Rapid quantitative and qualitative analysis of anthocyanins using UHPLC-3Q-MS/MS

A new method for quantifying anthocyanins using ultra-high performance liquid chromatography-triple quadrupole mass spectrometry (UHPLC-3Q-MS/MS) was developed. The method utilizes controlled and optimized in-source fragmentation, which enables group-specific detection and subsequent quantification of all anthocyanins derived from a single anthocyanidin using common MRM transitions (Figure 1). The method works for a wide variety of different sample types, including crude plant extracts, simplifying and speeding up the analysis of anthocyanins.

Fig. 1: Anthocyanidins included in the method (A) and example MRM chromatograms combined from two berry extracts (B).

Typical analysis methods for anthocyanins include methods based on (U)HPLC-UV/Vis and conventional compound-specific tandem mass spectrometry. For the UV/Vis-based methods, separation of all compounds is naturally needed, and the compound-specific MS/MS methods require several SRM or MRM transitions to comprehensively quantify all anthocyanins derived from one anthocyanidin aglycone. As described, the developed method bypasses the limitations of the latter by using in-source fragmentation, and different cone voltages are used in the method to ensure the efficient cleavage of different types of glycones.

The method complements similar earlier analysis methods published for several other groups of polyphenolic compounds [1,2]. These previous methods can be combined with our new methods allowing the quantification of all major polyphenolic compound groups in one UHPLC-3Q-MS/MS injection.

[1] Engström MT, Pälijärvi M, Fryganas C, Grabber JH, Mueller-Harvey I, Salminen JP. J Agric Food Chem 2014; 62: 3390 – 3399

[2] Engström MT, Pälijärvi M, Salminen JP. J Agric Food Chem 2015; 63: 4068 – 4079