{"id":1200,"date":"2016-12-06T22:04:53","date_gmt":"2016-12-06T21:04:53","guid":{"rendered":"http:\/\/www.newslab.sk\/2016\/12\/06\/stanovenie-stopovych-prvkov-v-krvnom-sere-2\/"},"modified":"2017-10-04T14:52:48","modified_gmt":"2017-10-04T12:52:48","slug":"establishing-trace-elements-in-blood-serum","status":"publish","type":"post","link":"https:\/\/www.newslab.sk\/en\/establishing-trace-elements-in-blood-serum\/","title":{"rendered":"Establishing Trace Elements in Blood Serum"},"content":{"rendered":"<p><span style=\"color: #ff0000;\"><strong>*All tables, charts, graphs and pictures that are featured in this article can be found in the .pdf<br \/>\nattachment at the end of the paper.<\/strong><\/span><\/p>\n<p>&nbsp;<\/p>\n<p><strong>\u00davod<\/strong><\/p>\n<p>V\u00e4\u010d\u0161ina stopov\u00fdch kovov s\u00fa esenci\u00e1lne v\u00fd\u017eivov\u00e9 prvky pre \u010dloveka ale aj pre zvierat\u00e1, patria sem sel\u00e9n, zinok, me\u010f, kobalt, mang\u00e1n, molybd\u00e9n, chr\u00f3m, nikel a \u017eelezo. Medzi v\u00fdznamn\u00e9 funkcie stopov\u00fdch prvkov patr\u00ed determin\u00e1cia, mobiliz\u00e1cia a kon\u0161tantn\u00e1 v\u00e4zba na biologick\u00e9 ligandy. Niektor\u00e9 s\u00fa vyu\u017eit\u00e9 pri elektrick\u00fdch impulzoch pozd\u013a\u017e nervov\u00fdch vl\u00e1kien in\u00e9 s\u00fa stabilnou s\u00fa\u010das\u0165ou komplexov s enz\u00fdmami alebo nukleov\u00fdmi kyselinami pr\u00edpadne ligandmi. V biologick\u00fdch procesoch sa uplat\u0148uj\u00fa ako sp\u00fa\u0161\u0165a\u010de, aktiv\u00e1tori ako aj kontrol\u00f3ri celej \u0161k\u00e1ly reakci\u00ed. In\u00e1 skupina stopov\u00fdch prvkov vytv\u00e1ra pevn\u00fd statick\u00fd komplex a st\u00e1vaj\u00fa sa integr\u00e1lnou \u010das\u0165ou prote\u00ednov a enz\u00fdmov. Mno\u017estvo biologick\u00fdch syst\u00e9mov s\u00fa z\u00e1visl\u00e9 na dennom pr\u00edjme stopov\u00fdch prvkov (me\u010f, zinok, mang\u00e1n).\u00a0 Oxidat\u00edvny stres je v\u00fdznamn\u00fdm faktorom v infek\u010dnom procese ak s\u00fa v deficite mikroprvky<sup>1<\/sup>. Stopov\u00e9 prvky a niektor\u00e9 ich zl\u00fa\u010deniny maj\u00fa antiv\u00edrusov\u00fa aktivitu prostredn\u00edctvom inaktiv\u00e1cie celul\u00e1rnych prote\u00ednov. Teda stopov\u00e9 prvky zohr\u00e1vaj\u00fa v\u00fdznamn\u00fa \u00falohu pri chorob\u00e1ch vyvolan\u00fdmi v\u00edrusmi<sup>2-5<\/sup>. Imunitn\u00fd syst\u00e9m predstavuje v\u00fdznamn\u00fa zlo\u017eku organizmu integrituj\u00faceho fyziologick\u00e9 funkcie , ktor\u00e9 eliminuj\u00fa cudzorod\u00e9 zlo\u017eky, infek\u010dn\u00e9 mikr\u00f3by ako aj odumret\u00e9 vlastn\u00e9 bunky. Tieto mechanizmy s\u00fa zabezpe\u010den\u00e9 prostredn\u00edctvom ne\u0161pecifickej (vrodenej) alebo \u0161pecifickej (z\u00edskanej) imunity so zlo\u017eit\u00fdmi procesmi, ktor\u00e9 s\u00fa koordinovan\u00e9 celou radou buniek (lymfocyty, makrof\u00e1gy, antig\u00e9n prezentuj\u00face bunky a pod.) a molekul\u00e1rnych l\u00e1tok (cytok\u00ednov, imunoglobul\u00ednov, imunohorm\u00f3nov). Makrof\u00e1gy predstavuj\u00fa bunky prvej l\u00ednie pri obrann\u00fdch procesoch vyzna\u010duj\u00face sa fagocyta\u010dnou, cytotoxickou a sekretorickou aktivitou. Mikroelementy ako zinok, sel\u00e9n, \u017eelezo, me\u010f a in\u00e9 v\u00fdznamne ovplyv\u0148uj\u00fa jednotliv\u00e9 zlo\u017eky ne\u0161pecifickej imunity. Vybran\u00e9 mikroelementy zohr\u00e1vaj\u00fa v\u00fdznamn\u00fa \u00falohu v ochrane buniek pred po\u0161koden\u00edm vyvolan\u00e9 oxidantmi. Fagocyty produkuj\u00face reakt\u00edvne oxidanty sa z\u00fa\u010dast\u0148uj\u00fa v ochrane proti infek\u010dn\u00fdm agens. Nedostatok zinku zni\u017euje funkciu NK buniek, pri\u010dom pr\u00eddavok tohto prvku v\u00fdrazne zlep\u0161\u00ed aktivitu NK buniek. Poznatky o \u0161pecifickom \u00fa\u010dinku mikroelementov na funkciu neutrofilov s\u00fa nejasn\u00e9<sup>6<\/sup>.