title: Fluorophotometric measurement of the buffering action of human tears in vivo.
abstract: OBJECTIVE
  The buffering action of human tears is thought to be important to keep its pH constant. We measured the change in pH in the precorneal tear film in vivo when the acidic solution is challenged, using a fluorophotometric technique.
  
  
  METHODS
  Twelve eyes from 6 healthy subjects were entered in this study. Each subject was pretreated with either one drop of 0.4% oxybuprocaine for once (light anesthesia), three times (deep anesthesia), or none (controls). The measurement was initiated by instilling 20 microl of 0.067 M phosphate buffer at pH 5.5 containing 2 mM bis-carboxyethyl-carboxyfluorescein free acid, a pH sensitive dye, into the subject's eye. The pH was determined by the ratio of fluorescent intensities at two excitation wavelengths (490 and 430 nm). pH recovery time (PHRT) as defined by the time required for pH to reach 95% of pH at equilibrium was used for the marker of tear buffering action. Tear turnover rate was also determined using the fluorescent decay curve at 430 nm, which was independent of pH, but dependent on dye concentration.
  
  
  RESULTS
  Immediately after the instillation, the pH value in the tear film was around 6.0-6.5 in all cases. The tear film rapidly became more alkaline, reaching its normal value in 2.3 +/- 0.5 min in untreated eyes. The pretreatment with 0.4% oxybuprocaine retarded the neutralization process. A single regression analysis revealed that the PHRT had a significant negative correlation with the tear turnover rate (r = -0.78).
  
  
  CONCLUSIONS
  Our results suggest that the neutralization process of tears largely depends on the tear turnover rate. The buffering action of tears in vivo consists of the tear turnover as well as its chemical buffering capacity.
authors.affiliation: Department of Ophthalmology, Keio University School of Medicine, Tokyo, Japan. yamadam@med.keio.ac.jp
authors.forename: M
authors.initials: M
authors.lastname: Yamada
authors.forename: M
authors.initials: M
authors.lastname: Kawai
authors.forename: H
authors.initials: H
authors.lastname: Mochizuki
authors.forename: Y
authors.initials: Y
authors.lastname: Hata
authors.forename: Y
authors.initials: Y
authors.lastname: Mashima
mesh_terms: D000136:Acid-Base Equilibrium / Q000502:physiology*; D000328:Adult; D000779:Anesthetics, Local / Q000008:administration & dosage; D002021:Buffers; D005260:Female; D005452:Fluoresceins / Q000378:metabolism; D005456:Fluorescent Dyes / Q000378:metabolism; D015590:Fluorophotometry; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008297:Male; D010710:Phosphates / Q000378:metabolism; D011343:Procaine / Q000008:administration & dosage / Q000031:analogs & derivatives; D013666:Tears / Q000502:physiology*
chemical_list: D000779:Anesthetics, Local; D002021:Buffers; D005452:Fluoresceins; D005456:Fluorescent Dyes; D010710:Phosphates; D011343:Procaine; C043829:2',7'-bis(carboxyethyl)-5(6)-carboxyfluorescein; C005298:benoxinate; C018279:sodium phosphate