高效液相色谱法测定烟酰胺注射液含量及检测有关物质  

Content Determination of Nicotinamide Injection and Detection of Related Substances by HPLC

作　　者：谭菊英 蒋洁 刘庄蔚 何虹 杨欣智 朱健萍 邓鸣 赵庄 TAN Juying;JIANG Jie;LIU Zhuangwei;HE Hong;YANG Xinzhi;ZHU Jianping;DENG Ming;ZHAO Zhuang(Guangxi Institute for Drug Control,Nanning,Guangxi,China 530021)

机构地区：[1]广西壮族自治区药品检验研究院,广西南宁530021

出　　处：《中国药业》2025年第1期78-82,共5页China Pharmaceuticals

基　　金：广西壮族自治区药品监督管理局药品安全科研项目[桂药监办函〔2020〕108号]。

摘　　要：目的建立测定烟酰胺注射液含量及检测有关物质的高效液相色谱(HPLC)法。方法色谱柱为WatersXselectHSST3柱(250 mm×4.6 mm,5μm);流动相为稀醋酸(12 g冰醋酸用水稀释至100 mL)5 mL,加水900 mL,加稀氨溶液(25.0%~30.0%浓氨水0.7 g,用水稀释至100 mL)30 mL、乙腈15 mL,用水稀释至1000 mL,再加四氢呋喃5 mL;流速为0.4,1.2 mL/min;检测波长为264 nm;柱温为30℃;进样量为10μL。结果含量测定中,烟酰胺的质量浓度在60.42~140.98μg/mL范围内与峰面积线性关系良好(r=0.9997,n=5);精密度、稳定性、重复性试验结果的RSD均低于2.0%;回收率为100.11%,RSD为0.76%(n=9);3批样品含量分别为100.61%,100.77%,99.60%。有关物质检测中,烟酰胺与杂质、各杂质间均能有效分离,辅料不干扰测定,供试品溶液在24 h内稳定性良好;烟酸(杂质A)、3-氰基吡啶、2-吡啶甲酰胺、4-吡啶甲酰胺、N-氧代烟酰胺在各自质量浓度范围内与峰面积线性关系均良好(r>0.9997,n=7);回收率分别为102.20%,102.00%,101.76%,101.96%,102.14%,RSD分别为0.99%,0.76%,0.76%,0.80%,1.03%(n=9);检测限分别为0.26,1.12,0.78,0.63,0.13 ng;定量限分别为0.86,3.74,2.59,2.09,0.43 ng;3批样品中均仅检出杂质A,含量分别为0.24%,0.41%,0.061%。结论该方法准确度高、灵敏度好,可用于烟酰胺注射液的含量测定及有关物质检测。Objective To establish a high-performance liquid chromatography(HPLC)method for the content determination of Nicotinamide Injection and detection of related substances.Methods The chromatographic column was a Waters Xselect HSS T3 column(250 mm×4.6 mm,5μm);the mobile phase consisted of 5 mL of dilute acetic acid(12 g of glacial acetic acid diluted to 100 mL with water),900 mL of water,30 mL of dilute ammonia solution(0.7 g of 25.0%-30.0%concentrated ammonia solution,diluted to 100 mL with water),15 mL of acetonitrile,diluted to 1000 mL with water,and 5 mL of tetrahydrofuran;the flow rate was 0.4 mL/min and 1.2 mL/min;the detection wavelength was 264 nm;the column temperature was 30℃;the injection volume was 10μL.Results In the content determination,the linear range of nicotinamide was 60.42-140.98μg/mL(r=0.9997,n=5).The RSDs of precision,stability,and repeatability test results were all lower than 2.0%.The recovery rate of nicotinamide was 100.11%with an RSD of 0.76%(n=9).The contents of nicotinamide in the three batches of samples were 100.61%,100.77%,and 99.60%,respectively.In the detection of related substances,nicotinamide could be effectively separated from impurities and impurities could also be well separated from each other,and excipients did not interfere with the determination.The stability of the test solution was good within 24 h.Niacinamide(impurities A),3-cyanopyridine,Pyridine-2-carboxamide,isonicotinamide,and nicotinamuide-N-oxide had a good linear relationship with peak area within their respective mass concentration ranges(r>0.9997,n=7).The recovery rates were 102.20%,102.00%,101.76%,101.96%,and 102.14%,with RSDs of 0.99%,0.76%,0.76%,0.80%,and 1.03%(n=9),respectively.The limits of detection were 0.26,1.12,0.78,0.63,and 0.13 ng,respectively.The limits of quantification were 0.86,3.74,2.59,2.09,and 0.43 ng,respectively.Only impurity A was detected in all three batches of samples,with contents of 0.24%,0.41%,and 0.061%,respectively.Conclusion This method has high accuracy and good sensitivity

关 键 词：高效液相色谱法 烟酰胺注射液 含量测定 有关物质检测 

分 类 号：R927.2[医药卫生—药学] R977.22