| 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. |
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