鼻炎片质量标准研究
摘要
目的:鼻炎片收载于《中国药典》2005年版一部,具有祛风宣肺,清热解毒之功效,多用于治疗急、慢性非特异性炎症和过敏引起的鼻炎等症。现行鼻炎片的质量标准不够完善,含量测定指标单一,已不能满足中药现代化质量控制的要求。本论文主要就鼻炎片的质量标准,进行了系统的研究,建立了合理的薄层鉴别、含量测定方法,整个操作简洁、合理,可以作为本品的质量控制方法。
方法:采用薄层色谱法对制剂中三味药材进行定性鉴别。甘草:采用硅胶G薄层板,以三氯甲烷-丙酮-氨水(2:7:1)的溶液为展开剂,10%H_2SO_4乙醇液为显色剂,在日光下检视。防风:采用硅胶G薄层板,以三氯甲烷-甲醇(4:1)的溶液为展开剂,10%H_2SO_4乙醇液为显色剂,置紫外光灯(365nm)下检视。苍耳子:采用硅胶GF254薄层板,以环己烷-乙酸乙脂-甲酸(1:3:0.2)的溶液为展开剂,置紫外光灯(254nm)下检视。
以升麻素苷为指标,建立臣药防风药材的含量测定方法。采用Diamonsil C_(18)分析柱(4.6*250mm,5μm),甲醇-乙腈-水(18:12:70)为流动相,检测波长为254nm,流速为1.0ml/min,柱温为室温,能较好的测定升麻素苷含量。以5-0-甲基维斯阿米醇苷为指标,建立臣药防风药材的含量测定方法。采用Diamonsil C_(18)分析柱(4.6*250mm,5μm),甲醇-乙腈-5%冰乙酸(13:12:75)为流动相,检测波长为254nm,流速为1.0ml/min,柱温为室温,能较好的测定5-0-甲基维斯阿米醇苷含量。
结果:在定性鉴别中,供试品色谱在与对照药材色谱相应的位置上,分别显示相同颜色的斑点或荧光斑点,阴性样品色谱在与对照药材色谱相应的位置上不显示任何颜色的斑点。
含量测定中,升麻素苷在0.118-2.360μg范围内与峰面积呈良好的线性关系,回归方程为Y=24.736X+0.7825,r=0.9998;精密度、重复性、回收率的RSD<3%;在分析浓度下,12小时内基本稳定(RSD=1.58%);本办法适合检测含量在3.9-28.4μg/ml浓度范围的供试品溶液。
此外,5-O-甲基维斯阿米醇苷在0.102-2.040μg范围内与峰面积呈良好的线性关系,回归方程为Y=29.379X+0.6648,r=0.9997;精密度、重复性、回收率的RSD<3%;在分析浓度下,12小时内基本稳定(RSD=1.52%);本办法适合检测含量在5.7μg/m-42.2μg/ml浓度范围的供试品溶液。
结论:薄层色谱法鉴别处方中三味药材,其鉴别方法快速简便,专属性强,斑点清晰,重现性好;高效液相色谱法测定臣药防风的含量,其方法专属性强,精密度好,准确度高、供试品溶液稳定。
Biyan tablet is recorded in Chinese Pharmacopoeia,it is helpful to cure chronic non-specific inflammation and allergic inflammation.The existing quality standards Biyan tablet is too simple to meet the requirements of modern traditional Chinese medicine quality control.The paper made systematic study on the quality standard for the Biyan tablet and established reasonal methods of analysis.The whole technological process were simple and rapid which can be used for quality control of the medicine.
Method:Uses the thin-layer chromatography to carry on the qualitative distinction of three medicinal materials in the preparation.Radix Glycyrrhizae:a silica gel G plate was adopted with developing solvent consisted of Chloroform,Acetone and Ammonia(2:7:1),an alcoholic solution of sulfuric acid(10%) was used as colouring agent,inspected at sunlight. Saposhnikovia divaricata:a silica gel G plate was adopted with developing solvent consisted of Chloroform and methanol(4:1),an alcoholic solution of sulfuric acid(10%) was used as colouring agent,inspected at 365nm.Fructus Xanthii:a silica gel GF254 plate was adopted with developing solvent consisted of Cyclohexane,ethyl acetate and formic acid(1:3:0.2),inspected at 254 nm.
