In biosecure composting, animal mortalities are so completely isolated during the degradation process that visual inspection cannot be used to monitor progress or the process status. One novel approach is to monitor the volatile organic compounds (VOCs) released by decaying mortalities and to use them as biomarkers of the process status. A new method was developed to quantitatively analyze potential biomarkers—dimethyl disulfide, dimethyl trisulfide, pyrimidine, acetic acid, propanoic acid, 3-methylbutanoic acid, pentanoic acid, and hexanoic acid—from field-scale biosecure mortality composting units. This method was based on collection of air samples from the inside of biosecure composting units using portable pumps and solid phase microextraction (SPME). Among four SPME fiber coatings, 85 μm CAR/PDMS was shown to extract the greatest amount of target analytes during a 1 h sampling time. The calibration curves had high correlation coefficients, ranging from 96 to 99%. Differences between the theoretical concentrations and those estimated from the calibration curves ranged from 1.47 to 20.96%. Method detection limits of the biomarkers were between 11 pptv and 572 ppbv. The applicability of the prepared calibration curves was tested for air samples drawn from field-scale swine mortality composting test units. Results show that the prepared calibration curves were applicable to the concentration ranges of potential biomaker compounds in a biosecure animal mortality composting unit.