Is it just me who thinks that biology researchers can be a bit sloppy when it comes to making and dispensing reagents accurately? Analytical chemists would laugh at our faith measuring cylinders for anything other than fairly rough work like making up media.
When it comes to more accurate work, like making up assay standards, measuring cylinders just just don’t cut it anymore. Here are a few ways to ensure that your stock solutions/standards are as accurate as possible, allowing you to get your experiments off to the best possible start.
1. Make up solutions and standards using volumetric flasks.
For accurate and reproducible stock solutions your tool of choice should be a volumetric flask. Volumetric flasks are far more accurate than measuring cylinders and pipettes, especially if you use class A flasks, which are manufactured to extremely stringent standards. Volumetric flasks are available in various sizes from 1 mL upwards and you can find a guide to using them here.
2. Use the correct balance, correctly.
Obviously there’s no point in using a highly accurate volumetric flask to measure the solvent volume, only to weigh out the solid using a balance that is too coarse for your needs. Make sure you use a balance that is made for weighing a mass in the range you require, and that it is calibrated and on a level surface.
For small masses, it is often quite difficult to weigh out exactly the amount you require. But normally , solutions don’t have to be made up to an exact concentration – you just need to know their concentration (whatever it is) exactly.
A good approach is to weigh out the reagent as close to the target weight as possible then, note down the weighed mass and make up the solution in the volumetric flask. The actual concentration of the solution you made up can then be calculated exactly..
3. Make reagents in large batches where possible
Another way to ensure that stock solutions are reproducible from experiment to experiment is to make them up in large batches where possible. This not only means that you are working with the same solution in each experiment, but also that you are using a larger volume when making up the solution, which should help with the accuracy.
4. Take into account the strength of reagents you are using.
When making up stock solutions, the strength of the reagent is often overlooked. If the reagent is less than 100% strength i.e. less than 100% of the mass is the actual reagent (the remainder being impurities), it will normally be stated by the manufacturer on the container and you should take this into account when calculating how much of it to use.
Say, for example, you need to weigh out 20g of a reagent to make a 1M solution, but the reagent is only manufactured to 97% percent strength. Only 97% (19.4g) of that 20g will be your reagent so you actually have to add 20.61g ([100/97]*20) of the solid to give you 20g of reagent… and 0.61g of the impurit(y/ies).
Do you agree that many biology researchers could/should do things more accuracy? Shout out your opinion in the comments.
Photo: Dublin Ninja