed\u00ed a\u00a0ze vzorku se odstran\u00ed pevn\u00e9 \u010d\u00e1stice filtrac\u00ed p\u0159es jednor\u00e1zov\u00e9 membr\u00e1nov\u00e9 filtry o\u00a0porozit\u011b 0,45 \u00b5m. Po p\u0159\u00eddavku sm\u011bsn\u00e9ho roztoku vnit\u0159n\u00edch standard\u016f je vzorek p\u0159ipraven k\u00a0LC-MS\/MS anal\u00fdze. P\u0159ed chromatogrfickou anal\u00fdzou se vzorky zakoncentruj\u00ed z\u00a0objemu 1\u20135\u00a0ml on-line SPE (drogy, nikotin a\u00a0jeho metabolity). K\u00a0on-line prekoncentraci vzork\u016f byl pou\u017eit autosampler Aspec GX-271(Gilson) a\u00a0kolonka Hypersil Gold (20\u00a0\u00d7 2,1 mm, 12 \u00b5m), na kterou bylo 1\u20135 ml vzorku vlo\u017eeno pr\u016ftokem 0,5 ml\/min. Potom byla kolonka je\u0161t\u011b promyta 1 ml UHQ vody, aby se dokon\u010dil transfer vzorku a\u00a0odstranily n\u011bkter\u00e9 interference. Eluce analyt\u016f na chromatografickou kolonu byla prov\u00e1d\u011bna mobiln\u00ed f\u00e1z\u00ed jej\u00edm pr\u016ftokem koncentra\u010dn\u00ed kolonkou v\u00a0protism\u011bru v\u00a0zapojen\u00ed podle obr.\u00a01<\/em>. Pouze anal\u00fdza ethylsulf\u00e1tu se vzhledem k\u00a0jeho vysok\u00fdm koncentrac\u00edm (des\u00edtky\u00a0\u00b5g\/l) prov\u00e1d\u00ed bez prekoncentrace s\u00a0n\u00e1st\u0159ikem 5 \u00b5l vzorku na kolonu.<\/p>\n V\u0161echny pou\u017e\u00edvan\u00e9 chemik\u00e1lie jsou \u010distoty p. a. nebo vy\u0161\u0161\u00ed. Pro kalibraci jsou pou\u017e\u00edv\u00e1ny certifikovan\u00e9 referen\u010dn\u00ed chemik\u00e1lie, nap\u0159. fy Merc, Lipomed, Chromservis. Jako vnit\u0159n\u00ed standardy jsou pou\u017e\u00edv\u00e1ny deuterovan\u00e9 l\u00e1tky, tak\u00e9 certifikovan\u00e9. Podm\u00ednky kapalinov\u00e9 chromatografie pro jednotliv\u00e9 postupy shrnuje tabulka 2<\/em>.<\/p>\n Identifikace a\u00a0kvantifikace se ve v\u0161ech p\u0159\u00edpadech prov\u00e1d\u00ed za pomoci vyhodnocovac\u00edho syst\u00e9mu Analyst\u00ae Software. Jednotliv\u00e9 l\u00e1tky se identifikuj\u00ed porovn\u00e1n\u00edm reten\u010dn\u00edch \u010das\u016f s\u00a0reten\u010dn\u00edmi \u010dasy standard\u016f a\u00a0shodnosti pom\u011bru intenzit dvou sledovan\u00fdch p\u0159echod\u016f dan\u00e9 l\u00e1tky ve vzorku a\u00a0v\u00a0kalibra\u010dn\u00edm roztoku. Kvantifikace se prov\u00e1d\u00ed na z\u00e1klad\u011b hodnot vypo\u010dten\u00fdch pomoc\u00ed kalibra\u010dn\u00edch k\u0159ivek metodou kalibrace na vnit\u0159n\u00ed standard (deuterovan\u00fd standard). Ve stanovovan\u00e9m rozsahu byly v\u0161echny kalibra\u010dn\u00ed k\u0159ivky line\u00e1rn\u00ed (korela\u010dn\u00ed koeficient 0,99 a\u00a0lep\u0161\u00ed).<\/p>\n Pro uk\u00e1zku jsou na obr. 2\u20138<\/em> uvedeny kalibra\u010dn\u00ed k\u0159ivky a\u00a0chromatogramy extrahovan\u00fdch iont\u016f standard\u016f. Je zobrazen prekurzorov\u00fd, produktov\u00fd ion a\u00a0vnit\u0159n\u00ed standard.