1 st fraction 1.5 1.0 0.5 (x10,000,000) 1:TIC (1.00) 2.5 Inten.(x10,000) 185.102 0.0 1.0 2.0 3.0 4.0 5.0 6.0 7.0 8.0 9.0 10.0 11.0 2 nd fraction 1.50 1.25 1.00 0.75 0.50 0.25 1.5 1.0 0.5 0.0 (x10,000,000) 1:TIC (1.00) 0.0 1.0 2.0 3.0 4.0 5.0 6.0 7.0 8.0 9.0 10.0 11.0 Inten.(x10,000) 3 th fraction 1.5 1.0 0.5 2.0 1.5 1.0 0.5 0.0 227.107 384.182 280.977 450.264 847.939 705.777 755.133 895.225 655.563 186.109 545.736 1004.459 1038.537 (x10,000,000) 1:TIC (1.00) Inten.(x10,000) 2.5 2.0 1.5 1.0 0.5 0.0 1190.750 1301.812 250 500 750 1000 1250 m/z 185.115 235.001 384.182 487.862 279.168 509.278 613.759 781.853 850.840 957.000 Inten.(x100) 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 250 500 750 1000 1250 m/z 311.221 406.206 14302.595 487.235 558.720 895.508 656.145 822.520 690.052 250 500 750 1000 1250 m/z 0.0 1.0 2.0 3.0 4.0 5.0 6.0 7.0 8.0 9.0 10.0 11.0 12.0 Fig.7: TIC traces (a), MS spectra (b) and deconvoluted MS spectrum (c) from HPLC/MS analysis <strong>of</strong> the fractions obtained during the preparative ITP isolation <strong>of</strong> saliva sample spiked with lysozyme standard. For details, see Experimental. 1114.782 0.0 14200 14225 14250 14275 14300 14325 14350 14375 mass <strong>Zborník</strong> <strong>príspevkov</strong> z 18. medzinárodnej <strong>vedeckej</strong> <strong>konferencie</strong> "Analytické metódy a zdravie loveka", ISBN 978-80-969435-7-9 - 131 - c) 1209.565 b) b) a) a) b) a) hotel Falkensteiner, Bratislava 11. - 14. 10. 2010
CONCLUSIONS This work studied the possibilities <strong>of</strong> preparative isotachophoresis (pITP) as a sample pretreatment technique before high performance liquid chromatography-mass spectrometric separation (HPLC-MS) in analysis <strong>of</strong> high molecular weight compounds present in complex biological matrix. The analysis <strong>of</strong> chosen analyte (human lysozyme) present in complex biological matrix (saliva) at different concentration levels was selected as an example <strong>of</strong> solving the analytical problem by using pITP-HPLC-MS combination. Without using pITP pretreatment it was impossible to detect lysozyme in saliva by direct HPLC-MS analysis while its using significantly simplified the saliva sample what is favourable for the minimizing <strong>of</strong> the matrix effects in HPLC-MS. Human lysozyme was found in the saliva samples <strong>of</strong> 4 volunteers using just deconvolution <strong>of</strong> data from MS mode after the background correction. Proper optimization <strong>of</strong> HPLC separation conditions as well as the data acquisition in MS and MS/MS modes can improve both the level <strong>of</strong> identification and the concentration limit <strong>of</strong> detection <strong>of</strong> lysozyme present in biological matrices. Acknowledgements This work was supported by a grant from the Slovak Research and Development Agency (No.VVCE-0070-07) and the grants <strong>of</strong> Slovak Grant Agency, No`s.1/0882/09 and 1/0546/10. REFERENCES [1] F. Amado, M. J. Lobo, P. Domingues, J. A. Duarte, R. Vitorino, Expert. Rev. Proteomics 7 (2010) 709 [2] D. Koh, Y. Yang, L. Khoo, S. Z Nyunt, V. Ng, C. L. Goh, Ann. Acad. Med. Singapore 33 (2004) 307 [3] F. M. Everaerts, J. L. Beckers, T. P. E. M. Verheggen, Isotachophoresis, Theory, Instrumentation and Applications, Elsevier, Amsterdam, 1976 [4] Kaniansky, D., Marák, J., J. Chromatogr. 