{"id":1204,"date":"2016-12-06T21:29:25","date_gmt":"2016-12-06T20:29:25","guid":{"rendered":"http:\/\/www.newslab.sk\/2016\/12\/06\/stanovenie-%ce%b1-tokoferolu-vitamin-e-v-sere-kvapalinovou-chromatografiou-s-trojitym-kvadrupolom-lcmsms\/"},"modified":"2017-10-05T09:04:44","modified_gmt":"2017-10-05T07:04:44","slug":"establishing-%ce%b1-tocopherol-e-vitamin-level-in-serum-by-way-of-triple-quadrupole-liquid-chromatography-lcmsms","status":"publish","type":"post","link":"https:\/\/www.newslab.sk\/en\/establishing-%ce%b1-tocopherol-e-vitamin-level-in-serum-by-way-of-triple-quadrupole-liquid-chromatography-lcmsms\/","title":{"rendered":"Establishing \u03b1-tocopherol (E vitamin) Level in Serum by Way of Triple Quadrupole Liquid Chromatography (LCMSMS)"},"content":{"rendered":"

*All tables, charts, graphs and pictures that are featured in this article can be found in the .pdf\u00a0<\/strong><\/span>
\nattachment at the end of the paper.<\/strong><\/span><\/p>\n

 <\/p>\n

Vitam\u00edny rozpustn\u00e9 v\u00a0tukoch s\u00fa l\u00e1tky potrebn\u00e9 pre spr\u00e1vne fungovanie organizmu. \u00a0Spolu s\u00a0prote\u00ednmi, tukmi a\u00a0sacharidmi patria k\u00a0z\u00e1kladn\u00fdm zlo\u017ek\u00e1m na\u0161ej potravy. Vitam\u00edny z\u00a0tejto skupiny p\u00f4sobia pri metabolick\u00fdch procesoch (katabolizmus, anabolizmus) ako katalyz\u00e1tory. V\u010faka ich rozpustnosti v\u00a0tukoch maj\u00fa spolo\u010dn\u00fd absorp\u010dn\u00fd mechanizmus z\u00a0GITu ako tuky. Ich nedostatok m\u00f4\u017ee by\u0165 sp\u00f4soben\u00fd zn\u00ed\u017een\u00fdm pr\u00edjmom potravy s\u00a0obsahom tukov alebo poruchou\u00a0 vstreb\u00e1vania (malabsorpcia) tukov.<\/p>\n

Vitam\u00edn E je tvoren\u00fd nieko\u013ek\u00fdmi anal\u00f3gmi tokoferolu (\u03b1-, \u03b2-, \u03b3-, \u03b4-) a\u00a0ich pr\u00edslu\u0161n\u00fdmi nenas\u00fdten\u00fdmi tokotrienolmi. Vitam\u00edn E je siln\u00fdm antioxidantom v\u00fdznamne \u00fa\u010dinn\u00fd v\u00a0kombin\u00e1ci\u00ed s\u00a0in\u00fdmi rastlinn\u00fdmi antioxidantmi. Je zn\u00e1me, \u017ee spoma\u013euje proces starnutia buniek, chr\u00e1ni tkanivo ko\u017ee, o\u010d\u00ed, pe\u010dene. Monitorovanie statusu vitam\u00ednu E\u00a0 v\u00fdznamne pom\u00e1ha pri odha\u013eovan\u00ed nedostato\u010dn\u00e9ho pr\u00edjmu vitam\u00ednov z\u00a0potravy. Taktie\u017e pri nedostatku \u03b1-tokoferol transportn\u00e9ho prote\u00ednu, ktor\u00fd napom\u00e1ha\u00a0 zabudovaniu \u03b1- tokoferolu do lipoprote\u00edn\u00fdch \u010dast\u00edc (VLDL)\u00a0 v\u00a0pe\u010deni.<\/p>\n

