Column volume, micro

A disposable micro-affinity avidin cartridge was prepared by slurry packing bulk B/A Poros resin into a 500 pm x 5 cm piece of peek tubing fitted with 0.062" X 0.028" 2 pm stainless steel frits at both ends of the colunm. Biotinyl-SH2 was expressed in e. coli cells using a T7 expression plasmid and purified as described by Consler et al. (10). An aliquot corresponding to approximately 2.5nmoles of BiotinyI-SH2 was loaded onto the avidin micro affinity column at a flow rate of 2 column volumes/min. 

Another significant factor that impacts column efficiency is the temperature mismatch of an eluent entering a column. This effect can be minimized by a preheating coil or a micro-thermostat however, it should be noted that such heating coils or tubing can lead to an increased extra-column volume and thus band-broadening, as previously discussed. 

Concerning function integration, for example, micro-flow membrane reactors can exhibit similar process intensification, as shown already for their large-scale counterparts [75]. Separation columns for proteomics, immobilizing enzymes, utilize the large surface-to-volume ratios. Surface tension differences can guide and transport liquids selectively. 

Figure 5.18 Comparison of space-time yields of direct fluorination of toluene for the falling film micro reactor (FFMR), micro bubble column (MBC) and laboratory bubble column (LBC) referred to the reaction volume (a) and referred to an idealized reactor geometry (b) [38],...

The above-mentioned space-time yields were referred solely to the reaction volume, i.e. the micro channel volume. When defining this quantity via an idealized reactor geometry, taking into accoimt the construction material as well, natarally the difference in space-time yield of the micro reactors from the laboratory bubble column becomes smaller. Still, the performance of the micro reactors is more than one order of magnitude better [38], The space-time yields for the micro reactors defined in this way ranged from about 200 to 1100 mol monofluorinated product... 

LC-NMR hyphenation consists of a liquid chromatograph (autosampler, pump, column and oven) and a classical HPLC detector. The flow of the detector is brought via an interface to the flow-cell NMR probe. Using commercial NMR flow-cells with volumes between 40 and 180 p,L, in connection with microbore columns or packed capillaries, complete spectra have been provided from 1 nmol of sample. These micro-cells allow expensive deuterated solvents to be used, and thus eliminate solvent interference without excessive cost. The HPLC eluent can be split in order to allow simultaneous MS detection. 

FIGURE 1.30 Micro-HPLC separation of all 4 stereoisomers of the dipeptide alanyl-alanine as FMOC derivatives (a) and DNP-derivatives (b), respectively, on a 0-9-(tert-butylcarbamoyl)quinine-based CSP. Experimental conditions Column dimension, 150 X 0.5 mm ID mobile phase (a) acetonitrile-methanol (80 20 v/v) containing 400 mM acetic acid and 4 mM triethylamine, and (b) methanol-0.5 M ammonium acetate buffer (80 20 v/v) (pHa 6.0) flow rate, 10 ixLmin temperature, 25 C injection volume, 250 nL detection, UV at 250 nm. (Reproduced fromC. Czerwenka et al., J. Pharm. Biomed. Anal., 30 1789 (2003). With permission.)... 

A 0.5-yL syringe-loaded micro injector (Rheodyne 7520) is used for sample injection. To minimize the dead volume (< 20 nL) the fused silica column is connected directly to the injector block. 

High-performance liquid chromatography (HPLC) has become a standard separation technique used in both academic and commercial analytical laboratories. However, there are several drawbacks to standard HPLC, including high solvent consumption, large sample quantity, and decreased detection sensitivity. Micro-HPLC (pHPLC) is a term that encompasses a broad range of sample volumes and column sizes (as shown in Table 3.1), but Saito and coworkers provided narrower definitions in their review based on the size of the columns. ... 

Figure 5.4A and B compares the uncorrected elution volumes, the elution volumes corrected by simple addition of and the elution volumes calculated using Equation 5.21 in RP gradient elution on a conventional and on a micro-bore C18 column. The effect of the gradient dwell volume is more important for separations on short columns and especially on narrow-bore columns with i.d. 2 mm. 

The effect of the dwell volume on the retention times of analytes increases with decreasing retention factor at the start of gradient elution and with increasing ratio VpIV, and becomes very significant in the instrumental setup with the dwell volume comparable to or larger than the column hold-up volume, which is more likely to occur in micro- or in capillary LC than in conventional analytical LC (see Figure 5.4) [12]. 

Activity Measurements. To test catalytic properties of various samples partial oxidation of methanol to formaldehyde was studied in a flow micro-reactor operating under normal atmospheric pressure (10). For each run about 0.2 g of catalyst sample was used and the activities were measured at 173 C in the absence of any diffusional effects. The feed gas consisted of 72, 2 and by volume of nitrogen, oxygen and methanol vapor respectively. Reaction products were analysed with a 10% Carbowax 20 M column (2m long) maintained at 60 C oven temperature. 

Controlled by the MS-MS software, the PE Series 200-micro LC pump operates iso-cratically using 80% acetonitrileiwater at 40 pl/min. A 50 x 1 mm C18 column (Keystone) is used between the pump and autosampler to provide back-pressure. The syringe/system flush solution is the 80% acetonitrileiwater used as the mobile phase. The autosampler is connected directly to the MS-MS Turboion Spray source. The injected sample volume is 20 pi. 

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