<\/p>\n<p>Stopov\u00e9 prvky sa v\u00a0tkaniv\u00e1ch nach\u00e1dzaj\u00fa v koncentr\u00e1cii\u00a0<strong>ni\u017e\u0161ej ako 50 ppm (&lt; 50\u00a0\u00d7\u200910<sup>\u22126<\/sup>\u00a0g\/g)<\/strong>. Ich potrebn\u00fd denn\u00fd pr\u00edjem je men\u0161\u00ed ne\u017e 50 mg denn\u011b. \u00a0Hlavn\u00e9 prvky \u2013\u00a0Fe, I, Cu, Zn, Co, Cr, Mo, Se, F, Mn, Ni, As, Sn, Si, V.<\/p>\n<ul>\n<li>s\u00fa esenci\u00e1lne \u2013 telo si ich nedok\u00e1\u017ee samo vytvori\u0165 a s\u00fa z\u00e1visl\u00e9 na ich pr\u00edjem potravou;<\/li>\n<li>ich nedostatok alebo nadbytok sp\u00f4sobuje zdravotn\u00e9 probl\u00e9my.<\/li>\n<\/ul>\n<p>Hlavnou biochemickou \u00falohou stopov\u00fdch prvkov je katalytick\u00e9 p\u00f4sobenie v enz\u00fdmoch a\u00a0modul\u00e1ci\u00e1ch enz\u00fdmov\u00fdch aktiv\u00edt. Taktie\u017e maj\u00fa ve\u013ek\u00fd v\u00fdznam v ochrane pred oxida\u010dn\u00fdm stresom (SOD\u00a0potrebuje Mn, Cu a Zn).<\/p>\n<p>Pri\u00a0reakcii ak\u00fatnej f\u00e1zy IL-1 a\u00a0IL-6\u00a0sp\u00f4sob\u00ed redistrib\u00faciu stopov\u00fdch prvkov (pokles Zn a Fe v s\u00e9re).<\/p>\n<p>Toxicita \u2212 u zdrav\u00fdch jedincov je ve\u013ek\u00e1 tolerancia v koncentra\u010dnom rozmedz\u00ed. U ren\u00e1lnych por\u00fach sa m\u00f4\u017ee hor\u0161ie vylu\u010dova\u0165 Se a Cr, Cu a Mn zasa pri chorob\u00e1ch pe\u010dene. Stanovuj\u00fa sa met\u00f3dou at\u00f3movej absorp\u010dnej\u00a0spektrometrie.<\/p>\n<p><strong>Me\u010f (Cu)<\/strong><\/p>\n<p>Me\u010f je s\u00fa\u010das\u0165ou akt\u00edvneho centra mnoh\u00fdch (predov\u0161etk\u00fdm oxidoreduk\u010dn\u00fdch) enz\u00fdmov. S\u00favis\u00ed to so schopnos\u0165ou medi prija\u0165 alebo odovzda\u0165 elektr\u00f3n (Cu<sup>+ &#8211;<\/sup>\/ Cu<sup>++<\/sup>). Pr\u00edkladom je cytochr\u00f3m \u2013 c oxid\u00e1za, posledn\u00fd \u010dl\u00e1nok prenosu elektr\u00f3nov v\u00a0termin\u00e1lnej oxid\u00e1cii. \u010eal\u0161\u00ed enz\u00fdm, lyzyloxid\u00e1za m\u00e1 v\u00fdznam v\u00a0synt\u00e9ze elast\u00ednu a\u00a0kolag\u00e9nu.<\/p>\n<p>V\u00a0\u013eudskom tele je 100 &#8211; 150 mg medi. Z\u00a0gastrointerstici\u00e1lneho traktu sa me\u010f dostane do krvi, kde sa via\u017ee na album\u00edn, aminokyseliny a\u00a0transcuprein. Tieto transportn\u00e9 formy sa dostan\u00fa do pe\u010dene, kde sa me\u010f zabuduje do ceruloplazm\u00ednu, ktor\u00fd je hlavn\u00fdm transportn\u00fdm prote\u00ednom medi.<\/p>\n<p><strong>Doporu\u010den\u00e1 denn\u00e1 d\u00e1vka <\/strong>je<strong> 2- 3 mg.<\/strong> Nutri\u010dn\u00fd nedostatok medi sa v\u00a0praxi nevyskytuje, v\u00a0experimente vedie k\u00a0an\u00e9mii.<\/p>\n<p><strong>Zinok (Zn)<\/strong><\/p>\n<p>Zinok je s\u00fa\u010das\u0165ou akt\u00edvneho centra viac ako 300 enz\u00fdmov a\u00a0pre in\u00e9 bielkoviny je potrebn\u00fd ako aktiv\u00e1tor alebo stabiliz\u00e1tor \u0161trukt\u00fary. Zinok je esenci\u00e1lny prvok pri synt\u00e9ze nukleov\u00fdch kysel\u00edn a\u00a0pre funkciu g\u00e9nov. Zvl\u00e1\u0161tnos\u0165 zinku v\u00a0porovnan\u00ed s\u00a0in\u00fdmi stopov\u00fdmi prvkami spo\u010d\u00edva v\u00a0tom, \u017ee nie je ve\u013emi akt\u00edvny v\u00a0oxida\u010dno-reduk\u010dn\u00fdch reakci\u00e1ch (nekatalyzuje vznik a premenu bioreakt\u00edvnych foriem kysl\u00edka). Je schopn\u00fd vytvori\u0165 4 koordina\u010dn\u00e9 v\u00e4zby s\u00a0at\u00f3mami s\u00edry v\u00a0cyste\u00ednoch alebo s\u00a0dus\u00edkmi histid\u00ednov.