In 2020, CARB adopted the low NOX omnibus ruling, which provided revisions to on-road heavy duty engine compliance standards and certification practices. As part of the updates to the regulation, CARB has introduced a new in-use vehicle testing process that broadens the operation modes tested and considers the manufacturer’s intended vehicle application. Compared to the previous method, or the Not-to-Exceed approach, cold start and low ambient temperature provisions were included as part of the updates. The inclusion of low temperature operation requires the OEMs to design a robust engine and aftertreatment package that extends NOX conversion performance. The following work discusses the NOX emissions performance impact in a low temperature ambient environment. The engine and aftertreatment system evaluated was designed to comply with CARB’s low NOX regulations. The cycles tested included the CARB Southern NTE cycle and an FTP-LLC protocol. Both test sequences were intended to replicate drive cycles observed in the field. Overall, results indicated higher emissions for the low ambient temperature conditions. Utilizing the 3-bin moving average window method, emissions results were calculated for idle, low load, and medium / high load bins. While the CARB Southern NTE cycle showed an increase in the idle and low load bins, the system was able to maintain compliance. The medium / high load bin, however, exceeded the compliance limit by ~40% due to changes in the exhaust conditions. For the FTP-LLC protocol, LLC segments also increased, but maintained compliance based on the 2031+ in-use NOX standards. Furthermore, low ambient temperature operation creates challenges for controlling emissions even with a low NOX system.