10.1 Introduction For the purposes of this chapter we will define small scale reactions as those involving re action mixture volumes of less than 5ml. When performing organie reactions on this sc ale special problems arise, most notably: 1. Difficulties in measuring out small quantities of sensitive reagents. 2. Significant losses of material due to apparatus design. 3. Difficulties in excluding trace amounts of water from moisture-sensitive reactions. Whenever reactions are performed, material los ses are obtained as a consequence of the above problems. Normally these los ses only account for a few percent of the total material, however this percentage can increase dramatically as the reaction sc ale decreases. For example if a moisture- sensitive reaction was carried out on a one mole scale, it would take 18g of water to completely stop the reaction occurring, however if the same reaction is carried out on a O.1mmol sc ale, then only 1.8mg of water would completely quench the reaction. The problem of weighing out small quantities of sensitive reagents is best solved by accurately weighing larger quantities, and making up solutions in an inert solvent, ideally the reaction solvent. Since the molarity of the solution is known, a quantitative aliquot of this solution can then be added to the reaction mixture using a syringe. This effectively reduces the problems of weighing out the material to those that exist in larger scale reactions, and discussed in earlier chapters. As a general rule many of the techniques used in setting up reactions discussed in the earlier chapters, can, with care, be applied to small sc ale reactions. Indeed there are a wide
M. Casey et al., Advanced Practical Organic Chemistry
variety of miniature chemical apparatus commercially available for just this
purpose. This chapter will outline some of the more specialized techniques that can be employed to alleviate the problems associated with small scale, and that can be carried out without the requirement of relatively expensive specialized glassware.
10.2 Reactions at or below room temperature
When carrying out small-scale reactions at or below room temperature it is quite possible to use a conventional apparatus set-up. Most reactions are carried out in round-bottomed flasks, which are available in sizes down to lrnl. The main problems arise when you come to work up the reaction mixture. If an aqueous or organic extraction is required, then the material must be transferred to a separating funnel. This will inevitably lead to some loss of material during the transfer, but this should be minimal. The main problem arises from the fact that separating funnels rarely come in sizes below lOml. In addition, because of their design there will always be some loss of material in the apparatus itself. This is mainly because the re action mixture necessarily comes into contact with a large surface area of glassware, and retrieving material coated over this large area tends to be difficult. A useful alternative for extractions involving 2ml or less is to use a glass sampIe vial or small test-tube (Fig. 10.1). The reaction mixture can be washed into the sampIe vial, and the extraction solvent added. The lid of the vial is then put on, and the mixture shaken. Removal of the lid allows any pressure build-up to be released. Particular care should be taken when removing the lid to avoid spillage. The required solvent layer is then removed using a Pasteur pipette. Three or four extractions are usually sufficient to recover the majority of the material. This technique is particularly straightforward for diethyl ether extractions of aqueous solutions in which the ether layer is required, since it can readily be pipetted from the top of the aqueous layer (Fig. 10.1). If you require the lower solvent layer, this can be recovered either by pipetting away the top layer, or by pipetting from the bottom of the vial. Because efficient separation of the two phases is required, it is preferable to use a 190 Small Scale Reactions
sampIe vial
/ extraction
- solvent --:::--: ~
reaction •: •: . :. -mixture
Pasteur pipette
:/
- or - ... _
~~~~~~~ - extracts
drying agent ""'" Figure 10.1 tall, thin vial rather than a short, fat one. A second vial containing drying agent can then be used to dry the extracts. Removal of the drying agent is normally achieved by filtration of the solvent mixture, and on a small scale this is best achieved using a Pasteur pipette fitted with a cotton wool plug as the filtration apparatus (Fig. 10.2). Once the solvent has been transferred into the filtration pipette, it can be forced through the plug by applying pressure with a pipette teat. Evaporation of the solvent in the normal way then yields the emde reaction product. As stated earlier, material is inevitably lost with each transfer of apparatus. 1t is possible to cut down the number of transfers by using the sample vial as the reaction vessel. Very small magnetic fleas are now commonly available, and will fit most small sample vials. Consequently, with magnetic stirring, the vials can serve as small reaction vessels (Fig. 10.3). They are conveniently attached to the top of a magnetic stirrer machine using plasticine. Small Scale Reactions 191
/ tcat- --- Pasteur -:- pipette
~ o o - cotton wool - plug - riltered Solulion
/ Figure 10.2 When capped, the vial is a sealed system, consequently this set-up is only useful for small scale reactions at room temperature that do not involve an increase or decrease in press ure inside the reaction vessel. For reactions at low temperature, or those requiring a positive pressure of an inert gas atmosphere, it is often more convenient to use a Pyrex test tube fitted with a septum (Fig. 10.3). In all other respects the arrangement is the same and, since Pyrex test tubes are available in a range of sizes, this apparatus can inert gas I eap j !- septum ::;
-.;
sampie via1--
- test tube reaclion _____ mi)tlure~
=. __ magnelic ficas - = Figure 10.3 Use of sampie via! or test tube as areaction vessel 192 Small Scale Reactions
cope with a range of reaction volumes down to about O.2ml. Again the reaction vessel can also be used for a subsequent extraction procedure simply by replacing the septum with a bung.
