Determination of voriconazole in aqueous humor by liquid chromatography-electrospray ionization-mass spectrometry.
A novel method based on liquid chromatography-mass spectrometry with electrospray ionization (LC-MS) has been developed for analysis of voriconazole in aqueous humor. The separation was achieved on a reversed-phase C(18) column eluted by 70% acetonitrile-30% water-0.01% TFA. The correlation between the concentration of voriconazole to peak area was linear (r(2)=0.9990) between 0.04 and 60 ng, with a coefficient of variance of less than 3%. Limit of quantitation (LOQ) was estimated to be 5 ng/ml voriconazole with an injection volume of 2 microl of aqueous humor. Both intra-day and inter-day imprecision were less than 3% over the whole analytical range. Parallel analyses of voriconazole samples by LC-MS and by high-performance liquid chromatography (HPLC)-UV showed that the two methods were highly correlated (r(2)=0.9985). LC-MS was used to the determine voriconazole levels achieved in the aqueous humor of the rabbit eye, following topical application of 5 or 10 microg voriconazole in the form of eyedrops for 11 days b.i.d. The lower dosage produced an aqueous humor concentration of 7.29+/-5.84 microg/ml, while the higher dosage produced a concentration of 14.56+/-12.90 microg/ml.
Singapore Eye Research Institute, Singapore National Eye Center, Singapore 168751, Singapore.
Lei
L
Zhou
Randolph D
RD
Glickman
Nancy
N
Chen
William E
WE
Sponsel
John R
JR
Graybill
Kwok-Wai
KW
Lam
D000818:Animals; D000935:Antifungal Agents / Q000032:analysis*; D001082:Aqueous Humor / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D011743:Pyrimidines / Q000032:analysis*; D011817:Rabbits; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D014230:Triazoles / Q000032:analysis*; D065819:Voriconazole
D000935:Antifungal Agents; D011743:Pyrimidines; D014230:Triazoles; D065819:Voriconazole