Influence of filtering on the effective concentration and sterility of a 2% cyclosporine ophthalmic solution: a quality improvement perspective.

BACKGROUND: Pharmaceutical companies do not sell formulations for all diseases; thus, healthcare workers have to treat some diseases by concocting in-hospital preparations. An example is the high-concentration 2% cyclosporine A (CyA) ophthalmic solution. Utilizing a filter in sterility operations is a general practice for concocting in-hospital preparations, as is the case for preparing a 2% CyA ophthalmic solution. However, whether filtering is appropriate concerning the active ingredient content and bacterial contamination according to the post-preparing quality control of a 2% CyA ophthalmic solution is yet to be verified. METHODS: We conducted particle size, preparation concentration, and bacterial contamination studies to clarify aforementioned questions. First, we measured the particle size of CyA through a laser diffraction particle size distribution. Next, we measured the concentration after preparation with or without a 0.45-µm filter operation using an electrochemiluminescence immunoassay. Finally, bacterial contamination tests were conducted using an automated blood culture system to prepare a 2% CyA ophthalmic solution without a 0.45 µm filtering. Regarding the pore size of the filter in this study, it was set to 0.45 µm with reference to the book (the 6th edition) with recipes for the preparation of in-hospital preparations edited by the Japanese Society of Hospital Pharmacists. RESULTS: CyA had various particle sizes; approximately 30% of the total particles exceeded 0.45 µm. The mean ± standard deviation of filtered and non-filtered CyA concentrations in ophthalmic solutions were 346.51 ± 170.76 and 499.74 ± 76.95ng/mL, respectively (p = 0.011). Regarding bacterial contamination tests, aerobes and anaerobes microorganisms were not detected in 14 days of culture. CONCLUSIONS: Due to the results of this study, the concentration of CyA may be reduced by using a 0.45-µm filter during the preparation of CyA ophthalmic solutions, and furthermore that the use of a 0.45-µm filter may not contribute to sterility when preparing CyA ophthalmic solutions.

Determination of the Optimal Wavelength of the Hemolysis Index Measurement.

Many biochemical auto-analyzers have methods that measure the hemolysis index (HI) to quantitatively assess the degree of hemolysis. Past reports on HI are mostly in vitro studies. Therefore, we evaluated the optimal wavelength of HI measurement ex vivo using clinical samples. Four different wavelengths (410/451 nm: HI-1, 451/478 nm: HI-2, 545/596 nm: HI-3 and 571/596 nm: HI-4) were selected for HI measurement, and correlations were examined from the measurement results of 3890 clinical samples. Another set of 9446 clinical samples was used to examine the correlation of HI with lactate dehydrogenase (LDH), aspartate aminotransferase (AST) and potassium (K). Strong correlations were found between HI-4 and HI-1 and between HI-4 and HI-3. HI-1 and HI-2 cannot correctly assess hemolysis for high bilirubin samples, and HI-3 cannot correctly assess hemolysis for high triglyceride samples. LDH, AST and K correlated positively with HI-4 in clinical samples. For every 1-unit increase in HI-4, LDH increased by 19.51 U/L, AST by 1.03 U/L and K by 0.061 mmol/L, comparable to reports of other studies. In clinical samples, HI-4 was less susceptible to bilirubin and chyle and reflected well the changes in LDH, AST and K caused by hemolysis. This suggested that the optimal wavelength for HI measurement is 571 nm.

What is the best wavelength for the measurement of hemolysis index?

BACKGROUND: Hemolysis is a common problem in the handling of serum specimens. The hemolysis index (HI) provides a warning of hemolysis in auto-analyzers. However, HI has not been standardized, and each laboratory's original method is applied. Especially, the wavelength used for HI measurement is different in each laboratory. Thus, we investigated the warning ability of HI at various wavelengths. METHODS: We selected 4 wavelength types, and each HI was measured and calculated (410 nm/HI-1, 451 nm/HI-2, 545 nm/HI-3, and 571 nm/HI-4). To compare the 4 HI types, we investigated the influence of 3 interference components using artificially hemolyzed specimens (AHSs). We also investigated both the relationship between HI and hemoglobin concentration (Hb) and that between HI and 31 biochemical test values in AHSs. RESULTS: In the interference assessment, only HI-4 showed no influence on the 3 interference components. The correlation between Hb and HI-4 was very strong (rS = 0.9987). A 1-unit increase in HI-4 corresponded to a 14.8-mg/dL increase in Hb. CONCLUSION: We found the best wavelength for HI to be at or near 571 nm.