<\/p>\n<p>Zinok je prvok, ktor\u00fd m\u00e1 vz\u0165ah k\u00a0vn\u00edmavosti k\u00a0niektor\u00fdm druhom zhubn\u00fdch n\u00e1dorov. Hr\u00e1 toti\u017e v\u00fdznamn\u00fa rolu v\u00a0tvorbe T-lymfocytov, ktor\u00e9 maj\u00fa schopnos\u0165 likvidova\u0165 n\u00e1dorov\u00e9 bunky.<\/p>\n<p><strong>Doporu\u010den\u00e1 denn\u00e1 d\u00e1vka<\/strong>\u00a0pre dospel\u00fdch\u00a0<strong>mu\u017eov<\/strong> je <strong>10 mg<\/strong> a pre <strong>\u017eeny<\/strong> <strong>7 mg.<\/strong><\/p>\n<p>Deficit zinku v\u00a0experimente vedie k\u00a0malform\u00e1ci\u00e1m, retard\u00e1cii rastu, k\u00a0hypogonadizmu a\u00a0k\u00a0zhor\u0161enej funkcii imunitn\u00e9ho syst\u00e9mu.<\/p>\n<p>Cie\u013eom tejto pr\u00e1ce bolo vyvin\u00fa\u0165 a\u00a0zvalidova\u0165 pracovn\u00fd postup pre met\u00f3du na stanovenie stopov\u00fdch prvkov me\u010f a\u00a0zinok v krvnom s\u00e9re pre rutinn\u00fa diagnostiku v\u00a0laborat\u00f3riu klinickej bioch\u00e9mie.<\/p>\n<p><strong>\u00a0<\/strong><\/p>\n<p><strong>Experiment\u00e1lna \u010das\u0165<\/strong><\/p>\n<p><em>Pou\u017eit\u00e9 pr\u00edstroje a\u00a0zariadenia<\/em><\/p>\n<p>Na stanovenie stopov\u00fdch prvkov bol pou\u017eit\u00fd at\u00f3mov\u00fd absorp\u010dn\u00fd spektrometer firmy Agilent Technologies 240Z AA (Austr\u00e1lia) s\u00a0elektrotermickou atomiz\u00e1ciou GTA 120 (Graphite Tube Atomizer) v\u00a0spojen\u00ed s\u00a0automatick\u00fdm pod\u00e1va\u010dom vzoriek PSD 120 (Programmable Sample Dispenser). Ako ochrann\u00fd plyn bol pou\u017eit\u00fd arg\u00f3n (0,3l\/min). Merania boli analyzovan\u00e9 na pyrolytick\u00fdch grafitov\u00fdch kyvet\u00e1ch bez platformy firmy Agilent Technologies (Nemecko). D\u00e1vkovan\u00fd objem vzoriek bol 20\u00b5l pre me\u010f a 10\u00b5l pre zinok. Pracovn\u00e9 podmienky pre stanovenie oboch stopov\u00fdch prvkov s\u00fa uveden\u00e9 v\u00a0tabu\u013eke I\u00a0a\u00a0teplotn\u00e9 programy s\u00fa uveden\u00e9 v\u00a0tabu\u013eke II.<\/p>\n<p><em>Tabu\u013eka I: Pracovn\u00e9 podmienky pre stanovenie Cu a Zn<\/em><\/p>\n<table width=\"75%\">\n<tbody>\n<tr>\n<td width=\"33%\"><strong>Parameter<\/strong><\/td>\n<td width=\"33%\"><strong>Cu<\/strong><\/td>\n<td width=\"33%\"><strong>Zn<\/strong><\/td>\n<\/tr>\n<tr>\n<td width=\"33%\">Vlnov\u00e1 d\u013a\u017eka (nm)<\/td>\n<td width=\"33%\">324,8<\/td>\n<td width=\"33%\">213,9<\/td>\n<\/tr>\n<tr>\n<td width=\"33%\">\u0160trbina (nm)<\/td>\n<td width=\"33%\">0,5<\/td>\n<td width=\"33%\">1,0<\/td>\n<\/tr>\n<tr>\n<td width=\"33%\">Korektor pozadia<\/td>\n<td width=\"33%\">bez korekcie<\/td>\n<td width=\"33%\">bez korekcie<\/td>\n<\/tr>\n<tr>\n<td width=\"33%\">Zdroj \u017eiarenia<\/td>\n<td width=\"33%\">kat\u00f3dov\u00e1 v\u00fdbojka<\/td>\n<td width=\"33%\">kat\u00f3dov\u00e1 v\u00fdbojka<\/td>\n<\/tr>\n<tr>\n<td width=\"33%\">Pr\u00fad lampy (mA)<\/td>\n<td width=\"33%\">4,0<\/td>\n<td width=\"33%\">5,0<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n<p>&nbsp;<\/p>\n<p><em>Tabu\u013eka II: Teplotn\u00fd\u00a0 a\u00a0\u010dasov\u00fd program pre stanovenie Cu a Zn<\/em><\/p>\n<table width=\"100%\">\n<tbody>\n<tr>\n<td colspan=\"2\" width=\"24%\"><strong>F\u00e1za<\/strong><\/td>\n<td width=\"21%\"><strong>Teplota<\/strong><\/p>\n<p><strong>(\u00b0C)<\/strong><\/td>\n<td width=\"25%\"><strong>\u010cas<\/strong><\/p>\n<p><strong>(s)<\/strong><\/td>\n<td width=\"28%\"><strong>Prietok arg\u00f3nu<\/strong><\/p>\n<p><strong>(l\/min)<\/strong><\/td>\n<\/tr>\n<tr>\n<td