\u00a0<\/strong><\/p>\n

Experiment\u00e1lna \u010das\u0165<\/strong><\/p>\n

\u00a0<\/em>Chemik\u00e1lie a vzorky<\/em><\/p>\n

V\u0161etky chemik\u00e1lie potrebn\u00e9 na pr\u00edpravu mobilnej f\u00e1zy a\u00a0pr\u00edpravu vzorky boli HPLC analytickej \u010distoty. Metanol, etanol a voda boli zak\u00fapen\u00e9 z\u00a0firmy Merck KGaA (Darmstadt, Nemecko). \u0160tandardn\u00fd roztok \u03b1-tokoferolu (vitam\u00edn E), butylovan\u00fd hydroxytolu\u00e9n (BHT) boli zak\u00fapen\u00e9 z firmy Sigma-Aldrich (St. Louis, USA), certifikovan\u00fd referen\u010dn\u00fd materi\u00e1l pre vitam\u00edn \u03b1-tokoferol- CRM ClinChek\u00ae<\/sup> Control bol zak\u00fapen\u00fd z\u00a0firmy Recipe (Nemecko).<\/p>\n

\u00a0<\/em><\/p>\n

Odber a\u00a0pr\u00edprava \u00a0vzoriek<\/em><\/p>\n

Vzorky pacientov boli po odbere centrifugovan\u00e9 a\u00a0s\u00e9rum \u00a0uskladnen\u00e9\u00a0 pri teplote \u2013 20 \u00b0C a\u017e do \u010fal\u0161ieho spracovania. Vzorky boli spracovan\u00e9 v\u00a0 polypropyl\u00e9nov\u00fdch sk\u00famavk\u00e1ch (eppendorf). 50\u00b5l s\u00e9ra bolo precipitovan\u00e9\u00a0 pridan\u00edn 100\u00b5l etanolu (vychladen\u00fd na 4 \u00b0C) obsahuj\u00faceho stabiliz\u00e1tor BHT (1%). Takto pripraven\u00e1 vzorka bola 5min. premie\u0161\u00e1van\u00e1 na trepa\u010dke a\u00a0n\u00e1sledne centrifugovan\u00e1 (8000x g, 5min.). Do HPLC bolo injekovan\u00fdch 20\u03bcl supernatantu.<\/p>\n

 <\/p>\n

Chromatografick\u00e1\u00a0 technika a\u00a0doplnkov\u00fd materi\u00e1l<\/em><\/p>\n

Merania boli realizovan\u00e9 na kvapalinovom chromatografe HPLC Agilent 1260 s\u00e9rie (Agilent Technologies, Singapur). Na detekciu bol pou\u017eit\u00fd 6410 Agilent trojit\u00fd kvadrup\u00f3l s\u00a0elektrosprejom (ESI).\u00a0 I\u00f3ny boli detekovan\u00e9 v\u00a0pozit\u00edvnom m\u00f3de vysoko \u0161pecifickou a\u00a0citlivou technikou mnohon\u00e1sobn\u00e9ho monitorovania odozvy (MRM \u2013 multiple reaction monitoring).\u00a0 Reten\u010dn\u00fd \u010das, prekurzorov\u00fd a\u00a0produktov\u00fd i\u00f3n, rovnako ako in\u00e9 parametre analytu s\u00fa uveden\u00e9 v\u00a0tabu\u013eke 1. D\u00e1ta na kvantitat\u00edvne stanovenie vitam\u00ednu boli vyhodnocovan\u00e9 cez MassHunter Workstation Software (Agilent). Na chromatografick\u00fa anal\u00fdzu bola pou\u017eit\u00e1 analytick\u00e1 kol\u00f3na Kinetex C18\u00a0100A 4,6 x 50mm, 2.6\u03bcm (Phenomenex, CA,USA) a\u00a0predkol\u00f3na Security Guard Cartridge C18 4,0 x 3.0mm.<\/p>\n

