The quantitation of sugars, including glucose, the primary fermentable sugar; maltose (DP2); and maltotriose (DP3), is a standard procedure during the corn-to-ethanol fermentation process. The quantitation of glucose by the Megazyme Assay utilizing glucose oxidase and peroxidase enzymes (GOPOD) and UV-Vis detection, high-performance liquid chromatography with refractive index detection (HPLC-RID), and liquid chromatography mass spectrometry (LC-MS) with electrospray ionization (ESI) and selected ion monitoring (SIM) was studied. Three biological flask fermentation replicates were analyzed every 12 h beginning at 14 h of fermentation (T14) until near completion of fermentation (T62). The method comparison results for glucose quantitation showed that the LC-MS SIM analysis had the lowest limit of quantitation (LOQ) at 2 ppm and the widest dynamic range of 2.7 orders of magnitude. The HPLC-RID analysis had a linear dynamic range (LDR) of 1.5 orders of magnitude with an LOQ of 1500 ppm. The Megazyme GOPOD analysis had an LDR of 0.9 orders of magnitude with an LOQ of 120 ppm. The HPLC-RID method was ideal for glucose quantitation when it was present in high concentrations. In contrast, maltose and maltotriose components were found to be present in lower concentrations, such that simultaneous quantitation of the three analytes is difficult during fermentation. The LC-MS method was the only method able to quantify the concentration of glucose successfully and simultaneously with DP2 and DP3 in all the fermentation broth samples collected from T14 through T62 during the corn-to-ethanol fermentation process.
Background and Objectives Antibiotics are used in the corn‐based ethanol production to control the proliferation of bacteria during fermentation of starch to ethanol. Dried distiller's grains with solubles (DDGS) is a by‐product of corn‐based fermentation that can effectively be used as feed for cattle, swine, chicken, and other livestock. High amounts of antibiotic residues in the feed may lead to antibiotic resistant bacteria. Findings Antibiotics were extracted from DDGS samples by liquid‐liquid extraction, purified by solid phase extraction, and quantified by liquid chromatography mass spectrometry in multiple reaction monitoring mode to quantify five antibiotics: penicillin G, erythromycin, tylosin, virginiamycin M1, and virginiamycin S1. DDGS was collected and analyzed quarterly over the course of ten months to obtain a representation of DDGS produced in various parts of the United States in different seasons. Conclusions Penicillin G was detected in a single sample. Tylosin was detected in 28%, erythromycin in 45%, virginiamycin M1 in 53%, and virginiamycin S1 in 83% of the total samples. Significance and Novelty Largest study to date to quantify antibiotics in DDGS (33 ethanol plants in 14 states).
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