width=\"12%\"><strong>\u00a0<\/strong><\/p>\n<p><strong>Su\u0161enie<\/strong><\/p>\n<p><strong>\u00a0<\/strong><\/td>\n<td width=\"10%\"><strong>1<\/strong><\/p>\n<p><strong>2<\/strong><\/p>\n<p><strong>3<\/strong><\/td>\n<td width=\"21%\">85<\/p>\n<p>95<\/p>\n<p>120<\/td>\n<td width=\"25%\">5<\/p>\n<p>40<\/p>\n<p>10<\/td>\n<td width=\"29%\">0,3<\/p>\n<p>0,3<\/p>\n<p>0,3<\/td>\n<\/tr>\n<tr>\n<td rowspan=\"3\" width=\"12%\"><strong>\u00a0<\/strong><\/p>\n<p><strong>Pyrol\u00fdza<\/strong><\/td>\n<td rowspan=\"3\" width=\"10%\"><strong>4<\/strong><\/p>\n<p><strong>\u00a0<\/strong><\/p>\n<p><strong>5<\/strong><\/p>\n<p><strong>\u00a0<\/strong><\/p>\n<p><strong>6<\/strong><\/td>\n<td width=\"21%\">300 (Zn)<\/p>\n<p>1100 (Cu)<\/td>\n<td width=\"25%\">5<\/td>\n<td width=\"29%\">0,3<\/td>\n<\/tr>\n<tr>\n<td width=\"21%\">300 (Zn)<\/p>\n<p>1100 (Cu)<\/td>\n<td width=\"25%\">2<\/td>\n<td width=\"29%\">0,3<\/td>\n<\/tr>\n<tr>\n<td width=\"21%\">300 (Zn)<\/p>\n<p>1100 (Cu)<\/td>\n<td width=\"25%\">2<\/td>\n<td width=\"29%\">0<\/td>\n<\/tr>\n<tr>\n<td rowspan=\"2\" width=\"12%\"><strong>\u00a0<\/strong><\/p>\n<p><strong>Atomiz\u00e1cia<\/strong><\/td>\n<td rowspan=\"2\" width=\"10%\"><strong>\u00a0<\/strong><\/p>\n<p><strong>7<\/strong><\/p>\n<p><strong>\u00a0<\/strong><\/p>\n<p><strong>8<\/strong><\/td>\n<td width=\"21%\">1900 (Zn)<\/p>\n<p>2200 (Cu)<\/td>\n<td width=\"25%\">1<\/td>\n<td width=\"29%\">0*<\/td>\n<\/tr>\n<tr>\n<td width=\"21%\">1900 (Zn)<\/p>\n<p>2200 (Cu)<\/td>\n<td width=\"25%\">2<\/td>\n<td width=\"28%\">0*<\/td>\n<\/tr>\n<tr>\n<td width=\"14%\"><strong>\u010cistenie<\/strong><\/td>\n<td width=\"10%\"><strong>9<\/strong><\/td>\n<td width=\"21%\">2000 (Zn)<\/p>\n<p>2300 (Cu)<\/td>\n<td width=\"25%\">2<\/td>\n<td width=\"28%\">0,3<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n<p>*- zaznamen\u00e1vanie sign\u00e1lu<\/p>\n<p>&nbsp;<\/p>\n<p><em>Chemik\u00e1lie a\u00a0roztoky<\/em><\/p>\n<p>Kyselina dusi\u010dn\u00e1 TraceSELECT<sup>\u00ae<\/sup> (Sigma -Aldrich Co., USA); Triton<sup>\u00ae<\/sup> X-100 p.a. (Merck, Nemecko), voda TraceSELECT<sup>\u00ae<\/sup> (Sigma-Aldrich Co., USA), lyofilizovan\u00fd s\u00e9rov\u00fd kalibr\u00e1tor ClinCal<sup>\u00ae<\/sup> &#8211; Serum Calibrator \u0161ar\u017ea 243 Recipe<sup>\u00ae<\/sup> Chemical + Instruments, Nemecko), \u0161tandardn\u00fd referen\u010dn\u00fd materi\u00e1l Seronorm<sup>\u2122<\/sup> Trace Elements Serum \u0161ar\u017ea 0903106( SERO AS, N\u00f3rsko), kontroln\u00e9 lyofilizovan\u00e9 krvn\u00e9 s\u00e9rum ClinChek<sup>\u00ae<\/sup> s\u00a0dvoma hladinami kontrol(Level I,II) \u0161ar\u017ea 347( Recipe<sup>\u00ae<\/sup> Chemical + Instruments, Nemecko).<\/p>\n<p><em>Pracovn\u00e9 postupy<\/em><\/p>\n<p>Pr\u00edprava s\u00e9rov\u00e9ho kalibr\u00e1tora, kontroln\u00e9ho krvn\u00e9ho s\u00e9ra a\u00a0\u0161tandardn\u00e9ho referen\u010dn\u00e9ho materi\u00e1lu K\u00a0lyofilizovan\u00e9mu s\u00e9rov\u00e9mu kalibr\u00e1toru, kontroln\u00e9mu krvn\u00e9mu s\u00e9ru a\u00a0\u0161tandardn\u00e9mu referen\u010dn\u00e9mu materi\u00e1lu sme pridali 3ml deionizovanej vody, ktor\u00e9 sme mie\u0161ali kr\u00fa\u017eiv\u00fdm pohybom, nechali sme st\u00e1\u0165 30 min\u00fat pri laborat\u00f3rnej teplote. Po 30 min\u00fatach bolo mo\u017en\u00e9 s\u00e9rov\u00fd kalibr\u00e1tor, kontroln\u00e9 s\u00e9rum a\u00a0\u0161tandardn\u00fd referen\u010dn\u00fd materi\u00e1l pou\u017ei\u0165 pre stanovenie.