Preparative pumps

Supervisor had requested him to prepare pump A for restart. Operator was unaware pump A had been worked on during day shift. 

Supervisor had requested pump A to be prepared for standby. Did this by asking plant operator to prepare pump and electrician to reset pump. This request was made at about 19 00. 

A modern solvent delivery system consists of one or more pumps, solvent reservoirs, and a degassing system. HPLC pumps can be categorized in several ways by flow range, driving mechanism, or blending method. A typical analytical pump has a flow range of 0.001-10 mL/min, which handles comfortably the flow rates required for most analytical work (e.g., 0.5-3 mL/min). Preparative pumps can have a flow range from 30 mL/min up to L/m. 

The outlook is good for applications of these picosecond methods to an increasing number of studies on reactive intermediates because of the limitations imposed by the time resolution of nanosecond methods and the generally greater challenges of the use of a femtosecond spectrometer. The pump-probe technique will be augmented in more widespread applications of the preparation-pump-probe method that permits the photophysics and photochemistry of reactive intermediates to be studied. 

Sample preparation Pump 125 mL of a solution of interferon in ethylene glycol 50 mM sodium phosphate buffer containing 1 M NaCl 50 50 onto the column (which was previously equilibrated with 1 M NaCl in ethylene glycol water 50 50) then start the gradient. 

Dual pumps deliver solvent at a constant flow rate to the sample and reference sides of the instrument. The standard pump heads deliver solvent at a rate of 0.1-9.9cm min" irrespective of system pressure, but work best at 0.1-3.0cm min" However, preparative pump heads are available which increase this range to 0.2-19.8 cm min A pre-pump ensures the consistency of the flow rate by supplying a head of solvent to the system. 

These limitations have recently been eliminated using solid-state sources of femtosecond pulses. Most of the femtosecond dye laser teclmology that was in wide use in the late 1980s [11] has been rendered obsolete by tliree teclmical developments the self-mode-locked Ti-sapphire oscillator [23, 24, 25, 26 and 27], the chirped-pulse, solid-state amplifier (CPA) [28, 29, 30 and 31], and the non-collinearly pumped optical parametric amplifier (OPA) [32, 33 and 34]- Moreover, although a number of investigators still construct home-built systems with narrowly chosen capabilities, it is now possible to obtain versatile, nearly state-of-the-art apparatus of the type described below Ifom commercial sources. Just as home-built NMR spectrometers capable of multidimensional or solid-state spectroscopies were still being home built in the late 1970s and now are almost exclusively based on commercially prepared apparatus, it is reasonable to expect that ultrafast spectroscopy in the next decade will be conducted almost exclusively with apparatus ifom conmiercial sources based around entirely solid-state systems. 

The most connnon commercially prepared amplifier systems are pumped by frequency-doubled Nd-YAG or Nd-YLF lasers at a 1-5 kHz repetition rate a continuously pumped amplifier that operates typically in the 250 kHz regime has been described and implemented connnercially [40]. The average power of all of the connnonly used types of Ti-sapphire amplifier systems approaches 1 W, so the energy per pulse required for an experiment effectively detennines the repetition rate. 

An interferometric method was first used by Porter and Topp [1, 92] to perfonn a time-resolved absorption experiment with a -switched ruby laser in the 1960s. The nonlinear crystal in the autocorrelation apparatus shown in figure B2.T2 is replaced by an absorbing sample, and then tlie transmission of the variably delayed pulse of light is measured as a fiinction of the delay This approach is known today as a pump-probe experiment the first pulse to arrive at the sample transfers (pumps) molecules to an excited energy level and the delayed pulse probes the population (and, possibly, the coherence) so prepared as a fiinction of time. 

The methods diseussed so far, fluoreseenee upeonversion, the various pump-probe speetroseopies, and the polarized variations for the measurement of anisotropy, are essentially eonventional speetroseopies adapted to the femtoseeond regime. At the simplest level of interpretation, the infonnation eontent of these eonventional time-resolved methods pertains to populations in resonantly prepared or probed states. As applied to ehemieal kineties, for most slow reaetions (on the ten pieoseeond and longer time seales), populations adequately speeify the position of the reaetion eoordinate intemiediates and produets show up as time-delayed speetral entities, and assignment of the transient speetra to ehemieal stnietures follows, in most oases, the same prinoiples used in speotrosoopio experiments perfomied with oontinuous wave or nanoseoond pulsed lasers. 

