title: Measurement of macular pigment: Raman spectroscopy versus heterochromatic flicker photometry.
abstract: OBJECTIVE
  There are several techniques for measuring macular pigment (MP) in vivo, of which Raman spectroscopy (RS) is a recently developed objective
  
  
  METHODS
  This study reports the reproducibility, test-retest variability, and validity of RS MP readings, by comparing them with heterochromatic flicker photometry (HFP).
  
  
  METHODS
  MP was measured with HFP and RS in 120 healthy subjects, and the latter technique was also used on two separate occasions in a sample of 20 subjects to investigate the intersessional variability of readings. Intrasessional reproducibility of RS MP measurements was also calculated. In addition, serum concentrations of lutein (L) and zeaxanthin (Z) were measured and correlated with both RS and HFP MP readings.
  
  
  RESULTS
  Mean (+/-SD) MP in the right eye was 0.279 +/- 0.145 and 0.319 +/- 0.155 with RS and HFP, respectively. The differences between corresponding MP readings taken on RS and HFP lay within the Bland-Altman 95% limits of agreement for the two instruments in 93.6% and 94.4% of cases in the right and left eyes, respectively. Intrasessional reproducibility of RS readings, expressed as the coefficient of variation, was 8.42% +/- 7.12%. Ninety-five percent of MP readings taken with RS on two separate occasions lay within the 95% limits of agreement for the two sessions. A positive, but insignificant, relationship was observed between RS and HFP MP readings and serum concentrations of L and Z (RS, P = 0.356; HFP, P = 0.540).
  
  
  CONCLUSIONS
  RS, an objective method of measuring MP levels in vivo, exhibits acceptable reproducibility and test-retest variability. The results demonstrated good correlation between RS and HFP measurements of MP, thus authenticating RS against a validated psychophysical technique of measuring MP. However, investigators should use only one of these instruments for the duration of any given study because of differences in the scientific rationale, and the factors that influence RS and HFP measurements of MP.
authors.affiliation: Department of Ophthalmology, Waterford Institute of Technology, Waterford, Republic of Ireland. neelamk@sehb.ie
authors.forename: Kumari
authors.initials: K
authors.lastname: Neelam
authors.forename: Nicholas
authors.initials: N
authors.lastname: O'Gorman
authors.forename: John
authors.initials: J
authors.lastname: Nolan
authors.forename: Orla
authors.initials: O
authors.lastname: O'Donovan
authors.forename: Hwee Bee
authors.initials: HB
authors.lastname: Wong
authors.forename: Kah Guan
authors.initials: KG
authors.lastname: Au Eong
authors.forename: Stephen
authors.initials: S
authors.lastname: Beatty
mesh_terms: D000328:Adult; D002851:Chromatography, High Pressure Liquid; D005260:Female; D006801:Humans; D014975:Lutein / Q000032:analysis* / Q000097:blood; D008297:Male; D008875:Middle Aged; D010783:Photometry / Q000379:methods*; D015203:Reproducibility of Results; D012160:Retina / Q000737:chemistry*; D012168:Retinal Pigments / Q000032:analysis* / Q000097:blood; D012680:Sensitivity and Specificity; D013059:Spectrum Analysis, Raman / Q000379:methods*; D024341:Xanthophylls; D065146:Zeaxanthins; D019207:beta Carotene / Q000031:analogs & derivatives* / Q000032:analysis* / Q000097:blood
chemical_list: D012168:Retinal Pigments; D024341:Xanthophylls; D065146:Zeaxanthins; D019207:beta Carotene; D014975:Lutein