[Modeling of thermal degradation of linarin during concentration process].

Ganmaoling granule, with annual sale of over one billion yuan, is the first brand of domestic cold medicine sales. As the only traditonal Chinese medicine（TCM） quality control indicator of Ganmaoling granule, linarin is thermally unstable. Its content will be changed significantly during the production process, which would then affect the quality of the finished product. In this paper, the law of degradation of linarin was investigated. The experimental results showed that degradation reaction of linarin belongs to the first reaction characteristics. The effective methods to reduce the loss of linarin would be realized fortunately by strictly controlling the heating temperature or shortening the heating time.

[Study on balance of process of alcohol precipitation of ganmaoling granules].

Ganmaoling granule is the first brand of domestic cold medicine sales, but its preparation method and process control parameters are relatively rough. Therefore it is urgent to upgrade the technologies of large varieties of traditional Chinese medicine (TCM). This paper focused on the balance between the remove of impurity and the retention of linarin during the process of alcohol precipitation of Ganmaoling granules. The effects of four factors on the process were investigated via single factor experiments. The results showed that the precipitating period, the initial ethanol concentration and the final ethanol concentration had a great effect on retention of linarin while the initial density of the extract has not. Similarly, the initial ethanol concentration, the final ethanol concentration and the initial extract density have a great effect on the yield of dry extract while the time of alcohol precipitation has not. The parameters of alcohol precipitation of Ganmaoling granules were optimized as 16 h of precipitating period, 95% ethanol as the initial reagent, 70% of the final ethanol concentration, and 1.10 of the initial extract density.

[On-line monitoring of extraction of coldrine by near-infrared spectrum].

Extraction of the four Chinese herbals is the beginning step of the production process of coldrine granules and influences on drug quality significantly. In this paper, the on-line near infrared spectrum was collected during the extraction process of coldrine and then pre-processed by the first derivative. Partial least square regression (PLSR) model was developed for the quantity indicators of linarin, chlorogenic acid and solid content, according to results of both HPLC and weight-loss as reference methods. The correlation coefficient, root mean square error of cross-validation (RMSECV), root mean square error of calibration (RMSEC) and root mean square error of prediction (RMSEP) were used to optimize model parameters and confirm their performance. Correlation coefficients of three quality control indicator models reached more than 0.95.Values of RMSEC of linarin, chloroenic acid and solid content were 0.010 4, 0.009 34 and 0.055 5, respectively. And the values of RMSEP were 0.009 47, 0.142 and 0.008 42, respectively. The models, built on-line analyze data, revealed that the correlation coefficients of predicted values and measured values were greater than 0.97 and values of RSEP of linarin, chloroenic acid and solid content were 8.14%, 8.17% and 9.86%, respectively. The results showed that the NIR method could achieve the on-line detection and real-time monitoring of multi-indexes during the extraction process of coldrine. The technology could be used for drug quality control in the process of practical production, reducing the batch differences and ensuring pharmaceutical quality stability. In addition, it could provide real-time production data for subsequent product quality backtracking.

[Study on main pharmacodynamics and underlying mechanisms of 999 Ganmaoling].

To observe synergistic effects of 999 Ganmaoling (GML) and its Chinese/Western materia medica (CMM and WMM) on pharmacodynamic action and to study underlying mechanisms, their anti-inflammatory, antipyretic effects were compared by assaying the increased capillary permeability induced by glacial acetic acid in mice, ear swelling induced by Xylene in mice, non-specific pleurisy induced by carrageenan in rats, and yeast induced fever in rats. Crystal violet (CV) and microbial activity (XTT) assay were used to evaluate the inhibition of GML and its CMM and WMM on KPN biofilm formation, and scanning electron microscopy (SEM) was applied for observing KPN biofilm morphology changes. The results showed that compared with control group, GML could reduce exudation amount of Evans-Blue and the degree of Ear swelling significantly, and CMM and WMM have no significant effects. The concentration of TNF-α and IL-1ß of rat pleural effusion in GML, CMM and WMM group decreased significantly. The concentration of TNF-α, IL-1ß and IL-8 in GML group, TNF-α, IL-8 in WMM group and IL-8 in CMM in rats serum decreased significantly. The body temperature in rats decreased significantly in GML and WMM group after 4-8 h of administration. CMM group showed no significant difference in rat body temperature compare with control. Compared with control group, GML (55-13.75 g•L⁻¹) could inhibit KPN biofilm formation and reduce number of viable cells in the KPN biofilm. CMM (45-22.5 g•L⁻¹) and WMM (10 g•L⁻¹) could also inhibit KPN biofilm formation and reduce number of viable cells (P<0.01). Result of SEM also showed that GML (55 g•L⁻¹) and its CMM (45 g•L⁻¹) and WMM (10 g•L⁻¹) could interfere the bacterial arrangement of KPN biofilm and extracellular matrix. GML and its CMM & WMM could inhibit the formation of KPN biofilm, CMM & WMM in GML showed synergism and complementation in inhibit KPN biofilm. Results showed that GML had obvious anti-inflammatory and antipyretic effects and could destruct KPN mature biofilm. WMM and CMM showed obvious synergistic effect against inflammation and inhibition of KPN biofilm formation and reduction of number of viable cells but no same effects against fever.

