桔苷灵糖浆及其原料药的质量控制与药效学的研究
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摘要
桔苷灵糖浆是本研究室以桔梗等中药有效组分为主要原料研发的现代中药新制剂,迄今为止,尚未见有相关报道。本研究以桔梗有效组分为质量控制指标,采用高效液相色谱与蒸发光散射检测器联用的方法与技术,对桔苷灵糖浆及其原料药进行了定性、定量研究,为本制剂及其原料药进一步的研发与应用建立了质量控制的方法与技术。同时对桔苷灵糖浆进行了镇咳、祛痰的研究,为桔苷灵糖浆新中药的开发及临床应用奠定了药效学的基础。
桔苷灵糖浆:1.指纹图谱定性方法与技术样品前处理:桔苷灵糖浆2mL, D101大孔吸附树脂柱(柱床:280mm×14mm),洗脱溶剂为水-乙醇系统,收集50%乙醇流分,浓缩至干,以甲醇定溶至5mL,即得样品液。色谱条件:Hanban Lichrospher C18色谱柱(4.6mm×250mm ,5μm),柱温为35℃;检测器为ELSD,漂移管温度为90℃,雾化温度为50℃,载气流速为1.4SLM,压力为0.40MPa;流动相水-乙腈,梯度洗脱,流速为1.0mL/min。在本色谱条件下,对10批次的桔苷灵糖浆样品进行了测定。结果:①样品各峰基本达到基线分离;②以桔梗皂苷D作为参照峰,归纳了9个共有峰,标准谱各峰相对保留时间为:0.0692(1)、0.5983(2)、0.6181(3)、0.7441(4)、0.9739(5)、1.0000(S)、1.0389(7)、1.1668(8)、1.2118(9);相对峰面积为:0.1186(1)、0.0424(2)、0.2314(3)、0.1716(4)、0.1691(5)、1.0000(S)、0.2513(7)、0.0381(8)、0.1313(9),建立了桔苷灵糖浆的标准指纹图谱。以相似度为指标,采用相关系数法和夹角余弦法,对指纹图谱进行了方法学考察,其结果:精密度的相似度≥0.9994、稳定性的相似度≥0.9993、重复性的相似度≥0.9969。经3批样品验证,其色谱图与标准指纹图谱的相似度均≥0.9399,表明,所建立的指纹图谱可信度高,可用于桔苷灵糖浆的质量控制。2.多指标体系的定量方法与技术样品前处理方法及其色谱条件同1,以桔梗皂苷D为标准品建立了以8种主要成分为质量控制指标的含量测定方法。对本方法进行了方法学考察,其结果:线性方程Y=1.4258X-12.449(r=0.9996),线性范围为3.96μg～19.8μg,精密度RSD=1.95%,稳定性RSD=1.15%,重复性RSD=2.47%,添加回收率平均为94.00%, RSD=2.36%。桔苷灵糖浆样品主要有效组分的平均含量为1.3633 mg/mL,RSD=6.47%。本方法灵敏、准确、重复性好、回收率高、操作简便。
原料药:1.指纹图谱定性方法与技术样品前处理:原料药10mg,精密称定,以甲醇定溶至5mL,即得样品液。色谱条件:同前。在本色谱条件下,对10批次的原料药样品进行了测定。结果:①样品各峰基本达到基线分离;②以桔梗皂苷D作为参照峰,归纳了8个共有峰,标准谱各峰相对保留时间为:0.5988(1)、0.6128(2)、0.7448(3)、0.9732(4)、1.0000(S) 1.0391(6)、1.1701(7)、1.2153(8);相对峰面积为:0.1425(1)、0.4004(2)、0.1783(3)、0.1406(4)、1.0000(S)、0.4296(6)、0.0985(7)、0.3361(8),建立了原料药的标准指纹图谱。以相似度为指标,采用相关系数法和夹角余弦法,对指纹图谱进行了方法学考察,其结果:精密度的相似度≥0.9998、稳定性的相似度≥0.9987、重复性的相似度≥0.9640。经3批样品验证,其色谱图与标准指纹图谱的相似度均≥0.9763,表明,所建立的指纹图谱可信度高,可用于原料药的质量控制。2.多指标体系的定量方法与技术样品前处理方法及其色谱条件同1,以桔梗皂苷D为标准品建立了以8种主要成分为质量控制指标的含量测定方法。对本方法进行了方法学考察,其结果:线性方程Y=1.5486X-12.073(r=0.9990),线性范围为3.96μg～19.8μg,精密度RSD=1.95%,稳定性RSD=2.47%,重复性RSD=2.44%,添加回收率平均为97.33%,RSD=2.59%。桔苷灵糖浆样品主要有效组分的平均含量为38.42%,RSD=3.33%。本方法灵敏、准确、重复性好、回收率高、操作简便,可用于原料药的质量控制。
