| 引用本文: | 康斯丹,黄巧玲.HPLC测定人血清中甲氨蝶呤浓度[J].中国现代应用药学,2014,31(6):738-741. |
| KANG Sidan,HUANG Qiaoling.Determination of Methotrexate Concentration in Human Plasma by HPLC[J].Chin J Mod Appl Pharm(中国现代应用药学),2014,31(6):738-741. |
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| 摘要: |
| 目的 建立高效液相色谱法测定人血清中甲氨蝶呤(methotrexate，MTX)浓度的方法。方法 以茶碱为内标，色谱柱为Agilent Eclipse XDB-C18(4.6 mm×250 mm，5 μm)，流动相为磷酸盐缓冲液(pH 6.6)-甲醇(82∶18)，检测波长为MTX 306 nm和茶碱254 nm，流速为1.0 mL·min-1，柱温为30 ℃。结果 MTX浓度分别在0.03~1.65 μmol·L-1(r=0.999 9)和1.65~66.08 μmol·L-1(r=0.995 8)内线性关系良好，平均方法回收率97.6%。平均萃取回收率68.5%，日内、日间RSD均<5%，血清最低检测浓度为0.03 μmol·L-1。结论 该方法灵敏、准确，线性范围宽，适用于临床甲氨蝶呤的血药浓度监测。 |
| 关键词: 甲氨蝶呤 高效液相色谱法 血药浓度 骨肉瘤 |
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| Determination of Methotrexate Concentration in Human Plasma by HPLC |
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KANG Sidan, HUANG Qiaoling
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The Third Hospital of Hangzhou, Hangzhou 310009, China
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| Abstract: |
| OBJECTIVE To develop an HPLC method to determine the concentration of methotrexate(MTX) in human plasma. METHODS Theophyline was used as the internal standard. The analytical column was Agilent Eclipse XDB-C18(4.6 mm×250 mm, 5 μm). The mobile phase was phosphate buffer solution with pH 6.6-methanol(82∶18). The UV determine wavelength of MTX was 306 nm and theophyline was 254 nm. The flow rate was 1.0 mL·min-1, and the column temperature was 30 ℃. RESULTS The calibration curves of MTX were linear at a concentration range from 0.03 to 1.65 μmol·L-1(r=0.999 9) and 1.65 to 66.08 μmol·L-1(r=0.995 8). The intra-day and inte-day precision were both less than 5%. The average method recovery was 97.6% and the average extraction recovery was 68.5%, with a lowest limit of 0.03 μmol·L-1. CONCLUSION The method is sensitive, accurate, wide linear range, suitable for plasma concentration monitoring of MTX. |
| Key words: methotrexate HPLC plasma concentration osteosarcoma |