Place 0 5 ml. of acetone, 20 ml. of 10% aqueous potassium iodide solution and 8 ml. of 10% aqueous sodium hydroxide solution in a 50 ml. conical flask, and then add 20 ml. of a freshly prepared molar solution of sodium hypochlorite. Well mix the contents of the flask, when the yellow iodoform will begin to separate almost immediately allow the mixture to stand at room temperature for 10 minutes, and then filter at the pump, wash with cold w ater, and drain thoroughly. Yield of Crude material, 1 4 g. Recrystallise the crude iodoform from methylated spirit. For this purpose, place the crude material in a 50 ml. round-bottomed flask fitted with a reflux water-condenser, add a small quantity of methylated spirit, and heat to boiling on a water-bath then add more methylated spirit cautiously down the condenser until all the iodoform has dissolved. Filter the hot solution through a fluted filter-paper directly into a small beaker or conical flask, and then cool in ice-water. The iodoform rapidly crystallises. Filter at the pump, drain thoroughly and dry. 

When the ij hours boiling is complete, preheat a Buchner funnel and flask by pouring some boiling water through the funnel with the filter-paper already in position, and then quickly filter the boiling solution. Transfer the filtrate to a beaker to cool, and then wash the insoluble residue of diphenylurea on the filter twice with hot water, and drain thoroughly. Cool the filtrate in ice-water the monophenylurea separates as colourless needles. Filter at the pump and drain well. Recrystallise the crude product from boiling water, as in the previous preparation. Yield of monophenylurea, 2 5-3 g. m.p. 147°. 

Carry out this preparation precisely as described for the a-compound, but instead of zinc chloride add 2 5 g. of anhydrous powdered sodium acetate (preparation, p. 116) to the acetic anhydride. When this mixture has been heated on the water-bath for 5 minutes, and the greater part of the acetate has dissolved, add the 5 g. of powdered glucose. After heating for I hour, pour into cold water as before. The viscous oil crystallises more readily than that obtained in the preparation of the a-compound. Filter the solid material at the pump, breaking up any lumps as before, wash thoroughly with water and drain. (Yield of crude product, io o-io 5 g.). Recrystallise from rectified spirit until the pure -pentacetylglucose is obtained as colourless crystals, m.p- 130-131° again two recrystallisations are usually sufficient for this purpose. 

Prepare two solutions, one containing i g. of diphenylamine in 8 ml. of warm ethanol, and the other containing 0-5 g. of sodium nitrite in i ml. of water, and cool each solution in ice-water until the temperature falls to 5°. Now add o 8 ml. of concentrated hydrochloric acid steadily with stirring to the diphenylamine solution, and then without delay (otherwise diphenylamine hydrochloride may crystallise out) pour the sodium nitrite solution rapidly into the weil-stirred mixture. The temperature rises at once and the diphenylnitrosoamine rapidly crystallises out. Allow the mixture to stand in the ice-water tor 15 minutes, and then filter off the crystals at the pump, drain thoroughly, wash with water to remove sodium chloride, and then drain again. Recrystallise from methylated spirit. Diphenylnitrosoamine is thus obtained as very pale yellow crystals, m.p. 67 68° yield, 0 9-1 o g. 

The recrystallisation of diazoaminobenzene has to be performed with care, as the substance is freely soluble in most liquids and tends moreover to decompose if its solution is not rapidly cooled. Place 2 g. of the crude, freshly prepared, well-drained material in a boiling-tube, add about 15-20 ml. of ethanol and 1-2 drops of 10% aqueous sodium hydroxide solution, and then heat rapidly until boiling if the solution should contain insoluble impurities, filter through a small fluted paper, and at once cool the filtrate in ice-water. The diazoaminobenzene should rapidly crystallise out from the cold and stirred solution filter the crystals rapidly at the pump whilst the solution is still cold, as... 

