Micro apparatus

Although apparatus employing ground-glass joints is excellent for work on a macro scale, it is not always suitable for very small-scale work as the joints are often disproportionately large compared with the rest of the assembly. The semi-micro apparatus described on pp. 59-72 can therefore be considered as being of general utility for this scale of work, especially as the use of corks has been reduced to a minimum. 

Alternatively the semi micro apparatus shown in Fig. 38 (p. 63) may be used. Heat the anilide and sulphuric acid under reflux for 15 minutes in such a manner that the vapour does not rise higher than half-way up the vertical condenser through which water is passed. Then dilute the solution in the flask with 5 ml. of water. Empty the vertical condenser and run cold water through the inclined condenser. Now increase the rate of heating and distil off 1-2 ml. of aqueous acetic acid. 

Rhoeadine crystallises from a mixture of chloroform and ether, sublimes at 215-25" /0-02 mm., and then melts at 256-57-5° (vac.), and has [a] / ° -f 232° (CHClj). According to Awe it has m.p. 272-4° (Kofler s micro-apparatus), and [a]] ° -f 243° (CHClj). A crystalline liydriodide, B. HI. 2H2O, was prepared by the addition of potassium iodide to a solution of the alkaloid in acetic acid. Rhoeadine contains one methoxyl group and a dioxymethylene group, but no reactive hydrogen (Zerewitinoff). 

Figure 8.3 Apparatus used for sample preparation involving distillation or sublimation. A, distillation apparatus for organic volatiles in water B, micro apparatus for continuous steam distillation with continuous llquid-llguid extraction of the distillate C, micro sublimation appeuratus.

The following micro apparatus will be found useful ... 

When an apparatus of this type is used, it is important to be sure that there are no leaks in the system which could cause a false value of hydrogen uptake. It is well to maintain the system under a small positive pressure without shaking for about fifteen minutes and to note if there has been a change in pressure as with the micro apparatus. This becomes particularly important if the compound is not easily reduced. 

So, since nobody actually has any microscale reduced-pressure distillations, and with certain sizes of equipment the O-rings can squeeze out from under the plastic caps, and since I don t use them for reduced-pressure (vacuum) distillation anyway (I switch to a semi-micro apparatus with real 14/20 joints and—gasp —grease), I don t know how vacuum tight these seals really are. 

Attention must also be paid to the distance between the condenser and the evaporator when subliming substances that have low vapor pressures. Apparatus that has a variable condenser position is thus extremely useful because this position can then be varied to suit the substance and the pressure to be used. The distance from the substance to the condenser is usually 7-25 mm in micro apparatus. 

The following micro apparatus will be found useful micro porcelain, silica, and platinum crucibles of 0.5 to 2 ml capacity micro beakers (5 ml) and micro conical flasks (5 ml) micro centrifuge tubes (0.5, 1.0 and 2.0 ml) micro test-tubes (40-50 x 8 mm) micro volumetric flasks (1, 2, and 5 ml)... 

A rotating enzyme-immobilized reactor and a flat pH electrode were incorporated into a sealed cell for use under continuous-flow/stopped-flow (SF) operation for the rapid determination of penicillins G and V in tablets and injectables [50]. A co-immobilization in a rotating bioreactor and amperometric detector resulted in a sensitive system for determination of succinylcholine and acetylcholine in pharmaceutical preparations [51]. A tandem system incorporating two rotating bioreactors into a continuous-flow/SF sample/reagentprocessing setup was apphed for the determination of alkaline phosphatase activity in serum samples [52]. By functional combination of the SF and flow-injection analysis (FIA), an automated micro apparatus was constructed resulting in significant reduction of the injection volumes of enzyme and substrate [53]. SF/continuous flow methods were apphed to acquire kinetic information also [54, 55]. 

The difficulties of working with small quantities of liquids are much greater than with small quantities of solids. For example a competent worker can, and does in fact, often work with 100 mg, of solid without any special apparatus. With liquids this is often not practicable because of the much greater losses entailed, particularly when it is realised that one ordinary-sized drop weighs about 50-100 mg. The account which follows gives details of modifications of standard apparatus suitable for the semi-micro scale defined above. 