\u00a0<\/strong><\/p>\n\n\n\n
\n\n\n\n
Tabu\u013eka 1<\/strong> Optimaliz\u00e1cia MS\/MS krokov na kvantitat\u00edvne stanovenie vitam\u00ednu\u00a0E<\/p>\n\n\n\n\n
Analyt<\/strong><\/td>\ntr <\/sub>(min)<\/strong><\/td>\nPrekurzorov\u00fd i\u00f3n MS1<\/strong><\/td>\nProduktov\u00fd i\u00f3n MS2<\/strong><\/td>\nCE (V)<\/strong><\/td>\nPolarita<\/strong><\/td>\n<\/tr>\n
\u03b1-<\/strong>tokoferol<\/strong><\/td>\n3,2<\/td>\n431<\/td>\n165<\/td>\n20<\/td>\npozit\u00edvna<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

tr <\/sub><\/strong>\u2013 <\/sub><\/strong>reten\u010dn\u00fd \u010das, CE \u2013 kol<\/strong>\u00edzna energia<\/strong><\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

\u00a0<\/strong><\/p>\n

V\u00fdsledky a\u00a0diskusia<\/strong><\/p>\n

Pr\u00edprava a\u00a0stabilita vzoriek<\/em><\/p>\n

\u0160tandardn\u00e9 roztoky boli stabilizovan\u00e9 BHT (1g.l-1<\/sup>) a skladovan\u00e9 pri -20 \u00b0C. Vzorky pacientov po odbere boli centrifugovan\u00e9 v\u00a0chladenej centrif\u00fage\u00a0 a\u00a0po centrifugovan\u00ed zmrazen\u00e9 pri -20\u00b0C. Stabilita vzoriek za tak\u00fdchto podmienok bola 3 mesiace.<\/p>\n

\u00a0<\/em><\/p>\n

Chromatografick\u00e1 anal\u00fdza<\/em><\/p>\n

Pri separ\u00e1cii vitam\u00ednu\u00a0E bola pou\u017eit\u00e1 analytick\u00e1 kol\u00f3na s\u00a0reverznou f\u00e1zou. Pri v\u00fdbere kol\u00f3ny, mobilnej f\u00e1zy a\u00a0in\u00fdch parametrov sa postupovalo tak, aby sme v\u00a0 kr\u00e1tkom \u010dase dosiahli \u010do najlep\u0161iu separ\u00e1ciu analytu. V\u00a0tabu\u013eke 2 s\u00fa uveden\u00e9 testovan\u00e9 parametre a\u00a0optim\u00e1lne podmienky separ\u00e1cie vitam\u00ednu E. Koncentr\u00e1cia vitam\u00ednu v\u00a0s\u00e9re bola vypo\u010d\u00edtan\u00e1 z\u00a0externej kalibra\u010dnej krivky. Zo z\u00e1sobn\u00e9ho roztoku 2,32 mmol.l-1<\/sup> \u03b1 – tokoferolu v\u00a0metanole bol pripraven\u00fd pracovn\u00fd roztok 232 \u03bcmol.l–<\/sup>1. Z\u00a0pracovn\u00e9ho roztoku boli rieden\u00edn s \u00a0BHT (1g.l-1<\/sup>) pripraven\u00e9 \u0161tandardn\u00e9 kalibra\u010dn\u00e9 roztoky\u00a0 \u03b1-tokoferolu v\u00a0rozsahu (7,25; 14,5; 29,0; 38,6 a 58,0 \u03bcmol.l–<\/sup>1) \u2013 obr\u00e1zok 2)). \u00a0V\u00a0tomto rozsahu kalibra\u010dn\u00fdch \u0161tandardn\u00fdch roztokov bola testovan\u00e1 linearita met\u00f3dy. Medza detekcia (LOD) bola stanoven\u00e1 ako koncentr\u00e1cia analytu vo vzorke, ktor\u00e1 v\u00a0pou\u017eitom detek\u010dnom syst\u00e9me vykazovala zvolen\u00fa hodnotu \u00a0pomer sign\u00e1l \/ \u0161um >5. Hodnota medze\u00a0 detekcie pre \u03b1-tokoferol bola 0,01 \u03bcmol.l-1<\/sup>. Medza stanovenia (LOQ) bola vypo\u010d\u00edtan\u00e1 ako dvojn\u00e1sobok LOD.<\/p>\n