<\/p>\n Meze stanovitelnosti (LOQ) dosa\u017een\u00e9 popsan\u00fdmi postupy v\u00a0matrici odpadn\u00ed voda jsou uvedeny v\u00a0tabulce 3<\/em>.<\/p>\n Podrobn\u011bji jsou pou\u017eit\u00e9 analytick\u00e9 metody pro stanoven\u00ed nikotinu a\u00a0jeho metabolit\u016f a\u00a0ethylsulf\u00e1tu pops\u00e1ny ve sborn\u00edku z\u00a0konference Hydroanalytika 2019 [36].<\/p>\n Pro skupinu syntetick\u00fdch kathinon\u016f byly hled\u00e1ny mo\u017enosti sn\u00ed\u017een\u00ed mez\u00ed stanovitelnosti, proto\u017ee nebyl zaznamen\u00e1n pozitivn\u00ed n\u00e1lez t\u011bchto l\u00e1tek (s\u00a0v\u00fdjimkou dvou hodnot mefedronu t\u011bsn\u011b na \u00farovni LOQ, obdob\u00ed hodnocen\u00ed 2018). Zkou\u0161en\u00e1 metodika vych\u00e1zela z\u00a0d\u0159\u00edv\u011bj\u0161\u00ed metodiky off-line SPE vyvinut\u00e9 v\u00a0projektu Program IGA 2013\u00a0\u2013 Projekt \u201eNov\u00e9 drogy\u00a0\u2013 anal\u00fdza trhu epidemiologie u\u017e\u00edv\u00e1n\u00ed a\u00a0identifikace mo\u017enosti pro sni\u017eov\u00e1n\u00ed \u0161kod\u201c [37]. Ani p\u0159i zpracov\u00e1n\u00ed 500\u00a0ml vzorku nebylo dosa\u017eeno lep\u0161\u00edch mez\u00ed stanovitelnosti ne\u017e p\u0159i on-line SPE metod\u011b [38].<\/p>\n Pro \u0159e\u0161en\u00ed projektu \u201e\u010cist\u00e1 voda\u00a0\u2013 zdrav\u00e9 m\u011bsto: Komun\u00e1ln\u00ed odpadn\u00ed voda jako diagnostick\u00e9 m\u00e9dium hlavn\u00edho m\u011bsta Prahy\u201c, ve\u00a0kter\u00e9m jsou monitorov\u00e1ny vedle nez\u00e1konn\u00fdch drog a\u00a0jejich metabolit\u016f (v\u010detn\u011b syntetick\u00fdch kathinon\u016f) tak\u00e9 n\u011bkter\u00e1 l\u00e9\u010diva a\u00a0dal\u0161\u00ed leg\u00e1ln\u00ed drogy, jako jsou ethanol a\u00a0nikotin, resp. jejich metabolity, byly vypracov\u00e1ny a\u00a0zvalidov\u00e1ny postupy pro stanoven\u00ed t\u011bchto l\u00e1tek. V\u00a0z\u00e1vislosti na chemick\u00fdch vlastnostech a\u00a0mo\u017enostech stanoven\u00ed jednotliv\u00fdch l\u00e1tek byly o\u00a0tyto l\u00e1tky roz\u0161\u00ed\u0159eny ji\u017e d\u0159\u00edve pou\u017e\u00edvan\u00e9 metody kapalinov\u00e9 chromatografie s\u00a0hmotnostn\u00ed detekc\u00ed za podm\u00ednek ionizace elektrosprejem v\u00a0pozitivn\u00edm m\u00f3du a\u00a0v\u00a0negativn\u00edm m\u00f3du. Metody byly akreditov\u00e1ny. Celkem jsou uveden\u00fdmi metodami stanovov\u00e1ny t\u0159i des\u00edtky l\u00e1tek. Metody rozvinut\u00e9 ve V\u00daV TGM, v. v. i., spl\u0148uj\u00ed po\u017eadavky na citlivost pro anal\u00fdzu v\u011bt\u0161iny zneu\u017e\u00edvan\u00fdch drog a\u00a0jejich metabolit\u016f i\u00a0dal\u0161\u00edch l\u00e1tek sledovan\u00fdch v\u00a0r\u00e1mci Projektu.