桔梗饮片:样品前处理:对提取溶剂、提取温度、提取次数和精制方法考察,确定前处理方法:精密称取桔梗粉末0.5g,石油醚脱脂,甲醇超声提取4次(提取温度65℃),合并提取液,浓缩至干后加5mL水溶解,过滤,滤液行D101大孔吸附树脂柱(柱床:280mm×14mm),洗脱溶剂为水-乙醇系统,梯度洗脱,收集50%乙醇流分,浓缩至干,以甲醇定溶至5mL,即得样品液。色谱条件:同前。以桔梗皂苷D为标准品建立了以8种主要成分为质量控制指标的含量测定方法。对本方法进行了方法学考察,其结果:线性方程Y=1.2706X-11.947(r=0.9991),线性范围为4.168μg～20.84μg,精密度RSD=0.958%,稳定性RSD=1.33%,重复性RSD=2.98%,添加回收率平均为95.60%,RSD=1.91%。桔苷灵糖浆样品主要有效组分的平均含量为0.8005%,RSD=2.02%。本方法灵敏、准确、重复性好、回收率高、操作简便,可用于桔梗饮片的质量控制。
桔苷灵糖浆药效学:采用氨水诱发小鼠咳嗽法和大鼠毛细管祛痰法研究桔苷灵糖浆的镇咳和祛痰作用。实验结果:①氨水诱发小鼠咳嗽法,桔苷灵糖浆高、中、低剂量组潜伏期分别为50.08士12.61s、37.77士7.69s、31.00士5.85s,与正常组比较(26.08士8.68s)均能使小鼠咳嗽潜伏期延长,其中高、中剂量组差异有极显著性(P<0.0l),两者镇咳率分别达到92.02%和44.82%,低剂量组无差异,镇咳率为18.87%;与磷酸可待因组(43.25士11.60s)比较,高剂量组和中剂量组均无差异、低剂量组差异有极显著性(P<0.0l)。②大鼠毛细管祛痰法,桔苷灵糖浆高、中、低剂量组排痰量分别为:15.80士1.36cm、13.18士1.48cm、11.75士0.46cm,与生理盐水组(10.12士1.54cm)比较均能使大鼠120min内排痰量明显增加,其中高、中剂量组差异有极显著性(P<0.0l),低剂量组无差异;与氯化铵组(15.17士1.14cm)比较,高剂量组和中剂量组无差异、低剂量组差异有极显著性(P<0.0l)。上述结果表明桔苷灵糖浆具有镇咳、祛痰的作用,有开发研究价值。
桔苷灵糖浆是一正在研发中的现代中药新制剂,本文的相关研究均属首次,其成果具有创新性和重要的理论意义及实际应用价值。
A new preparation Jieganling syrup is researched in this article. It’s main material is active ingredients of platycodonis. So far, there are no relevant reports about it. This research use active ingredients of platycodonis as standards, adopt HPLC-ELSD method to carry through qualitative and quantitative study on Jieganling syrup and it’s material.Establish quality control method and technology for further study and application of Jieganling syrup and it’s material. At the same time, carry through antitussive and expectorant research of Jieganling syrup, hoping to establish a foundation for the pharmacodynamics of Jieganling syrup’s exploitation and clinical application.
