Studies On The Extraction And Purification Of Saponins In Seeds Of Nigella Glandulifera On Part Of The Respiratory Efficacy

Objective: To study the extraction and purification of saponins in Uyghur Medicine ofthe seeds of Nigella Glandulifera on relieving cough, antiasthma, expelling phlegm andanti-inflammatory. Methods:1. The preliminary study of the main chemical constituentsof Nigella Glandulifera: by way of in vitro pre-test method and special test, to study themain chemical constituents of Nigella Glandulifera preliminarily. To assay the contents oftotal saponins from Nigella Glandulifera by HPLC with hederagenin and oleanic acid asreference substances.2. Extraction and purification of total Saponins: to determine thecontent of total Saponins by UV and to optimize the extraction technology of totalsaponins from the seeds of Nigella Glandulifera by Central Composite Design/ResponseSurface Methodology. Study the separation and enrichment elution conditions of AB-8type macroporous resin: the loading concentration, pH, the ethanol concentration andusage, the elution flow rate.3. Investigated the effects of saponins extracts on coughfrequency, incubation period induced by strong aqua, spray induced asthma, the influenceof phenol red secretory volume in trachea, external ear swelling of mice induced bydimethyl benzene, voix pedis swelling of rats induced by carrageenan and tube doubledilution method. Through investigating the weight growth and index changes, expound theeffect of total saponins from Nigella Glandulifera on relieving cough, antiasthma,expelling phlegm, anti-inflammatory and bacteriostasis. Results:1. Preliminary studieshave shown that the seed of Nigella Glandulifera mainly consists of volatile oil, grease,alkaloids, flavonoids, saponins, carbohydrate, organic acids, amino acids, peptides orproteins and so on. A high performance liquid chromatography method for determinationof the content of hederagenin and oleanic acid was established in seeds of NigellaGlandulifera at the same time. The linear ranges of hederagenin and oleanic acid were respectively22.76μg/mL~227.6μg/mL (r=0.9998) and40.28μg/mL~201.4μg/mL(r=0.9997). Their average recoveries were respectively94.6%and93.8%.The totalsaponins accounted for4.76%.2. The optimum conditions for ultrasonic extraction were12times the amount of50%ethanol, and90minutes every once under60℃, the loadingconcentration was determined as3.93mg/mLand eluded with50%ethnol with8BV. Theelution flow rate was determined at4BV/h and the rate of total saponins was62.1%withthe methods as above.3. The results showed that saponins extracts could prolong coughincubation period obviously, the high dose group significantly reduced the number of micecough within2minutes and increased the amount of phenol red excretion, agonizedexternal ear swelling of mice induced by dimethyl benzene and voix pedis swelling of ratsinduced by carrageenan. Conclusion: The initial results showed that the total saponinswhich accounted for62.1%from the purification of seeds of Nigella Glanduliferapossessed the relieving cough, expelling phlegm and anti-inflammatory effects, amongwhich the expelling phlegm is the strongest actions followed by anti-inflammation.