Factors Retention time

Occasionally, samples are run that adsorb onto the packing material. Generally, if one of the performance characteristics of the column changes by 10% or more, it is prudent to clean the column. These performance characteristics are (1) asymmetry factor, retention time, resolution, and theoretical plates. 

A tentative list of factors that may be investigated in the robustness test is presented in Table 8. This list is not complete and additional factors may be added. The limits for the factor levels are proposals and should be evaluated case by case. The significance of the effects of the factors on the responses such as the resolution of all peak pairs, the tailing factor, retention times, analysis time, etc., is evaluated. 

Briefly, the method involves determining the capacity factors (retention time corrected for an unretained substance) for a suitable set of reference substances (having known K(k values) using RP-HPLC. The relationship between the capacity factors and Kol for the reference or calibration compounds is determined from regression analysis of a log-log plot of the two properties. The capacity factors of compounds having unknown Koc values then are determined using the identical experimental conditions, and Koc values then are calculated from the regression expression. 

The lipophilicity of compounds can be measured with any commercially available HPLC equipment most commonly UV absorption is used as detection system for the chromatography. The retention factor (retention time) of a compound is the analytical parameter needed to measure lipophilicity. To measure the retention factor, the retention time of the compound under investigation has to be determined reproducibly together with the dead time. The dead time is determined using the retention time of a not retained substance on reversed phase columns like buffer salts. 

Despite quite different retention factors (bars), which depend on the hydro-phobic character of the phases, one observes very similar separation factors (line). Under such chromatographic conditions, the stationary phase decreases to a secondary factor in the optimization process (see Fig. 8) there is a large range of retention factors (retention time), and a very small range of separation factors (selectivities). 

Recovery factor Reduced column length Reduced plate height Reduced velocity Relative retention ratio Retardation factor d Retention time Retention volume Selectivity coefficient Separation factor... 

A solute s capacity factor can be determined from a chromatogram by measuring the column s void time, f, and the solute s retention time, (see Figure 12.7). The mobile phase s average linear velocity, m, is equal to the length of the column, L, divided by the time required to elute a nonretained solute. 

In a chromatographic analysis of low-molecular-weight acids, butyric acid elutes with a retention time of 7.63 min. The column s void time is 0.31 min. Calculate the capacity factor for butyric acid. 

The identities of the solutes are defined such that solute A always has the smaller retention time. Accordingly, the selectivity factor is equal to 1 when the solutes elute with identical retention times, and is greater than 1 when is greater than fr A-... 

In the same chromatographic analysis for low-molecular-weight acids considered in Example 12.2, the retention time for isobutyric acid is 5.98 min. What is the selectivity factor for isobutyric acid and butyric acid ... 

Now that we have defined capacity factor, selectivity, and column efficiency we consider their relationship to chromatographic resolution. Since we are only interested in the resolution between solutes eluting with similar retention times, it is safe to assume that the peak widths for the two solutes are approximately the same. Equation 12.1, therefore, is written as... 

The retention times for solutes A and B are replaced with their respective capacity factors by rearranging equation 12.10... 

Any improvement in resolution obtained by increasing ki generally comes at the expense of a longer analysis time. This is also indicated in Figure 12.11, which shows the relative change in retention time as a function of the new capacity factor. Note that a minimum in the retention time curve occurs when b is equal to 2, and that retention time increases in either direction. Increasing b from 2 to 10, for example, approximately doubles solute B s retention time. 

Adjusting the capacity factor to improve resolution between one pair of solutes may lead to an unacceptably long retention time for other solutes. For example, improving resolution for solutes with short retention times by increasing... 

Using the data from Problem 1, calculate the resolution and selectivity factors for each pair of adjacent compounds. For resolution, use both equations 12.1 and 12.21, and compare your results. Discuss how you might improve the resolution between compounds B and C. The retention time for an unretained solute is 1.19 min. 

Internal standards at a known concentration are added to the sample after its preparation but prior to analysis to check for GC retention-time accuracy and response stability. If the internal standard responses are in error by more than a factor of two, the analysis must be stopped and the initial calibration repeated. Only if all the criteria have been met can sample analysis begin. 

The definition of polymer thermal stabiUty is not simple owing to the number of measurement techniques, desired properties, and factors that affect each (time, heating rate, atmosphere, etc). The easiest evaluation of thermal stabiUty is by the temperature at which a certain weight loss occurs as observed by thermogravimetric analysis (tga). Early work assigned a 7% loss as the point of stabiUty more recentiy a 10% value or the extrapolated break in the tga curve has been used. A more reaUstic view is to compare weight loss vs time at constant temperature, and better yet is to evaluate property retention time at temperature one set of criteria has been 177°C for 30,000 h, or 240°C for 1000 h, or 538°C for 1 h, or 816°C for 5 min (1). 