Prepare a mixture of 30 ml, of aniline, 8 g. of o-chloro-benzoic acid, 8 g. of anhydrous potassium carbonate and 0 4 g. of copper oxide in a 500 ml. round-bottomed flask fitted with an air-condenser, and then boil the mixture under reflux for 1 5 hours the mixture tends to foam during the earlier part of the heating owing to the evolution of carbon dioxide, and hence the large flask is used. When the heating has been completed, fit the flask with a steam-distillation head, and stcam-distil the crude product until all the excess of aniline has been removed. The residual solution now contains the potassium. V-phenylanthrani-late add ca. 2 g. of animal charcoal to this solution, boil for about 5 minutes, and filter hot. Add dilute hydrochloric acid (1 1 by volume) to the filtrate until no further precipitation occurs, and then cool in ice-water with stirring. Filter otT the. V-phcnylanthranilic acid at the pump, wash with water, drain and dry. Yield, 9-9 5 g. I he acid may be recrystallised from aqueous ethanol, or methylated spirit, with addition of charcoal if necessary, and is obtained as colourless crystals, m.p. 185-186°. 

Filter at the pump, and wash well with water. (Yield, about 20 g.) Recrystallise a portion from hot methylated spirit, reserving the remainder for the following preparation. Benzoin is a very pale yellow (almost colourless) crystalline substance, m.p. 137°. 

For this preparation, the cinnamaldehyde must first be purified by careful redistillation at the water-pump, and a fraction of steady b.p. (e.g., i26°/i5 mm.) collected. 

Dichloramine-T. Dilute 80 ml, of freshly prepared 2N sodium hypochlorite soluticMi (preparation, p. 525) with 80 ml. of w ter, and then add with stirring 5 g. of finely powdered toluene-p-sulphonamide, a clear solution being rapidly obtained. Cool in ice-water, and then add about 50 ml. of a mixture of equal volumes of glacial acetic acid and water slowly with stirring until precipitation is complete the dichloro-amide separates at first as a fine emulsion, which rapidly forms brittle colourless crystals. Filter off the latter at the pump, wash well with... 

Mix 6 2 ml. (6 4 g.) of pure ethyl acetoacetate and 5 ml. of pure phenylhydrazine in an evaporating-basin of about 75 ml. capacity, add 0 5 ml. of acetic acid and then heat the mixture on a briskly boiling water-bath (preferably in a fume-cupboard) for I hour, occasionally stirring the mixture with a short glass rod. Then allow the heavy yellow syrup to cool somewhat, add 30-40 ml. of ether, and stir the mixture vigorously the syrup may now dissolve and the solution shortly afterwards deposit the crystalline pyrazolone, or at lower temperatures the syrup may solidify directly. Note. If the laboratory has been inoculated by previous preparations, the syrup may solidify whilst still on the water-bath in this case the solid product when cold must be chipped out of the basin, and ground in a mortar with the ether.) Now filter the product at the pump, and wash the solid material thoroughly with ether. Recrystallise the product from a small quantity of a mixture of equal volumes of water and ethanol. The methyl-phenyl-pyrazolone is obtained... 

Preparation of REAOENTS.t It is essential for this preparation that the zinc powder should be in an active condition. For this purpose, it is usually sufficient if a sample of ordinary technical zinc powder is vigorously shaken in a flask with pure ether, and then filtered off at the pump, washed once with ether, quickly drained and without delay transferred to a vacuum desiccator. If, however, an impure sample of zinc dust fails to respond to this treatment, it should be vigorously stirred in a beaker with 5% aqueous sodium hydroxide solution until an effervescence of hydrogen occurs, and then filtered at the pump, washed thoroughly with distilled water, and then rapidly with ethanol and ether, and dried as before in a vacuum desiccator. The ethyl bromoacetate (b.p. 159 ) and the benzaldehyde (b.p. 179 ) should be dried and distilled before use. 

Prepare a mixture of 4 g. of. V phenylanthranilic acid and 5 ml. of ethanol, and boil the solution under reflux for 20 minutes. Cool the mixture, when the 2,3-diphenylquinoxaline will rapidly crystallise. Filter off the quinoxaline at the pump, and recrystallise it from ethanol. It forms colourless crystals, m.p. 125° Yield, 1 0 g. 

Meanwhile set up the ether distillation apparatus as used in the preparation of triethyl phosphite (p. 308). Distil off the ether and then fractionally distil the residue at water-pump pressure. The di-isopropyl hydrogen phosphite distils at 79Vi4 mm. other b.ps. are 8o°/i5 mm., 82-5°/i7 mm. Yield, 25 g., 89%. 

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