用于乳糖含量测定的新型氨基键合硅胶色谱固定相的制备及评价
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摘要
基于合成的氨基键合硅胶色谱固定相,按照2015版《中华人民共和国药典》乳糖项下含量测试方法,建立了高效液相色谱-示差折光检测(HPLC-RI)分离乳糖与蔗糖的分析方法。考察了3种不同类型的氨基色谱固定相制备的色谱柱(300 mm×4.6 mm,5μm)对乳糖和蔗糖的保留时间、分离度和峰面积稳定性等色谱行为的影响。以乙腈-水(70∶30,v/v)为流动相进行等度洗脱,流速为1.0 mL/min,进样量为10μL。结果表明:使用异丙基侧链保护的氨基色谱柱时,乳糖和蔗糖的分离度为3.03,实现了二者的良好分离,且各目标物峰形良好;乳糖峰面积的RSD仅为1.14%,小于药典规定的2.0%。该法满足2015版《中华人民共和国药典》方法中乳糖含量测定的要求,适合作为乳糖含量测定的质控色谱柱。
A method for the separation and determination of lactose and sucrose using high performance liquid chromatography-refractive index(HPLC-RI)detection base on the prepared amino-bonded silica gel chromatographic stationary phase was established. The method was accorded to the content determination method in Pharmacopoeia of the People's Republic of China(2015). Then,a series of chromatographic behaviors of lactose and sucrose such as the retention times,resolutions and peak area stabilities were investigated on three different types of amino-bonded silica columns(300 mm×4. 6 mm,5 μm). The mobile phase was acetonitrilewater(70 ∶30,v/v)with isocratic elution,the flow rate was 1. 0 mL/min and the injection volume was 10 μL. The resolution of lactose and sucrose was 3. 03 by isopropyl side chain protected amino column,and the peak shape of each target compound was good. The relative standard deviation(RSD)of peak area of lactose was 1. 14%,less than 2. 0% prescribed in the Pharmacopoeia of the People 's Republic of China(2015). Therefore,this method meets the requirements of lactose content determination,and is suitable as the quality control method in the above mentioned Pharmacopoeia.
A method for the separation and determination of lactose and sucrose using high performance liquid chromatography-refractive index(HPLC-RI)detection base on the prepared amino-bonded silica gel chromatographic stationary phase was established. The method was accorded to the content determination method in Pharmacopoeia of the People's Republic of China(2015). Then,a series of chromatographic behaviors of lactose and sucrose such as the retention times,resolutions and peak area stabilities were investigated on three different types of amino-bonded silica columns(300 mm×4. 6 mm,5 μm). The mobile phase was acetonitrilewater(70 ∶30,v/v)with isocratic elution,the flow rate was 1. 0 mL/min and the injection volume was 10 μL. The resolution of lactose and sucrose was 3. 03 by isopropyl side chain protected amino column,and the peak shape of each target compound was good. The relative standard deviation(RSD)of peak area of lactose was 1. 14%,less than 2. 0% prescribed in the Pharmacopoeia of the People 's Republic of China(2015). Therefore,this method meets the requirements of lactose content determination,and is suitable as the quality control method in the above mentioned Pharmacopoeia.
引文
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