Microtox technology is one of quick toxicity test methods for determining the poisonous or toxic substances in the environment by luminous bacteria as a indicator organism.This paper reviewed the appearance and development of microtox technology.Besides,we expounded the methods and stepwise technique map and addressed key questions of microtox technology for the evaluation of the comprehensive toxicity of Chinese medicinal materials.In conclusion,microtox technology is a promising method from a vantage point of the toxicity detection of Chinese medicinal materials.

In this research,Microtox technology was used to evaluate the comprehensive toxicity of Shen-Fu injection,which was an exclusive product of the pharmaceutical company.Firstly,vibrio fischeri was used as test strain.The best test system and methodology were identified.Under the best test conditions,vibrio fischeri was firstly used in the luminescent bacteria comprehensive toxicity of the exclusive product Shen-Fu injection.The results showed that in the 2 mL reaction system,the best recovery liquid volume was 0.9 mL/ freeze-dried vial,50 μL bacterial suspensions/ sample,the detection time was 10 min after adding bacterial suspensions,the pH was 5-10,the luminous intensity was 800 000-12 000 000 for 10 rin.The RSD of replication experiment and intermediate precision test was ? and ?,respectively,which fitted to the requirements.There was no significant difference on EC50 values among 9 batches of tested Shen-Fu injection (41.07％,RSD ＜ 5％).It was concluded that there was significant concentration-effect relationship between Shen-Fu injection and the toxicity of vibrio fischeri.There was no significant difference on EC50 values among different batches.It indicated that there was small quality volatility among different batches of Shen-Fu injection.The control on product manufacturing was strong.

This study aimed at exploring the application of microtox technology to the evaluation of comprehensive toxicity of Sheng Mai injections.Characteristic parameters of vibrio fischeri (CS234) were measured by methodology inspection under different conditions to optimize the reaction condition with reliable technology.It was the first experiment to accomplish the comprehensive toxicity of Sheng Mai injections from various manufacturers using vibrio fischeri under the target condition.It was found that parameters of the optimum condition contained 0.9 mL recovery liquid volume in each freeze-dried vial and 50 μ L bacteria suspension of each sample,being included in 2 mL solution in aggregate under 10 mins' detection time after adding bacteria suspension with the favorable pH value ranging from 5 to 10 and luminous intensity from 0.8 to 1.2 million within 10 mins.The relative standard deviation values of replication experiment and precision test were all below 15％.Under this optimum detection condition,it was found that the EC50 values were 22.10％,34.10％ and 46.04％,respectively,presenting significant statistical differences (P＜0.05).These results demonstrated that the growth of biological detection standards of Sheng Mai injection was highlighted a long way to go.Besides,the microtox-based fast test for the evaluation of the comprehensive toxicity of Sheng Mai injection showed a prospective application to controlling the quality of different products from manufacturers.

In this research,we chiefly explored a new fast test method--microtox technology to evaluate the comprehensive toxicity of Shen Mai injection.We investigated characteristic parameters of vibrio fischeri (CS234) under different conditions in the pursuit of the optimum test parameters of the reliable method;and then locked the best parameters for Shen Mai injection assay with the products from different manufacturers by microtox fast test.As a result,the optimum reaction condition included 0.9 mL recovery liquid in each freeze-dried vial and 50 μL bacteria suspension of each sample within 2 mL solution in aggregate with 10-mins detection after adding bacteria suspension which pH value ranging from 5 to 10 and luminous intensity from 0.8 to 1.2 million in 10 mins.The relative standard deviation values of replication experiment and precision test were all below 15％.Under this optimum detection condition,it was found that the EC50 values were 35.60％,92.34％ and 146.57％,differing among the samples from three representative drug manufacturers,respectively (P ＜ 0.01).In conclusion,the concentration-effect relationship of Shen Mai injection existed in the toxicity assessment of vibrio fischeri with various ECs0 values from the three manufacturers.These results suggested that biological detection standards of Shen Mai injection presented great growth potential.Microtox-based fast test in the evaluation of comprehensive toxicity of ShenMai injection showed favorable application prospects over controlling the quality of different sources of products.

