When replicating these methods, special attention should be paid to the system pressure profiles produced with a new column. In our experience, despite the emphasis placed by column manufacturers on consistency of performance, different column and packing batches can result in columns that perform considerably better (albeit it with higher system pressure) or worse for profiling applications. In some cases, the pressure profiles for a column can be prohibitive for large project use. We routinely evaluate column pressure profiles and select columns with average (for their type) performance, ensuring that similar performing columns can be regularly procured. Representative pressure traces are provided with the experimental protocols. Furthermore, if replicating these methods on different instrumentation, attention should be paid to the extra-column volume of the LC-MS system and consequential holdup time (see quality control section Table ), as this will affect the accuracy of retention times, with a knock-on effect to method performance and the accuracy of method-specific analyte databases.