摘要
目的建立高效液相色谱法测定血浆中亚甲蓝浓度的方法。方法血浆样品经Varian C2柱固相萃取后采用高效液相-紫外检测法(HPLC-UV)测定。色谱条件:分析柱:Kromasil-C18(4.6 mm×250 mm,5μm);柱温:40℃;流动相:乙腈-0.3%三乙胺溶液(磷酸调pH至2.07)=31∶69;流速:1.0 mL.min-1;检测波长:在同一通道中采用可变波长,0～6.7 min中检测波长设定为668 nm来检测亚甲蓝;6.7～10 min检测波长设定为254 nm来检测内标非那西丁。采用内标法定量,以非那西丁为内标物。结果亚甲蓝浓度在8.55～855μg.L-1内线性关系良好(r=0.999 5),最低定量限为8.55μg.L-1。萃取回收率为81.08%～93.57%,批内精密度为3.93%～6.75%,批间精密度为10.16%～13.36%。结论本方法简便、灵敏、准确、所需血浆量少,为临床开发亚甲蓝的新应用提供了研发基础。
OBJECTIVE To establish a HPLC method for the determination of methylene blue in plasma. METHOD The drug was extracted from plasma with C2 column by SPE method. Separation was performed on a Kromasil C18 reverse-phase analytical column （4. 6 mm × 250 mm, 5 um） maintained at 40 ℃. The mobile phase consisted of acetonitrile-0. 3% triethylamine of aqueous solution with the pH adjusted to 2. 07 by phosphoric acid （31： 69）, at a flow rate of 1 mL · min -1. The UV detector used a variable wavelength that was set at 668 nm for the first 6.7 min to detect methylene blue and then switched to 254 nm until 10 min to detect phenacetin. Phenacetin was used as internal standard for the quantitation. RESULTS The good linearity of methylene blue was obtained in the range of 8.55 - 855 ug . L- 1. The correlation coefficient was 0. 999 5 and the lower limit of quantification was 8.55 ug. L-1 . The extraction recoveries were in the range of 81.08% -93.57%. The preeisions of intra-batch and inter-batch were 3.93% -6. 75% and 10. 16% - 13.36% , respectively. CONCLUSION The method is simple, sensitive, accurate and provides a research basis for the development of new application of methylene blue in medicine.
出处
《中国药学杂志》
CAS
CSCD
北大核心
2011年第3期231-234,共4页
Chinese Pharmaceutical Journal