Jieganling syrup:1. Fingerprint qualitative method and technology: Sample pre- treatment: Jieganling syrup 2mL, D101 macroporous resin column (column bed: 280 mm×14mm), mobile phase: water-ethanol system, collect 50% ethanol fraction, and concentrate, then dissolved with methanol, and got sample. Chromatographic conditions: Hanban Lichrospher C18 column (4.6 mm×250mm, 5μm) was used with the mobile phase consisted of water and acetonitrile at a flow rate of 1.0mL/min, gradient elution; the column temperature was set at 35℃; the detector of ELSD: the drift tube temperature to 90℃, nebulizer temperature to 50℃, air speed is 1.4SLM, the air pressure is set at 0.40MPa. In this chromatography conditions, 10 batches of Jieganling syrup samples were determined. Results:①all the peaks are basic separated;②use platycodin D as a reference peak, summed up nine common peaks, their relative retain times are as follows: 0.0692 (1), 0.5983 (2), 0.6181 (3 ), 0.7441 (4), 0.9739 (5), 1.0000 (S), 1.0389 (7), 1.1668 (8), 1.2118 (9); relative peak area: 0.1186 (1), 0.0424 (2), 0.2314 (3 ), 0.1716 (4), 0.1691 (5), 1.0000 (S), 0.2513 (7), 0.0381 (8), 0.1313 (9), and establish a Jieganling syrup reference fingerprint . Use similarity value which is counted by cosine ratio and correlation methods as the standards to carry through methodological study of the fingerprint .Results: the similarity value of precision≥0.9998, the similarity value of the stability≥0.9987, the similarity value of reproducibility≥0.9964. The similarity values of the three samples are≥0.9763, show that the fingerprint has high credibility and can be used for quality control of Jieganling syrup. 2. More standards system quantitative method and technology: sample pre-treatment method and chromatographic conditions are same to1.Use platycodin D as the reference to establish quality control method of eight major components. The result of methodological study is that: linear equation Y = 1.4258X-12.449 (r = 0.9996), the range of 3.96μg ~ 19.8μg, precision RSD = 1.95%, stability RSD = 1.15%, reproducibility RSD = 2.48%, average recovery rate of 94.00%, RSD = 2.36%, Jieganling syrup samples content 1.3633 mg / mL main active ingredients, RSD = 6.47%. This method is sensitive, accurate, reproducible, high recovery rate, easy to operate.
Material: 1. Fingerprint qualitative method and technology: Sample pre- treatment: material 10 mg , dissolved with methanol, and got sample. Chromato- graphic conditions are same to before. In this chromatography conditions, 10 batches of material samples were determined. Results:①all the peaks are basic separated;②use platycodin D as a reference peak, summed up eight common peaks, their relative retain times are as follows: 0.5988(1), 0.6128(2), 0.7448(3), 0.9732(4), 1.0000(S), 1.0391(6), 1.1701(7), 1.2153 (8); relative peak area:0.1425(1), 0.4004(2), 0.1783(3), 0.1406(4), 1.0000(S), 0.4296(6), 0.0985(7), 0.3361(8), and establish a material reference fingerprint . Use similarity value which is counted by cosine ratio and correlation methods as the standards to carry through methodological study of the fingerprint .Results: the similarity value of precision≥0.9998, the similarity value of the stability≥0.9987, the similarity value of reproducibility≥0.9964. The similarity values of the three samples are≥0.9763, show that the fingerprint has high credibility and can be used for quality control of material. 2. More standards system quantitative method and technology: sample pre-treatment method and chromatographic conditions are same to1.Use platycodin D as the reference to establish quality control method of eight major components. The result of methodological study is that: linear equation Y=1.5486X-12.073 (r=0.9990), the range of 3.96μg ~ 19.8μg, precision RSD = 1.95%, stability RSD=2.47%, Reproducibility RSD = 2.44%, average recovery rate of 97.33%, RSD=2.59%, material samples content 38.42% main active ingredients, RSD=3.33%. This method is sensitive, accurate, reproducible, high recovery rate, easy to operate.