Hold the tube horizontally and quickly seal this end in a micro-burner. Attach the tube (with the open end upwards) to a thermometer in the melting-point apparatus (Fig. i(c), p. 3) so that the trapped bubble of air in the capillary tube is below the surface of the bath-liquid. Now heat the bath, and take as the b.p. of the liquid that temperature at which the upper level of the bubble reaches the level of the surface of the batn liquid. 

Distillation under reduced pressure. The student should first read details of this operation on a macro-scale (p. 28). For micro-scale work the apparatus shown in Fig. 40 is very convenient. A small pear-... 

A very suitable apparatus for semi-micro steam-distillation, particularly for suspensions that are likely to bump badly, is showm in Fig. 44. This consists of a 50 ml. Kjeldahl flask, clamped at an angle of 45°, and fitted with a long glass tube for the inlet of steam. The Outlet-tube is bent twice, first at 135° and then at 45° as shown, and fitted into a small water-condenser. 

Preparations, By employing suitable apparatus as described above, the student is enabled to carry out the following semi-micro preparations. 

Dissolve 1 g. of anthracene in 10 ml. of glacial acetic acid and place in 50 ml. bolt head flask fitted with a reflux water-condenser. Dissolve 2 g. of chromium trioxide in 2 ml. of water and add 5 ml. of glacial acetic acid. Pour this solution down the condenser, shake the contents of the flask and boil gently for 10 minutes. Cool and pour the contents of the flask into about 20 ml. of cold water. Filter off the crude anthraquinone at the pump, wash with water, drain well and dry. Yield, 1 g. Purify by re crystallisation from glacial acetic acid or by sublimation using the semi-micro sublimation apparatus (Fig. 35, p. 62, or Fig. 50, p. 70). 

The fiimace E is now switched on, and takes about 20 minutes before the combustion tube packing has reached the required temperature. After 15 minutes, carbon dioxide is again passed for 5 minutes, and the apparatus tested as before for the appearance of micro-bubbles. If these are not at first obtained, the sweeping-out process must be continued until they do appear. Now that the combustion-tube packing is heated up to the required temperature and the apparatus filled with carbon dioxide, all air being swept out, the combustion may be started. 

Two simple methods for the semi-micro estimation of halogens are available, (a) the Carius method, and (b) the Parr bomb method. It should be emphasised that there are other methods available for these estimations on the true micro scale, but they do not lend themselves, by virtue of the balances, apparatus and manipulation required, to semi-micro work, or to the intermittent usage which class-work necessarily entails. 

First, considerably greater emphasis has been placed on semimicro techniques and their application to preparations, separations, analysis and physical determinations such as those of molecular weight. We have therefore greatly expanded the section on Manipulation on a semi-micro scale which was in the Third Edition, and we have described many more preparations on this scale, some independent and others as alternatives to the larger-scale preparations which immediately precede them. Some 40 separate preparations on the semi-micro scale are described in detail, in addition to specific directions for the preparation of many classes of crystalline derivatives required for identification purposes. The equipment required for these small-scale reactions has been selected on a realistic basis, and care has been taken not to include the very curious pieces of apparatus sometimes suggested as necessary for working on the semi-micro scale. 

Fig. 11,10, 3, depicts a housing for the apparatus of Fig. 11,10, 2, a it is easily constructed from lengths of angle iron and asbestos board. If desired, torch bulbs may be fixed in the appropriate holders in the walls, and connected with a dry battery or accumulator in order to provide illumination of the melting point apparatus. The electric bulb immediately behind the apparatus should be connected to an inde pendent micro-switch. Alternatively,... 

For heating baths, see Table 4. For distillation apparatus on a micro or semi-micro scale see Aldrich and other glassware catalogues. Alternatively, some useful websites for suppliers of laboratory glassware are www.wheatonsci.com,www.sigmaaldrich.com and www.kimble-kontes.com. 

The basic apparatus for polarographic analysis is depicted in Fig. 16.1. The dropping mercury electrode is here shown as the cathode (its most common function) it is sometimes referred to as the working or micro-electrode. The... 

A simple apparatus suitable for this procedure is shown in Fig. 16.17. B is a 3 volt torch battery or 2 volt accumulator, M is a micro-ammeter, R is a 500 ohm, 0.5 watt potentiometer, and E,E are platinum electrodes. The potentiometer is set so that there is a potential drop of about 80-100 millivolts across the electrodes. 

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