论文部分内容阅读
目的考察HPLC中不同进样溶剂对醋酸奥曲肽色谱行为的影响,并分析其产生原因。方法采用HPLC法及紫外分光光度法。色谱条件:色谱柱为Diamonsil C18柱(4.6 mm×250 mm,5μm),流动相为体积分数0.1%三氟乙酸-乙腈(体积比7:3),检测波长为215 nm,柱温为30℃,流速为1.0 mL.min-1,进样量为20μL。结果同一质量浓度醋酸奥曲肽溶于不同的进样溶剂中进行HPLC分析发现,进样溶剂不同导致醋酸奥曲肽色谱峰面积不同。当进样溶剂为乙醇-水体系时,同一质量浓度醋酸奥曲肽色谱峰面积随乙醇体积分数的增大而减小,且当乙醇体积分数小于40%时峰面积变化不明显,大于40%时变化显著。此外,低(20 mg.L-1)、中(40 mg.L-1)、高(60 mg.L-1)3种质量浓度的醋酸奥曲肽其色谱峰面积随进样溶剂中乙醇体积分数变化而变化的趋势相一致。当进样溶剂分别为体积分数0%、40%、80%及100%的乙醇时,醋酸奥曲肽在质量浓度为10～60 mg.L-1内与峰面积均呈现良好的线性关系,相关系数r分别为0.999 9、0.999 9、0.999 3、0.999 2。紫外检测结果显示,在2种检测波长(215、280 nm)下,随着溶剂中乙醇体积分数的升高,醋酸奥曲肽紫外吸光度值增大。结论相同色谱条件下,醋酸奥曲肽色谱峰面积随进样溶剂的极性降低而减小,造成这种现象的原因是当进样溶剂强度大于流动相时,部分药物随着溶剂流出色谱柱而未被保留,从而导致药物主峰面积减小。
Objective To investigate the influence of different injection solvents on the chromatographic behavior of octreotide acetate and to analyze its causes. Methods HPLC and UV spectrophotometry. Chromatographic conditions: Diamonsil C18 column (4.6 mm × 250 mm, 5 μm) was used as the mobile phase. The mobile phase consisted of 0.1% trifluoroacetic acid-acetonitrile with the volume ratio of 7: 3 and the detection wavelength was 215 nm. , The flow rate of 1.0 mL.min-1, injection volume of 20μL. Results The same mass concentration of octreotide acetate dissolved in different injection solvents for HPLC analysis found that injection of different solvents leading to different octreotide acetate chromatographic peak area. When the injection solvent was ethanol-water system, the peak area of octreotide acetate with the same mass concentration decreased with the increase of ethanol volume fraction, and the change of peak area was not obvious when the volume fraction of ethanol was less than 40% Significant. In addition, the chromatographic peak areas of octreotide acetate (20 mg.L-1), medium (40 mg.L-1) and high (60 mg.L-1) The trend of change is consistent. When the injection solvent was 0%, 40%, 80% and 100% ethanol respectively, octreotide acetate showed a good linear relationship with the peak area in the concentration range of 10-60 mg.L-1, and the correlation coefficient r are 0.999 9,0.999 9,0.999 3,0.999 2, respectively. UV detection results showed that the UV absorbance of octreotide acetate increased with the increase of ethanol volume fraction in the two detection wavelengths (215 and 280 nm). Conclusion Under the same chromatographic conditions, the peak area of octreotide acetate decreases with the decreasing polarity of the sample solvent. The reason for this phenomenon is that when the injection solvent intensity is higher than that of the mobile phase, part of the drug flows out of the column with the solvent Be retained, resulting in reduced peak area of the drug.