A novel strategy for the quality control of carbonized Typhae pollen using colorimeter, liquid chromatography-mass spectrometry, and efficacy evaluation coupled with multivariate statistical analysis.

In this study, a novel quality control strategy was proposed, aiming to establish a multivariate specification for the processing step by exploring the correlation between colors, chemical components, and hemostatic effects of the carbonized Typhae pollen (CTP) using multivariate statistical analysis. The CTP samples were stir-fried at different durations. Afterward, the colorimeter and LC-MS techniques were applied to characterize the CTP samples, followed by the determination of bleeding time and clotting time using mice to evaluate their hemostatic effect. Then, principal component analysis, hierarchical cluster analysis, and multi-block partial least squares were used for data analysis on colors, chemical components, and their correlation with the hemostatic effect. Consequently, 13 critical quality attributes (CQAs) of CTP were identified via multivariate statistical analysis-L*, a*, b*, 3,4-dihydroxybenzoic acid, 4-hydroxybenzoic acid, 3-hydroxybenzoic acid, quercetin-3-O-glucoside, azelaic acid, kaempferol-3-O-glucoside, quercetin, naringenin, kaempferol, and isorhamnetin. The multivariate specification method involving the 13 CQAs was developed and visualized in the latent variable space of the partial least squares model, indicating that the proposed method was successfully applied to assess the quality of CTP and the degree of carbonization. Most importantly, this study offers a novel insight into the control of processing for carbonized Chinese herbal medicines.

Study on the Hepatoprotective Effects of Hugan Tablets Based on Serum and Liver Metabonomics / 中国药房

OBJECTIVE:To elucidate the efficacy and mechanism of Hugan tablets in hepatoprotective effects from perspective of metabolic pathways. METHODS:36 male rats were randomly divided into normal group (0.5%sodium carboxymethyl cellu-lose),model group(0.5%sodium carboxymethyl cellulose)and Hugan tablets group(1.7 g/kg),12 in each group,intragastrically administrated once a day,for 9 d. After 1 h of last administration,rats in model group and Hugan tablets group were intraperitone-ally injected 50%CCl4 peanut oil solution 1 mL/kg to induce liver injury. After 24 h of modeling,malondialdehyde(MDA),super-oxide dismutase(SOD),glutathione peroxidase(GSH-Px)levels in liver tissue of rats were detected. Nuclear magnetic resonance spectroscopy(1H-NMR)metabolomics technique was adopted to establish the serum and liver metabolite profiles of rats,and the ef-fects of Hugan tablets on changes of metabolic profile and potential biomarkers in serum and liver of rats with CCl4-induced acute liver injury were analyzed. RESULTS:Compared with normal group,MDA level in liver tissue of rats in model group was signifi-cantly increased(P<0.05),SOD and GSH-Px levels were significantly reduced(P<0.05). Both body physiology and material me-tabolism of rats were obviously changed,and levels of 11 metabolic potential biomarkers in serum and 14 metabolic potential bio-markers in liver were significantly increased/decreased (P<0.05). Compared with model group,MDA level in liver tissue in Hugan tablets group was significantly reduced(P<0.05),SOD and GSH-Px levels were significantly increased(P<0.05). Serum and liver metabolism tended to be normal,6 metabolic potential biomarkers(isoleucine,leucine,3-hydroxybutyrate,acetone,ace-toacetate,choline) in serum and 8 metabolic potential biomarkers (3-hydroxybutyrate,alanine,glutamate,pyruvate,succinate, choline,lactate,glucose)in liver got significant callback(P<0.05). CONCLUSIONS:The hepatoprotective mechanism of Hugan tablets may be associated with antioxidative stress and regula-tion of lipid metabolism,glucose metabolism and amino acid metabolism.

Study on HPLC Fingerprints of Naodesheng Solid Dispersion Capsules / 中国药房

OBJECTIVE:To establish HPLC fmgerprints of Naodesheng solid dispersion capsules.METHODS:HPLC method was adopted.The determination was performed on Hyspersil ODS2 column with mobile phase consisted of acetonitrile-0.05％ phosphoric acid (gradient elution) at the flow rate of 1.0 mL/min.The detection wavelengths were 210 nm (puerarin),345 nm (hydroxfsaffior yellow A).The column temperature was 30 ℃,and sample size was 10 μL.Using puerarin and hydroxysaffior yellow A as reference,HPLC fingerprints of 10 batches of sample were determined.Similarity evaluation,common peak identification and chemical components confirmation were performed by using Similarity Evaluation System of TCM Chromatographic Fingerprints (2012 edition).RESULTS:There were 29 and 23 common peaks in HPLC fingerprints of 10 batches of samples at 210 nm and 345 nm,respectively.The similarity was higher than 0.90.The medicinal material attribute of common peaks were Panax notoginseng,Ligusticum chuanxiong,Carthamus tinctorius,Pueraia lobata and Crataegus pinnatifida.Moreover,7 chemical components were identified at 210,345 nm,respectively.CONCLUSIONS:Established HPLC fingerprints can provide reference for identification and quality evaluation of Naodesheng solid dispersion capsules.

