Abstract. Nitrogen oxides, emitted from air traffic, are of concern due to their impact on climate by changing atmospheric ozone and methane levels. Using the DLR research aircraft Falcon, total reactive nitrogen (NOy) in-flight measurements were carried out at high altitudes to characterize emissions in the fresh aircraft exhaust from the latest-generation Rolls-Royce Trent XWB-84 engine aboard the long-range Airbus A350-941 aircraft during the ECLIF3 (Emission and CLimate Impact of alternative Fuels 3) experiment. The impact of different engine thrust settings, monitored in terms of combustor inlet temperature, pressure and engine fuel flow, was tested for two different fuel types: Jet A-1 and, for the first time, a 100 % sustainable aviation fuel (SAF) under similar atmospheric conditions. In addition, a range of combustor temperatures and an additional blended SAF were tested during ground-based emission measurements. For the data measured during ECLIF3, we confirm that the NOx emission index increases with increasing combustion temperature, pressure and fuel flow. We find that as expected, the fuel type has no measurable effect on the NOx emission index. These measurements are used to compare to cruise NOx emission index estimates from three engine emission prediction methods. Our measurements thus help to understand the ground to cruise correlation of current engine emission prediction methods while serving as input for climate modelling and extending the extremely sparse data set on in-flight aircraft nitrogen oxide emissions to newer engine generations.