ADAM, Vojtech, Ondrej ZITKA, Petr DOLEZAL, Ladislav ZEMAN, Ales HORNA, Jaromir HUBALEK, Jan SILENY, Sona KRIZKOVA, Libuše TRNKOVÁ a Rene KIZEK. Lactoferrin isolation using monolithic column coupled with spectrometric or micro-amperometric detector. IEEE Sensors Journal. USA, 2008, roč. 8, č. 1, s. 464-487. ISSN 1530-437X. Dostupné z: https://dx.doi.org/10.3390/s8010464. |
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@article{1662606, author = {Adam, Vojtech and Zitka, Ondrej and Dolezal, Petr and Zeman, Ladislav and Horna, Ales and Hubalek, Jaromir and Sileny, Jan and Krizkova, Sona and Trnková, Libuše and Kizek, Rene}, article_location = {USA}, article_number = {1}, doi = {http://dx.doi.org/10.3390/s8010464}, keywords = {lactoferrin; milk protein; linear sweep voltammetry; screen-printed carbon electrode; electrochemistry; liquid chromatography; monolithic column; UV-VIS spectrometry}, language = {eng}, issn = {1530-437X}, journal = {IEEE Sensors Journal}, title = {Lactoferrin isolation using monolithic column coupled with spectrometric or micro-amperometric detector}, url = {https://www.mdpi.com/1424-8220/8/1/464}, volume = {8}, year = {2008} }
TY - JOUR ID - 1662606 AU - Adam, Vojtech - Zitka, Ondrej - Dolezal, Petr - Zeman, Ladislav - Horna, Ales - Hubalek, Jaromir - Sileny, Jan - Krizkova, Sona - Trnková, Libuše - Kizek, Rene PY - 2008 TI - Lactoferrin isolation using monolithic column coupled with spectrometric or micro-amperometric detector JF - IEEE Sensors Journal VL - 8 IS - 1 SP - 464-487 EP - 464-487 SN - 1530437X KW - lactoferrin KW - milk protein KW - linear sweep voltammetry KW - screen-printed carbon electrode KW - electrochemistry KW - liquid chromatography KW - monolithic column KW - UV-VIS spectrometry UR - https://www.mdpi.com/1424-8220/8/1/464 L2 - https://www.mdpi.com/1424-8220/8/1/464 N2 - Lactoferrin is a multifunctional protein with antimicrobial activity and others to health beneficial properties. The main aim of this work was to propose easy to use technique for lactoferrin isolation from cow colostrum samples. Primarily we utilized sodium dodecyl sulphate - polyacrylamide gel electrophoresis for isolation of lactoferrin from the real samples. Moreover we tested automated microfluidic Experion electrophoresis system to isolate lactoferrin from the collostrum sample. The well developed signal of lactoferrin was determined with detection limit ( 3 S/N) of 20 ng/ml. In spite of the fact that Experion is faster than SDS-PAGE both separation techniques cannot be used in routine analysis. Therefore we have tested third separation technique, ion exchange chromatography, using monolithic column coupled with UV-VIS detector (LC-UV-VIS). We optimized wave length ( 280 nm), ionic strength of the elution solution (1.5 M NaCl) and flow rate of the retention and elution solutions (0.25 ml/min and 0.75 ml/min. respectively). Under the optimal conditions the detection limit was estimated as 0.1 mu g/ml of lactoferrin measured. Using LC-UV-VIS we determined that lactoferrin concentration varied from 0.5 g/l to 1.1 g/l in cow colostrums collected in the certain time interval up to 72 hours after birth. Further we focused on miniaturization of detection device. We tested amperometric detection at carbon electrode. The results encouraged us to attempt to miniaturise whole detection system and to test it on analysis of real samples of human faeces, because lactoferrin level in faeces is closely associated with the inflammations of intestine mucous membrane. For the purpose of miniaturization we employed the technology of printed electrodes. The detection limit of lactoferrin was estimated as 10 mu g/ml measured by the screen-printed electrodes fabricated by us. The fabricated electrodes were compared with commercially available ones. It follows from the obtained results that the responses measured by commercial electrodes are app. ten times higher compared with those measured by the electrodes fabricated by us. This phenomenon relates with smaller working electrode surface area of the electrodes fabricated by us ( about 50%) compared to the commercial ones. The screen-printed electrodes fabricated by us were utilized for determination of lactoferrin faeces. Regarding to fact that sample of faeces was obtained from young and healthy man the amount of lactoferrin in sample was under the limit of detection of this method. ER -
ADAM, Vojtech, Ondrej ZITKA, Petr DOLEZAL, Ladislav ZEMAN, Ales HORNA, Jaromir HUBALEK, Jan SILENY, Sona KRIZKOVA, Libuše TRNKOVÁ a Rene KIZEK. Lactoferrin isolation using monolithic column coupled with spectrometric or micro-amperometric detector. \textit{IEEE Sensors Journal}. USA, 2008, roč.~8, č.~1, s.~464-487. ISSN~1530-437X. Dostupné z: https://dx.doi.org/10.3390/s8010464.
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