Study on HPLC Fingerprints of Naodesheng Solid Dispersion Capsules / 中国药房

OBJECTIVE:To establish HPLC fmgerprints of Naodesheng solid dispersion capsules.METHODS:HPLC method was adopted.The determination was performed on Hyspersil ODS2 column with mobile phase consisted of acetonitrile-0.05％ phosphoric acid (gradient elution) at the flow rate of 1.0 mL/min.The detection wavelengths were 210 nm (puerarin),345 nm (hydroxfsaffior yellow A).The column temperature was 30 ℃,and sample size was 10 μL.Using puerarin and hydroxysaffior yellow A as reference,HPLC fingerprints of 10 batches of sample were determined.Similarity evaluation,common peak identification and chemical components confirmation were performed by using Similarity Evaluation System of TCM Chromatographic Fingerprints (2012 edition).RESULTS:There were 29 and 23 common peaks in HPLC fingerprints of 10 batches of samples at 210 nm and 345 nm,respectively.The similarity was higher than 0.90.The medicinal material attribute of common peaks were Panax notoginseng,Ligusticum chuanxiong,Carthamus tinctorius,Pueraia lobata and Crataegus pinnatifida.Moreover,7 chemical components were identified at 210,345 nm,respectively.CONCLUSIONS:Established HPLC fingerprints can provide reference for identification and quality evaluation of Naodesheng solid dispersion capsules.

Content Determination of 4 Flavonoids Constituents in Yinqiao Capsules by HPLC / 中国药房

OBJECTIVE:To establish a method for the determination of 4 flavonoids constituents in Yinqiao capsules. METH-ODS:HPLC method was adopted. The Hypersil ODS C18 column was used with the mobile phase A of methanol-water-acetic acid (10∶88∶2,V/V/V)and B of methanol-water-acetic acid(88∶10∶2,V/V/V)in gradient elution at the flow rate of 1.0 ml/min;the de-tection wavelength was 327 nm,the column temperature was 25 ℃,and the volume was 10 μl. RESULTS:There was a good linear relationship between the amount of quercetin and peak area in the range of 0.050 9-1.018 0 μg(r=0.999 8),kaempferide in the range of 0.050 2-1.004 0 μg(r=0.999 5),isorhamnetin in the range of 0.051 0-1.020 0 μg(r=0.999 4)and rutin in the range of 0.050 4-1.007 0 μg(r=0.999 8). RSDs of precision,stability and repeatability tests were <2%. The average recoveries were 100.09%(RSD=0.93%,n=9),99.83%(RSD=0.75%,n=9),100.51%(RSD=1.17%,n=9) and 101.19%(RSD=1.08%,n=9). CONCLUSIONS:The method is amount specific,stable and reproducible and can be used for the quality control of Yinqiao capsules.

Optimization of Preparation Technology of Stachydrine in Fructus Leonuri Based on Back-propagation Neural Network and Genetic Algorithm / 广州中医药大学学报

Objective To optimize the preparative procedure for stachydrine in Fructus Leonuri. Methods The preparation was screened by orthogonal experiment, and a mathematical model of relationship of extraction time, methanol concentration, and solid-liquid ratio with the content of stachydrine hydrochloride was established by using back-propagation (BP) neural network. And the process parameters were optimized with genetic algorithm (GA) . Results The optimum process parameters were as follows: extraction with 69% of methanol concentration and with solid-liquid ratio being 11 times for 62 min. The content of stachydrine obtained by BP neural network modeling and GA was higher than that achieved by orthogonal experiment. Conclusion The optimum preparative procedure could be achieved by combining BP modeling with GA. The model developed in this study was proved to be predictable and feasible for the optimization of process parameters of multi-dimension nonlinear system.

