Analysis of lipoproteins by capillary zone electrophoresis in microfluidic devices: Assay development and surface roughness measurements

Bruce H. Weiller, Laura Ceriotti, Takayuki Shibata, Dietrich Rein, Matthew A. Roberts, Jan Lichtenberg, J. Bruce German, Nico F. De Rooij, Elisabeth Verpoorte

Research output: Contribution to journalArticle

52 Scopus citations

Abstract

The development of a new assay for lipoproteins by capillary electrophoresis in fused-silica capillaries and in glass microdevices is described in this paper. The separation of low-density (LDL) and high-density (HDL) lipoproteins by capillary zone electrophoresis is demonstrated in fused-silica capillaries with both UV absorption and laser-induced fluorescence detection. This separation was accomplished using Tricine buffer (pH 9.0) with methylglucamine added as a dynamic coating. With UV detection, LDL eluted as a relatively sharp peak with a migration time of ∼11 min and HDL eluted as a broad peak with a migration time of 12.5 min. Fluorescence detection of lipoproteins stained with NBD-ceramide was used with the same buffer system to give comparable results. Furthermore, fluorescence staining of human serum samples yielded results similar to the fluorescently stained LDL and HDL fractions, showing that this method can be used to quantify lipoproteins in serum samples. The method was also used to detect lipoproteins in glass micro-CE devices. Very similar results were obtained in microdevices although with much faster analysis times, LDL eluted as a sharp peak at ∼25 s and HDL as a broad peak at slightly longer time. In addition, higher resolution was obtained on chips. To our knowledge, these results show the first separation and detection of lipoproteins in a microfluidic device using native serum samples. Atomic force microscopy was used to characterize the rms surface roughness (Rq) of microfluidic channels directly. Devices with different surface roughness values were fabricated using two different etchants for Pyrex wafers with a polysilicon masking layer. Using 49% HF, the measured roughness is Rq q = 10.9 ± 1.6 nm and with buffered HF (NH4F + HF) the roughness is Rq = 2.4 ± 0.7 nm. At this level of surface roughness, there is no observable effect on the performance of the devices for this lipoprotein separation.

Original languageEnglish (US)
Pages (from-to)1702-1711
Number of pages10
JournalAnalytical Chemistry
Volume74
Issue number7
DOIs
StatePublished - Apr 1 2002
Externally publishedYes

ASJC Scopus subject areas

  • Analytical Chemistry

Fingerprint Dive into the research topics of 'Analysis of lipoproteins by capillary zone electrophoresis in microfluidic devices: Assay development and surface roughness measurements'. Together they form a unique fingerprint.

  • Cite this

    Weiller, B. H., Ceriotti, L., Shibata, T., Rein, D., Roberts, M. A., Lichtenberg, J., German, J. B., De Rooij, N. F., & Verpoorte, E. (2002). Analysis of lipoproteins by capillary zone electrophoresis in microfluidic devices: Assay development and surface roughness measurements. Analytical Chemistry, 74(7), 1702-1711. https://doi.org/10.1021/ac011096y