<\/p>\n<p><em>Priame stanovenie<\/em><\/p>\n<p>\u0160tandardn\u00fd referen\u010dn\u00fd materi\u00e1l a kontroln\u00e9 krvn\u00e9 s\u00e9rum pre priame stanovenie boli rieden\u00e9 20x 0,2%(v\/v) Tritonom X-100 a\u00a01%HNO<sub>3 <\/sub>a\u00a0s\u00e9rov\u00fd kalibr\u00e1tor bol rieden\u00fd 20x 0,2%HNO<sub>3 <\/sub>v\u00a0pr\u00edpade medi a\u00a0v\u00a0pr\u00edpade zinku bol \u0161tandardn\u00fd referen\u010dn\u00fd materi\u00e1l a kontroln\u00e9 krvn\u00e9 s\u00e9rum rieden\u00e9 100x 0,2%(v\/v) Tritonom X-100 a\u00a01%HNO<sub>3 <\/sub>a\u00a0s\u00e9rov\u00fd kalibr\u00e1tor 100x 0,2%HNO<sub>3 <\/sub>priamo v\u00a02ml autosamplerov\u00fdch n\u00e1dobkach,z\u00a0ktor\u00fdch boli n\u00e1sledne d\u00e1vkovan\u00e9 do grafitovej kyvety(20\u00b5l pre stanovenie medi a 10 \u00b5l pre stanovenie zinku). Pre stanovenie obsahu Cu a\u00a0Zn v\u00a0s\u00e9re sa pou\u017eila met\u00f3da kalibra\u010dnej krivky, na vyhodnotenie sa pou\u017eila v\u00fd\u0161ka sign\u00e1lu.<\/p>\n<p>&nbsp;<\/p>\n<p><strong>V\u00fdsledky a diskusia<\/strong><\/p>\n<p><em>Optimaliz\u00e1cia teplotn\u00e9ho programu<\/em><\/p>\n<p>D\u00f4le\u017eit\u00fdm krokom pri stanoven\u00ed stopov\u00fdch prvkov technikou ETAAS je optimaliz\u00e1cia teploty a\u00a0\u010dasu v\u00a0teplotnom programe. K\u00a0tejto optimaliz\u00e1cii n\u00e1m posl\u00fa\u017eili krivky termick\u00e9ho rozkladu(pyrol\u00fdzy) a\u00a0krivky atomiz\u00e1cie pre me\u010f a\u00a0zinok(Graf \u010d.1 a 2). Z\u00a0tejto optimaliz\u00e1cie sme stanovili pre me\u010f teplotu pyrol\u00fdzy na 1100\u00b0C a\u00a0teplotu atomiz\u00e1cie na 2200\u00b0C a\u00a0v\u00a0pr\u00edpade zinku bola stanoven\u00e1 teplota pyrol\u00fdzy 300\u00b0C a\u00a0teplota atomiz\u00e1cie 1900\u00b0C.<\/p>\n<p>\u010eal\u0161\u00edm d\u00f4le\u017eit\u00fdm krokom bolo pou\u017eitie Tritonu X-100 na riedenie, aby sme dosiahli reprodukovate\u013en\u00e9 su\u0161enie v\u0161etk\u00fdch analyzovan\u00fdch vzoriek. V\u00a0tabu\u013eke II s\u00fa uveden\u00e9 teplotn\u00e9 a \u010dasov\u00e9 programy pre stanovenie medi a\u00a0zinku.<\/p>\n<p>&nbsp;<\/p>\n<p><em>Linearita a\u00a0citlivos\u0165<\/em><\/p>\n<p>Kalibra\u010dn\u00e9 grafy boli line\u00e1rne pre cel\u00fd koncentra\u010dn\u00fd rozsah 5-30 \u00b5mol\/l pre Cu a\u00a04-35 \u00b5mol\/l pre Zn, korela\u010dn\u00fd koeficient kalibra\u010dnej krivky Cu bol 0,9992 a\u00a0kalibra\u010dnej krivky Zn bol 0,9995. Citlivos\u0165 definovan\u00e1 ako koncentr\u00e1cia, ktor\u00e1 je potrebn\u00e1 na dosiahnutie sign\u00e1lu 0,0044 AU.s. (tzv. charakteristick\u00e1 hmotnos\u0165) bola 12,7 pg pre Cu a\u00a016,3 pg pre Zn.<\/p>\n<p>&nbsp;<\/p>\n<p><em>Spr\u00e1vnos\u0165<\/em><\/p>\n<p>Spr\u00e1vnos\u0165 navrhnut\u00e9ho postupu bola overen\u00e1 anal\u00fdzou \u0161tandardn\u00e9ho referen\u010dn\u00e9ho materi\u00e1lu krvn\u00e9ho s\u00e9ra (Seronorm<sup>\u2122<\/sup> Trace Elements Serum \u0161ar\u017ea 0903106) a\u00a0\u010fal\u0161\u00edm kontroln\u00fdm materi\u00e1lom krvn\u00e9 s\u00e9rum ClinChek<sup>\u00ae<\/sup> s\u00a0dvoma hladinami kontrol(Level I,II) \u0161ar\u017ea 347 (Recipe<sup>\u00ae <\/sup>Chemical + Instruments, Nemecko). Po\u010det meran\u00ed nasleduj\u00facich za sebou bol rovn\u00fd 10.\u00a0\u00a0\u00a0 Kompletn\u00e9 vyhodnotenie v\u00fdsledkov kontrol je uveden\u00e9 v\u00a0tabu\u013eke III.<\/p>\n<p>&nbsp;<\/p>\n<p><em>Presnos\u0165<\/em><\/p>\n<p>Presnos\u0165 bola vypo\u010d\u00edtan\u00e1 z\u00a0nameran\u00fdch hodn\u00f4t pre \u0161tandardn\u00fd referen\u010dn\u00fd materi\u00e1l krvn\u00e9ho s\u00e9ra Seronorm<sup>\u2122<\/sup> a\u00a0kontroln\u00fd materi\u00e1l krvn\u00e9 s\u00e9rum ClinChek<sup>\u00ae<\/sup>. Presnos\u0165 sme vyjadrili opakovate\u013enos\u0165ou v po\u010dte 10 za sebou nasleduj\u00facich meran\u00ed a reprodukovate\u013enos\u0165ou v po\u010dte 15 meran\u00ed z\u00edskan\u00fdch za nieko\u013eko dn\u00ed (tabu\u013eka III).