OBJECTIVE:To develop a method for simultaneous determination of harpagide,harpagoside,chlorogenic acid, caffeic acid and phillyrin in Xiaoer qingyan granules. METHODS:HPLC method was adopted. The determination was performed on Zorbax SB-C18 column with mobile phase consisted of acetonitrile-0.1% phosphoric acid solution (gradient elution) at a flow rate of 1.0 mL/min. The detection wavelength was set at 210 nm (0-13 min,harpagide),327 nm (13-25 min,chlorogenic acid and caffeic acid),277 nm(25-29 min,phillyrin),210 nm(29-40 min,harpagosid);the column temperature was 30℃,and sam-ple size was 10 μL. RESULTS:The linear ranges of harpagide,harpagosid,chlorogenic acid,caffeic acid and phillyrin were 8.400-168.0 ng(r=0.9996),11.30-226.0 ng(r=0.9998),128.8-257.6 ng(r=0.9993),8.110-162.2 ng(r=0.9996),29.69-593.8 ng(r=0.9994),respectively. LOQs of the 5 components were 33.39,451.2,515.2,324.5,1188 ng/mL;LODs were 8.348, 112.8,128.8,81.12,297.0 ng/mL,respectively;RSDs of precision,stability and reproducibility tests were all lower than 2.0%. The recoveries were 96.39%-98.64%(RSD=0.83%,n=6),96.60%-98.89%(RSD=0.89%,n=6),96.28%-99.22%(RSD=1.25%,n=6),96.49%-99.54%(RSD=1.16%,n=6),96.26%-99.70%(RSD=1.30%,n=6),respectively. CONCLUSIONS:The method is simple,accurate and suitable for simultaneous determination of 5 components in Xiaoer qingyan granules

Objective: To develop a RP-ion pair HPLC with wavelength switching for the simultaneous determination of 4 components (marine, oxymarine, salicylic acid, benzoic acid) in Fufang Kusheng Shuiyangsuan powder.Methods: An Agilent ZORBAX SB-C18 column(150 mm× 4.6 mm, 5 μm) was used;the mobile phase was acetonitrile (A)-0.1% phosphoric acid solution (0.2 g sodium heptanesulfonate was added to 100 ml solution) at a flow rate of 1.0 ml·min-1;the detection wavelength was 220 nm in 0-12 min and 280 nm in 12-25 min;the column temperature was 30℃.Results: The linear range of marine, oxymarine, salicylic acid and benzoic acid was 0.006 030-0.120 6 μg (r=0.999 4), 0.016 56-0.331 2 μg (r=0.999 9), 0.717 1-14.34 μg (r=0.999 9) and 0.512 0-10.24 μg (r=0.999 9), respectively;the average recovery was 98.14%, 97.20%, 97.05% and 98.39% with the RSDs of 1.38%, 0.32%, 0.81% and 1.26%(n=6) , respectively.Conclusion: The method is simple and rapid, and can be applied in the simultaneous determination of 4 components in Fufang Kusheng Shuiyangsuan powder.

AIM: To study the application of near infrared spectroscopy for quality control of in-process Wuji Baifeng Pill(Gallus Domesticus,Colla Cornus cervi,Carapax Trionycis,Otheca Mantidis,Radix et Rhizoma Ginseng,etc.). METHODS: Qualitative and quantitative methods for analysis of in-process product was developed by diffuse reflectance near infrared spectroscopy combined with chemometrics.Spectra of 95 samples of Tongren Wuji Baifeng Pill in-process and 19 raw material negative in-process products were collected.Similarity match model was built on the basis of spectra of 75 batches of qualified in-process products.Using this model,the similarity match values of other samples were calculated.Multivariate calibration models for paeonin and water content in in-process product were developed with partial least square algorithm. RESULTS: The results showed that the proposed similarity match model could reflect the quality characteristic of Tongren Wuji Baifeng Pills in-process and reveal the absence and presence of raw materials.The correlation coefficients of the calibration model were within 0.965 7 and 0.992 1 while the values of root mean square error of prediction were 0.005 5% and 0.45%,respectively. CONCLUSION: The experiment shows that the established method is accurate,fast and non-destructive,and can be applied to on-line detection.

