摘要
【目的】设计并论证检测玻璃体内苏拉明质量浓度的双波长等吸收紫外分光法?【方法】 25只白兔摘除眼球冷冻后获取玻璃体,经过匀浆?沉淀?稀释等系列预处理后首先检验正常兔眼玻璃体个体差异性及紫外分光法测定玻璃体内苏拉明浓度的方法学特异性?接着建立标准曲线方程,考察检测方法的精密度和准确度,确立最低检测定量限,最后检测苏拉明在玻璃体内的样品稳定性?【结果】选取261 nm和269 nm两波长,全部6个个体及混合玻璃体的吸光值差ΔA(A261-A269)介于±0.002之间;外加高?低浓度苏拉明的曲线以及实际注药的高?低浓度苏拉明曲线与空白玻璃体的曲线走势一致且位于其上,各曲线的波峰波谷位置未见偏移;标准曲线方程为y = 4 234 x + 2,r = 0.999 1;次低?中浓度?次高标准浓度点的日内相对标准差(RSD)分别为13.16%?9.67%?10.35%,日间RSD分别为17.59%?10.09%?11.11%,相对回收率分别为:(95.89±0.08) %?(96.69±0.07) %?(97.43±0.01) %;设计标准曲线的最低质量浓度45 μg/mL;其日内?日间相对标准差分别为14.14%?15.94%,检验回收率为(94.92±0.01) %;常温组样品,8 h及以内稳定性尚好,之后不稳定?低温组及3次冻融组稳定性良好?【结论】在给定条件下,白兔玻璃体的个体差异可以忽略。
Objective To establish and demonstrate the two-wavelength ultraviolet spectrophotome-try (TWUS) on the assay of suramin in the vitreous of rabbits. Methods 25 albino rabbits were sacri-ficed and enucleated eyeballs. After a series of procedures, including frozen, ground, added acetoni-trile, centrifuged and diluted, the vitreous were measured for theirs continuous photo spectrum at wave-length range from 350 nm to 190 nm, and the absorbance was measured just at wavelength 261 nm and 269 nm. The verified items included individual difference of rabbits, specificity of the assay, stan-dard curve, precision ( relative standard deviation ), accuracy ( relative rate of recovery ) and sensitivi-ty ( the minimum detective concentration, ρmin ) of the assay, as well as the stability of the detected simple. Results When wavelength of 261 nm and 269 nm was chosen as a pair of equal point of ab-sorbance, all of the 6 rabbits had no difference in absorbance. There were similar photo spectrums a-mong the blank vitreous, drug -injected vitreous and drug-added vitreous. The wave crest of all spec-trum localized at 265nm or about showed no marked shift. The standard equation of the line was y = 4 234 x + 2 (r = 0.999 1 ). During one day, the relative standard deviation of 3 different concentra-tion were 13.16%, 9.67% and 10.35%, with theirs relative rate of recovery ( 95.89 ± 0.08 )%, ( 96.69 ± 0.07 )% and ( 97.43 ± 0.01 ) % respectively. The ρmin was 45 μg/mL. Although the samples could not be stored at room temperature over 8 h, those stored at 4 ℃ and -20 ℃ or less than 8 h at room temperature were very stable for measurement. Conclusion On given conditions, the individual difference of rabbits can be neglected. All the data of each verified item are in accor-dence with the national relevant rules. Therefore, two-wavelength ultraviolet spectrophotometry is a reli-able method for measurement of drug mass concentration in vitreous.
出处
《中山大学学报（医学科学版）》
CAS
CSCD
北大核心
2004年第3期225-230,共6页
Journal of Sun Yat-Sen University:Medical Sciences
基金
广东省重点科技攻关基金资助项目(97-45)
中山眼科中心创新基金资助项目(20009)
