2. Materials and methods2.1. Materi

2. Materials and methods
2.1. Materials and apparatus
Methanol was purchased from Anaoua Chemical Supplies, USA and distilled before use. Lipid was prepared by fungal hydrolysis of bakery waste collected from the local Starbucks Coffee(Pleissner et al., 2014a,b). The obtained waste lipid was heated to 100 _C for 8 h to remove moisture. Candida antarctica lipase-B (Novozyme-435), standard fatty acid methyl esters and CDCl3-TMS were purchased from Sigma–Aldrich, St. Louis, Missouri,USA. Potassium hydroxide and anhydrous sodium sulfate were purchased from ACROS Organics, Geel, Belgium. Ethyl acetate was purchased from HK Labware, Hong Kong. The conversion of
lipid to FAME produced in each reaction was calculated using 1HNMR (Knothe, 2000; Karmee and Chadha, 2005). A Bruker 400 MHz instrument was used for recording of 1H NMR spectra.
The reaction was monitored qualitatively by thin layer chromatography (TLC). The TLC plates were purchased from Macherey–Nagel, Dren, Germany. All reactions were performed in 50 mL round bottomed flasks. Each reaction was carried out in duplicate and the results presented here were averaged.