反相高效液相色谱法定量检测重组人心钠素

目的 :建立反相高效液相色谱法 (RP HPLC)测定重组人心钠素 (rhANP)含量的方法。方法 :在WatersAlliance 2 6 90高效液相色谱仪上 ,以WatersYMC PackC18柱 (2 5 0mm× 4 .6mm ,5μm)为分离柱 ,996紫外可见分光光度检测器 ,检测波长 2 15nm ,流动相A :0 .1%三氟乙酸 水溶液 ;流动相B :0 .1%三氟乙酸 乙腈。进行线性梯度洗脱 ,B相 0 ,30 ,35min比例依次为 0 % ,6 0 % ,10 0 %。结果 :rhANP在 2 .12～ 2 1.2 0mg·L- 1浓度范围内呈现良好的线性关系 (r =0 .9996 ,n =5 ) ,平均回收率大于 95 %。结论 :RP HPLC法测定rhANP的方法准确、快速、可行。

Determination of recombination human atrial natriuretic polypeptide by reverse phase high performance liquid chromatography

AIM: To develop a reverse phase high performance liquid chromatography(RP-HPLC) method to determine the contents of recombination human atrial natriuretic polypeptide( rhANP) . METHODS: The rhANP was assayed by RP-HPLC with a Waters YMC-Pack C_ 18 (250 mm×4.6 mm,5 μm). The mobile phase A was 0.1 % trifluoracetic-water,and the mobile phase B was 0.1 % trifluoracetic-acetonitrile. The ratio of phase B was 0 %, 60 %,100 % in 0,30,35 min,respectively. The detecting wavelength was 215nm.The flow rate was 0.5 mL·min -1 . RESULTS: The calibration cure was linear in the concentrations ranged from 2.12 to 21.20 mg· L -1 ( r =0.9996, n =5). CONCLUSION: RP-HPLC is simple and reliable, which can be applied to the determination of rhANP.