Studies On The Quality Control Of Tongren Wuji Baifeng Wan

Tongren Wuji Baifeng Wan consists of nineteen Chinese Materia Medicaincluding pullus cum Osse Nigko, Radix Ginseng, Radix Paeoniae Alba, Radix andRhizoma Salviae Miltiorrhizae and Radix Agelicae Sinesis, etc. It' s commonly used fortreating emaciation, general feebleness, aching and limpness of loins and knees,disorder of menstruation with abnormal uterine bleeding and excessive leucorrhea. Toimprove the quality of Tongren Wuji Baifeng Wan (water-honeyed pills) in anall-around way, standards for quality control were improved and established bydifferent methods. Near infrared spectroscopy (NIRS) is a rapid, cost effective andnon-destructive method allowing, besides the identification of drugs, the simultaneousdetermination of components in a mixture by multivariate data analysis. It has beenwidely used in pharmaceutical area, but few studies have reported application of NIRSto Chinese traditional patent medicine until now. In this study, a near infraredspectroscopy method was developed for qualitative and quantitative analysis of rawmaterials, excipients, semi-product and finished product of Tongren Wuji Baifeng Wan.In all, this paper provided some useful references for total quality control of large TCMformula.The main works can be summarized as follows:1. To establish the quality standard for Tongren Wuji Baifeng Wan through qualitativeand quantitative methods.Radix Paeoniae Alba, Radix and Rhizoma Salviae Miltiorrhizae, Radix Ginseng,Radix and Rhizoma Glycyrrhizae, Radix Rehmanniae Praeparata, RhizomaChuanxiong and Radix Agelicae Sinesis were identified by thin layer chromatography(TLC) method. The results showed that all the medical materials mentioned abovecould be distinguished distinctly and the procedure was selective, precise and reliable.To establish an HPLC method for the determination of paeonin and danshensu inTongren Wuji Baifeng Wan. Chromatography was carried out on a C₁₈ column. For determination of paeonin, the mobile phase consisted of methanol-water (30:70),UV-detection was accomplished at 232 nm; for determination of danshensu, the mobilephase consisted of methanol-0.5% acetic acid (8:92), UV-detection was accomplishedat281 nm.Amino acids were determined by automatic amino acid analyzer(AAAA).Chromatography was carried out on a column of cation exchange resin. AAswere separated with five citrate buffers. Detection was by spectrophotometry at 570and 440 nm with the ninhydrin reaction.Accurate and repeatable results were achieved by simple procedures. The proposedmethods were suitable for the quality control of Tongren Wuji Baifeng Wan.2. To establish methods for the determination of parts of harmful residues in TongrenWuji Baifeng Wan.Atomic absorption spectrophotometry was used for determination of Lead, Cadmiumand Copper; atomic fluorescence spectrometry for Arsenic and Mercury; gaschromatography for organochlorine pesticides and organophosphorus pesticidesresidues; gas chromatography mass spectrometry for pyrethrins pesticides residue.The methods were accurate, reliable and applicable for control of harmful residues inTongren Wuji Baifeng Wan. Meanwhile, the proposed methods provide reference fordetermination of harmful residues in TCM formulae.3. To establish the fingerprints of the semi-product and finished product of TongrenWuji Baifeng Wan by NIRS method.Wuji Baifeng Wan is a TCM having similar name with Tongren Wuji Baifeng Wanbut different ingredients. To distinguish Tongren Wuji Baifeng Wan with other product,NIR diffuse reflectance spectra of semi-products, finished products, negativesemi-product samples and Wuji Baifeng Wan samples produced by differentpharmaceutical factories were colleted. The spectra of 75 semi-products were used as the fingerprint database, based on which the similarity match model was built. Usingthis model, the similarity match values of the rest semi-products and negativesemi-product samples were computed. Another similarity match model was establishedon fingerprint database formed by spectra of 70 batches of Tongren Wuji Baifeng Wan.Then the similarity match values of the rest Tongren Wuji Baifeng Wan and WujiBaifeng Wan samples were computed.The results showed that the proposed method could distinguish the semi-productsfrom negative samples, Tongren Wuji Baifeng Wan from Wuji Baifeng Wan,respectively. NIRS fingerprinting was proved to be a simple, fast and non-destructiveway for integrated quality analysis of TCM.4. To develop qualitative and quantitative methods for analysis of raw materials,excipients, semi-product and finished product of Tongren Wuji Baifeng Wan by NIRSin combination with chemometrics.A standard NIR spectral library of 18 kinds of raw materials was constructed, basedon which the unknown materials were verified by QC compare search.Discriminant analysis based on principle component analysis (PCA) andMahalanobis distance was used in identification of pullus cum Osse Nigko, thesemi-product and finished product, respectively.Calibration models determining paeonin in Radix Paeoniae Alba, salvianolic acid Bin Radix and Rhizoma Salviae Miltiorrhizae, total content of ginsenoside Rg₁ and Re,ginsenoside Rb₁ in Radix Ginseng, paeonin and water content in semi-product as wellas total amino acids, paeonin and water content in finished product were built bypartial least squares (PLS) regression, respectively.It was proved that the qualitative methods could distinguish all samples accuratelywhile the quantitative methods were precise, reliable, simple, fast andenvironmental-friendly, and could be applied to at-spot detection.