Nanoflow gradient generator for capillary high-performance liquid chromatography.
A novel method of generating a nanoflow gradient elution for a capillary high-performance liquid chromatography (HPLC) system has been developed. An important feature of this system is that any gradient (GR) profile generated by a conventional microflow GR pump can be asymptotically traced and converted as a corresponding nanoflow GR profile simply by using a 10-port switching valve with two injection loops installed. Consequently, it has been called an "asymptotic trace 10-port valve" (AT10PV) nanoflow GR generator. Performance of the AT10PV nanoflow GR generator was tested in the range of flow rates from 50 to 500 nL/min. The test demonstrated that the AT10PV nanoflow GR generator can asymptotically trace the original gradient profile with good reproducibility. A capillary HPLC system using the AT10PV nanoflow GR generator provides reasonably good repeatability of peak retention times on the chromatogram of the tryptic digest of a BSA sample, RSD of less than 0.3% at a flow rate of 200 nL/min. It also enables sequential running of a series of sample injections in the same manner as conventional analysis at microflow rates.
Division of Biological Sciences, Graduate School of Science, Hokkaido University, Sapporo 001-0021, Japan. deguchi@glyco.sci.hokudai.ac.jp
Kisaburo
K
Deguchi
Shinya
S
Ito
Shinji
S
Yoshioka
Izumi
I
Ogata
Akihiro
A
Takeda
D000096:Acetone; D000097:Acetonitriles; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation* / Q000379:methods*; D044085:Microfluidics; D036103:Nanotechnology; D015203:Reproducibility of Results; D012710:Serum Albumin, Bovine / Q000032:analysis / Q000378:metabolism; D012997:Solvents; D013997:Time Factors; D014357:Trypsin / Q000378:metabolism
D000097:Acetonitriles; D012997:Solvents; D000096:Acetone; D012710:Serum Albumin, Bovine; D014357:Trypsin; C032159:acetonitrile