Highly Efficient and Ultra-small Volume Separation by Pressure-Driven Liquid Chromatography in Extended Nanochannels

• Published in: Small (Weinheim an der Bergstrasse, Germany) Vol. 8; no. 8; pp. 1237 - 1242
• Main Authors: Ishibashi, Ryo, Mawatari, Kazuma, Kitamori, Takehiko
• Format: Journal Article
• Language: English
• Published: Weinheim WILEY-VCH Verlag 23.04.2012; WILEY‐VCH Verlag
• Subjects: chip-based separation; Chromatography, Liquid - methods; extended nanospace; liquid chromatography; nanofluidics; Nanotechnology - methods; Pressure; pressure-driven flow
• ISSN: 1613-6810; 1613-6829
• DOI: 10.1002/smll.201102420

The rapidly developing interest in nanofluidic analysis, which is used to examine liquids ranging in amounts from the attoliter to the femtoliter scale, correlates with the recent interest in decreased sample amounts, such as in the field of single‐cell analysis. For general nanofluidic analysis, the fact that a pressure‐driven flow does not limit the choice of solvents (aqueous or organic) is important. This study shows the first pressure‐driven liquid chromatography technique that enables separation of atto‐ to femtoliter sample volumes, with a high separation efficiency within a few seconds. The apparent diffusion coefficient measurement of the unretentive sample suggests that there is no increase in the viscosity of toluene in the extended nanospace, unlike in aqueous solvents. Evaluation of the normal phase separation, therefore, should involve only the examination of the effect of the small size of the extended nanospace. Compared to a conventionally packed high‐performance liquid chromatography column, the separation here results in a faster separation (4 s) by 2 orders of magnitude, a smaller injection volume (100 fL) by 9 orders, and a higher separation efficiency (440 000 plates/m) by 1 order. Moreover, the separation behavior agrees with the theory showing that this high efficiency was due to the small and controlled size of the separation channel, where the diffusion through the channel depth direction is fast enough to be neglected. Our chip‐based platform should allow direct and real‐time analysis or screening of ultralow volume of sample. Pressure‐driven liquid chromatography in a 200 nm deep, open‐column channel enables separation of atto‐ to femtoliter sample volume, with a high separation efficiency of 440 000 plates/m within a few seconds. This high efficiency is due to the small and controlled size of the separation channel where the diffusion through the channel depth direction is fast enough to be neglected.