Eluents

In SEC the concentration by weight of polymer in the eluting solvent may be monitored continuously with a detector measuring refractive index, UV absorption, or infrared (IR) absorption [17]. The resulting chromatogram is therefore a weight distribution of the polymer as a function of retention volume, VR. 

Experimental SEC conditions require highly sensitive concentration detectors giving a detector response, which is linearly related to polymer concentration. The most common detector for monitoring polymer concentration in the eluent is the differential refractometer (DRI). The response of the detector to polymer concentration does not depend on polymer molecular weight except for very low polymers. 

The most sensitive detector is the differential UV photometer, which is appropriate for a polymer with a significant UV absorbance with a nonabsorbing eluent. This detector is not appreciably affected by flow pulsations, flow rate changes, and temperature fluctuations. 

When characterizing copolymers, it is necessary to have two detectors in series, e.g., a refractometer with either a UV detector or an IR detector. An IR detector is preferred for the detection of polyalkenes at elevated temperatures because baseline noise and drift are much less than for the refractometer detector. 

If the universal calibration is valid, then at a given VR the following relationship will apply  

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Liquid phase chromatography can use a supercritical fluid as an eluent. The solvent evaporates on leaving the column and allows detection by FID. At present, there are few instances in the petroleum industry using the supercritical fluid technique. 

The value of a graded series of eluents is that if one member of the series succeeds in desorbing a portion of an adsorbate, then another more power ful eluent should remove a further portion of the adsorbate. By mixing the eluents in various proportions, a finer gradation may be obtained, e.g., petroleum ether benzene and petroleum ether benzene and ether ether and acetone, etc. 

The average linear velocity u of the mobile phase in terms of the column length L and the average linear velocity of eluent (which is measured by the transit time of a nonretained solute) is... 

The concentrations of benzoic acid, aspartame, caffeine, and saccharin in a variety of beverages are determined in this experiment. A Gig column and a mobile phase of 80% v/v acetic acid (pH = 4.2) and 20% v/v methanol are used to effect the separation. A UV detector set to 254 nm is used to measure the eluent s absorbance. The ability to adjust retention times by changing the mobile phase s pH is also explored. 

Fig. 5. Anion-exchange separation of insulin and insulin A- and B-chains, over diethylaminoethyl (DEAF) in a 10.9 x 200 mm column having a volume of 18.7 mL. Sample volume is 0.5 mL and protein concentration ia 16.7 mAf Tris buffer at pH 7.3 is 1 mg/mL for each component ia the presence of EDTA. Eluent (also 16.7 mAf Tris buffer, pH 7.3) flow rate is 1.27 ml,/min, and protein detection is by uv absorbance at 280 nm. The straight line depicts the salt...

Fig. 11. Separation of P-labeled A, adenovims Type 5, and B, poHovims Type 1 on Sepharose 2B where the column is 2.1 x 56 cm eluent, 0.002 M...

This reversed-phase chromatography method was successfully used in a production-scale system to purify recombinant insulin. The insulin purified by reversed-phase chromatography has a biological potency equal to that obtained from a conventional system employing ion-exchange and size-exclusion chromatographies (14). The reversed-phase separation was, however, followed by a size-exclusion step to remove the acetonitrile eluent from the final product (12,14). 

Uranium ores are leached with dilute sulfuric acid or an alkaline carbonate [3812-32-6] solution. Hexavalent uranium forms anionic complexes, such as uranyl sulfate [56959-61-6], U02(S0 3, which are more selectively adsorbed by strong base anion exchangers than are other anions in the leach Hquors. Sulfate complexes are eluted with an acidified NaCl or ammonium nitrate [6484-52-2], NH NO, solution. Carbonate complexes are eluted with a neutral brine solution. Uranium is precipitated from the eluent and shipped to other locations for enrichment. Columnar recovery systems were popular in South Africa and Canada. Continuous resin-in-pulp (RIP) systems gained popularity in the United States since they eliminated a difficult and cosdy ore particle/leach hquor separation step. 

