Structural profiling of wax biopolymer from Pinus roxburghii Sarg. needles using spectroscopic methods

Pinus roxburghii Sarg. is the most abundant species in Himalayan region. The needles of the species largely contribute to the forest biomass and remain the major cause of forest fires leading to climate change, biodiversity loss, etc. Intriguingly, the layer of needles contains wax, a biomacromolecule with potential chemical functionalities for value addition. In the present study, a distinctive approach towards complete structural analysis of the isolated wax in its native state has been done using 1H, 13C, HSQC, HMBC, COSY, TOCSY along with GC-MS of the methyl esters of constituent fatty acids. The wax was isolated in a quantitative yield of 1.64% and analyses suggest that it is a polymer of linearly attached fatty acid esters which on hydrolysis yielded three types of ω-hydroxy fatty acids viz. 12-hydroxydodecanoic acid, 14-hydroxytetradecanoic acid and 16-hydroxyhexadecanoic acid in a ratio of 1:1:2 respectively. Complete assignments for a carbonyl group, α-, β- and other methylenes present in wax were achieved; corroborating the presence of polyester. In particular, identification of wax structure was accomplished through NMR; thereby providing a lead towards future structural analysis of waxes in their native form. The study would also be helpful to generate commercially important compounds derived from pine needle wax. This will offer an opportunity for utilisation of pine needle biomass: a root cause of Himalayan forest fires.