<\/p>\n","protected":false},"excerpt":{"rendered":" The wastewater-based epidemiology approach to the analysis of illicit drugs was conceptualized in 2001 and for the first time was applied in 2005 for cocaine estimation in Italy. Further, other illicit drugs as heroine, cannabis and amphetamine have been estimated.<\/p>\n","protected":false},"author":8,"featured_media":7943,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":{"_acf_changed":false,"footnotes":""},"categories":[90,89],"tags":[1105,1890,663,1107,1889,213],"coauthors":[1855,327,599,1856,1857],"class_list":["post-7990","post","type-post","status-publish","format-standard","has-post-thumbnail","hentry","category-waste-management","category-water-technology-water-supply-waste-water-treatment","tag-alcohol","tag-ethylsulfate","tag-illicit-drugs","tag-nicotine","tag-nicotine-metabolites","tag-wastewater"],"acf":[],"_links":{"self":[{"href":"https:\/\/www.vtei.cz\/en\/wp-json\/wp\/v2\/posts\/7990","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/www.vtei.cz\/en\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/www.vtei.cz\/en\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/www.vtei.cz\/en\/wp-json\/wp\/v2\/users\/8"}],"replies":[{"embeddable":true,"href":"https:\/\/www.vtei.cz\/en\/wp-json\/wp\/v2\/comments?post=7990"}],"version-history":[{"count":4,"href":"https:\/\/www.vtei.cz\/en\/wp-json\/wp\/v2\/posts\/7990\/revisions"}],"predecessor-version":[{"id":30580,"href":"https:\/\/www.vtei.cz\/en\/wp-json\/wp\/v2\/posts\/7990\/revisions\/30580"}],"wp:featuredmedia":[{"embeddable":true,"href":"https:\/\/www.vtei.cz\/en\/wp-json\/wp\/v2\/media\/7943"}],"wp:attachment":[{"href":"https:\/\/www.vtei.cz\/en\/wp-json\/wp\/v2\/media?parent=7990"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/www.vtei.cz\/en\/wp-json\/wp\/v2\/categories?post=7990"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/www.vtei.cz\/en\/wp-json\/wp\/v2\/tags?post=7990"},{"taxonomy":"author","embeddable":true,"href":"https:\/\/www.vtei.cz\/en\/wp-json\/wp\/v2\/coauthors?post=7990"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}Tabulka 2. Podm\u00ednky kapalinov\u00e9 chromatografie
\nTable 2. Liquide chromatography settings<\/h5>\n<\/a>\nTabulka 3. Meze stanovitelnosti (LOQ) v\u00a0matrici odpadn\u00ed voda
\nTable 3. Limits of quantification for wastewater<\/h6>\n<\/a>\n<\/a>\nObr. 7. Uk\u00e1zka chromatogramu extrahovan\u00fdch iont\u016f v\u00a0re\u017eimu ESI- (nor-THC)
\nFig. 7. Example of extracted ion chromatograms in ESI- (nor-THC)<\/h6>\n<\/a>\nObr. 8. Uk\u00e1zka chromatogramu extrahovan\u00fdch iont\u016f v\u00a0re\u017eimu ESI- (ethylsulf\u00e1t)
\nFig. 8. Example of extracted ion chromatograms in ESI- (ethylsulfate)<\/h6>\nZ\u00e1v\u011br<\/h2>\n