498 (1990) 191 [5] Kaniansky, D., Marák, J., Madajová, V., Šimuniová, E., J. Chromatogr. 638 (1993) 137 [6] T. Hirokawa, Y. Kiso, J. Chromatogr. A 658 (1994) 343 [7] E. Kenndler, D. Kaniansky, J. Chromatogr. 209 (1981) 306 [8] M. Hutta, D. Kaniansky, E. Kovalíková, J. Marák, M. Chalányová, V. Madajová, E. Šimuniová, J. Chromatogr. A 689 (1995) 123 [9] Kaniansky, D., Madajová, V., Hutta, M., Žilková, I., J. Chromatogr. 286 (1984) 395 [10] Schoots, A. C., Everaerts, F. M., J. Chromatogr. 227 (1983) 328 [11] R. Sladkovský, M. Urbánek, P. Solich, Chromatografia 58 (2003) 187 [12] Nielen M. W., Bovee T. F., van Engelen M. C., Rutgers P., Hamers A. R., van Rhijn J. H., Hoogenboom L. R., Anal. Chem. 78 (2006) 424 <strong>Zborník</strong> <strong>príspevkov</strong> z 18. medzinárodnej <strong>vedeckej</strong> <strong>konferencie</strong> "Analytické metódy a zdravie loveka", ISBN 978-80-969435-7-9 - 132 - hotel Falkensteiner, Bratislava 11. - 14. 10. 2010
- Page 2 and 3:
Zborník príspevkov z 18. medziná
- Page 4 and 5:
Konferencia bola venovaná pamiatke
- Page 6 and 7:
Obsah zborníka príspevkov z 18. m
- Page 8 and 9:
ANALÝZA BENZÉNU, TOLUÉNU, ETYLBE
- Page 10 and 11:
Obr. 2. Schéma uzatvoreného syst
- Page 12 and 13:
Reálne vzorky Na obr. 5 je znázor
- Page 14 and 15:
Modernizovaný oblúkový výboj (o
- Page 16 and 17:
ICP-Torch Transport-Tube Bypass (Zu
- Page 18 and 19:
Tabuka V : Výsledky kalibrácie -
- Page 20 and 21:
LITERATÚRA [1] A. M.Ure, C. M. Dav
- Page 22 and 23:
2 EXPERIMENTÁLNA AS 2.1 Použité
- Page 24 and 25:
3.2 Optimalizácia experimentálnyc
- Page 26 and 27:
tenzidu. CPT Tritonu X-114 je okolo
- Page 28 and 29:
MOŽNOSTI VYUŽITIA KOMBINÁCIE TEC
- Page 30 and 31:
Pre iónovo-výmennú chromatografi
- Page 32 and 33:
Obr. 7: Skúmanie vplyvu koncentrá
- Page 34 and 35:
Po týchto skúsenostiach sme sa ro
- Page 36 and 37:
IZOELEKTRICKÁ FOKUSACE VE SKOKOVÉ
- Page 38 and 39:
Obr.2. Schéma neutralizaního reak
- Page 40 and 41:
Tab 1: Závislost délky zóny na d
- Page 42 and 43:
BIOAKUMULÁCIA TOXICKÝCH PRVKOV MI
- Page 44 and 45:
Tabuka 4 Bioakumulovaný Zn jednotl
- Page 46 and 47:
Úbytok kovu v % 100 80 60 40 20 0
- Page 48 and 49:
DEVELOPMENT OF THE SOLID SAMPLING E
- Page 50 and 51:
3. Results and discussion 3.1. Meth
- Page 52 and 53:
different atomization signal profil
- Page 54 and 55:
Financial support from the Scientif
- Page 56 and 57:
Zhydrolyzované ftaláty boli deriv
- Page 58 and 59:
UVONENIE PRCHAVÝCH ORGANICKÝCH ZL
- Page 60 and 61:
a 3B) v headspace bunkách CALU-1 v
- Page 62 and 63:
koncentrácia niekokých alkánov a
- Page 64 and 65:
Obrázok 4: A a B: Porovnanie konce
- Page 66 and 67:
TEPLOTNE PROGRAMOVANÉ PLYNOVO CHRO
- Page 68 and 69:
metyl x-metyl-y-oát OV 1 I P s OV
- Page 70 and 71:
metyl x-metyl-y-oát OV 1 I P s OV
- Page 72 and 73:
Fig. 1. Chromatogram GC separácie
- Page 74 and 75:
OV 1 Obr. 3. Závislos homomorfnýc
- Page 76 and 77:
Záver Namerali sa teplotne-program
- Page 78 and 79:
modifier solutions for ensure homog
- Page 80 and 81:
applied to attain a stabilization o
- Page 82 and 83:
against aqueous standards with a pr
- Page 84 and 85:
Ludrová (6). Maslo sa vyrobilo zmi
- Page 86 and 87:
Tabuka 2. Zloženie mastných kysel
- Page 88 and 89: Tabuka 3. Obsah jednotlivých izom
- Page 90 and 91: [19] J. Blaško, R. Kubinec, I. Ost
- Page 92 and 93: 2. Experimental Instrumentation Flo
- Page 94 and 95: Analysis of real samples The boiler