Urine metabonomic study of intervention effects of Morinda officinalis how. on 'kidney-yang deficiency syndrome' / 药学学报

To investigate the intervention effects of Morinda officinalis How. on 'Kidney-yang deficiency syndrome' induced by hydrocortisone in rats, the metabolic profiles of rat urine were characterized using proton nuclear magnetic resonance and principal component analysis (PCA) was applied to study the trajectory of urinary metabolic phenotype of rats with 'Kidney-yang deficiency syndrome' under administration of M. officinalis at different time points. Meanwhile, the intervention effects of M. officinalis on urinary metabolic potential biomarkers associated with 'Kidney-yang deficiency syndrome' were also discussed. The experimental results showed that in accordance to the increased time of administration, an obvious tendency was observed that clustering of the treatment group moved gradually closed to that of the control group. Eight potential biomarkers including citrate, succinate, alpha-ketoglutarate, lactate, betaine, sarcosine, alanine and taurine were definitely up- or down-regulated. In conclusion, the effectiveness of M. oficinalis on 'Kidney-yang deficiency syndrome' is proved using the established metabonomic method and the regulated metabolic pathways involve energy metabolism, transmethylation and transportation of amine. Meanwhile, the administration of M. officinalis can alleviate the kidney impairment induced by 'Kidney-yang deficiency syndrome'.

Metabonomic study of intervention effects of Morinda officinalis on 'kidney-yang deficiency syndrome' / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the metabolic profile of hydrocortisone-induced 'Kidney-yang deficiency syndrome'in rats and the intervention effects of Morinda officinalis.</p><p><b>METHOD</b>Proton nuclear magnetic resonance (1H-NMR) technique was used to analyze the rat metabonome in serum. Orthogonal partial least squares discriminant analysis (OPLS-DA) were processed to analyze the metabonome difference between the control and hydrocortisone treated samples. Twelve potential biomarkers were selected, via the parameter of variable importance in the projection (VIP). Principal components analysis (PCA) was employed to process the data from the M. officinalis. treatment group and the intervention effects of M. officinalis, was investigated through the selected potential biomarkers.</p><p><b>RESULT</b>After hydrocortisone treatment, the energy metabolism, amino acids metabolism and gut microflora environment were seriously disturbed and transmethylation was surpressed. M. officinalis could effectively alleviate the disturbance of energy and amino acids metabolism and enhance transmethylation, but could not modulate the gut microflora environment.</p><p><b>CONCLUSION</b>The results obtained suggested that metabonomic studies could better reflect the whole status of metabolism in bio-systems, and could be treated as a potential powerful approach in pharmacological studies and investigation of the essence of 'syndrome' in traditional Chinese medicine.</p>

Effects of rhubarb aglycone combined with thrombolysis on brain microvascular basement membrane impairment in rats with thrombus-occluded cerebral ischemia / 中国中药杂志

<p><b>OBJECTIVE</b>To explore the protective effects of rhubarb aglycone combined with urokinase (UK) thrombolysis on brain microvascular basement membrane impairment in rats with thrombus-occluded cerebral ischemia by regulating the expression of IgG, CoLIV and LN in rats brain, by which the level of injury of brain microvascular basement membrane could be detected.</p><p><b>METHOD</b>Rats were randomly divided into sham-operated, model, thrombolysis, rhubarb aglycone and combination (rhubarb aglycone combined with thrombolysis) groups. Moreover, rats in model, thrombolysis, rhubarb aglycone and combination groups were randomly divided into 3, 6, and 9 h groups respectively. Model of thrombus-occluded cerebral ischemia was duplicated by using the combination of rats' auto-thrombus with inserting the nylon thread. Rats were administrated with thrombolysis therapy through artery at 3, 6, and 9 h after cerebral ischemia. At 24 h of administration through artery, intracranial hemorrhage ratio (ICHR) and mortality of rats were observed, and then the brain of rats was taken. In the study, expression of IgG, CoLIV and LN in rats brain were measured.</p><p><b>RESULT</b>Thrombolysis at 9 h of cerebral ischemia made rats mortality and BHR increase, administration of combined therapy could make them decrease. Expression of IgG level in rats brain of 9 h and 6 h model groups increased, while CoLIV and LN expression decreased significantly. In each administration 9 h group, IgG level was lower, and CoLIV and LN were higher, such changes appeared significantly in rhubarb aglycone and association groups.</p><p><b>CONCLUSION</b>Brain microvascular basement membrane impairment could be caused by the therapy of delayed thrombolysis, which made the mortality and BHR increase. Rhubarb aglycone combined with the therapy of thrombolysis could perform the protective effects on brain microvascular basement membrane and then decrease the ICHR and mortality caused by thrombolysis after cerebral ischemia.</p>

Study of Lvfukang Capsules on experimental arrhythmia induced by aconitine in rats / 中华中医药杂志