Platycodonis: sample pre-treatment: the operating parameter: solvent, temperature, the times of extraction and refining method were optimized. Platycodonis 0.5 g, degrease, ultrasonic extraction(65℃) with methanol 4 times, merger extract, then concentrate , dissolved with water, D101 macroporous resin column (column bed: 280 mm×14mm), mobile phase: water-ethanol system, collect 50% ethanol fraction , concentrate, then dissolved with methanol, now got the sample. Chromatographic conditions are same to before.Use platycodin D as the reference to establishment quality control method of the eight major components. The results of methodological study is that: linear equation Y=1.2706X-11.947 (r=0.9991), the range of 4.168μg～20.84μg, precision RSD = 0.958%, stability RSD=1.33%, Reproducibility RSD = 2.98%, average recovery rate of 95.60%, RSD=1.91%. material samples content 0.8005% main active ingredients, RSD=2.02%. This method is sensitive, accurate, reproducible, high recovery rate, easy to operate.
Jieganling syrup Pharmacodynamics: Research on ammonia induced mice coughing and rats expectorant experiments. Results:①ammonia induce: Jieganling syrup at high, medium and low dosage groups incubation period were 50.08士12.61 s, 37.77士7.69 s, 31.00士5.85 s. Compared with the normal group (26.08士8.68 s ) , it can extend the incubation period. Among them, high and medium-dose groups had significant differences (P <0.0l), and their antitussive rate reached 92.02 percent and 44.82 percent. Low-dose group had no difference, its antitussive rate was 18.87 %. Compared with codeine phosphate group (43.25士11.60 s), the high and medium- dose groups had no difference. Low-dose group had significant difference(P <0.0l).②rats capillary expectorant experiment: Jieganling syrup at high, medium and low-dose group expectoration respectively: 15.80士1.36 cm, 13.18士1.48 cm, 11.75士0.46 cm.Compared with the normal group (10.12士1.54 cm), each of these three dose groups can increase the expectoration in 120 minutes. Among them, high and medium-dose groups had significant differences (P <0.0l), but low-dose has no difference. Compared with ammonium chloride group (15.17士1.14 cm), the high and medium-dose groups had no difference. Low-dose group had significant difference (P <0.0l). Results show that Jieganling syrup has antitussive effect and expectorant effect, and it has the value of further research.
Jieganling syrup is a new traditional Chinese medicine preparation in research. It was studied for the first time in this article. The results have innovation, important theoretical meaning and practical value.