[Study on toxicity of 999 Ganmaoling grain and influence of diet on hepatic toxicity].

This paper was aimed to compare the acute toxicity of 999 Ganmaoling grain and its different ingredients, and investigate the influence of routine diet on the hepatic toxicity induced by Ganmaoling in mice, so as to provide experimental basis for the clinical safety evaluation. Mice were given a single dose of Ganmaoling grain or its different ingredients respectively by gavage, and then observed for 14 days. LD50 values of Ganmaoling grain or its chemical ingredient and the maximal tolerated dose of its herb ingredient were determined. Mice were divided into starvation and diet group, a single dose of Ganmaoling grain was administered by gavage. LD50 values were estimated after 14 day observation. Mice were divided into starvation and diet group. At the same time,control group was set up for each. A single dose of Ganmaoling grain was given. Serum biochemical indexes were detected, liver weight index was calculated and liver tissue morphological change was observed after 6 h. LD50 values were 4.42, 0.64 g•kg⁻¹ for Ganmaoling grain group and chemical ingredient group, respectively. The maximal tolerated dose of the herb ingredient group was close to 24.24 g•kg⁻¹. The toxic symptom was basically similar in the Ganmaoling grain and the chemical ingredient group. The body weight and food intake were decreased to a certain extent in both groups. There were pathological changes of liver and heart tissue in some of the surviving animals. The animals in the Ganmaoling grain group exhibited a lighter toxicity and recovered faster than that in the chemical ingredient group. LD50 values of Ganmaoling grain were 2.56, 6.93 g•kg⁻¹ for starvation and diet group respectively. TD50 values were 1.29, 6.31 g•kg⁻¹ for starvation and diet group respectively. The toxicity of 999 Ganmaoling was less, which may be related to the reduction of toxicity after the combination of herb and chemical ingredients. Compared with starvation group, the values of LD50 and TD50 of diet group was significantly increased, and toxicity was decreased. From the point of view of safety, it is safer to use Ganmaoling in the absence of hunger or after meal. The above tests provide experimental basis for the clinical safety use of Ganmaoling.

[Separation and purification of total flavonids of Astragalus membranaceus with ethanol/phosphate aqueous two-phase system].

OBJECTIVE: To study separation and purification of flavonids with ethanol/phosphate aqueous two-phase system. METHOD: The diversity of phase separation ability and the distribution of target products in various systems were taken as indicators to analyze aqueous two-phase extraction systems and phase diagrams formed by ethanol and some common salts, screen out EtOH/ K2HPO4 system as the optimla system for extracting total flavonids, and study the impact of proportion of components in EtOH/K2 HPO4 system on the partition coefficient and phase ratio of flavonids. RESULT AND CONCLUSION: The EtOH/K2 HPO4 system with omegaEtOH 36.05% and omegaKHPO4 18.20% has been proved as the optimal conditions for separating and purifying total flavonoids of Astragalus (TFA). Under this optimal condition, the partition coefficient and the extraction yield of TFA reached 10.33 and 96.6%, respectively. After extraction, the contents of A. membranaceus saponins and A. membranaceus polysaccharides in top and bottom phases were determined at the same time, showing that A. membranaceus saponins in the removal rate reached 92.01%, and A. membranaceus polysaccharides were totally concentrated in bottom water phase, indicating a removal rate of 100%. Therefore, this is beneficial to separate and purify total flavonids from A. membranaceus crude extracts.

[Separation and purification of flavonoids from Smilax glabra by macroporous adsorption resin].

