Refractive index and osmolality of human tears

Tear osmolality measurement, as a basic index of lacrimal physiology, is technically difficult. We investigated a simple, digital nanoliter technique of critical angle refractive index measurement. After initial comparison of the techniques with aqueous NaCl (0.5 to 1.4%), 3 successive tear samples were obtained from each of 40 normal, noncontact lens wearing subjects, aged 17 to 63 years, for analysis by nanoliter osmometry and digital refractometry. In addition, tear lactoferrin concentration was estimated with the Lactoplate immunoassay. For the range of NaCl concentrations tested, osmolality and refractive index measurements were highly correlated with concentration, and with each other. For 0.9% NaClaq the mean value of osmolality was 292.0 mOsm/kg and refractive index was 1.33419. For human tears average values were found to be: for osmolality, 303.7 +/- 22.9 mOsm/kg; for refractive index, 1.33698 +/- 0.00110; and for lactoferrin level, 1.64 +/- 0.47 mg/ml. No statistically significant correlation was obtained between either refractive index or lactoferrin level and osmolality, but refractive index and lactoferrin level were significantly correlated (r = 0.64, p < 0.001). The high correlation of refractometry and osmolality for aqueous NaCl indicates comparability of the techniques for this solution, but this does not apply to human tears. Unlike tear osmolality, tear refractive index is dependent to a greater extent upon composition, as is suggested by its correlation with lactoferrin level. Therefore, refractometry is not acceptable as a direct alternative to osmometry in the assessment of lacrimal function.