Taking prim-O-glucosylcimifugin as the target,A reversed-phase high performance liquid chromatography method was developed to determine Saposhnikovia divaricata in the preparation.A Diamonsil C_(18)column(4.6*250mm, 5μm) was used.The mobile phase was consisted of methanol,acetonitrile and water(18:12:70),detected at 254nm,the velocity of flow was 1.Oml.min~(-1), temperature of the column was Room' s temperature.Taking 4' -O-β-D-glucosyl -5-O-methylvisamminol as the target,A reversed-phase high performance liquid chromatography method was developed to determine Saposhnikovia divaricata in the preparation.A Diamonsil C_(18) column(4.6*250mm,5μm) was used.The mobile phase was consisted of methanol,acetonitrile and 5%acetic acid(13:12:75) ,detected at 254nm,the velocity of flow was 1.Oml.min~(-1),temperature of the column was Room' s temperature.
Result:In the identification,the sample displayed the same spotes or fluorescence spots as the reference substance did in the Chromatography,while the negative sample did not.
In the essay,the liner ranges of prim-O-glucosylcimifugin was 0.118-2.360μg,liner regression equation was Y=24.736X+0.7825,r=0.9998.The RSD of precision was less than 3%,of repeatability was less than 3%,of recovery was less than 3%.The sample solution was stable within 12 hours(RSD=1.58%).The content of prim-O-glucosylcimifugin was can be determined by this method in the concentration of 3.9-28.4μg/ml;The liner ranges of 4' -O-β-D-Glucosyl -5-0-methylvisamminol was 0.102 - 2.040μg,liner regression equation was Y=29.379X+0.6648,r=0.9997.The RSD of precision was less than 3%,of repeata bility was less than 3%,of recovery was less than 3%.The sample solution was stable within 12 hours(RSD=1.52%).The content of 4' -O-β-D-glucosyl-5-O -methylvisamminol was can be determined by this method in the concentration of 5.7-42.2μg/ml.
Conclusion:The TLC methods for identification and HPLC method for assay have been developed and validated.The methods are simple,rapid,specific,sensitive and accurate.
方法:采用薄层色谱法对制剂中三味药材进行定性鉴别。甘草:采用硅胶G薄层板,以三氯甲烷-丙酮-氨水(2:7:1)的溶液为展开剂,10%H_2SO_4乙醇液为显色剂,在日光下检视。防风:采用硅胶G薄层板,以三氯甲烷-甲醇(4:1)的溶液为展开剂,10%H_2SO_4乙醇液为显色剂,置紫外光灯(365nm)下检视。苍耳子:采用硅胶GF254薄层板,以环己烷-乙酸乙脂-甲酸(1:3:0.2)的溶液为展开剂,置紫外光灯(254nm)下检视。
以升麻素苷为指标,建立臣药防风药材的含量测定方法。采用Diamonsil C_(18)分析柱(4.6*250mm,5μm),甲醇-乙腈-水(18:12:70)为流动相,检测波长为254nm,流速为1.0ml/min,柱温为室温,能较好的测定升麻素苷含量。以5-0-甲基维斯阿米醇苷为指标,建立臣药防风药材的含量测定方法。采用Diamonsil C_(18)分析柱(4.6*250mm,5μm),甲醇-乙腈-5%冰乙酸(13:12:75)为流动相,检测波长为254nm,流速为1.0ml/min,柱温为室温,能较好的测定5-0-甲基维斯阿米醇苷含量。
结果:在定性鉴别中,供试品色谱在与对照药材色谱相应的位置上,分别显示相同颜色的斑点或荧光斑点,阴性样品色谱在与对照药材色谱相应的位置上不显示任何颜色的斑点。
含量测定中,升麻素苷在0.118-2.360μg范围内与峰面积呈良好的线性关系,回归方程为Y=24.736X+0.7825,r=0.9998;精密度、重复性、回收率的RSD<3%;在分析浓度下,12小时内基本稳定(RSD=1.58%);本办法适合检测含量在3.9-28.4μg/ml浓度范围的供试品溶液。
此外,5-O-甲基维斯阿米醇苷在0.102-2.040μg范围内与峰面积呈良好的线性关系,回归方程为Y=29.379X+0.6648,r=0.9997;精密度、重复性、回收率的RSD<3%;在分析浓度下,12小时内基本稳定(RSD=1.52%);本办法适合检测含量在5.7μg/m-42.2μg/ml浓度范围的供试品溶液。
结论:薄层色谱法鉴别处方中三味药材,其鉴别方法快速简便,专属性强,斑点清晰,重现性好;高效液相色谱法测定臣药防风的含量,其方法专属性强,精密度好,准确度高、供试品溶液稳定。
Biyan tablet is recorded in Chinese Pharmacopoeia,it is helpful to cure chronic non-specific inflammation and allergic inflammation.The existing quality standards Biyan tablet is too simple to meet the requirements of modern traditional Chinese medicine quality control.The paper made systematic study on the quality standard for the Biyan tablet and established reasonal methods of analysis.The whole technological process were simple and rapid which can be used for quality control of the medicine.