10.3 Reactions above room temperature
Carrying out small scale reactions above room temperature, particularly those involving solvent at reflux, is very difficult. The problems are associated with preventing the evaporation of very sm all amounts of solvent, and materiallosses in the apparatus. This is usually caused by losses of material through ground-glass joint attachments of condensers to the reaction fiask, and by inefficient condensation of the solvent vapour. One solution is to use a sealed tube (see Chapter 8) as the reaction vessel, and this is probably the equipment of choice for reactions involving volatile solvents (:::; 50°C) on scales below Iml. For higher boiling solvents it is possible to use a one-piece apparatus, incorporating an air condenser system (Fig. 10.4). This apparatus can be conveniently constructed at the required size from a piece of Pyrex tubing. The air condenser is adequate enough to prevent the evaporation of higher boiling solvents (~100°C). Alternatively,
, a one-piece apparatus incorporating a water condenser can also be used inert gas , inert gas
_ - - septum _ __ : 11 i
air condenser _ water condenser
reaction mixt ure
~/~~ Figure 10.4 Small scale air condenser and water condenser systems. Small Scale Reactions 193
(Fig. 10.4). In this case the condenser system is more efficient, however construction of the apparatus is correspondingly more complex. In both cases the reflux apparatus can be easily constructed to allow re action volumes down to about O.5m1.
10.4 Reactions in nmr tubes
In many cases it is necessary to monitor c10sely the progress of a sma11 scale re action by methods other than tlc; one very useful alternative is nuc1ear magnetic resonance (nmr). With larger scale reactions this is simply done by removing an aliquot of the reaction mixture and recording its nmr spectrum. Obviously this approach cannot be applied to reactions involving relatively small quantities of material. The answer is to carry out the reaction in an nmr tube. Both 5mm and lOmm diameter nmr tubes can conveniently be used as reaction vessels (Fig. 10.5) in the same way as test- tubes were employed in Section 10.2. inert gas J
septum_ iii
nmrtube- inert gas supply
must be removed I prior to running the nmr spectrum
reaction mix.ture~ -.
Figure 10.5 The reaction can then be monitored by recording the spectrum of the reaction mixture direct1y. There are however several important points to note when using nmr tubes as reaction vessels: 194 Small Scale Reactions
1. The reaction solvent should be chosen carefully to ensure that it does
not obscure the nmr region to be observed. 2. Magnetic stirring cannot be used because the magnetic flea in the nmr tube would interact with the nmr field causing severe broadening of the spectrum. Similarly the presence of paramagnetic material in the reaction mixture will lead to broadening of the spectrum. 3. For high-quality nmr spectra to be recorded, the reaction mixture should be homogeneous. The presence of solids in the reaction mixture will lead to poorly resolved spectra. As a general rule you should use the reaction solvent that is normally employed for the type of reaction being carried out, however its deuterated equivalent must be used if IH nmr spectra are to be observed. Agitation of the reaction mixture is probably best achieved using sonication in an ultrasonic bath (Chapter 8), although periodie shaking will often suffice. If you require to carry out the reaction at elevated temperatures then it is usually advisable to use a sealed nmr tube. Thick-walled nmr tubes are commercially available if the reaction mixture is required to withstand increased reaction pressures typical of those obtained in sealed tube experiments.
10.5 Purification of materials
The purification of small quantities of materials (~ 50mg) also poses certain problems. A number of simple techniques used are outlined below.
105.1 Distillation By far the most convenient method of carrying out distillation on a small sc ale is to use a Kugelrohr apparatus (see Chapter 9). In order to cut down on los ses through ground glass joint connections it is often necessary to use a one piece Kugelrohr bulb set (Fig. 10.6). This can be conveniently made to the size required from a piece of Pyrex tubing. After distillation is complete the apparatus is left to cool, and the purified material recovered by cutting up the apparatus into three seetions using a glass knife. Small Scale Reactions 195
G)()(j~ ./' erude sampIe 1
c ~ ~ ~
Figure 10.6
10.5.2 Crystallization Crystallizations on a small scale are most conveniently carried out using a Craig tube apparatus as described in Chapter 9.
10.5.3 Chromatography All the normal chromatography techniques (see Chapter 9) can be used to purify small quantities of material. Indeed preparative hplc and glc are often more successful when small quantities of material are involved. In the case of flash chromatography however, it is often impractical to simply scale down the equipment. A useful alternative is to employ a Pasteur pipette as the column. Such a column is easily constructed using a pipette containing a cotton wool plug (Fig. 1O.7a). The pipette is then filled with the required adsorbent (typically silica gel). The amount of adsorbent used depends upon the quantity of cmde sampie to be purified, however it is inadvisable to fill the pipette more than three-quarters full, otherwise there is insufficient room for the solvent. Next, the eluting solvent is added to the top of the column and allowed to run through the column under gravity. More solvent is added to the top of the column as required. Once the solvent starts to appear at the bottom of the column, pressure can be applied 196 Small Scale Reactions
using a pipette teat, forcing the solvent through at a faster rate. After about
lcal -
PaSleur pipette_
- solvenl
- ad,orbent -adsorbent
cotton wool plug_
(a) (b)
Figure 10.7 two column-volumes of solvent have been passed through the silica it should be ready for use. The sampie is applied to the top of the column in the usual way and pressure is applied using a pipette te at. The main difference between this arrangement and the more usual flash chromatography set-up is that the pressure applied to the top of the colurnn is not constant. The teat is constantly being removed to a110w the addition of more solvent to the top of the column. Consequently the solvent is not passing through the column at a constant rate. In most instances this does not appear to significantly affect the separations achieved. If, however, this proves to be a problem, a miniature chromatography column with solvent reservoir can easily be constructed from Pyrex glass tubing (Fig. 1O.7b). With a cotton wool plug in the bottom this can be used in exactly the same way as the larger columns described in Chapter 9.
