两种方法测定复方氨基酸注射液(18AA-Ⅴ)中亚硫酸氢钠含量

Determination of sodium bisulfite in compound amino acid injection (18AA-V) by HPLC and colorimetry

目的:建立高效液相色谱(HPLC)法和比色法两种方法用于测定复方氨基酸注射液(18AA-Ⅴ)中的抗氧剂亚硫酸氢钠,并比较两种方法是否存在显著性差异。方法:HPLC法色谱条件为采用Shoedx Sugar SH1011色谱柱(300 mm×8 mm,6μm),以0.04 mol/L磷酸溶液为流动相,流速0.8 ml/min,柱温为30℃,检测波长为200 nm;比色法原理为亚硫酸氢钠在酸性条件下与甲醛和碱性品红可生成紫红色络合物,通过测定该络合物在556 nm波长处的吸光度,可计算出亚硫酸氢钠的含量。结果:HPLC法亚硫酸氢钠在41.8~313.9μg/ml范围内峰面积与浓度呈良好线性关系(r=0.999 5),平均回收率为102.4%,RSD为0.65%(n=9);比色法亚硫酸氢钠在0.8~2.5μg/ml浓度范围内呈良好的线性关系(r=0.999 0),平均回收率为99.7%,RSD为0.83%(n=9)。结论:两种方法均可用于测定复方氨基酸注射液(18AA-Ⅴ)中的抗氧剂亚硫酸氢钠,且无显著性差异,但HPLC法操作更简便,适合批量样品的测定。

Objective：To develop HPLC and colorimetry methods for determination of sodium bisulfite in compound amino acid injection （ 18AA -V）. Methods： Shoedx Sugar SH1011（8 mm ×300 mm, 6 μm）was adopted for HPLC, and 0.04 mol/L phos- phoric acid solution was used as mobile phase. The flow rate was 0.8 ml/min. The temperature was at 30℃. The detection wavelength was at 200 nm. Sodium bisulfite can be determined by colorimetry based on the colorimetric reaction between sodium bisul- rite, basic violet and formaldehyde in the acidic environment. Results： The linear range of sodium bisulfite was between 41.8 μg/ml and 313.9μg/ml （ r = 0. 9995 ） by HPLC, and average recovery was 102.4% with an RSD of 0.65% （ n = 9, while the linear range of sodium bisulfite was between 0.8 μg/ml and 2.5μg/ml （ r = 0. 999 0） by colorimetry, and the average recover3, was 99.7% with an RSD of 0.83% （ n = 9}. Conclusions： Both the methods can determine sodium bisulfite in compound amino acid injection （ 18AA -V） and with no significant difference, but the HPLC is more easy to operate, which is suitable to determine the batch samples.