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Objective:To further confirm the anti-endotoxin active parts of Lidanpaidu prescription through observing the anti-en-dotoxin intensity based on a half-in vivo method. Methods:The extracting solution of Lidanpaidu prescription was extracted by the sol-vents with different polarity to obtain the active part. The anti-endotoxin activity of the samples was determined by using the Azo color matrix method, and the amount of inactivated endotoxin was compared among the various extraction parts with different dosages. Re-sults:Compared with that of the control group, the amount of inactivated endotoxin by the rat plasma in each administration group was all higher, and the differences were statistically significant (P0. 05). The rat plasma of the water soluble fraction administration groups also showed endotoxin inactivation, and the difference be-tween the high dosage group and the low dosage group was statistically significant (P<0. 05). Conclusion:The results of the half-in vivo method showed the consistent anti-endotoxin effect of the whole prescription and the water soluble fraction, furthermore, the dose-effect relationship is preliminarily appeared, suggesting that the water soluble fraction remains the effective components of the whole prescription to a great extent. Therefore, the water soluble fraction can be further confirmed as the active anti-endotoxin part of Lidan-paidu prescription.
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Objective: To quantitatively analyze the content of glucose in glucose injection by near infrared spectroscopy to control the quality of the product.Methods: A quantitative model was established by near infrared reflectance spectroscopy and the injection came from pharmaceutical enterprises with different concentrations of glucose and the solution samples came from laboratories with different concentrations of glucose.The liquid sample accessories were selected, a quantitative model was established by a partial least squares method, and the effect of environmental temperature on the model was studied as well.Results: According to the quantitative model, the correlation coefficient reached up to 0.999 3, and RMS deviation (RMSECV) was 0.077 4.In the verification test, the results of various liquid formulas containing glucose were similar to those of the laboratory results.The prediction error was less than 5% within the temperature range of 20-35℃.Conclusion: The model of near-infrared partial least squares (PLS) can accurately predict the content of glucose in glucose products without sodium chloride, and can be used for the quality control of glucose intermediates and the finished products in the large infusion manufacturers
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Objective:To assess the genetic diversity and phylogenetic relationship of Desmodium styraeifolium from fifteen regions of Guangdong and Guangxi Zhuang autonomous region. Methods:The molecular technique ISSR((inter-simple sequence repeat))was applied to investigate the genetic diversity of Desmodium styraeifolium from fifteen regions of Guangdong and Guangxi Zhuang autonomous region. The data was analyzed with Popgene 1. 32,and a cluster diagram was presented by UPGMA. Results:Totally 51 amplified fragments were obtained by 7 ISSR primers. The results analyzed by Popgene 1. 32 showed that the Shannon diversity index(I)was 0. 3,the NEI’s genetic diversity coefficient(H)was 0. 246 4,the coefficient of genetic differentiation (GST)was 0. 123 8,and the gene flow(Nm)was 3. 539 7. Conclusion:The above mentioned results exhibit that Desmodium styraeifolium from Guangdong,Guangxi and some wild herbal populations has high genetic diversity. The clustering results illustrate that the genetic distance of Desmodium styraeifolium originated from Guangdong and Guangxi is related with geographic distance.
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Compared with that for botanical drugs and animal-derived drugs, the identification study for mineral traditional Chi-nese medicines is relatively weak. The traditional identification methods can’ t meet the quality control requirements of mineral tradi-tional Chinese medicines, and the application of modern analysis techniques are needed urgently in the systematic research of mineral traditional Chinese medicines. In the paper, the identification of traditional methods combined with some modern analysis techniques such as X-ray diffraction, near infrared spectroscopy and Raman spectroscopy for mineral traditional Chinese medicines was summarized and analyzed to provide basic idea and methods for the systematic identification construction of mineral traditional Chinese medicines.
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This study was aimed to establish X-ray diffraction (XRD) Fourier fingerprint of mineral Chinese medicine Actinolitum, in order to provide a new method for evaluating the quality of Actinolitum. Actinolitum samples were analyzed by the technology of powder XRD. And the XRD Fourier fingerprint was determined. Accord to the fingerprints and the intensity of each characteristic peak in XRD patterns of Actinolitum, the similarity of different samples were calculated using the law of cosines and the correlation coefficient method. Systematic cluster analysis was also used for the data. The results showed that XRD patterns of 10 certified products, 3 doping products and 2 counterfeits of Actinolitum were obtained. The geometric and topological characteristics of 10 certified products were consistent. XRD fingerprint of Actinolitum from 10 certified products had 18 common characteristic peaks. The similarity analysis showed that the similarity of XRD patterns common peak of certified products were higher among 15 samples (> 0.98). The similarity of doping products was slightly lower (0.85-0.97). And the counterfeits had the lowest similarity (< 0.2). These three had significant differences which can be distinguished. The results of cluster analysis were consistent with the similarity analysis results. It was concluded that XRD fingerprint had good specificity and feasible. It was accurate and reliable, which can be used to distinguish and evaluate Actinolitum.
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This study aimed to identify Inulae Herba, Inulae Flos and their closely related species using the ITS2 bar-code, and secure the quality and clinical curative effect of these medicinal materials. DNA was extracted from Inula linariifolia, I. japonica, I. britanica, which are the original species of Inulae Herba and Inulae Flos, together with their closely related species. The ITS2 sequence was amplified by PCR and sequenced bidirectionally. Sequence assembly and generation of consensus sequence were conducted by the CodonCode Aligner. The genetic distances were comput-ed using MEGA 5.0 in accordance with the Kimura-2-parameter (K2P) model, and the phylogenetic tree constructed by the neighbor-joining (NJ) method. The results showed that the ITS2 sequences of the various species have stable variable sites. The intraspecific genetic distances among Inulae Herba and Inulae Flos were obviously lower than the interspecific genetic distance among the above two medicinal materials and its adulterants. The NJ tree based on ITS2 sequences can clearly differ from Inulae Herba, Inulae Flos and their closely related species. It is concluded that ITS2 sequence can be used as DNA barcode to identify Inulae Herba, Inulae Flos and their closely related species.