Method:Uses the thin-layer chromatography to carry on the qualitative distinction of three medicinal materials in the preparation.Radix Glycyrrhizae:a silica gel G plate was adopted with developing solvent consisted of Chloroform,Acetone and Ammonia(2:7:1),an alcoholic solution of sulfuric acid(10%) was used as colouring agent,inspected at sunlight. Saposhnikovia divaricata:a silica gel G plate was adopted with developing solvent consisted of Chloroform and methanol(4:1),an alcoholic solution of sulfuric acid(10%) was used as colouring agent,inspected at 365nm.Fructus Xanthii:a silica gel GF254 plate was adopted with developing solvent consisted of Cyclohexane,ethyl acetate and formic acid(1:3:0.2),inspected at 254 nm.
Taking prim-O-glucosylcimifugin as the target,A reversed-phase high performance liquid chromatography method was developed to determine Saposhnikovia divaricata in the preparation.A Diamonsil C_(18)column(4.6*250mm, 5μm) was used.The mobile phase was consisted of methanol,acetonitrile and water(18:12:70),detected at 254nm,the velocity of flow was 1.Oml.min~(-1), temperature of the column was Room' s temperature.Taking 4' -O-β-D-glucosyl -5-O-methylvisamminol as the target,A reversed-phase high performance liquid chromatography method was developed to determine Saposhnikovia divaricata in the preparation.A Diamonsil C_(18) column(4.6*250mm,5μm) was used.The mobile phase was consisted of methanol,acetonitrile and 5%acetic acid(13:12:75) ,detected at 254nm,the velocity of flow was 1.Oml.min~(-1),temperature of the column was Room' s temperature.
Result:In the identification,the sample displayed the same spotes or fluorescence spots as the reference substance did in the Chromatography,while the negative sample did not.
In the essay,the liner ranges of prim-O-glucosylcimifugin was 0.118-2.360μg,liner regression equation was Y=24.736X+0.7825,r=0.9998.The RSD of precision was less than 3%,of repeatability was less than 3%,of recovery was less than 3%.The sample solution was stable within 12 hours(RSD=1.58%).The content of prim-O-glucosylcimifugin was can be determined by this method in the concentration of 3.9-28.4μg/ml;The liner ranges of 4' -O-β-D-Glucosyl -5-0-methylvisamminol was 0.102 - 2.040μg,liner regression equation was Y=29.379X+0.6648,r=0.9997.The RSD of precision was less than 3%,of repeata bility was less than 3%,of recovery was less than 3%.The sample solution was stable within 12 hours(RSD=1.52%).The content of 4' -O-β-D-glucosyl-5-O -methylvisamminol was can be determined by this method in the concentration of 5.7-42.2μg/ml.
Conclusion:The TLC methods for identification and HPLC method for assay have been developed and validated.The methods are simple,rapid,specific,sensitive and accurate.
引文
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[2]中华耳鼻喉科头颈外科杂志编辑委员会,中华医学会耳鼻喉科分会.变应性鼻炎的治疗原则和推荐方案(2004年,兰州)[J].中华耳鼻喉科头颈外科杂志,2005;40(3):166-167.
[3]Bousquet J,Van Cauwenberge P,Khaltaev N.Allergic rhinitis and its impact on asthma[J].Allergy Clin Immunol,2001;108:147-334.
[4]AsherMI,Weilland SK.The International study on Asthma and Allergy Asthma in Childhood(ISAAC),ISAAC steering Committee[J].Clin Exp Allergy,1998;5:52.
[5]顾云燕,赵邠兰,刘青.花粉症普查资料分析[J].中华耳鼻咽喉科杂志,1997;5(3):108.
[6]邹红云,李力,仇东辉等.乌鲁木齐市变应性鼻炎患病率调查及发病因素分析[J].新疆预防医学,2000;16(3):141.
[7]杜尊民.泰安市学龄前儿童变应性鼻炎流行病学调查[J].泰山医学院学报,2003;24(1):79.
[8]程雷,三好彰,殷敏.儿童青少年常年性变应性鼻炎的流行现状[J].中国中西医结合耳鼻咽喉科杂志,2000;8(2):55-57.
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