Pri valid\u00e1cii parametrov met\u00f3dy (opakovatete\u013enos\u0165, reprodukovate\u013enos\u0165, v\u00fd\u0165a\u017enos\u0165) bol pou\u017eit\u00fd referen\u010dn\u00fd materi\u00e1l CRM ClinChek\u00ae<\/sup> Control od Recipe (Nemecko). Opakovate\u013enos\u0165 bola stanoven\u00e1 p\u00e4\u0165n\u00e1s\u0165n\u00e1sobn\u00fdm opakovan\u00fdm stanoven\u00edm kontroln\u00e9ho s\u00e9ra pri dvoch hladin\u00e1ch v\u00a0jeden de\u0148. Z\u00a0tabu\u013eky 3 vypl\u00fdva, \u017ee opakovate\u013enos\u0165 pre\u00a0\u03b1-tokoferol vyjadren\u00e1 varia\u010dn\u00fdm koeficientom (CV) \u00a0neprekro\u010dila 5,0%. Reprodukovate\u013enos\u0165 bola\u00a0 stanoven\u00e1 opakovan\u00fdm stanoven\u00edm kontroln\u00e9ho s\u00e9ra pri dvoch hladin\u00e1ch po\u010das \u0161iestich dn\u00ed. Z\u00edskan\u00e9 hodnoty varia\u010dn\u00e9ho koeficientu boli 5,4% a\u00a05,1%.<\/p>\n

V\u00fd\u0165a\u017enos\u0165 (spr\u00e1vnos\u0165 ) met\u00f3dy bola overen\u00e1 tak, \u017ee k referen\u010dn\u00e9mu materi\u00e1lu (CRM ClinChek\u00ae<\/sup> Control 1 a\u00a03 ) boli prid\u00e1van\u00e9\u00a0 \u0161tandardy so zn\u00e1mou koncentr\u00e1ciou \u03b1-tokoferolu (0,68; 1,29 \u03bcmol.l-1<\/sup>) . V\u00fd\u0165a\u017enos\u0165 bola n\u00e1sledne vypo\u010d\u00edtan\u00e1 pod\u013ea \u00a0vz\u0165ahu:<\/p>\n\n\n\n
\n\n\n\n
\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0 [c] nameran\u00e1 \u00a0– [c] CRM ClinChek\u00ae<\/sup> Control<\/p>\n

V\u00fd\u0165a\u017enos\u0165 = \u00a0 \u00a0 _____________________________________________________ \u00a0 \u00a0 \u00a0x 100%<\/p>\n

[c] pridan\u00e9ho \u0161tandardu<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

V\u00fd\u0165a\u017enos\u0165 \u03b1-tokoferolu pri n\u00edzkej koncentra\u010dnej hladine bola 95%, pri vy\u0161\u0161ej koncentra\u010dnej hladine 99%. Met\u00f3da bola otestovan\u00e1 na\u00a0 skupine pacientov (n=30) n\u00e1hodne vybrat\u00fdch z\u00a0denn\u00e9ho s\u00faboru analyzovan\u00fdch vzoriek. Vekov\u00e9 rozp\u00e4tie vybrat\u00fdch pacientov bolo 43-69 rokov, z\u00a0toho 40% \u017eien. Stanoven\u00e9 koncentr\u00e1cie vitam\u00ednu E boli v\u00a0rozsahu deklarovan\u00fdch referen\u010dn\u00fdch hodn\u00f4t 11,6 \u2013 34,8 \u03bcmol.l-1<\/sup><\/p>\n\n\n\n\n
<\/td>\n<\/td>\n<\/td>\n<\/td>\n<\/tr>\n
<\/td>\n<\/td>\n\n\n\n\n
Tabu\u013eka 2<\/strong> Optimaliz\u00e1cia chromatografickej met\u00f3dy stanovenia vitam\u00ednu\u00a0E<\/p>\n\n\n\n\n\n\n\n\n
Parametre<\/strong><\/td>\nRozsah testovania<\/strong><\/td>\nOptimum<\/strong><\/td>\n<\/tr>\n
prietok<\/strong><\/td>\n0,5 \u2013 1,0 ml.min-1<\/sup><\/td>\n0,8 ml.min-1<\/sup><\/td>\n<\/tr>\n
teplota kol\u00f3ny<\/strong><\/td>\n40 \u2013 55 \u00b0C<\/td>\n48 \u00b0C<\/td>\n<\/tr>\n
mobiln\u00e1 f\u00e1za<\/strong><\/td>\nmetanol, acetonitril, voda, etanol<\/td>\nmetanol:voda (97:3,v\/v)<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

