| 高效液相色谱法测定细胞内罗丹明123的浓度 |
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| 引用本文： | 宋剑萍. 高效液相色谱法测定细胞内罗丹明123的浓度[J]. 中南大学学报(医学版), 2006, 31(4): 610-612 |
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| 作者姓名： | 宋剑萍 |
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| 作者单位： | 湖南省肿瘤医院药剂科,长沙,410006 |
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| 摘 要： | 目的:建立高效液相色谱法细胞内罗丹明123浓度的测定方法。方法:采用Zirchrom反相色谱柱(KromasilC18,250mm×4.6mm,5μm),柱温40℃,流动相为乙腈-水相(65∶35,v/v),水相为1.2mmol/L四丁基溴化铵和15mmol/L醋酸钾,检测波长为390nm,样品经沉淀蛋白后进样,内标为罗丹明B。结果:罗丹明123在3~300μg/L范围内线性良好,最低检测浓度为1μg/L,日内、日间RSD均小于5%。结论:本方法专一性强、灵敏度高、简单可靠,可用于研究细胞摄取能力。
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| 关 键 词： | 罗丹明123 浓度 高效液相色谱法 |
| 文章编号： | 1672-7347（2006）04-0610-03 |
| 收稿时间： | 2006-07-02 |
| 修稿时间： | 2006-07-02 |
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| SONG Jian-ping.
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| Authors: | SONG Jian-ping |
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| Affiliation: | Department of Pharmacy, Hunan Tumor Hospital, Changsha 410006, China. sjping333@126.com |
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| Abstract: | OBJECTIVE: To establish a high-performance liquid chromatography with visible wavelength detection method for rhodamine 123 in cell lysate. METHODS: The HPLC separation was performed on a Kromasil C(18) (5 microm, 4.6 mm x 250 mm) column, using water (15 mmol/L potassium acetate and 1.2 mmol/L tetrabutylammonium bromide) - acetonitrile (65:35) as the mobile phase. The wave length was 390 nm, the internal standard was rhodamine B. Protein in the sample was precipitated by trichloroacetic acid. RESULTS: The calibration curve was linear in the range of 3 - 300 microg/L. The intra-day and inter-day RSDs were less than 5%. CONCLUSION: The method is accurate,sensitive, simple, and reliable for determining rhodamine 123 in cell lysate. |
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| Keywords: | rhodamine 123 concentration high-performance liquid chromatography |
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