<\/p>\n<p><em>Tabu\u013eka III:<\/em><\/p>\n<table width=\"102%\">\n<tbody>\n<tr>\n<td rowspan=\"2\" width=\"15%\"><strong>\u00a0<\/strong><\/td>\n<td rowspan=\"2\" width=\"22%\"><strong>Presnos\u0165- opakovate\u013enos\u0165ou<\/strong><\/p>\n<p><strong>RSD (%)<\/strong><\/td>\n<td rowspan=\"2\" width=\"16%\"><strong>Presnos\u0165-<\/strong><\/p>\n<p><strong>reproduk.<\/strong><\/p>\n<p><strong>RSD(%)<\/strong><\/td>\n<td colspan=\"2\" width=\"26%\"><strong>Spr\u00e1vnos\u0165-<\/strong><\/p>\n<p><strong>Priem.hodnota \u00b1 SD<\/strong><\/td>\n<td rowspan=\"2\" width=\"19%\"><strong>Spr\u00e1vnos\u0165-<\/strong><\/p>\n<p><strong>v\u00fd\u0165a\u017enos\u0165ou<\/strong><\/p>\n<p><strong>(%)<\/strong><\/td>\n<\/tr>\n<tr>\n<td width=\"13%\"><strong>certifik.<\/strong><\/td>\n<td width=\"13%\"><strong>nameran\u00e1<\/strong><\/td>\n<\/tr>\n<tr>\n<td rowspan=\"2\" width=\"15%\"><strong>Seronorm<sup>TM<\/sup><\/strong><\/p>\n<p><strong>(\u0161ar\u017ea 0903106)<\/strong><\/td>\n<td width=\"22%\">0,99(<strong>Cu<\/strong>)<\/td>\n<td width=\"16%\">1,61<\/td>\n<td width=\"13%\">29,6\u00b11,4<\/td>\n<td width=\"13%\">29,5\u00b10,3<\/td>\n<td width=\"19%\">99,7<\/td>\n<\/tr>\n<tr>\n<td width=\"22%\">1,97(<strong>Zn<\/strong>)<\/td>\n<td width=\"16%\">1,22<\/td>\n<td width=\"13%\">26,6\u00b11,3<\/td>\n<td width=\"13%\">25,9\u00b10,5<\/td>\n<td width=\"19%\">97,5<\/td>\n<\/tr>\n<tr>\n<td rowspan=\"2\" width=\"15%\"><strong>ClinChek<sup>\u00ae<\/sup>-Level I<\/strong><\/p>\n<p><strong>(\u0161ar\u017ea 347)<\/strong><\/td>\n<td width=\"22%\">1,41(<strong>Cu<\/strong>)<\/td>\n<td width=\"16%\">2,51<\/td>\n<td width=\"13%\">12,6\u00b11,9<\/td>\n<td width=\"13%\">12,7\u00b10,2<\/td>\n<td width=\"19%\">101<\/td>\n<\/tr>\n<tr>\n<td width=\"22%\">1,86(<strong>Zn<\/strong>)<\/td>\n<td width=\"16%\">2,37<\/td>\n<td width=\"13%\">20,2\u00b13,0<\/td>\n<td width=\"13%\">20,3\u00b10,4<\/td>\n<td width=\"19%\">100,5<\/td>\n<\/tr>\n<tr>\n<td rowspan=\"2\" width=\"15%\"><strong>ClinChek<sup>\u00ae<\/sup>-Level II<\/strong><\/p>\n<p><strong>(\u0161ar\u017ea 347)<\/strong><\/td>\n<td width=\"22%\">1,23(<strong>Cu<\/strong>)<\/td>\n<td width=\"16%\">2,46<\/td>\n<td width=\"13%\">21,1\u00b13,1<\/td>\n<td width=\"13%\">21,3\u00b10,3<\/td>\n<td width=\"19%\">100,9<\/td>\n<\/tr>\n<tr>\n<td width=\"22%\">2,35(<strong>Zn<\/strong>)<\/td>\n<td width=\"16%\">2,91<\/td>\n<td width=\"13%\">31,2\u00b14,7<\/td>\n<td width=\"13%\">30,8\u00b10,7<\/td>\n<td width=\"19%\">98,7<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n<p>&nbsp;<\/p>\n<p><em>Medza detekcie (LOD) a stanovenia (LOQ)<\/em><\/p>\n<p>Pre ur\u010denie medz\u00ed bolo pou\u017eit\u00e9 3\u03c3 krit\u00e9rium, ktor\u00fdm je ur\u010den\u00e1 medza detekcie analytu ako trojn\u00e1sobok smerodajnej odch\u00fdlky \u0161umov\u00e9ho sign\u00e1lu (LOD). Medza stanovenia bola vypo\u010d\u00edtan\u00e1 ako desa\u0165n\u00e1sobok smerodajnej odch\u00fdlky \u0161umov\u00e9ho sign\u00e1lu (LOQ). V pr\u00edpade Cu bola medza detekcie 0,18\u00b5mol\/l a medza stanovenia 0,61\u00b5mol\/l, v pr\u00edpade Zn bola medza detekcie \u00a0\u00a00,16\u00b5mol\/l a medza stanovenia 0,53\u00b5mol\/l.<\/p>\n<p>&nbsp;<\/p>\n<p><strong>Z\u00e1ver<\/strong><\/p>\n<p>V tejto pr\u00e1ci bola vypracovan\u00e1 analytick\u00e1 met\u00f3da pre kvantitat\u00edvne stanovenie stopov\u00fdch prvkov me\u010f a\u00a0zinok v\u00a0krvnom s\u00e9re pre rutinn\u00fa diagnostiku v\u00a0laborat\u00f3riu klinickej bioch\u00e9mie. Validovan\u00e1 analytick\u00e1 met\u00f3da je prev\u00e1dzkovo realizovate\u013en\u00e1 s\u00a0presnos\u0165ou(opakovate\u013enos\u0165, reprodukovate\u013enos\u0165) do 5% a\u00a0spr\u00e1vnos\u0165ou(v\u00fd\u0165a\u017enos\u0165) v\u00a0intervale 85-115% v\u00a0celom kalibra\u010dnom rozsahu.