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Objective:To develop a qualitative analysis model for the fast identification of Dens Draconis and its adulterants by NIR correlation coefficient method. Methods:On the basis of the traditional morphological identification, the spectra were collected u-sing the fiber accessory of a near-infrared spectroradiometer. The reference spectra were set up using the NIR spectra of certified Dens Draconis. The characteristic spectral section was chosen and the appropriate threshold was set to establish a qualitative analysis model for the rapid identification of Dens Draconis and its adulterant. Results:The spectral section of 5 000-4 200 cm-1 was selected as the characteristic spectral section, the correlation coefficient of Dens Draconis and its adulterant was calculated in training set samples, and 92. 67% was used as the threshold. Totally 10 batches of validation set samples were validated the qualitative analysis model, and the prediction accuracy was 90%. Conclusion:The method has good prediction ability, and can be used in the rapid identification of Dens Draconis and its adulterant.
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The misuse of toxic drugsisseriousone of the threats of public health. In this study, toxic Hyoscyami Semen and its adulterants were identified by DNA barcoding. The genomic DNA was extracted from 61 samples including Hyoscyami Semen and its adulterants by reagent kit method. Their ITS2 sequences were amplified, and purified PCR products were sequenced. Sequence assembly and consensus sequence generation were performed using CodonCode Aligner v 4.25. The genetic distances, variable sites and the neighbor-joning (NJ) phylogenetic tree were computed by MEGA 6.0 in accordance with the Kimura 2-parameter(K2P) model. The results showed that the intra-specific genetic distances of Hyoscyamusniger were 0.005 which were smaller than inter -specific ones (0.360) of H. niger and their adulterants. The NJ tree showed that H. niger was clustered into one monophyletic branch, and clearly separated with other species. Therefore, ITS2 sequence was able to identify Hyoscyami Semen and its adulterants to ensure the safty of medicines.
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To establish seven kinds of minerals containing sulfate kind of near infrared spectral identification method of traditional Chinese medicine (TCM). 7 species of mineral medicine containing sulfate after crushing sieving, measure all the samples in 12 000-4 000 cm-1 section within the scope of the near infrared spectrum, spectrum signal by different pretreatment methods, after the screening of the different characteristics of the spectrum to extract the effective information, using cluster analysis method for qualitative identification. In 8 600-8 100 cm-1, 5 843-4 245 cm-1, 7 096-6 337 cm -1 section within the scope of the atlas signal after the vector normalization and multiple scattering correction, K-average clustering analysis to 20 batches sulfate kind of mineral medicine is divided into seven categories, the results of the analysis method and the traditional traits identification results are basically identical. This method is simple, fast, and can be used for these minerals containing sulfate class the qualitative identification and quality control of Chinese traditional medicine.
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This study was aimed to establish a qualitative model of near-infrared spectroscopy in order to accurately and rapidly identify several mineral Chinese medicinals from fossil including Os Draconis, Dens Draconis, Fossil Shell of Spirifer, and Fossil Crabs. The near-infrared diffuse reflectance spectroscopy combined with OPUS software was used to analyze the spectral characteristics of these samples. The pattern recognition method was explored through cluster analysis. And the accuracy of the model was verified. The results showed that these mineral Chinese medicinals from fossil had their characteristics absorption so that they can be quickly and accurately differentiated from each other through pattern recognition method. It was concluded that based on near-infrared spectroscopic mod-eling, these mineral Chinese medicinals from fossil including Os Draconis, Dens Draconis, Fossil Shell of Spirifer, and Fossil Crabs can be quickly and accurately identified.
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Objective To isolate the anti-tumor protein groups from Pinellia ternata rhizome and investigate the anti-tumor activity of these protein groups on Bel-7402 cell. Methods The total raw protein was isolated with sepharose column chromatography. Methyl-thiazolyl-tetrazolinm (MTT) was used to analyze the effect of Pinellia ternata Protein on inhibiting growth of tumor cells. Apoptosis was detected by flow cytometry (FCM). Results The 30% and 60% (NH4)2SO4 deposition part of total proteins from Pinellia ternata rhizome have no certain relationship between quantity and inhibitory action, but protein peak 3 eluted with 0.05 mol/L or 0.1 mol/L NaCl from 30% (NH4)2SO4 deposition part showed a effect of concentration depending. FCM analysis showed that the protein of 30% (NH4)2SO4 deposition part could induce apoptosis. Conclusion The 30% (NH4)2SO4 deposition part of total proteins from Pinellia ternata rhizome could significantly inhibit Bel-7402 growth and induce its apoptosis.
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Objective To discriminate Rhizoma Pinelliae from its relative species by electrophoro-gram of protein.Methods Using isoelectrofocusing(IEF) technology to compare the electrophorogram of tuber of Pinellia ternata produced in different habitats and from its relative plants then determining the pH values.Results The IEF electrophorogram of Rhizoma Pinelliae was stable with eight clear characteristic protein bands and had obvious differences from the electrophorogram of the tubers of its relative plants.Conclusion The IEF electrophoresis of Rhizoma Pinelliae can be used as the distinguishing basis.The PI value of the characteristic protein of P.ternana offers the basic parameter for separation and purification of the protein in Rhizoma Pinelliae.