Ten participants attending elective anterior segment eye surgery received 0.5% caspofungin eye drops either 1 drop hourly for 4 h or 1 drop an hour before surgery. The eye drops were generally well tolerated. In the absence of inflammation or corneal abrasion, topical caspofungin does not achieve clinically relevant concentrations.Fungal keratitis is a potentially blinding condition, with Candida, Aspergillus, and Fusarium spp. (2, 21) being the common pathogens. Current treatment for fungal keratitis is inadequate (8); thus, alternatives are needed.Caspofungin has activity against a wide range of fungi, with MIC 90 values ranging from 60 to 2,000 ng/ml (15, 23). Caspofungin eye drops are effective for treatment of fungal keratitis in rabbits (11, 18) and have good penetration into animal eyes with inflammation or previous corneal abrasion (10, 23). Clinically, caspofungin is administered mostly intravenously (1,4,6,9,19,20) or intravitreally (24) for ocular mycosis. The successful use of topical caspofungin alone (13) or in combination with voriconazole (22) has been reported for two patients. However, the utility of topical caspofungin in fungal keratitis remains unknown, partly due to limited data on its penetration into human aqueous humor after topical administration. This study aimed to investigate the ocular penetration of 0.5% caspofungin eye drops into human aqueous humor.
A rapid, precise, and sensitive liquid chromatography/mass spectrometry (LC/MS) method to quantify the caspofungin concentration in human aqueous humor was developed and validated. Sample preparation involved simple dilution of aqueous humor samples with acetonitrile. Azithromycin was the internal standard. Good linearity over 10 to 5,000 ng/ml was observed. The lower limit of quantification was 10 ng/ml. The intra-and interday accuracies (percent bias) were within 11%, while the intra-and interday precisions were within 6%.Current topical treatment for fungal keratitis is inadequate (13) given the lack of favorable outcomes with existing antifungal eye drops (i.e., fluconazole, amphotericin B, and natamycin) (7,14). Accordingly, it is important to investigate the topical application of newer antifungal agents that are available only as injections. In rabbit models (5, 8), caspofungin eye drops were effective in inhibiting the progression of fungal keratitis. It remains unknown if caspofungin eye drops are able to penetrate the human eye. Studies in this area are impeded by the absence of a simple and sensitive analytical assay to quantify caspofungin in human aqueous humor.Currently, high-performance liquid chromatography (HPLC) with fluorescence or amperometric detection (4,11,12,15) and liquid chromatography/tandem mass spectrometry (LC/ MS/MS) (1-3, 9, 16) are used to quantify caspofungin in biological samples. The majority of these methods, however, involve liquid-liquid extraction and require large sample volumes. Because aqueous humor samples usually have small volumes (Ͻ150 l), these methodologies are not suitable. This study aimed to develop a rapid and sensitive LC/MS assay for caspofungin that involves simple preparation and low sample volumes.Caspofungin diacetate was provided by Merck Sharp & Dohme. Azithromycin dihydrate (internal standard [IS]) was purchased from Kopran Ltd. (Maharashtra, India). Calibration standard and quality control (QC) stock solutions of caspofungin (1 mg/ml) were prepared separately in water and stored at Ϫ80°C. IS stock solution (1 mg/ml) was freshly prepared in acetonitrile for each analysis. Working solutions of caspofungin and IS were freshly prepared by diluting the stock solutions with water and acetonitrile, respectively.The calibration curve was constructed using water due to the limited availability of blank aqueous humor (6, 10). Calibration standards were prepared by adding 30 l IS working solution to 30 l caspofungin working solutions. The final concentrations of calibration standards were 10, 50, 100, 500, 1,000, 2,000, and 5,000 ng/ml with the IS (500 ng/ml). All QC samples (30, 300, and 4,000 ng/ml) were freshly prepared. The blank aqueous humors were pooled to prepare aqueous humorbased low (30 ng/ml; n ϭ 3) and high (4,000 ng/ml; n ϭ 3) QC samples (30 l each) to validate the water-based calibration curve.The Shimadzu LC system (Shimadzu, Japan) comprised a high-pressure gradient unit (LC-20AD pump and LC-20ADsp pump), DGU-20A 3 degasser, and CTO-10A column ...
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.
hi@scite.ai
10624 S. Eastern Ave., Ste. A-614
Henderson, NV 89052, USA
Copyright © 2024 scite LLC. All rights reserved.
Made with 💙 for researchers
Part of the Research Solutions Family.