Drug testing plays a pivotal role in workplace safety and compliance particularly in safety-sensitive roles where direct observation protocols are sometimes required. Current DOT guidelines allow employees to self-identify their gender to determine who observes the collection, providing flexibility for transgender and nonbinary individuals. However, the recent executive order 14168 aiming to define gender strictly as male or female based on biological sex could upend this approach. If enforced, the DOT may be required to revise its current practices—leaving employers uncertain about how to navigate compliance requirements while continuing to uphold respectful, inclusive workplace policies.