 <\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n<\/td>\n<\/tr>\n

<\/td>\n<\/tr>\n
<\/td>\n\n\n\n\n
Tabu\u013eka 3<\/strong>\u00a0 Opakovate\u013enos\u0165 a\u00a0reprodukovate\u013enos\u0165 stanovenia vitam\u00ednu\u00a0E met\u00f3dou LC-MS-MS<\/p>\n\n\n\n\n
\u00a0<\/strong><\/p>\n

analyt<\/strong><\/td>\n

v\u00a0s\u00e9rii (n=15)<\/strong>\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0 \u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0<\/strong>medzi s\u00e9riami \u00a0(n=6)<\/strong><\/p>\n

priemer\u00b1SD\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0 \u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0CV<\/strong>%\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0 <\/strong>priemer\u00b1SD\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0 \u00a0\u00a0\u00a0\u00a0CV%<\/strong><\/p>\n

\u03bcmol.l-1\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0 \u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0\u00a0<\/sup>\u03bcmol.l-1<\/sup><\/strong><\/td>\n<\/tr>\n

\u00a0<\/strong><\/p>\n

\u03b1-tokoferol<\/strong><\/td>\n

 <\/p>\n

25,3\u00b10,21<\/p>\n

57,1\u00b1<\/strong>0,04<\/td>\n

 <\/p>\n

3,6<\/p>\n

4,0<\/td>\n

 <\/p>\n

26,7\u00b1<\/strong>0,61<\/p>\n

50,2\u00b1<\/strong>1,11<\/td>\n

 <\/p>\n

5,4<\/p>\n

5,1<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

 <\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

\u00a0<\/strong>\u00a0<\/strong>Z\u00e1ver<\/strong><\/p>\n

Vyvinut\u00e1 met\u00f3da stanovenia vitam\u00ednu E v\u00a0s\u00e9re je zalo\u017een\u00e1 na princ\u00edpe LCMSMS. Met\u00f3da sp\u013a\u0148a po\u017eadovan\u00e9 valida\u010dn\u00e9 charakteristiky, je \u00a0spo\u013eahliv\u00e1, citliv\u00e1 a \u0161pecifick\u00e1. Vzh\u013eadom na uveden\u00e9 skuto\u010dnosti je \u00a0mo\u017en\u00e9 met\u00f3du zavies\u0165 v\u00a0klinick\u00fdch laborat\u00f3ri\u00e1ch ako alternat\u00edvu stanovenia vitam\u00ednu E v\u00a0rutinnej diagnostike.<\/p>\n

