In this study, fractionation by sequential anti-solvent precipitation was applied to ionoSolv lignins for the first time. Pretreatment with the aqueous low-cost protic ionic liquid N,Ndimethylbutylammonium hydrogen sulfate ([DMBA][HSO 4 ], 80 wt% in water) was applied to Miscanthus (herbaceous), willow (hardwood) and pine (softwood) to extract lignin.Then, lignin was sequentially precipitated by addition of water as anti-solvent.Fractionation appeared to be controlled by molecular weight of lignin polymers. Fractions isolated with minimal water volumes were shown to have high molecular weight, polydispersity, thermal stability and T g (178 °C). Later precipitates were more monodisperse, had high phenolic and total hydroxyl content, lower thermal stability and T g (136 °C). Addition of 1 g of water per g of dry IL was able to precipitate up to 90 wt% Page 2 of 52 ACS Paragon Plus Environment ACS Sustainable Chemistry & Engineering 3 of lignin. Fractional precipitation represents a novel lignin isolation technique that can be performed as part of the lignin recovery procedure enabling a high degree of control of lignin properties. The effect of the fractionation on lignin structural, chemical and thermal properties was thoroughly examined by 2D HSQC NMR, GPC, TGA and DSC and compared to the unfractionated lignin precipitate obtained by addition of an excess of water.