OBJECTIVE: To investigate the process of separating and purifying flavonoids from Smilax glabra. METHOD: With the yield of flavonoids as index, the optimum process of separating and purifying flavonoids from S. glabra Roxb was screened by static and dynamic adsorption tests. RESULT: The static saturated adsorption capacity of D101 macroporous resin to flavonoids of S. glabra was 45.6 mg x g(-1) (dry resin). The optimum conditions of dynamic adsorption and elution were as that the pH, the concentration, the adsorption velocity of the extracting solution, and the adsorption capacity were 6.00 +/- 0.20, 4.2 mg x mL(-1), 2 mL x min(-1) and 15 mL, respectively. The adsorbed resin column was washed by 100 mL 60% ethanol with pH value of 8.00 +/- 0.20 at the eluting velocity of 3 mL x min(-1) after washed by 100 mL distilled water. CONCLUSION: The flavonoids of S. glabra was able to be easily separated and purified by D101 macroporous resin under the optimum conditions above, and the recovery rate was higher than 90%. The content of obtained flavonoids reached 62.6%, which was 2 times of the content before purification.

[Extracting flavonoids from Smilax glabra by microwave-assisted method].

OBJECTIVE: To optimize the process of extracting flavonoids from Smilax glabra. METHODS: Flavonoids were extracted from Smilax glabra by microwave-assisted method, and the extracting time, microwave power, ethanol concentration, solid-solvent ratio and extracting temperature were optimized through single factor experiment and orthogonal test. RESULTS: The optimun process parameters of extracting flavonoids were as follows: the extracting time, microwave power, ethanol concentration, solid-liquid ratio and extracting temperature were 5 minutes, 600 W, 60%, 1:30 and 80 degrees C, respectively. CONCLUSION: The proposed method has been applied stably and reliably to the extraction of flavonoids from Smilax glabra which has the advantages of high recovery and short extraction time. The recovery, the yield and the content of falvonoids are 93.2%, 2.66% and 32.6%, respectively.

[Experimental study on the pharmacology of 999 ganmaoling, a compound recipe of Chinese and Western materia medica].

OBJECTIVE: To study the pharmacologic characteristics in synergism and complementation of 999 Ganmaoling (GML), a compound recipe composed of Chinese and Western materia medica (CMM & WMM), as well as its theoretical basis of matching of Chinese and Western materia medica. METHODS: The torsion response induced by glacial acetic acid in mice, toe swelling induced by carrageenanin rats, delayed hypersensitive response in mice and fever induced by endotoxin in rats and rabbits were used to comparatively study the actions of CMM & WMM in GML. The effect of CMM in antagonizing liver damage caused by WD (acetaminophen) in mice was also studied. RT-PCR method was used to analyze the expression of related cytokines. RESULTS: GML showed a significant antipyretic and analgesic effect, it could inhibit the carrageenan induced inflammation, antagonize the endotoxin induced fever, and promote the amount for expression of cytokines in rats' splenic tissue with pneumococci infection to some extent. The CMM in GML showed certain protective effect on acetaminophen induced liver damage. CONCLUSION: GML has a potent antipyretic, analgesic and anti-inflammatory effects, CMM & WMM in GML showed markedly synergism and complementation, and CMM in it has liver protective effect.

[Clinical observation on effect of taizhi'an capsule in treating 150 patients with hyperlipidemia].

OBJECTIVE: To grasp the therapeutic effect of Taizhi'an (TZA) in lowering blood lipid level. METHODS: Three hundred patients with hyperlipidemia were randomly divided adopting numerical table method into 7 groups, the Taizhi'an group (A, n = 90), the half-dose Fenofibrate plus Taizhi'an group (B, n = 30), the full-dose Fenofibrate group (C, n = 30), the half-dose Simvastatin plus Taizhi'an group (D, n = 30), the full-dose Simvastatin group (E, n = 30), the Zhibituo group (F, n = 60) and the Xuezhikang group (G, n = 30). The effect in different groups were compared after 8 weeks treatment. RESULTS: In group A, the total cholesterol (TC) lowered by 12.7%, triglyceride (TG) lowered by 22.1% and the high density lipoprotein cholesterol (HDL-C) increased by 13.1%, the total effective rate being 82%. The therapeutic effect in group B was similar to that in group C, and that in group D was similar to that in group E (P > 0.05). The therapeutic effect of Taizhi'an was similar to that of Xuezhikang and Zhibituo, but was better than Zhibituo in lowering TG, LDL-C and increasing HDL-C, and better than Xuezhikang in lowering TG and increasing HDL-C (P < 0.05). CONCLUSION: When Taizhi'an used in combination with half-dose Simvastatin, it could not only enhance the blood lipid regulatory effect of Simvastatin but also reduce the dosage used and alleviate its adverse reaction. Compared with Xuezhikang and Zhibituo, Taizhi'an got the similar therapeutic effect, but was superior in regulating blood lipids.