\u00a0<\/strong><\/p>\n

Po\u010fakovanie<\/strong><\/p>\n

T\u00e1to pr\u00e1ca vznikla za podpory projektu ITMS 26220220197.<\/p>\n

\u00a0<\/strong><\/p>\n

Literat\u00fara<\/strong><\/p>\n

    \n
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  6. JShuing ding. Inez Schoenmakers, Kerry Jones at.al :Quantitative determination of vitamin D metabolites in plasma using UHPLC-MS\/MS<\/em>, Anal Bioanal Chem 2010 (398):779-789, ISSN:1618-2642<\/li>\n
  7. Moreno P., Salvad\u00f3 V.: Determination of eight water- and fat-soluble vitamins in multi-vitamin pharmaceutical formulations by high-performance liquid chromatography<\/em>, Journal of Chromatography A, 2000 (870): 207-215, ISSN: 0021-9673<\/li>\n
  8. Abahusain M. A., Wright J., Dickerson J.W.T., at.al: determination of Retinol, \u03b1- Tocopherol, and \u03b2 \u2013 Carotene by direct Extraction of Human Serum Using High Performance Liquid Chromatography,<\/em> Biomedical Chromatography, 1998 (12): 89-93, ISSN: 1099-0801<\/li>\n
  9. Capote F.P., Jimnez J.R., Granados J.M. Castro M.L.: Identification and determination of fat-soluble vitamins and metabolites in human serum by liquid chromatography\/triple quadrupole mass spectrometry with multiple reaction monitoring<\/em>, Rapid Commun. Mass Spectrom. 2007 (21): 1745-1754, ISSN: 1097-0231<\/li>\n
  10. Midttun \u00d8., Ueland P.M.: Determination of vitamins A, D, E in a\u00a0small volume of human plasma by high-throughput method based on liquid chromatography\/tandem mass spectrometry<\/em>, Rapid Commun. Mass Spectrom. 2011 (25): 1942-1948, ISSN:1097-0231<\/li>\n
  11. Grimmelt D.H., Zheng Q., at.al: Comprehensive MRM Analysis of Fat Soluble Vitamins including 25 (OH)- Vitamin D3: Sensitivity and Selectivity by ESI LC\/MS\/MS, <\/em>Ionics mass Spectrometry Group Inc.<\/em><\/li>\n
  12. David A, Armbruster TP. : Limit of Blank, Limit of Detection and Limit of Quantitation<\/em>, Clin Biochem Rev, 2008 (29): S49-52, ISSN: 0159-8090<\/li>\n
  13. International Conference on Harmonization (ICH) of Technical Requirements for the Registration of Pharmaceuticals for Human Use, Validation of analytical procedures: Text and Methodology. ICH-Q2B, Geneva; 1996.<\/li>\n
  14. Siong T.E, Choo K.S.: Simultaneous determination of retinol and \u03b1- tocopherol by high pressure liquid chromatography in micro-volumes of serum, <\/em>Mal J Nutr. 1999 (5): 21-30, ISSN: 1394-035<\/li>\n
  15. Talwar D., Ha T.KK., Cooney J., at. al: Routine method for the simultaneous measurment of retinol, \u03b1- tocopherol, and five carotenoids in human plasma by reverse phase HPLC,<\/em> Clinica Chimica Acta 1998 (270): 85-100, ISSN: 1873-3492<\/li>\n
  16. Li J., Wang Y.: Fat-Soluble Vitamins Analysis on an Agilent ZORBAX Eclipse PAH Polymeric C18 Bonded Column, <\/em>Application Note from Agilent Technologies (Shanghai) Co. Ltd.<\/li>\n
  17. Szczesniewski A.: Rapid Analysis of Vitamin D in Serum Using Triple Quadrupole LC\/MS, <\/em>\u00a0Application Note from Agilent Technologies, Inc. (Santa Clara), CA USA<\/li>\n
  18. Khan, A., Khan, M.I., Iqbal, Z., Shah, Y., Ahmad, L. and Watson, D. G.\u00a0An optimized and validated RP-HPLC\/UV detection method for simultaneous determination of all-trans-retinol (vitamin A) and alpha-tocopherol (vitamin E) in human serum, Comparison of different particulate reversed-phase HPLC columns.<\/em> Chromatogr. B 2010 (878):2339-2347, ISSN: 1570-0232<\/li>\n<\/ol>\n

    \u00a0<\/em><\/p>\n

    \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":"