Objective: To observe the anti- arrhythmic effects of Lvfukang Capsules on the experimental arrhythmic models induced by aconitine in rats, and provid accordance for clinical medication. Methods: 50 rats were divided into model control group, positive control group, high, middle and low dosage groups of Lvfukang Capsules, respectively. All the dosage groups were treated with successive medication 3 days, arrhythmic models induced by aconitine for 30minutes after the last dosage. To observe and record the time of ventricular premature beat (VP) and ventricular fibrillation (VF). Results: All the dosage groups of Lvfukang Capsules significantly delayed the time of ventricular premature beat (VP) and ventricular tachycardia (VT) of arrhythmic models of rats (P

Adsorption of Active Components in Naomaitong Granules by Different Macroporous Adsorptive Resins / 中国药房

OBJECTIVE: To optimize the macroporous absorbing resin which were of best action in adsorption and desorption on the active components in Naomaitong granules. METHODS: UV spectrophotometry and HPLC was employed to determine the adsorbability and desorption capacity of different macroporous absorbing resins on total anthraquinones, total ginsenosides, total alkaloids and Puerarin. RESULTS: There were differences in adsorption and desorption capacity on active components in Naomaitong granules among different macroporous absorbing resins. Considering the general adsorbability and desorption capacity of different macroporous absorbing resins, AB-8 turned out to be of the best purification effect on Naomaitong granules. CONCLUSION: The results serve as a theoretical basis for the production of Naomaitong granules.

Pharmacodynamic fingerprint of Flos Lonicerea by HPLC / 中草药

Objective Looking for the chemical components which have positive correlations with anti-inflammatiory activity of Flos Lonicerea to set up pharmacodynamic-fingerprint. Methods To gain the fingerprints of different parts by HPLC and to detect the pharmacological activities of anti-inflammation, then the correlationship between chemical components and pharmacological activities were detected by linear regression. Results Pharmacological activities of the methanol extracted part were best of all, so the fingerprint of the methanol part could represent the pharmacodynamic-fingerprint of Flos Lonicerea. Conclusion The fingerprint established by this way is more scientific and reasonable.

Studies on "floating sugar" mechanism in root of Achyranthes bidentata / 中草药

Object To study the "floating sugar" mechanism in the root of Achyranthes bidentata Bl. Methods The important influent factors were analyzed by orthogonal test. Results The primary and secondary orders of influent factors were surrounding temperature, relative humidity, raw drug moisture. The best preservative condition: surrounding temperature is 25 ℃, relative humidity 60% and drugs moisture 11%. Conclusion Raw drugs can be stored safely when surrounding temperature is below 35 ℃, relative humidity below 70%, raw drug moisture between 9% and 13%.

Variation of Ferulic Acid Content in Naomaitong by Different Extraction Process / 中药新药与临床药理

Objective To compare the variation of ferulic acid contents in Naomaitong by different extraction processes.Methods With ethanol or water as extracting solvent,the four herbal medicines of Rhizoma Chuanxiong,Radix Ginseng,Radix Puerariae,Radix et Rhizoma Rhei,which consisted of Naomaitong prescription,were decocted together or decocted separately firstly then mixed together were evaluated.With ferulic acid extraction rate as the index,the above extraction processes were evaluated.Results The extraction rate of ferulic acid extracted by alcohol was higher than that by water,but the ferulic acid content showed no obvious difference by decocting together or decocted separately firstly then mixed together.Conclusion It is suggested that proper extraction solvents and extraction methods should be adopted according to the different physicochemical characteristics of chemical contents in herbal medicine during extraction.

Correlation between fingerprint peaks and Compound Naomaitong 's effective fraction and its relevant herbs / 中成药

AIM:To establish the HPLC fingerprint of Compound Naomaitong effective parts,and to study the correlation analysis between fingerprint peaks and the effective fraction and its relevant herbs. METHODS:The chromatographic fingerprints of the effective fraction and the relevant fractions of its herbs were configured by HPLC/PDAD analysis. The relative deviation of retention time was utilized as indices to evaluate the correlation, the wavelength was set at 203 nm. RESULTS:The fingerprint of Compound Naomaitong effective parts was established and 36 copossessing fingerprint peaks were indicated. The assignment results of 14 peaks effective parts of fraction were indicated. CONCLUSION:The quality of Compound Naomaitong effective parts can be controlled by the HPLC fingerprint.

Determination of pueraria isoflavones content in effective parts of Naomaitong / 中成药

AIM: To establish an HPLC method for determing content of puerarin isoflavones in effective parts of Naomaitong Granules.METHODS: Gradient elution with methanol-water-glacial acetic acid was used as the mobile phase.The flow rate was 1.0 mL/min,and the detection wavelength was set at 250 nm.RESULTS: The average recoveries of puerarin,daizein-8-capiosy(1→6) glucoside,3-methoxyl-puerarin,daidzin,daidzein were 100.89%(RSD = 2.1%),100.23%(RSD = 1.06%),101.04%(RSD = 1.92%),99.82%(RSD = 2.02%),102.06%(RSD = 1.34%) respectively.CONCLUSION: The method is simple,accurate and can be used for quality control of the effective parts in Naomaitong Granules.

