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Background: Health-related quality of life (HRQOL) is defined as perceived physical and mental health over time. Lower HRQOL has previously been shown to be associated with elevated adverse health events and mortality. Leukocyte telomere length (LTL), which is a biomarker of aging at the cellular level, has also been associated with age-related diseases and mortality. However, the association of HRQOL with LTL in the general population is poorly understood. Objective: We examined the relationship between different measures of HRQOL and LTL by ethnicity among a nationally representative sample of US adults using the data of the National Health and Nutrition Examination Survey (NHANES, 2001-2002). Method: HRQOL were assessed through a brief set of survey-based questions that asked participants about their self-rated general health and the number of recent days when the person was physically unwell, mentally unwell, or limited in usual activities. Telomere length was assessed using the quantitative polymerase chain reaction method of telomere length relative to standard reference DNA. Multiple linear regression was used to estimate the relationship, where we sequentially controlled for demographic (age, sex, education, marital status), health risk indicator (cancer, hypertension, diabetes, obesity, heart failure) and health behavior (smoking, physical activity, and alcohol intake) variables. Log transformed value of LTL was used for regression modeling and all the analyses were estimated with adjustment for sample weights for the genetic sub-sample and design effects. Results: The current study included 2090 White, 788 Mexican American, and 669 African American participants with mean age of 46.89 (Standard Error, SE=0.56), 37.57 (SE=0.73) and 41.99 (SE=0.74) years respectively. After controlling for demographic factors, health risk indicators, and health behaviors, recent days of unwell physical health was associated with shorter telomere length for Whites (beta=-0.005, 95% Confidence Interval, 95% CI=-0.01 to -0.001, p value 0.03). Among African Americans, those who perceived their general health condition as “good” had shorter telomere length compared to those who perceived general health status as “excellent” (beta=-0.02, 95% CI=-0.03 to -0.01, p value 0.001). Mental health related quality of life showed no significant association with LTL in any race. For Mexican Americans none of the HRQOL measures had any association with LTL. Conclusion: Lower physical HRQOL and lower general HRQOL were associated with shorter LTL or faster cellular aging for Whites and African Americans in our study. The results endorse the value of assessing HRQOL,specially in relation to to physical and general health, and the importance of using this measure for evaluating age and disease related consequences.