桔苷灵糖浆:1.指纹图谱定性方法与技术样品前处理:桔苷灵糖浆2mL, D101大孔吸附树脂柱(柱床:280mm×14mm),洗脱溶剂为水-乙醇系统,收集50%乙醇流分,浓缩至干,以甲醇定溶至5mL,即得样品液。色谱条件:Hanban Lichrospher C18色谱柱(4.6mm×250mm ,5μm),柱温为35℃;检测器为ELSD,漂移管温度为90℃,雾化温度为50℃,载气流速为1.4SLM,压力为0.40MPa;流动相水-乙腈,梯度洗脱,流速为1.0mL/min。在本色谱条件下,对10批次的桔苷灵糖浆样品进行了测定。结果:①样品各峰基本达到基线分离;②以桔梗皂苷D作为参照峰,归纳了9个共有峰,标准谱各峰相对保留时间为:0.0692(1)、0.5983(2)、0.6181(3)、0.7441(4)、0.9739(5)、1.0000(S)、1.0389(7)、1.1668(8)、1.2118(9);相对峰面积为:0.1186(1)、0.0424(2)、0.2314(3)、0.1716(4)、0.1691(5)、1.0000(S)、0.2513(7)、0.0381(8)、0.1313(9),建立了桔苷灵糖浆的标准指纹图谱。以相似度为指标,采用相关系数法和夹角余弦法,对指纹图谱进行了方法学考察,其结果:精密度的相似度≥0.9994、稳定性的相似度≥0.9993、重复性的相似度≥0.9969。经3批样品验证,其色谱图与标准指纹图谱的相似度均≥0.9399,表明,所建立的指纹图谱可信度高,可用于桔苷灵糖浆的质量控制。2.多指标体系的定量方法与技术样品前处理方法及其色谱条件同1,以桔梗皂苷D为标准品建立了以8种主要成分为质量控制指标的含量测定方法。对本方法进行了方法学考察,其结果:线性方程Y=1.4258X-12.449(r=0.9996),线性范围为3.96μg～19.8μg,精密度RSD=1.95%,稳定性RSD=1.15%,重复性RSD=2.47%,添加回收率平均为94.00%, RSD=2.36%。桔苷灵糖浆样品主要有效组分的平均含量为1.3633 mg/mL,RSD=6.47%。本方法灵敏、准确、重复性好、回收率高、操作简便。
原料药:1.指纹图谱定性方法与技术样品前处理:原料药10mg,精密称定,以甲醇定溶至5mL,即得样品液。色谱条件:同前。在本色谱条件下,对10批次的原料药样品进行了测定。结果:①样品各峰基本达到基线分离;②以桔梗皂苷D作为参照峰,归纳了8个共有峰,标准谱各峰相对保留时间为:0.5988(1)、0.6128(2)、0.7448(3)、0.9732(4)、1.0000(S) 1.0391(6)、1.1701(7)、1.2153(8);相对峰面积为:0.1425(1)、0.4004(2)、0.1783(3)、0.1406(4)、1.0000(S)、0.4296(6)、0.0985(7)、0.3361(8),建立了原料药的标准指纹图谱。以相似度为指标,采用相关系数法和夹角余弦法,对指纹图谱进行了方法学考察,其结果:精密度的相似度≥0.9998、稳定性的相似度≥0.9987、重复性的相似度≥0.9640。经3批样品验证,其色谱图与标准指纹图谱的相似度均≥0.9763,表明,所建立的指纹图谱可信度高,可用于原料药的质量控制。2.多指标体系的定量方法与技术样品前处理方法及其色谱条件同1,以桔梗皂苷D为标准品建立了以8种主要成分为质量控制指标的含量测定方法。对本方法进行了方法学考察,其结果:线性方程Y=1.5486X-12.073(r=0.9990),线性范围为3.96μg～19.8μg,精密度RSD=1.95%,稳定性RSD=2.47%,重复性RSD=2.44%,添加回收率平均为97.33%,RSD=2.59%。桔苷灵糖浆样品主要有效组分的平均含量为38.42%,RSD=3.33%。本方法灵敏、准确、重复性好、回收率高、操作简便,可用于原料药的质量控制。
桔梗饮片:样品前处理:对提取溶剂、提取温度、提取次数和精制方法考察,确定前处理方法:精密称取桔梗粉末0.5g,石油醚脱脂,甲醇超声提取4次(提取温度65℃),合并提取液,浓缩至干后加5mL水溶解,过滤,滤液行D101大孔吸附树脂柱(柱床:280mm×14mm),洗脱溶剂为水-乙醇系统,梯度洗脱,收集50%乙醇流分,浓缩至干,以甲醇定溶至5mL,即得样品液。色谱条件:同前。以桔梗皂苷D为标准品建立了以8种主要成分为质量控制指标的含量测定方法。对本方法进行了方法学考察,其结果:线性方程Y=1.2706X-11.947(r=0.9991),线性范围为4.168μg～20.84μg,精密度RSD=0.958%,稳定性RSD=1.33%,重复性RSD=2.98%,添加回收率平均为95.60%,RSD=1.91%。桔苷灵糖浆样品主要有效组分的平均含量为0.8005%,RSD=2.02%。本方法灵敏、准确、重复性好、回收率高、操作简便,可用于桔梗饮片的质量控制。
桔苷灵糖浆药效学:采用氨水诱发小鼠咳嗽法和大鼠毛细管祛痰法研究桔苷灵糖浆的镇咳和祛痰作用。实验结果:①氨水诱发小鼠咳嗽法,桔苷灵糖浆高、中、低剂量组潜伏期分别为50.08士12.61s、37.77士7.69s、31.00士5.85s,与正常组比较(26.08士8.68s)均能使小鼠咳嗽潜伏期延长,其中高、中剂量组差异有极显著性(P<0.0l),两者镇咳率分别达到92.02%和44.82%,低剂量组无差异,镇咳率为18.87%;与磷酸可待因组(43.25士11.60s)比较,高剂量组和中剂量组均无差异、低剂量组差异有极显著性(P<0.0l)。②大鼠毛细管祛痰法,桔苷灵糖浆高、中、低剂量组排痰量分别为:15.80士1.36cm、13.18士1.48cm、11.75士0.46cm,与生理盐水组(10.12士1.54cm)比较均能使大鼠120min内排痰量明显增加,其中高、中剂量组差异有极显著性(P<0.0l),低剂量组无差异;与氯化铵组(15.17士1.14cm)比较,高剂量组和中剂量组无差异、低剂量组差异有极显著性(P<0.0l)。上述结果表明桔苷灵糖浆具有镇咳、祛痰的作用,有开发研究价值。
桔苷灵糖浆是一正在研发中的现代中药新制剂,本文的相关研究均属首次,其成果具有创新性和重要的理论意义及实际应用价值。
