柱前衍生液相色谱法测定藻酸双酯钠的含量

Determination of propylene glycol alginate sodium sulfate by pre-column derivatization high performance liquid chromatography

目的建立测定海洋药物藻酸双酯钠(PSS)含量的柱前衍生化高效液相色谱方法。方法利用0.1mol/L稀硫酸将PSS降解为D-甘露糖醛酸(M)和L-古罗糖醛酸(G),并与1-苯基-3-甲基-5-吡唑啉酮(PMP)进行柱前衍生化反应,采用高效液相色谱法测定PSS降解液中M和G的峰面积,以葡萄糖醛酸(GlcUA)为内标,以PSS对照品绘制标准曲线计算含量。色谱柱:Zorbax KP-C18(150mm×4.6mm,5μm),流动相:磷酸盐缓冲液(0.1mol/L,pH 7.0)/CH3CN(83/17,体积分数);流速:0.8mL/min;柱温30℃,检测器:可变波长检测器(VWD),检测波长:245nm。结果PSS在0.3～15mg/mL浓度范围内与峰面积比值呈良好的线性关系(R2=0.9966);在低、中、高3个浓度的日内精密度平均RSD为0.78%(n=6),日间精密度平均RSD为3.08%(n=5),重复性RSD为3.24%(n=5),24h内稳定性良好,RSD为2.93%,3个浓度平均回收率为115.77%,方法的最低检测限为0.25μg/mL。结论该方法专属性好,灵敏度高,重复性好,适合PSS的质量控制。

Objective To estabish a high performance liquid chromatography （HPLC） with pre-column derivatization technique for determining the content of propylene glycol alginate sodium sulfate （PSS）. Method PSS was hydrolyzed to ManUA （M） and GulUA（G） by 0. lmol/L sulphuric acid firstly, the hydrolysates were derived with 1-（4-isopropyl） phenyl-3-methyl-5-pyrazolone （PMP） and determined by a reversed phase high performance liquid chromatography （HPLC） method. Peak area ratio of M and G to GlcUA （internal standard） was used for calculation of PSS content according to PSS standard curve. Chromatographic column： Zorbax KP-C18 （150 mm）〈4.6 mm,5μm）; mobile phase： phosphate buffer （0. 1 mol/L, pH 7.0）/CH3CN （83/17, V/V）; Flow rate： 0. 8 mL/mim Column temperature 30 ℃, detector, variable wavelength detector （VWD）, detection wave length. 245 nm. Result The cali-bration curve was linear in the range of 0. 3-15 mg/mL with correlation coefficient of 0. 9966. The in-tra-day average RSD for PSS in three PSS concentrations （high, medium and low） was 0. 78% （n=6） and the inter-day average RSD was 3.08% （n=5）. The RSD for repeatability is 3.24% （n=5） and for stability is 2.93%. The average recovery was 115.77% and the low deetion limit was 0.25 g/mL. Con-clusion This method had a good specificity, high sensitivity, good repeatability and was suitable for quality control of PSS.