复方苦玄参口服液的制备工艺与质量控制

Preparation Process and Quality Control of Compound Ku Xuan Shen Oral Liquid

【目的】丰富复方苦玄参颗粒剂型,用于防控畜禽细菌性腹泻,研制复方苦玄参口服液并对其进行质量控制,为后期临床试验奠定药理学基础。【方法】基于前期试验基础,评价联合使用0.25%苯甲酸和0.075%山梨酸作为防腐剂的适用性,筛选0.025%~0.100%亚硫酸氢钠、焦亚硫酸钠和抗坏血酸中适宜的抗氧化剂;建立处方中苦玄参、救必应、桃金娘根、南刘寄奴四味药材的薄层色谱(TLC)鉴别分析方法和君药苦玄参指标性成分苦玄参苷IA高效液相色谱(HPLC)含量测定分析方法,并参照《中华人民共和国兽药典》2020版口服液相关规定对口服液进行质量控制。【结果】添加0.25%苯甲酸和0.075%山梨酸作为防腐剂,复方苦玄参口服液中大肠埃希菌、铜绿假单胞菌、金黄色葡萄球菌、黑曲霉和白色念珠菌活菌数在第14和28天时均符合抑菌效力标准;与阴性样品相比,添加0.075%焦亚硫酸钠作为抗氧化剂10 d时苦玄参苷IA降解量由7%下降到1.38%,显著增强了复方苦玄参口服液的稳定性;TLC结果显示,复方苦玄参口服液中苦玄参、救必应、桃金娘根、南刘寄奴与对照品或对照药材在相应位置上显示相同颜色的荧光斑点,表明建立的TLC方法专属性良好;HPLC结果显示,苦玄参有效成分苦玄参苷IA在14.063~450μg/mL(R 2=0.9996)范围内线性关系良好,精密度试验、重复性试验、稳定性试验结果相对标准偏差(RSD)分别为1.02%、0.44%和0.60%;苦玄参苷IA平均加样回收率为98.50%,RSD(n=6)为0.56%,表明所建立的复方苦玄参口服液HPLC方法专属性、精密度、重复性、稳定性等均符合方法学要求。测得3批复方苦玄参口服液中苦玄参苷IA的含量平均值依次为214.97、213.85、214.82μg/mL,RSD为0.38%。其他质控指标,如外观性状呈黄褐色,相对密度为1.039~1.055 g/mL、RSD为0.88%,pH为5.50~5.66、RSD为0.72%,微生物限度等均符合《中华人民共和国兽药典》2020版口服液相关规定。【结论】本研究制备的复方苦玄参口服液工艺简单,质量控制方法准确、可靠,可用于后续中试生产与质量控制。

【Objective】The experiment was aimed to enrich the dosage form of compound Ku Xuan Shen granules for the prevention and control of bacterial diarrhea in livestock and poultry,develop compound Ku Xxuan Shen oral liquid,and control its quality,lay a pharmacological foundation for later clinical trials.【Method】Based on the basis of the previous experiments,the applicability of the joint use of 0.25%benzoic acid and 0.075%sorbic acid as preservatives was evaluated,and the suitable antioxidants among 0.025%-0.100%sodium bisulfite,sodium metabisulfite and ascorbic acid were screened.TLC identification and analysis method for the four medicinal materials of Picria felterrae Lour.,Cortex ilicis Rotundae,root of Rhodomyrtus tomentosa and Artemisia anomala S.Moore in the prescription and a HPLC method for the content determination and analysis of the standard component of picroside IA in the royal medicine Picria felterrae Lour.were established.The quality control of oral liquid was carried out according to the relevant provisions of the 2020 edition of the Veterinary Pharmacopoeia of the People’s Republic of China.【Result】Adding 0.25%benzoic acid and 0.075%sorbic acid as preservatives,the viable counts of Escherichia coli,Pseudomonas aeruginosa,Staphylococcus aureus,Aspergillus niger and Candida albicans in the compound Ku Xuan Shen oral liquid met the criteria for bacteriostatic efficacy on the 14th and 28th day,which indicated that the combination of the preservative had a good anticorrosive effect.Compared with the negative samples,the degradation of picroside IA decreased from 7%to 1.38%at 10 d with the addition of 0.075%sodium metabisulphite as antioxidant,which significantly enhanced the stability of the compound Ku Xuan Shen oral liquid.TLC results showed that the fluorescent spots of Picria felterrae Lour.,Cortex ilicis Rotundae,root of Rhodomyrtus tomentosa and Artemisia anomala S.Moore in the oral liquid of compound Kuan Xuan Shen and the control or control herbs were in the same colour in the corresponding positions,which indicated that the specificity of the established TLC method was good.The HPLC results showed good linearity in the range of 14.063 to 450μg/mL(R 2=0.9996)for picroside IA,the active ingredient of Picria felterrae Lour.,and the relative standard deviation(RSD)of the results of the precision test,reproducibility test,stability test were 1.02%,0.44%and 0.60%,respectively.The average recovery was 98.50%with the RSD of 0.56%,which indicated that the established HPLC method of compound Ku Xuan Shen oral liquid met the methodological requirements in terms of specificity,linearity,stability and accuracy.The average values of the content of picroside IA in the three batches of compound Ku Xuan Shen oral liquid were 214.97,213.85 and 214.82μg/mL,respectively,and the RSD was 0.38%.Other quality control indicators such as yellow-brown appearance and properties,relative density was 1.039 to 1.055 g/mL,with RSD of 0.88%,pH was 5.50 to 5.66,with RSD of 0.72%,microbiological limits,etc.were in accordance with the relevant provisions of the 2020 edition of the Veterinary Pharmacopoeia of the People’s Republic of China for oral liquids.【Conclusion】The compound Ku Xuan Shen oral liquid prepared in this study had a simple process,accurate and reliable quality control method,and could be used for subsequent pilot production and quality control.