HPLC法测定注射用脱氧核苷酸钠的含量

Determination of sodium deoxyribonucleotide for injection by HPLC

目的建立高效液相色谱法测定注射用脱氧核苷酸钠的含量。方法采用Whatman TMPartisil-10 SAX（250mm×4.6mm,10μm）色谱柱,流动相为50mmol·L^-1磷酸二氢钾溶液（取磷酸二氢钾6.80 g,加水800mL,用磷酸调节pH至3.3,加水稀释至1 000mL）,柱温为30℃,检测波长为258 nm,流速为1.0mL·min^-1。结果注射用脱氧核苷酸钠中脱氧核糖胞嘧啶核苷酸钠（d CMPNa）、脱氧核糖腺嘌呤核苷酸钠（d AMP-Na）、脱氧核糖胸腺嘧啶核苷酸钠（d TMP-Na）和脱氧核糖鸟嘌呤核苷酸钠（d GMP-Na）的质量浓度分别在0.900 0-90.30mg·L^-1、0.940 0-94.70mg·L^-1、0.880 0-87.60mg·L^-1和0.740 0-74.00mg·L^-1内与峰面积呈良好的线性关系,且r均大于0.999 5。4种脱氧核糖核苷酸钠的平均加样回收率分别为99.3%（RSD=1.2%）、99.1%（RSD=0.8%）、98.2%（RSD=1.1%）和100.1%（RSD=1.5%）（n=9）。结论该方法简便、专属,分离效果好,灵敏度高,可用于测定注射用脱氧核苷酸钠中4种脱氧核糖核苷酸钠的含量。

Objective To develop a method for the quantitative analysis of sodium deoxyribonucleotide by HPLC.Methods The analysis was performed by using a Whatman TMPartisil-10 SAX（250mm × 4.6mm,10μm） at elution of 50mmol·L^-1KH2PO4 with ultraviolet detector（258 nm）,the flowrate was1.0mL·min^-1 and the column temperature was maintained at 30 ℃.Results All calibration curves showed good linearity（r 0.999 0） within test ranges.The recoveries were 98.2%-100.1% and the RSD for intraday was less than 2.0% at nine levels.Conclusions This method is simple,exclusive and of effective separation and high sensitivity.It is suitable for the determination of sodium deoxyribonucleotide.