<\/p>\n<p>&nbsp;<\/p>\n<p><strong>Po\u010fakovanie<\/strong><\/p>\n<p>T\u00e1to\u00a0 pr\u00e1ca vznikla za podpory projektu V\u00fdskumn\u00e9 centrum modern\u00fdch technol\u00f3gi\u00ed monitorovania a diagnostiky ochoren\u00ed ohrozuj\u00facich verejn\u00e9 zdravie s ITMS k\u00f3dom projektu 26220220197. Tento projekt je s\u00fa\u010das\u0165ou opera\u010dn\u00e9ho programu V\u00fdskum a v\u00fdvoj, ktor\u00fd je spolufinancovan\u00fd Eur\u00f3pskou \u00faniou z Eur\u00f3pskeho fondu region\u00e1lneho rozvoja.<\/p>\n<p><strong>\u00a0<\/strong><strong>\u00a0<\/strong><\/p>\n<p><strong>Literat\u00fara<\/strong><\/p>\n<ol>\n<li>Evans, P. and Halliwell, B., Micronutrients: oxidant\/antioxidant status. Br. J. Nutr. (Suppl.), 2001, 85, 67\u201374.<\/li>\n<li>van der Strate, B. W., Beljaars, L., Molema, G., Harmsen, M. C. and Meijer, D. K., Antiviral activities of lactoferrin. Antiviral Res., 2001, 52, 225\u2013239.<\/li>\n<li>Sherrington, C. A. and Olynyk, J. K., Iron as a cofactor in chronic hepatitis C infection. Liver, 2002, 22, 187\u2013189.<\/li>\n<li>Semba, R. D., Iron-deficiency anemia and the cycle of poverty among human immunodeficiency virus-infected women in the inner city. Clin. Infect. Dis. (Suppl.), 2003, 37, 105\u2013111.<\/li>\n<li>Bhaskaram, P., Micronutrient malnutrition, infection and immunity: An overview. Nutr. Rev., 2002, 60, 40\u201345.<\/li>\n<li>Erickson, K. L., Medina, E. A. and Hubbard, N. E., Micronutrients and innate immunity. J. Infect. Dis. (Suppl.), 2000, 182, 5\u2013 10.<\/li>\n<li>R\u00e1cz O., \u0160ipulov\u00e1 A., Metabolizmus stopov\u00fdch prvkov, 19-24<\/li>\n<\/ol>\n<p>&nbsp;<\/p>\n<p><strong><em>\u201eTento \u010dl\u00e1nok vznikol na z\u00e1klade v\u00fdskumn\u00fdch aktiv\u00edt v projekte, ktor\u00fd bol podporen\u00fd v r\u00e1mci OP V\u00fdskum a v\u00fdvoj, s n\u00e1zvom: V\u00fdskumn\u00e9 centrum modern\u00fdch technol\u00f3gi\u00ed monitorovania a diagnostiky ochoren\u00ed ohrozuj\u00facich verejn\u00e9 zdravie, ITMS 26220220197, spolufinancovan\u00fd zo zdrojov Eur\u00f3pskeho fondu region\u00e1lneho rozvoja\u201c.<\/em><\/strong><\/p>\n","protected":false},"excerpt":{"rendered":"<p>*All tables, charts, graphs and pictures that are featured in this article can be found in the .pdf attachment at the end of the paper. &nbsp; \u00davod V\u00e4\u010d\u0161ina stopov\u00fdch kovov s\u00fa esenci\u00e1lne v\u00fd\u017eivov\u00e9 prvky pre \u010dloveka ale aj pre zvierat\u00e1, patria sem sel\u00e9n, zinok, me\u010f, kobalt, mang\u00e1n, molybd\u00e9n, chr\u00f3m, nikel a \u017eelezo. Medzi v\u00fdznamn\u00e9 funkcie<\/p>\n","protected":false},"author":7,"featured_media":0,"comment_status":"open","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"_mi_skip_tracking":false,"footnotes":""},"categories":[289],"tags":[796,795,797,793,794],"class_list":["post-1200","post","type-post","status-publish","format-standard","hentry","category-biochemistry","tag-accuracy","tag-calibration-curve","tag-correctness","tag-electrothermic-atomisation","tag-heat-and-time-schedule","typ_clanku-review-article"],"acf":{"abstrakt":"<p>The objective of this paper was to develop and validate working procedures for the methods of establishing the levels of trace elements (Copper and Zinc) in blood serum for routine diagnosis in a clinical biochemistry laboratory. Validated analytical methods can be performed with preciseness (repeatability, reproducibility) up to 5 % and correctness (yield) in the 85 \u2013 115 % interval in the total calibration range.