The basic operations in dust collection by any device are (1) separation of the gas-borne particles from the gas stream by deposition on a collecting surface (2) retention of the deposit on the surface and (3) removal of the deposit from the surface for recovery or disposal. The separation step requires (1) application of a force that produces a differential motion of a particle relative to the gas and (2) a gas retention time sufficient for the particle to migrate to the coUecting surface. The principal mechanisms of aerosol deposition that are apphed in dust collectors are (1) gravitational deposition, (2) flow-line interception, (3) inertial deposition, (4) diffusional deposition, and (5) electrostatic deposition. Thermal deposition is only a minor factor in practical dust-collectiou equipment because the thermophoretic force is small. Table 17-2 lists these six mechanisms and presents the characteristic... 

An anaerobic digester is a no-recycle complete mix reactor. Thus, its performance is independent of organic loading but is controlled by hydraulic retention time (HRT). Based on kinetic theoiy and values of the pseudo constants for methane bac teria, a minimum HRT of 3 to 4 days is required. To provide a safety factor and compensate for load variation as indicated earlier, HRT is kept in the range 10 to 30 days. Thickening of feed sludge is used to reduce the tank volume required... 

If the mobile phase is a liquid, and can be considered incompressible, then the volume of the mobile phase eluted from the column, between the injection and the peak maximum, can be easily obtained from the product of the flow rate and the retention time. For more precise measurements, the volume of eluent can be directly measured volumetrically by means of a burette or other suitable volume measuring vessel that is placed at the end of the column. If the mobile phase is compressible, however, the volume of mobile phase that passes through the column, measured at the exit, will no longer represent the true retention volume, as the volume flow will increase continuously along the column as the pressure falls. This problem was solved by James and Martin [3], who derived a correction factor that allowed the actual retention volume to be calculated from the retention volume measured at the column outlet at atmospheric pressure, and a function of the inlet/outlet pressure ratio. This correction factor can be derived as follows. 

Analytical information taken from a chromatogram has almost exclusively involved either retention data (retention times, capacity factors, etc.) for peak identification or peak heights and peak areas for quantitative assessment. The width of the peak has been rarely used for analytical purposes, except occasionally to obtain approximate values for peak areas. Nevertheless, as seen from the Rate Theory, the peak width is inversely proportional to the solute diffusivity which, in turn, is a function of the solute molecular weight. It follows that for high molecular weight materials, particularly those that cannot be volatalized in the ionization source of a mass spectrometer, peak width measurement offers an approximate source of molecular weight data for very intractable solutes. 

It is important that records are not destroyed before their useful life is over. There are several factors to consider when determining the retention time for quality records ... 

A standard test probe is not absolutely necessary to monitor the column. Any well-resolved peak in the sample may be used. To use a sample component, baseline data must be established when the column is new and performing well. After establishing that the column is performing properly using the manufacturer s standard test procedure, calculate the assymetry factor, theoretical plates, and resolution of one or more of the sample components. Also note the retention time. This will become the baseline test mix, which will be used for later comparison. 

Table 17.2 shows good agreement between the retention times from the TSK PEO standards analyzed in groups of two or three and the retention times of the TSK PEO standards analyzed individually. ASTM-D5296 requires that for standards to be run as a group, the molecular weight must differ by a factor of 10 (11). The results in Table 17.2 showed that a difference in molecular weight by a factor of 6 is adequate to obtain consistent flow times for standards for the modem linear columns. 

Several factors can contribute to the difference in retention times for PEO in different mobile phases the viscosity of a mobile phase, the hydrodynamic volume of a PEO, and the swelling or void volume of a column. Shodex and TSK columns should swell more in water than in water/methanol, and PEO should therefore come out later in water than in water/methanol. PEO should also elute later in water than in water/methanol because water/methanol is a better solvent for PEO than water. The viscosity of the 50 50 water/methanol mobile phase is higher than the viscosity of water. PEO should therefore elute later in water/methanol than in water due to the difference in viscosity. The results in Table 17.9 indicate that the difference in retention time for PEO in water and in water/methanol depends more on the swelling of columns and the hydrodynamic volumes of PEO than the viscosities of mobile phases. 

---