Validation of methodology for quantifying methanol in crude glycerine used in animal nutrition

Author(s)

Carina Nazato

• Document type: Master's Dissertation
• Press: Piracicaba.
• Institution: Universidade de São Paulo (USP). Centro de Energia Nuclear na Agricultura (CENA/STB)
• Defense date: 2015-03-27

Examining board members

Adibe Luiz Abdalla; Ives Cláudio da Silva Bueno; Anderson Schwingel Ribeiro

• Advisor: Adibe Luiz Abdalla

Abstract

Studies with the inclusion of crude glycerin (CG) in animal feed have demonstrated benefits on the energy intake achieved. However, there is a variation between the levels of methanol found in CG, not always analyzed, but often passed on to producers and may pose risks for using it as animal feed. The objective of this study was to optimize and to validate the official methodology \"EN 14110\" for determining methanol in biodiesel for CG matrix; to apply the method for analysis in commercial samples and samples produced in the laboratory; and to evaluate the effect of methanol in ruminant feed through the in vitro gas production technique (PG). It was used a Shimadzu gas chromatograph model 2014 GC equipped with a flame ionization detector (FID). The validation parameters were linearity, limit of detection (LOD) and quantification (LOQ), precision, accuracy and matrix effect. There were analyzed two industrial\'s samples and a sample obtained in the laboratory by transesterification of the oil from Jatropha curcas. The semi-automatic in vitro gas production technique was used to evaluate the use of MeOH inclusion in the ruminant diet via CG replacing corn grain at 0, 25, 50, 75 and 100%, corresponding to MeOH doses of 0, 0.018, 0.035, 0.053 and 0.071 mg respectively. As a positive control treatment was added with the inclusion of 8.3 mg pure MeOH. The variables evaluated were total gas production (GP), degradability of organic matter (DMO), efficiency in methane conversion (CH4 effic) and the partition factor (PF). The bottles containing the diets were incubated at 39 ° C for 24h. The method showed good linearity (r2 >= 0.99), the LOD and LOQ were 0.015 and 0.031%, respectively. There was no significant matrix effect (4.36%). The accuracy of the concentration tested, 0.05% e 0.5% was 2.09% and 2.39%, respectively. The concentrations 0.05%, 0,2% and 0.5% (w/w) showed recovery level of 88%, 95% e 91% respectively. The industry samples concentrations were 0.047 and <LOQ, while the laboratory one showed 7.8% of methanol. The in vitro gas production showed that the addition of CG increased GP, DMO and CH4 effc and reduced PF. It was concluded that the method is suitable for analysis in CG MeOH, with good potential for analysis of the feed array after optimization. CG contaminated with MeOH influence ruminal fermentation, requiring further studies to elucidate the mechanisms involved (AU)

• FAPESP's process: 13/10359-6 - Validation of methodology for methanol quantification in crude glycerin used in animal feed
• Grantee: Carina Nazato
• Support Opportunities: Scholarships in Brazil - Master