    *All tables, charts, graphs and pictures that are featured in this article can be found in the .pdf\u00a0 attachment at the end of the paper.   Vitam\u00edny rozpustn\u00e9 v\u00a0tukoch s\u00fa l\u00e1tky potrebn\u00e9 pre spr\u00e1vne fungovanie organizmu. \u00a0Spolu s\u00a0prote\u00ednmi, tukmi a\u00a0sacharidmi patria k\u00a0z\u00e1kladn\u00fdm zlo\u017ek\u00e1m na\u0161ej potravy. Vitam\u00edny z\u00a0tejto skupiny p\u00f4sobia pri metabolick\u00fdch procesoch (katabolizmus, anabolizmus) ako<\/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":[469,470,471,472,805],"class_list":["post-1204","post","type-post","status-publish","format-standard","hentry","category-biochemistry","tag-lc-ms-ms-en","tag-lod-en","tag-lqd-en","tag-serum-en","tag--tocopherol-e-vitamin","typ_clanku-review-article"],"acf":{"abstrakt":"

    The objective of this paper was to develop a method for establishing \u03b1-tocopherol (E vitamin) level on a triple quadruple liquid chromatograph. Initial sample volume used for the analysis was 50\u03bcl of serum. Ions were detected in the positive mode with high-specificity and sensitive technology of multiple reaction monitoring (MRM).\u00a0The detection limit for \u03b1-tocopherol was 0.01 \u03bcmol.l-1<\/sup>.<\/p>\n

    Key words:<\/strong> \u03b1-tocopherol (E vitamin), serum, LC-MS-MS, LOD, LQD<\/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":"

    GENETICS<\/strong><\/h4>\r\n
      \r\n \t
    • The laboratory age<\/li>\r\n \t
    • The Odyssey of DNA reading<\/li>\r\n \t
    • Comparative genomic hybridisation: a methodological introduction<\/li>\r\n \t
    • Next generation sequencing and its application in clinical genetics<\/li>\r\n<\/ul>\r\n

      <\/h4>\r\n

      BIOCHEMISTRY<\/strong><\/h4>\r\n
        \r\n \t
      • Determining of the trace elements in blood serum<\/li>\r\n \t
      • Determining of \u03b1-tocopherol (vitamin E) in serum by way of liquid chromatography with tandem mass spectrometry (LC\/MS\/MS)<\/li>\r\n \t
      • Analysis of urinary calculi and its path to Europe<\/li>\r\n<\/ul>\r\n \r\n

        IMMUNOLOGY <\/strong><\/h4>\r\n
          \r\n \t
        • New autoantibodies in diagnosis of autoimmunity myopathies<\/li>\r\n \t
        • 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 \r\n

          CYTOLOGY AND PATHOLOGY <\/strong><\/h4>\r\n
            \r\n \t
          • 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":"28","upload_clanok":{"ID":1205,"id":1205,"title":"medirex_1_2015-varhacova","filename":"MEDIREX_1_2015-\u2013-Varhacova.pdf","filesize":149098,"url":"https:\/\/www.newslab.sk\/wp-content\/uploads\/2016\/12\/MEDIREX_1_2015-\u2013-Varhacova.pdf","link":"https:\/\/www.newslab.sk\/en\/establishing-%ce%b1-tocopherol-e-vitamin-level-in-serum-by-way-of-triple-quadrupole-liquid-chromatography-lcmsms\/medirex_1_2015-varhacova\/","alt":"","author":"7","description":"","caption":"","name":"medirex_1_2015-varhacova","status":"inherit","uploaded_to":1204,"date":"2016-12-06 20:26:20","modified":"2016-12-06 20:26:20","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\/1204","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=1204"}],"version-history":[{"count":0,"href":"https:\/\/www.newslab.sk\/en\/wp-json\/wp\/v2\/posts\/1204\/revisions"}],"wp:attachment":[{"href":"https:\/\/www.newslab.sk\/en\/wp-json\/wp\/v2\/media?parent=1204"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/www.newslab.sk\/en\/wp-json\/wp\/v2\/categories?post=1204"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/www.newslab.sk\/en\/wp-json\/wp\/v2\/tags?post=1204"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}