AIM:To improve the quality control standard based on 3 kinds of Radix Isatidis preparations-Radix Isatidis syrup,chewable tablet and granules. METHODS: A TLC method was established for iden tification of with Isatidis indigotica RS,Vleucine CRS and Arginine CRS as the references.A HPLC method was developed for the determination of adenosine and igoitrin in Radix Isatidis Preparations,with DIKMA Diamonsil C_(18) column(5 ?m,200 mm?4.6 mm id),a mixure of acetonitrile-water-triethylamine(10∶90∶0.2,using glacial acetic acid to adjust to pH 5.2)-water(85∶15) as the mobile phase.The detection wavelength was set at 245 nm. RESULTS: Isatidis indigotica,Vleucine and Arginine were detected in all the preparations.The contents of adenosine and igoitrin varied greatly in tested samples from different pharmaceutical enterprises. CONCLUSION: The above methods are simple,accurate,and suitable for the quality control of Radix Isatidis Preparations.

BACKGROUND: It was uncertain that the migration, differentiation and survival of transplanted bone marrow mesenchymal stem cells (BMSCs) into myocardium after the acute myocardial infarction. OBJECTIVE: To investigate the migration, differentiation and survival of rabbit transplanted autologous BMSCs in myocardium after the acute myocardial infarction. METHODS: Rabbit BMSCs were isolated and labeled by DAPI in vitro. Rabbit left anterior descending branch was ligated to establish acute myocardial infarction models. Following successful model establishment, 30 New Zealand rabbits were assigned to BMSC and control groups (n = 15). In the BMSC group, autologous BMSCs were infused into the surrounding sites of the infracted region by 4 points 1 hour following coronary artery ligation. In the control group, the same region was injected with an equal volume of saline. Injection volume was 30 μL in each point. Five animals from each group were sacrificed 10 minutes, 3 days and 4 weeks following transplantation. The heart was obtained to undergo frozen sections. The distribution of DAPI-labeled BMSCs was observed using fluorescence microscope. Immunofluorescence method was used to examine the troponin Ⅰ and α-actin. RESULTS AND CONCLUSION: DAPI-labeled BMSCs with blue nuclei were distributed extensively in the myocardium of the cell transplantation group, ovoid in shape and arranged in parallel with the cardiac muscle fibers. Troponin Ⅰ and α-actin were positive immunofluorescently in the cytoplasm of the labeled BMSCs. Results indicated that transplanted BMSCs in the ischemic myocardium could differentiate into myocardial cells under stimulation of local microenvironment.

Objective To further prove the multipotentiality of rabbit bone mesenchymal stem cells (MSCs) by inducing it into cardiomyocytes with 5-azacytidine. Methods MSCs from rabbit were abstracted, isolated, cultured. Then MSCs were induced by 5-azacytidine in vitro for 24 hours. After cultured for 4 weeks, MSCs’ differentiation was studied by immunohistochemistry and electron microscopy technique. Results Some of MSCs induced by 5-azacytidine expressed troponinT . Electron microscopy showed myofilaments. Conclusion MSCs which exist in bone marrow are mutipotential stem cells, and they can be induced into cardiomyocytes by 5-azacytidine in vitro.

OBJECTIVETo apply the near infrared spectroscopy in quality control for excipients of traditional Chinese medicine (TCM).

METHODA new type of transmittance and reflectance integration sphere accessory and a transmittance accessory were used to collect the spectra of processed honey and rice wine, respectively. Determination method of reducing sugar and ethanol in processed honey were established with partial least squares (PLS).

RESULTThe correlation coefficients (r), the root mean square error of calibration (RMSEC) and the root mean square error of prediction (RMSEP) of the proposed models were 0.9593, 1.00% and 1.19% for reducing sugar; 0.9529, 0.17% and 0.24% for ethanol.