Separation and purification of Naomaitong Granules by AB-8 macroporous absorption resin / 中成药

AIM: To study the purification of Naomaitong Granules(Radix et Rhizoma rhei,Rhizoma chuanxiong,Radix Puerariae lobatae.etc) by macroporous absorption resin. METHODS: Naomaitong Granule was purificated by macroporous absorption resin AB-8.UV spectrothotometry was used to determine the contents of total anthraquinones,total ginsenosides,total alkaloids,and the content of puerarin was determined by HPLC.The technic of purification was optimized according to the content above. RESULTS: The optimized technological conditions consisted of eoncentranon of original sample 120 mg/mL,the diameter and height was in proportion of 1∶10,the ratio of maximum adsorption to resin volume was 1∶6,water elution was 2B multiple of resin volume,8B multiple of resin volume 50% alcohol was the elution. CONCLUSION: AB-8 macroporous absorption resin can be used to purify Naomaitong Granules.

Quality control of Jiangzhi Jianfei Tablets / 中成药

AIM:To establish the methods of determining 2,3,5,4′-tetrahydroxystilbene-2-O-?-D-glycoside,puerain;ginsengnoside Rg_1,ginsengnoside Rb_1,notoginsengnoside R_1,and salvianolic B in Jiangzhi Jianfei Tablets(Radix Polygoni multiflori,Radix Puerariae lobatae,Radix et Rhizoma Notoginseng,Radix et Rhizoma Salviae miltiorrhizae,etc) with HPLC. METHODS: All chromatogram columns used were ODS columns.The mobile phase of acetonitrile-wate(25(∶)75) was chosen for 2,3,5,4′-tetrahydroxystilbene-2-?-D-glycoside.The flow rate was 1.0 mL/min.The detection wavelength was at 320 nm.The mobile phase of methanol-water(25(∶)75) was chosen for puerain.The flow rate was 1.0 mL/min.The detection wavelength was at 250 nm.The mobile phase of acetonitrile-water was chosen for ginsengnoside Rg_1,ginsengnoside Rb_1 and notoginsengnoside R_1.The flow rate was 1.0 mL/min.The detection wavelength was at 203 nm.The mobile phase of methanol-acetonitrile-methane acid-water(30(∶)10(∶)1(∶)59) was chosen for salvianolic B.The flow rate was 1.0 mL/min.The detection wavelength was 286 nm. RESULTS: The linear rangers were 0.476 ?g-4.760 ?g(r=0.999 6) for 2,3,5,4′-tetrahydroxystilbene-2-?-D-glycoside,0.168 96 ?g-1.689 6 ?g(r=0.999 8) for puerain,0.724 ?g-7.240 ?g(r=0.999 8) for ginsengnoside Rg_1,0.728 ?g-7.280 ?g(r=0.999 9) for ginsengnoside Rb_1,0.23 ?g-2.30 ?g(r=0.999 8) for notoginsengnoside R_1,0.306 ?g-3.060 ?g(r=0.999 9) for salvianolic B.Their average recoveries were 99.43%(RSD=1.08%,n=6),99.43%(RSD=1.66%,n=6),101.17%(RSD=2.08%,n=6),and 102.39%(RSD=2.60%,n=6),respectively. CONCLUSION: These methods of determining components in four kinds of herbs are simple,accurate,reproducible and can be used for quality control of Jiangzhi Jianfei Tablets.

Anthraquinones variation in extractive processes of Naomaitong extract / 中成药

AIM: To compare the contents variation of anthraquinones by 2 kinds of extractive process of Naomaitong extract (Radix et Rhizoma rhei, Rhizoma chuanxiong, Radix et Rhizoma qinseng, Radix puerariae lobatae, etc). METHODS: In application to RP-HPLC, Hypersil ODS2 C_ 18 column, the mobile phase was a mixture of methanol- 0.1% phosphate water (75 ∶25), the wavelength of detecting five aglycons in R. officinale was at 254 nm. RESULTS: The extraction rate of anthraquinones contents extracted by alcoholic extraction was higher than water extraction, the chrysophanol content in separated decoction was higher than that in the mixed decoction. CONCLUSION: There are dynamic variations in the extraction of Chinese medicine mixture. It is necessary to handle samples appropriately according to physichemical attributes of chemical contents.