Study On Quality Control Methods Of Qingkailing Tablet

Qingkailing tablet is one of extensively used Chinese patent drugs on clinic.The formula consists of cholic acid,hyodeoxycholic acid,baicalin,bubali cornu,gardenia fructus,lonicerae japonicae flos,isatidis radix and margaritifera concha.It is mainly used to treat colds,fever caused by exogenous pathogenic wind heat,upper respiratory tract infection and so on.The current standard of Qingkailing tablet in Chinese Pharmacopoeia includes qualitative analysis of chlorogenic acid,qualitative and quantitative analyses of cholic acid,hyodeoxycholic acid,baicalin and geniposide as well as determination of total nitrogen.Bad specificity,complicated operations and lack of comprehensiveness are disadvantages of these current methods.The study was started with analyzing the formula of Qingkailing tablet to make the standard more complete and accurate.Based on the index components of each crude drug,the research has been developing a diversified strategy as follows: 1.A qualitative method of bubali cornu in Qingkailing tablet based on Taq Man Prober real-time polymerase chain reaction(PCR)was established.DNA was amplified with specific primers of buffalo and then fluorescence signal was determined.The identification was specific that showing positive result in positice control group while showing no amplification in black and negative control group.In the detection system,minimum detectable concentration of buffalo DNA was up to 0.2ng/m L,and the result came to positive when the mass fraction of bubali cornu reach 0.2%.2.A pre-column derivation and HPLC method to determine amino acids in Qingkailing tablet was established.Phenyl isothiocyanate(PITC)was used as the deviatization reagent,and a gradient elution program was used to separate 18 kinds of amino acids.The new detection system separaterd amino acids well,and showed good linear.The average recoveries(n=6)ranged from 95.2% to 106.6%(RSD<3.0%).3.A HPLC method for simultaneous determination of chlorogenic acid,gardenia,and baicalin was established.A gradient elution program was used to separate the three components at detection wavelength of 238 nm.Specificity and linearity test were both fine,and the average recovery(n=9)of chlorogenic acid,gardenia,and baicalin were 100.7%,100.1% and 96.0%(RSD<2.0%),respectively.All the feasible methods above were of high specificity,sensitivity and accuracy.They can be applied to identify bubali cornu in Qingkailing tablet and to determine the indicative components of the crude drugs of bubali cornu,gardenia fructus,lonicerae japonicae flos,isatidis radix and margaritifera concha.The new methods that the study established may facilitate the quality control of Qingkailing tablet and lead to a more comprehensive and convenient pharmaceutical standard.