Another big advance in the appHcation of ms in biotechnology was the development of atmospheric pressure ionization (API) techniques. There are three variants of API sources, a heated nebulizer plus a corona discharge for ionization (APCl) (51), electrospray (ESI) (52), and ion spray (53). In the APCl interface, the Ic eluent is converted into droplets by pneumatic nebulization, and then a sheath gas sweeps the droplets through a heated tube that vaporizes the solvent and analyte. The corona discharge ionizes solvent molecules, which protonate the analyte. Ions transfer into the mass spectrometer through a transfer line which is cryopumped, to keep a reasonable source pressure. 

Sugar analysis by hplc has advanced greatly as a result of the development of columns specifically designed for carbohydrate separation. These columns fall into several categories. (/) Aminopropyl-bonded siHca used in reverse-phase mode with acetonitrile—water as the eluent. (2) Ion-moderated cation-exchange resins using water as the eluent. Efficiency of these columns is enhanced at elevated temperature, ca 80—90°C. Calcium is the usual counterion for carbohydrate analysis, but lead, silver, hydrogen, sodium, and potassium are used to confer specific selectivities for mono-, di-, and... 

Chromatographic methods including thin-layer, hplc, and gc methods have been developed. In addition to developments ia the types of columns and eluents for hplc appHcations, a significant amount of work has been done ia the kiads of detectioa methods for the vitamin. These detectioa methods iaclude direct detectioa by uv, fluoresceace after post-column reduction of the quiaone to the hydroquinone, and electrochemical detection. Quantitative gc methods have been developed for the vitamin but have found limited appHcations. However, gc methods coupled with highly sensitive detection methods such as gc/ms do represent a powerful analytical tool (20). 

Magnesium, calcium, barium, and strontium can also be deterrnined by ion chromatography with y -phenylenediamine in perchloric acid as the eluent. Ion chromatography by conductimetric detection has been described, and appHcations to environmental waters have been discussed (1,22—23). 

Ion chromatography can be used to determine chloride concentrations of 2—1000 ppb with a carbonate—bicarbonate eluent (23). Eluoride, nitrite, phosphate, bromide, nitrate, and sulfate do not interfere and can be measured simultaneously with a total analysis time of <30 min. 

An important publication by Kost et al. (63JGU525) on thin-layer chromatography (TLC) of pyrazoles contains a large collection of Rf values for 1 1 mixtures of petroleum ether-chloroform or benzene-chloroform as eluents and alumina as stationary phase. 1,3- and 1,5-disubstituted pyrazoles can be separated and identified by TLC (Rf l,3>i y 1,5). For another publication by the same authors on the chromatographic separation of the aminopyrazoles, see (63JGU2519). A-Unsubstituted pyrazoles move with difficulty and it is necessary to add acetone or methanol to the eluent mixture. Other convenient conditions for AH pyrazoles utilize silica gel and ethyl acetate saturated with water (a pentacyanoamine ferroate ammonium disodium salt solution can be used to visualize the pyrazoles). 

In HIC, the hydrophobic interactions are relatively weak, often driven by salts in moderate concentration (I to 2 M), and depend primarily on the exposed residues on or near the protein surface preservation of the native, biologically active state of the protein is an important feature of HIC. Elution can be achieved differentially by decreasing salt concentration or increasing the concentration of polarity perturbants (e.g., ethylene glycol) in the eluent. 

II) On-line eoupling of SPE to LC-MS/MS witli luixing of the organie SPE effluent witli aqueous eluent for optiiuuiu band foeussing on tlie EC eolumn... 

In reeent years, tire use of elevated temperatures has been reeognised as a potential variable in method development. Witlr inereased temperature, aqueous-organie mobile phases separations ean improve, viseosity deereases and diffusion inereases so baek pressures are redueed. At higher temperatures (usually with superheated water > 100 °C under modest pressures) water alone ean be used as the mobile phase and eair provide unique separation opportunities. The absenee of an organie solvent enables the use in HPLC of alternative deteetors sueh as FID or on-line LC-NMR using deuterium oxide as the eluent. 

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