OBJECTIVETo establish a HPLC-PDA method for the determination of baicalin, wogonoside, baicalein, wogonin and glycyrrhizic acid in Xiaochaihu Tang.

METHODA Symmetry Shield RP18 (4.6 mm x 250 mm, 5.0 microm) was used with a mobile phase of acetonitrile-0.01% H3PO4 in gradient elution. The detection wavelength was 251 nm,the flow rate was 0.45 mL x min(-1) and the column temperature was maintained at 30 degrees C.

RESULTThe accuracy, precision, sensitivity, specificity and linearity of this method met the requirements. The contents of the five effective fractions were determined simultaneously.

CONCLUSIONThe method is rapid,simple and accurate and it can be suitable for the determination of baicalin, wogonoside, baicalein, wogonin and glycyrrhizic acid in Xiaochaihu Tang simultaneously.

Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; Flavanones ; analysis ; Flavonoids ; analysis ; Glucosides ; analysis ; Glycyrrhizic Acid ; analysis ; Indicators and Reagents ; analysis ; Medicine, Chinese Traditional ; Rehmannia ; chemistry ; Spectrophotometry, Ultraviolet ; methods ; Technology, Pharmaceutical

OBJECTIVETo comparative study the acute toxicity of four extracts from Xanthii Fructus in mice.

METHODObserved the toxic manifestations in mice which were given the four extracts by intragastric administration and calculated the LD50 of the four extracts from Xanthii Fructus.

RESULTThe toxic manifestations of the mice given water extract by intragastric administration were repose, pilo-erection, cyanosis, intention tremor, respiratory inhibition, loss of righting reflex and convulsion . The toxic manifestations of the mice given ethanol extract by intragastric administration were repose, abdominal respiration, intention tremor, intermittent convulsions, incontinence. The LD50 of Xanthii Fructus processed water extract, processed ethanol extract, crude water extract, crude ethanol extract were material drug 155.93, 317.80, 167.6, 275.41 g x kg(-1), respectively.

CONCLUSIONThe acute toxicity of water extract is distinctly stronger than that of ethanol extract, but there is no marked distinguish between crude and processed extract.

Animals ; Chemistry, Pharmaceutical ; methods ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; toxicity ; Female ; Fruit ; chemistry ; Lethal Dose 50 ; Male ; Mice ; Reflex, Righting ; drug effects ; Respiration ; drug effects ; Xanthium ; chemistry

OBJECTIVETo study the transportation of Xiaochaihu Tang in Caco-2 cell model.

METHODThe safety concentration of Xiaochaihu Tang in Caco-2 cells was determined by MTT assay. Then the Caco-2 cell model was used to investigate the bi-directional transportation of Xiaochaihu Tang. The multicomponents of Xiaochaihu Tang and the influence of time were measured by high performance liquid chromatography (HPLC).

RESULTThe P(app) values of wogonoside and wogonin were (1.23 +/- 0.09) x 10(-6), (1.07 +/- 0.89) x 10(-5) cm x s(-1) from the AP side to BL side, and (2.12 +/- 0.19) x 10(-6) and (7.12 +/- 1.02) x 10(-6) cm x s(-1) from the BL side to AP side, respectively. The P(appAP --> BL)/P(app BL --> AP) ratio of wogonoside and wogonin were 0.58 and 1.49, respectively. Baicalin, baicalein and glycyrrhizic acid could not permeate the Caco-2 cell model.

CONCLUSIONThe transportation of wogonoside and wogonin in Caco-2 cell model may be a passive transportation.

Biological Transport, Active ; Caco-2 Cells ; metabolism ; Cell Line, Tumor ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; pharmacokinetics ; Flavanones ; analysis ; metabolism ; Flavonoids ; analysis ; metabolism ; Glucosides ; analysis ; metabolism ; Glycyrrhizic Acid ; analysis ; metabolism ; Humans