Study On The Preparation And Quality Standards Of Anti-hyperostosis Oral Liquid

Hyperostosis as modern medicine name,also known as spur,osteophyte,it is a middle-aged and elderly chronic disease. As growth of age,its incidence is increasing,clinically also known as hypertrophic arthritis,degenerative arthritis,bone arthritis,joint diseases and so on.With entering the aging socie- ty,the incidence of hyperostosis have an upward trend year after year,emphasizing by medical specialists.The prescription of anti-hyperostosis oral liquid was a empirical formula summarized by traditional Chinese physician in our hospital in years of clinical practice,which had been prooved effective by a great quantity of clinical observations.The prescription was composed of Radix et Rhizoma Glycyrrhizae,Radix Rehman- niae,Herba Cistanches,Flos Carthami,Radix Paeoniae Rubra, Caulis Spatholobi,Rhizoma Cibotii,Rhizoma Drynariae, Semen Raphani,Semen Persicae,Herba Pyrolae and Herba Epimedii, which has the function of anti-hyperostosis.It had being made into pellet by manufacturing laboratory of our hospital.Because the different active substance had the different physico- chemical property,the previous preparation was less reasonable. As to keep as many effective components as possible,and reduce the dosage and enhance the effectiveness and conv- enience of administration at the same time,the experiment was carried out to make it into oral liquid by optimizing the extraction and forming condition,and then make the quality standard of oral liquid.The principal elements of experiment were as follows:Part one:Study on the optimal preparation condition of anti-hyperostosis oral liquidObjective:To optimize the extraction,impurity removal technique,and then add appropriate adjuvant to the intermediate product as oral liquid.Methods:uniform design was carried out for choosing the preparation process.With PH of first decoction,PH of second decoction,PH of third decoction and three decoction time as evaluation factors,and the content of Glycyrrhizin,glucose, naringin,icariin,and the yield of powdered extract as indexes, the optimal extracting condition for Radix et Rhizoma Glycy- rrhizae,RadixRehmanniae,Herba Cistanches,Flos Carthami, Radix Paeoniae Rubra,Caulis Spatholobi,Rhizoma Cibotii, Rhizoma Drynariae,Semen Raphani,Semen Persicae,Herba Pyrolae and Herba Epimedii.Result:1The optimal extracting condition was PH of first decoction 6.0,PH of second decoction 6.5,PH of third decoction 9.0 and three decoction time 3.7h.2After high-speed centrifugation,the supernatant approp- riately add honey, flavor,add sodium benzoate 3g,add water to 1000ml, filling,sterilization.Conclusion:Through the uniform design,the optimal extracting and forming process of anti-hyperostosis oral liquid was determined.The process was stabile with a high content of active substance.Part two:Studies on the quality standards of anti- hyperostosis oral liquidObjective:1To establish methods for identifying radix rehmann- iae,radix paeoniae rubra,herba epimedii and radix et rhizo- ma glycyrrhizae by TLC.2To develop methods for determining the contents of Naringin and Icariin in Anti- hyperostosis oral liquid by HPLC. Methods:1Identification of Radix Rehmanniae preparata:silica gel G was used as the coating material and cyclohexane-ethyl acetate (8:1) as the mobile phase, examining under ultraviolet light(365nm). Identification of radix paeoniae rubra:silica gel G was used as the coating material and the upper solution of chloroform-ethyl acetate-methanol- concentrated ammonia test solution (8:1:3:0.5) as the mobile phase,spray with 5% vanillin solution, heat at the temperature of 105℃for five minutes and examine in daylight. Identification of Radix Rehmanniae preparata:silica gel GF254 was used as the coating material and Methanol- chloroform-formic acid (1:4:0.2) as the mobile phase, examining under ultraviolet light(254nm).Identification of radix et rhizoma glycyrrhizae:silica gel G prepared 1% NaOH solution was used as the coating material and chloroform-methanol-water (6:2:0.3)as the mobile spray phase,with 10% sulphuric acid–alcohol solution,heat at the temperature of 105℃for several minutes,examining under ultraviolet light(365nm).2Determination of the contents of active substance2.1Chromatogram condition:choose appropriate stationary phase and adjust the composition,proportion and flow rate of mobile phase to separate the peak of Naringin,or Aspero- saponin well and meet the requirements of number of theoretical plates.UV spectrophotometry:choose a suitable coloration method and the wavelengths of maximal absorption.2.2The investigation of linear range:a series of the reference solutions were prepared and the regression equation was obtained with the contents of reference as abscissa and the peak area(absorbability in UV absorption spectrophotometry)as ordinate.2.3The investigation of technology:investigate the precision,reproducibility,stability and recovery.2.4Determination of the sample:take three batch of samples for six portion and prepare for the test solution, determine the content of active substance in samples.Results:1Radix rehmanniae,radix paeoniae rubra,herba epimedii and radix et rhizoma glycyrrhizae all had clear map of TLC.Sample solution showed the same color of dots in the corresponding position with reference solution,and negative control solution showed no interference.2Naringin chromatographic conditions Diamonsil (250m- x4.6mm,5um) column. mobile phase:methanol-5% acetic acid (33:67),flow rate 1.0ml/min,column temperature: 30°C,The detection wavelength was set at 283nm.Regression equation: y=1625.1x-9.6231,r=0.9997.Naringin showed good linear relationship in 0.0664ug～1.0624ug.The RSD of the precision, reproducibility and stability in twenty four-hour period were 0.76%,0.43% and 2.09% respectively.The average recovery and RSD were 100.12% and 0.64%.3Icariin chromatographic conditions Diamonsil (250m- x4.6mm,5um) column.mobile phase:acetonitrile-water (27:73), flow rate 1.0ml/min,column temperature: 25°C,The detection wavelength was set at 270nm. Regression equation: y=1617.4x+53.123,r=0.9976.Icariin showed good linear relationship in 0.0948ug～1.5168ug.The RSD of the précis- ion,reproducibility and stability in twenty four-hour period were 0.50%,1.75% and 0.97% respectively. The average recovery and RSD were 100.03% and 0.16%.Conclusion:The TLC methods had good specificity and clear spots which can be use for the accurate identification of the traditional Chinese medicine in preparation;HPLC were stable,accurate,specific.They can effectively control the quality of the preparation.