A new preparation Jieganling syrup is researched in this article. It’s main material is active ingredients of platycodonis. So far, there are no relevant reports about it. This research use active ingredients of platycodonis as standards, adopt HPLC-ELSD method to carry through qualitative and quantitative study on Jieganling syrup and it’s material.Establish quality control method and technology for further study and application of Jieganling syrup and it’s material. At the same time, carry through antitussive and expectorant research of Jieganling syrup, hoping to establish a foundation for the pharmacodynamics of Jieganling syrup’s exploitation and clinical application.
Jieganling syrup:1. Fingerprint qualitative method and technology: Sample pre- treatment: Jieganling syrup 2mL, D101 macroporous resin column (column bed: 280 mm×14mm), mobile phase: water-ethanol system, collect 50% ethanol fraction, and concentrate, then dissolved with methanol, and got sample. Chromatographic conditions: Hanban Lichrospher C18 column (4.6 mm×250mm, 5μm) was used with the mobile phase consisted of water and acetonitrile at a flow rate of 1.0mL/min, gradient elution; the column temperature was set at 35℃; the detector of ELSD: the drift tube temperature to 90℃, nebulizer temperature to 50℃, air speed is 1.4SLM, the air pressure is set at 0.40MPa. In this chromatography conditions, 10 batches of Jieganling syrup samples were determined. Results:①all the peaks are basic separated;②use platycodin D as a reference peak, summed up nine common peaks, their relative retain times are as follows: 0.0692 (1), 0.5983 (2), 0.6181 (3 ), 0.7441 (4), 0.9739 (5), 1.0000 (S), 1.0389 (7), 1.1668 (8), 1.2118 (9); relative peak area: 0.1186 (1), 0.0424 (2), 0.2314 (3 ), 0.1716 (4), 0.1691 (5), 1.0000 (S), 0.2513 (7), 0.0381 (8), 0.1313 (9), and establish a Jieganling syrup reference fingerprint . Use similarity value which is counted by cosine ratio and correlation methods as the standards to carry through methodological study of the fingerprint .Results: the similarity value of precision≥0.9998, the similarity value of the stability≥0.9987, the similarity value of reproducibility≥0.9964. The similarity values of the three samples are≥0.9763, show that the fingerprint has high credibility and can be used for quality control of Jieganling syrup. 2. More standards system quantitative method and technology: sample pre-treatment method and chromatographic conditions are same to1.Use platycodin D as the reference to establish quality control method of eight major components. The result of methodological study is that: linear equation Y = 1.4258X-12.449 (r = 0.9996), the range of 3.96μg ~ 19.8μg, precision RSD = 1.95%, stability RSD = 1.15%, reproducibility RSD = 2.48%, average recovery rate of 94.00%, RSD = 2.36%, Jieganling syrup samples content 1.3633 mg / mL main active ingredients, RSD = 6.47%. This method is sensitive, accurate, reproducible, high recovery rate, easy to operate.
