反相HPLC法测定烟酸占替诺及有关物质的含量

Determination of xanthinol nicotinate and its related substances by RP-HPLC

目的:测定烟酸占替诺及有关物质的含量。方法:采用反向高效液相色谱法,梯度洗脱。色谱柱为迪马公司DiamonsilTM钻石C18(250mm×4.6mm,5μm);流动相A、B均为甲醇-水(12∶88、43∶57);每100ml流动相A、B中各加5滴三乙胺,并用1mol/LH3PO4溶液调pH=3.1±0.1;梯度条件为9minB浓度为0,9.1minB浓度为40%,30min停止。紫外检测器,波长为267nm,流速0.9ml/min,进样量20μl,柱温为室温,峰面积外标法和主成分自身对照法。结果:烟酸占替诺与茶碱分别在59.88～1197.6μg/ml(r=0.9999)和0.4～8μg/ml(r=0.9999)浓度范围内呈良好线性关系。茶碱平均回收率为99.6%,平均RSD=0.47%(n=3)。结论:本法选择性高,重现性好,准确,适合于烟酸占替诺与有关物质的含量测定。

Object: To develop an HPLC method for the assay of the content of xanthinol nicotinate and its related substances.Methods: The chromatographic separation was performed on a Diamonsil^(TM) C_(18) column (250 mm×4.6 mm ID,5 μm) with gradient elution. The mobile phase consisted of methanol-water (12∶88 and (43∶57,)v/v, for gradient A and B). 5 drops of thriethylamine were added in 100 ml of the mobile phase pH was adjusted to (3.1±0.1) using 1 mol/L H_(3)PO_(4) solution. Gradient condition were as follows: The content of the mobile phase B was 0 at 9 min, increased 40% at 9.1 min and retained to 45 min.The detective wavelength was set at 267 nm. The flow rate was 0.9 ml/min and the temperature of column was kept at room temperature. Results: There were good linearity over the range of 59.88～1 197.6 μg/ml (r=0.999 9) and 0.4～8 μg/ml (r=0.999 9) for xanthinol nicotinate and theophylline, respectively.The average recovery of theophylline was 99.6% with RSD of 0.47%(n=3). Conclusion:The established method was high selective, simple and accurate.It was suitable applied for the determination of xanthinol nicotinate and its related substances.