HPLC法测定劳拉西泮片有关物质的含量

HPLC determination of the related substances in iorazepam tablets

目的建立劳拉西泮片的含量及有关物质的HPLC方法。方法以十八烷基硅烷键合硅胶作为填充剂的ZORBAX SB-C18柱(250 mm×4.6 mm,5μm)为色谱柱,流动相:0.05 mol·L^(-1)的磷酸二氢铵(含0.5%的三乙胺,用磷酸调p H至2.5)∶甲醇∶乙腈=35∶35∶30;流速:1.0 m L·min^(-1);检测波长:235 nm;进样量:10μL;柱温:30℃。结果劳拉西泮峰及各杂质峰均能良好分离。杂质A浓度与峰面积在1.5μg·L^(-1)~8.048 mg·L^(-1)内线性关系良好。杂质B浓度与峰面积在15.0μg·L^(-1)~8.224 mg·L~(^(-1))内线性关系良好。杂质C浓度与峰面积在1.7μg·L^(-1)~8.144 mg·L^(-1)内线性关系良好;杂质D浓度与峰面积在2.5μg·L^(-1)~8.032 mg·L^(-1)内线性关系良好;杂质E浓度与峰面积在3.0μg·L^(-1)~8.032 mg·L^(-1)内线性关系良好。测定样品含量平均值为97.9%,有关物质均符合要求。结论该方法快速、简单,稳定,重现性好,可作为劳拉西泮片的含量及有关物质的检测方法。

Objective To determine the related substances and content in iorazepam tablets by HPLC. Methods The chromatographic separation was performed on ZORBAX SB-C18column( 4. 6 mm × 250 mm,5 μm) that was made by alkyl-bonded silica gels. The mobile phase was 0. 05 mol·L^(-1)ammonium dihydrogen orthophosphate( including 0. 5% trimethylamine and p H adjusted to 2. 5 with phosphoric acid) ∶ methanol∶ acetonitrile = 35∶ 35∶ 30. The flow rate was 1. 0 m L·min^(-1),the detection wavelength was 235 nm,the sample size was 10 μL,and the column temperature was set at 30℃. Results The resolution between iorazepam and the other peaks met the requirements. The linearity of related substance A concentration and peak area was 1. 5 μg·L^(-1)~ 8. 048 mg·L^(-1). The linearity of related substance B concentration and peak area was 15. 0 μg·L^(-1)~ 8. 224 mg·L^(-1). The linearity of related substance C concentration and peak area was 1. 7 μg·L^(-1)~ 8. 144 mg·L^(-1). The linearity of related substance D concentration and peak area was 2. 5 μg·L^(-1)~ 7. 952 mg·L^(-1). The linearity of related substance E concentration and peak area was 3. 0 μg·L^(-1)~ 8. 032 mg·L^(-1). Determination of the sample average content was 97. 9%,and the related substance met the requirements. Conclusions The method was quick,simple,stable,and well repeatable,which could be used to determine the content of iorazepam tablets and its related substances.