Measurement of macular pigment: Raman spectroscopy versus heterochromatic flicker photometry.
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.
Neelam
Kumari
K
Department of Ophthalmology, Waterford Institute of Technology, Waterford, Republic of Ireland. neelamk@sehb.ie
O'Gorman
Nicholas
N
Nolan
John
J
O'Donovan
Orla
O
Wong
Hwee Bee
HB
Au Eong
Kah Guan
KG
Beatty
Stephen
S
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
D012168:Retinal Pigments; D024341:Xanthophylls; D065146:Zeaxanthins; D019207:beta Carotene; D014975:Lutein