- Page 96 and 97: a) b) 2 2 1 1 3 3 3 3 Fig. 1 The ex
- Page 98 and 99: Conclusion remarks The identificati
- Page 100 and 101: screening, possible interactions am
- Page 102 and 103: The design consists of a factorial
- Page 104 and 105: elative area 15 10 5 -1,68 -1,00 0,
- Page 106 and 107: It can be seen that the programms d
- Page 108 and 109: nevodíkovými atómami izotropne s
- Page 110 and 111: O22—C6—C7 109.57 (15) C10—C11
- Page 112 and 113: [8] S. Teklu, L.L. Gundersen, T. La
- Page 114 and 115: iónom kovu môže by ovplyvnená a
- Page 116 and 117: V alšej asti práce sme študovali
- Page 118 and 119: vzorky, pomer hmotností vzorky a s
- Page 120 and 121: literatúre nenašli informácie. M
- Page 122 and 123: Záver Pri úprave vzorky pôdy met
- Page 124 and 125: galactose-4-epimerase [6, 7]. First
- Page 126 and 127: Abundance Abundance Abundance 1x10
- Page 128 and 129: Galactitol [mmol/mol creatinine] 60
- Page 130 and 131: References [1] J.T.R. Clarke, Clini
- Page 132 and 133: structural information of the analy
- Page 134 and 135: (x10,000,000) 1.5 1:TIC (1.00) 1.4
- Page 136 and 137: 1 st fraction 1.5 1.0 0.5 (x10,000,
- Page 140 and 141: RAPID LIQUID CHROMATOGRAPHY-MASS SP
- Page 142 and 143: the LC-ESI-IT-TOF MS analyzer. HPLC
- Page 144 and 145: (x10,000,000) 1:TIC (1.00) 4:TIC (1
- Page 146 and 147: Figure 5 is showing MS and MS/MS sp
- Page 148 and 149: Figure 7 present MS and MS/MS spect
- Page 150 and 151: MS and MS/MS spectra of potential m
- Page 152 and 153: DVOUROZMĚRNÁ KAPALINOVÁ CHROMATO
- Page 154 and 155: řřů ěř Pnčč ěě : ččěP2D
- Page 156 and 157: řěůčťě čě č ěě Obr. 6.
- Page 158 and 159: Obr. 9. Separaci metabolitů indolo
- Page 160 and 161: STANOVENIE OXIDANÝCH PRODUKTOV OXI
- Page 162 and 163: e d c b a G NO - 3 NO - 2 NO - 2 NO
- Page 164 and 165: Na obr. 4b je elektroforeogram z IT
- Page 166 and 167: Vhodnos navrhnutého analytického
- Page 168 and 169: Experimentálna as CZE separácie b
- Page 170 and 171: Séria kalibraných meraní modelov
- Page 172 and 173: Výsledky a diskusia Na obr. 2 sú
- Page 174 and 175: Tab. 2: Parametre regresných rovn
- Page 176 and 177: Tab. 3: Opakovatenosti migraných a
- Page 178 and 179: chloridu, typickej makrozložky v C
- Page 180 and 181: kationického roztoku elektrolytu b
- Page 182 and 183: Tab. 2: Stanovenie aniónov a kati
- Page 184 and 185: CHARAKTERIZÁCIA RP-HPLC FRAKCIONOV
- Page 186 and 187: Obr. 2. RP-HPLC profil vzorky HK Al
- Page 188 and 189:
Obr. 6. RP-HPLC profily frakcií vz
- Page 190 and 191:
Zoznam použitej literatúry [1] Ha
- Page 192 and 193:
a druhá kolóna kontaktným vodivo
- Page 194 and 195:
c b a 1000 1500 tm [s] 2000 Obr. 3:
- Page 196 and 197:
Záver ITP-CZE uskutonená použit
- Page 198 and 199:
Automatizácia je založená na po
- Page 200 and 201:
e d c b a 20mAu t CZE (min) 2 4 6 8
- Page 202 and 203:
kyselina; mandlová kyselina (3,75
- Page 204 and 205:
Literatúra [1] Th.P.E.M. Verheggen
- Page 206 and 207:
Z uvedeného vyplýva, že je stál
- Page 208 and 209:
2a.) 2b.) Obrázok 2.: RP - HPLC z
- Page 210 and 211:
Porovnaním chromatogramov frakcií
- Page 212 and 213:
Práca vznikla za podpory grantov V
- Page 214 and 215:
Vznik a funkcia humínových látok
- Page 216 and 217:
celkového náboja jedného gramu p
- Page 218 and 219:
Vzorky pôd a ich charakterizácia
- Page 220 and 221:
Zborník príspevkov z 18. medziná
- Page 222 and 223:
Obrázok 11: Kruhové diagramy perc
- Page 224:
[23] D. Gondar, R. Lopez, S. Fiol,