Adduct formation in LC-MS Analysis (esp. ESI)

Almost everything you analyze by Electrospray ionization mass spectrometry will create an adduct with something in the system. Normally, hydrogen is the most common adduct formed (M+1), but other chemicals, often in trace amounts may form adducts with your sample too. Sometimes we can take advantage of this fact and introduce our own adduct into the system (post column) to increase signal sensitivity or help us isolate one signal from another (the addition of an adduct can sometimes increase the signal seen for one species, but not the other). 

One of my favorite elements to form an adduct with is sodium (Na+). Two common forms are; sodium citrate and sodium acetate. Both have PKA’s between 3 and 6 so a variety of buffered solutions can be prepared for use. However, it is very important that we keep the concentration of sodium as low as possible so as to not clog the mass detector or suppress ionization completely (and see nothing BUT Sodium for weeks …). My suggestion is to initially prepare the buffers such that the solution is less than or equal to 3 mM in concentration. The lowest concentration should be used that yields reproducible results. Ranges from 0.1 mM to 5 mM are common. Only use the highest purity, volatile buffers (some manufacturer’s use names such as “ultra” to describe them) when preparing these ‘doping’ solutions for post-column addition and be sure and filter them through a 0.2 micron filter before use. A syringe pump can be used to deliver the solution during the run. A low flow rate should be used to infuse the adduct solution into the main inlet of the detector. Make sure you have a simple way of controlling the pump through the system (e.g. ‘On’ / ‘Off’, contact closure) so the flow can be turned off when you are not acquiring data. Be sure to not only monitor the baseline, but also measure true peak S/N values of a standard when evaluated the results (decreasing baseline noise may also mean the signal is decreasing too).

Ammonium (NH4) is another popular adduct to add to the system, often in the form of ammonium acetate. It reduces the chances of adding more sodium ions to the system (from glassware). Whichever adduct you use in your system, always start off testing as low a concentration as possible. Monitor the baseline carefully for noise and also to see if the addition of the compound is suppressing or enhancing the signal generated for your compound. Careful use of adducts in your system can provide you with another means to selectively enhance the signal of some compounds without changing the original chromatography method.

I must again emphasize to use the lowest concentration of doping agent. Proper pH control and mode choice are also very important. Use of a syringe pump for infusion, post column can help you to quickly optimize the fragmentor settings in real time.

• For additional information you may want to take a look at this useful Adduct Calculator webpage: "Mass Spectrometry Adduct Calculator(ESI)".