CONCLUSIONThe proposed method is fast, non-destructive, simple and accurate, and can be applied to at-spot detection,

Excipients ; chemistry ; Honey ; analysis ; Medicine, Chinese Traditional ; Oryza ; chemistry ; Quality Control ; Spectroscopy, Near-Infrared ; methods ; Wine ; analysis

Objective To establish limit test method of camphor in Niuhuang qingxin pills (prescription of the bureau) . Methods The separation was performed on an HP-INNOWAX capillary column (0.25 mm×30 m, 0.25 μm) and the column temperature was 110 ℃ . The temperatures of the inlet and the FID detector were 200 and 230 ℃ , respectively. The flow rate of the carrier gas was 1.8 mL·min-1 and the split ratio was 10:1. The injection volume was 1 μL. Results Good linearity was obtained for camphor within the range of 1.65 to 165 μg·mL-1, and the average recovery of low, medium, high concentration were 99.12％, 99.56％ and 99.56％, respectively,with RSD were 1.01％,0.69％ and 0.38％, respectively. Trace camphor was found in 7 samples out of 38 samples from 9 manufactures. Conclusion The proposed method was accurate, sensitive and simple, and was suitable for quality evaluation and safety control of Niuhuang qingxin pills (prescription of the bureau) .

Objective: To establish a GC method for the determination of borneol and isoborneol in Niuhuang Qingxin pills ( pre-scription of the bureau). Methods: A capillary column HP-INNOWAX(30 m×0. 25 mm,0. 25 μm) was used and the column tem-perature was kept at 110 ℃. The temperature of the inlet and the FID was 200 ℃ and 230 ℃, respectively. The carrier gas was N2 and the flow rate of the carrier gas was 1. 8 ml·min-1. The split ratio was 10: 1, and the injection volume was 1 μl. Results: The linear response range of borneol and isoborneol was 0. 01-5. 09 μg·ml-1(r=0. 999 5) and 0. 01-5. 03 μg·ml-1(r=0. 999 1), re-spectively. And the average recovery was 99. 34% and 99. 24% with the RSD of 0. 59% and 0. 62% , respectively (n=6). Conclu-sion: The proposed method is accurate, sensitive and simple, and can be used for the quality control of Niuhuang Qingxin pills (pre-scription of the bureau).

OBJECTIVE： To develop a method for simultaneous determination of 7 components of Niuhuang qingwei pills as chlorogenic acid， geniposide， forsythoside A， narirutin， baicalin， ammonium glycyrrhetate， chrysophanol. METHODS： HPLC-wavelength switching method was adopted. The determination was performed on Agilent ZORBAX SB-C18 column with mobile phase consisted of acetonitrile （A）-0.1％ phosphoric acid （B） gradient elution at the flow rate of 1.0 mL/min. The detection wavelengths were set at 348 nm （chlorogenic acid）， 238 nm （geniposide）， 330 nm （forsythoside A）， 280 nm （narirutin and baicalin）， 237 nm （ammonium glycyrrhetate）， 254 nm （chrysophanol）. The column temperature was 30 ℃， and sample size was 10 μL. RESULTS： The linear ranges of chlorogenic acid， geniposide， forsythoside A， narirutin， baicalin， ammonium glycyrrhetate， chrysophanol were 0.011 67-0.233 4 μg （r＝0.999 4）， 0.042 91-0.858 1 μg （r＝0.999 4）， 0.125 0-2.500 μg （r＝0.999 9）， 0.118 0- 2.360 μg （r＝0.999 9）， 0.119 6-2.392 μg （r＝0.999 7）， 0.030 57-0.611 4 μg （r＝0.999 6）， 0.006 201-0.124 0 μg（r＝0.999 4）， respectively； the limits of quantitation were 1.167， 0.858， 1.250， 1.180， 1.196， 0.611， 0.620 μg/mL， respectively； RSDs of precision tests were 0.98％， 1.04％， 0.59％， 1.50％， 0.83％， 1.24％ and 1.32％ （n＝6）， respectively. RSDs of stability tests were 1.21％， 0.97％， 1.42％， 0.71％， 0.98％， 1.87％ and 1.63％ （n＝6， 12 h）， respectively. Average recoveries were 98.32％， 98.11％， 98.81％， 98.50％， 98.30％， 98.16％ and 97.83％， and the RSDs were 1.37％， 1.41％， 0.64％， 1.01％， 1.18％， 1.16％ and 1.16％ （n＝6）， respectively. CONCLUSIONS： Established method is easy and reproducible. It can be used for the quality control of Niuhuang qingwei pills.