Material: 1. Fingerprint qualitative method and technology: Sample pre- treatment: material 10 mg , dissolved with methanol, and got sample. Chromato- graphic conditions are same to before. In this chromatography conditions, 10 batches of material samples were determined. Results:①all the peaks are basic separated;②use platycodin D as a reference peak, summed up eight common peaks, their relative retain times are as follows: 0.5988(1), 0.6128(2), 0.7448(3), 0.9732(4), 1.0000(S), 1.0391(6), 1.1701(7), 1.2153 (8); relative peak area:0.1425(1), 0.4004(2), 0.1783(3), 0.1406(4), 1.0000(S), 0.4296(6), 0.0985(7), 0.3361(8), and establish a material reference fingerprint . Use similarity value which is counted by cosine ratio and correlation methods as the standards to carry through methodological study of the fingerprint .Results: the similarity value of precision≥0.9998, the similarity value of the stability≥0.9987, the similarity value of reproducibility≥0.9964. The similarity values of the three samples are≥0.9763, show that the fingerprint has high credibility and can be used for quality control of material. 2. More standards system quantitative method and technology: sample pre-treatment method and chromatographic conditions are same to1.Use platycodin D as the reference to establish quality control method of eight major components. The result of methodological study is that: linear equation Y=1.5486X-12.073 (r=0.9990), the range of 3.96μg ~ 19.8μg, precision RSD = 1.95%, stability RSD=2.47%, Reproducibility RSD = 2.44%, average recovery rate of 97.33%, RSD=2.59%, material samples content 38.42% main active ingredients, RSD=3.33%. This method is sensitive, accurate, reproducible, high recovery rate, easy to operate.
Platycodonis: sample pre-treatment: the operating parameter: solvent, temperature, the times of extraction and refining method were optimized. Platycodonis 0.5 g, degrease, ultrasonic extraction(65℃) with methanol 4 times, merger extract, then concentrate , dissolved with water, D101 macroporous resin column (column bed: 280 mm×14mm), mobile phase: water-ethanol system, collect 50% ethanol fraction , concentrate, then dissolved with methanol, now got the sample. Chromatographic conditions are same to before.Use platycodin D as the reference to establishment quality control method of the eight major components. The results of methodological study is that: linear equation Y=1.2706X-11.947 (r=0.9991), the range of 4.168μg～20.84μg, precision RSD = 0.958%, stability RSD=1.33%, Reproducibility RSD = 2.98%, average recovery rate of 95.60%, RSD=1.91%. material samples content 0.8005% main active ingredients, RSD=2.02%. This method is sensitive, accurate, reproducible, high recovery rate, easy to operate.
Jieganling syrup Pharmacodynamics: Research on ammonia induced mice coughing and rats expectorant experiments. Results:①ammonia induce: Jieganling syrup at high, medium and low dosage groups incubation period were 50.08士12.61 s, 37.77士7.69 s, 31.00士5.85 s. Compared with the normal group (26.08士8.68 s ) , it can extend the incubation period. Among them, high and medium-dose groups had significant differences (P <0.0l), and their antitussive rate reached 92.02 percent and 44.82 percent. Low-dose group had no difference, its antitussive rate was 18.87 %. Compared with codeine phosphate group (43.25士11.60 s), the high and medium- dose groups had no difference. Low-dose group had significant difference(P <0.0l).②rats capillary expectorant experiment: Jieganling syrup at high, medium and low-dose group expectoration respectively: 15.80士1.36 cm, 13.18士1.48 cm, 11.75士0.46 cm.Compared with the normal group (10.12士1.54 cm), each of these three dose groups can increase the expectoration in 120 minutes. Among them, high and medium-dose groups had significant differences (P <0.0l), but low-dose has no difference. Compared with ammonium chloride group (15.17士1.14 cm), the high and medium-dose groups had no difference. Low-dose group had significant difference (P <0.0l). Results show that Jieganling syrup has antitussive effect and expectorant effect, and it has the value of further research.
