High throughput mass spectrometry assay for early chick gender determination: less than 3s total analysis time per sample.

Chicken eggs are one of the most consumed foods around the world. However, male chickens from egg-producing species are considered unprofitable as they produce too little meat and no eggs. For this reason, day-old male chicks are discarded, lead-ing to animal welfare concerns. As an alternative for chick culling, we have recently reported 3-[(2-aminoethyl)sulfanyl]butanoic acid (ASBA) as a biomarker for early in ovo gender determination. In the present work, we describe the optimization of acoustic droplet ejection-mass spectrometry (ADE-MS) conditions and automated sample prepa-ration compatibility for the high throughput quantification of ASBA from allantoic fluid. Special attention is given to the optimization of ADE-MS compatible liquid handling and the development of the data processing to ensure a reliable gender prediction. We have been able to accurately determine the gender of day-9 eggs with a prediction accuracy of 96%, with a throughput of 1800 samples per hour.