<\/p>\n<p><strong>Key words:<\/strong> electrothermic atomisation, heat and time schedule, calibration curve, accuracy, correctness.<\/p>\n","casopis":[{"ID":1000,"post_author":"7","post_date":"2015-11-21 14:33:24","post_date_gmt":"2015-11-21 13:33:24","post_content":"<h4><strong>GENETICS<\/strong><\/h4>\r\n<ul>\r\n \t<li>The laboratory age<\/li>\r\n \t<li>The Odyssey of DNA reading<\/li>\r\n \t<li>Comparative genomic hybridisation: a methodological introduction<\/li>\r\n \t<li>Next generation sequencing and its application in clinical genetics<\/li>\r\n<\/ul>\r\n<h4><\/h4>\r\n<h4><strong>BIOCHEMISTRY<\/strong><\/h4>\r\n<ul>\r\n \t<li>Determining of the trace elements in blood serum<\/li>\r\n \t<li>Determining of \u03b1-tocopherol (vitamin E) in serum by way of liquid chromatography with tandem mass spectrometry (LC\/MS\/MS)<\/li>\r\n \t<li>Analysis of urinary calculi and its path to Europe<\/li>\r\n<\/ul>\r\n&nbsp;\r\n<h4><strong>IMMUNOLOGY <\/strong><\/h4>\r\n<ul>\r\n \t<li>New autoantibodies in diagnosis of autoimmunity myopathies<\/li>\r\n \t<li>Taking advantage of flow-based cytometry in determining prognostic markers<\/li>\r\n<\/ul>\r\nCD38 and ZAP-70 in patients with B-CLL\r\n\r\n&nbsp;\r\n<h4><strong>CYTOLOGY AND PATHOLOGY <\/strong><\/h4>\r\n<ul>\r\n \t<li>Mucinous ovarian carcinoma \u2013 news in diagnosis from the pathologist\u2019s perspective<\/li>\r\n<\/ul>","post_title":"newsLab","post_excerpt":"","post_status":"publish","comment_status":"closed","ping_status":"closed","post_password":"","post_name":"newslab-12015","to_ping":"","pinged":"","post_modified":"2017-08-16 21:43:10","post_modified_gmt":"2017-08-16 19:43:10","post_content_filtered":"","post_parent":0,"guid":"http:\/\/www.newslab.sk\/casopis\/newslab-12015\/","menu_order":0,"post_type":"casopis","post_mime_type":"","comment_count":"0","filter":"raw"}],"strana":"24","upload_clanok":{"ID":1201,"id":1201,"title":"medirex_1_2015-hauks","filename":"MEDIREX_1_2015-\u2013-Hauks.pdf","filesize":210913,"url":"https:\/\/www.newslab.sk\/wp-content\/uploads\/2016\/12\/MEDIREX_1_2015-\u2013-Hauks.pdf","link":"https:\/\/www.newslab.sk\/en\/establishing-trace-elements-in-blood-serum\/medirex_1_2015-hauks\/","alt":"","author":"7","description":"","caption":"","name":"medirex_1_2015-hauks","status":"inherit","uploaded_to":1200,"date":"2016-12-06 20:57:06","modified":"2016-12-06 20:57:06","menu_order":0,"mime_type":"application\/pdf","type":"application","subtype":"pdf","icon":"https:\/\/www.newslab.sk\/wp-includes\/images\/media\/document.png"}},"_links":{"self":[{"href":"https:\/\/www.newslab.sk\/en\/wp-json\/wp\/v2\/posts\/1200","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/www.newslab.sk\/en\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/www.newslab.sk\/en\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/www.newslab.sk\/en\/wp-json\/wp\/v2\/users\/7"}],"replies":[{"embeddable":true,"href":"https:\/\/www.newslab.sk\/en\/wp-json\/wp\/v2\/comments?post=1200"}],"version-history":[{"count":0,"href":"https:\/\/www.newslab.sk\/en\/wp-json\/wp\/v2\/posts\/1200\/revisions"}],"wp:attachment":[{"href":"https:\/\/www.newslab.sk\/en\/wp-json\/wp\/v2\/media?parent=1200"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/www.newslab.sk\/en\/wp-json\/wp\/v2\/categories?post=1200"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/www.newslab.sk\/en\/wp-json\/wp\/v2\/tags?post=1200"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}