Jieganling syrup is a new traditional Chinese medicine preparation in research. It was studied for the first time in this article. The results have innovation, important theoretical meaning and practical value.
引文
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[2]赵树春,富丽,刘敏莉,等.桔梗等3种植物营养成分的测定[J].食品科学,1994,(4):47.
[3] Saeki T,Koike K,Nikaido T.A comparative study on commercial, botanical gardens and wild samples of the roots of Platycodon grandiflorum by HPLC analysis[J].PlantaMedica,1999,65:428～431.
[4]徐丽萍.桔梗皂苷提取方法的比较[J].食品工业科技,2007,2(04):133～134.
[5]卢朝国,赵俊廷,黄雪,等.超临界CO2萃取桔梗有效成分的工艺研究[J].河南工业大学学报(自然科学版),2007,28(1):43～45.
[6]杨献文,李苏梅,刘墨祥,等.桔梗总皂苷的制备工艺研究[J].沈阳药科大学学报,2002,19(6):441～442.
[7]杨献文,刘墨祥,刘云刚,等.大孔吸附树脂柱色谱法分离桔梗总皂苷[J].吉林农业大学学报,2001,23(2):50～52.
[8] TOSHIYUKI AKIYAMA et al.Chemical Studies on the Oriental Plant Drugs.Sapogenins of the Roots of Platycodin grandiflorum A. De,Candolle[J].Chem. Pharm.Bull,1972,20(9):1945～1956.
[9]郑毅男,刘可越,徐宝军,等.桔梗抗肥胖机理试验研究[J].吉林农业大学学报,2002,24(6):42～46,53.
[10]徐丽萍,桔梗总皂苷降血脂的研究[J].食品工业科技,2007,08:224～225.
[11]宋杨,齐云,刘彬,等.甘草桔梗皂苷对酪氨酸酶抑制的合并效应研究[J].中国实验方剂学杂志,2007,13(3):7～10.
[12]齐云,刘彬,宋杨,等.甘草、桔梗总皂苷对体外磷脂酶A2活性的影响[J].中药药理与临床,2006; 22(5):19～21.
[13] Shin CY,Lee WJ,Lee EB,et al·Platycodin D and D3 increase airway mucin release in vivo and in vitro in rats and hamsters[J].Planta Med,2002,68(3): 221.
[14]弓晓杰,陈丽荣,孙印石,等.桔梗中抑制酪氨酸酶活性成分的研究[J].中药材,2004,27(4): 257～259.
[15]严华,粟晓黎,马双成,等.重量法测定桔梗中总皂苷含量的不确定度评定[J].中国药事,2006,20 (9 ):546～548.
[16]赵耕先,黄泉秀,彭国平等,影响桔梗皂甙含量有关因素的研究[J].中草药,1987,18(6):17.
[17]许传莲,杨腊虎,郑毅男,等.应用RP-HPLC法测定不同产地中桔梗皂甙D[J].吉林农业大学学报, 1999, 21 (4):35～38.
[18]朱丹妮,舒娈,郅慧,等.HPLC-ELSD法测定桔梗中桔梗皂苷D的含量[J].植物资源与环境学报,2001,10(4):11～13.
[19]夏泉,董婷霞,詹华强,等.HPLC—ELSD法测定不同来源桔梗中桔梗皂苷E的含量[J].药物分析杂志,2006,26(5):637～639.
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