Background: Mapping can be performed to predict utility values from condition-specific measures when preference-based measures are absent. A previously developed algorithm that predicts EQ-5D-3L index values from the Multiple Sclerosis Impact Scale (MSIS-29) has not yet been externally validated. Aim: To examine the external validity of a previously developed mapping algorithm by testing the accuracy of predicting EQ-5D-3L index values from MSIS-29 among multiple sclerosis (MS) patients in Sweden. Methods: Cross-sectional individual-level data were collected from population-based Swedish registers between 2011 and 2014. Health-related quality of life was assessed through MSIS-29 and EQ-5D-3L at one point in time among 767 individuals with known disability level of MS. A previously developed mapping algorithm was applied to predict EQ-5D index values from MSIS-29 items, and the predictive accuracy was assessed through mean absolute error and root mean square error. Results: When applying the algorithm, the predicted mean EQ-5D-3L index value was 0.77 compared to the observed mean index value of 0.75. Prediction error was higher for individuals reporting EQ-5D values <0.5 compared to individuals reporting EQ-5D values ≥0.5. Mean absolute error (0.12) and root mean square error (0.18) were smaller or equal to the prediction errors found in the original mapping study. Conclusion: The mapping algorithm had similar predictive accuracy in the two independent samples although results showed that the highest predictive performance was found in groups with better health. Varied predictive accuracy in subgroups is consistent with previous studies and strategies to deal with this are warranted.