Scientific Characterization of Traditional Softening Method of Corydalis Rhizoma / 中国实验方剂学杂志

ObjectiveModern scientific methods and techniques were used to scientifically characterize the traditional softening process of Corydalis Rhizoma， so as to clarify the scientificity and rationality of the traditional process， and provide reference for inheriting the processing methods and experience of traditional Chinese medicine. MethodLow-field nuclear magnetic resonance imaging （LF-NMR/MRI） was used to characterize the water types and distribution in the softening process of Corydalis Rhizoma. Samples during the softening process was cut into thick slices and its section was observed by stereoscopic microscope. High performance liquid chromatography （HPLC） was employed to determine the content change of tetrahydropalmatine during the softening process with the mobile phase of methanol-0.1% phosphoric acid solution （60∶40， triethylamine regulated to pH 6.5） and detection wavelength at 280 nm. The determination method of softening endpoint of Corydalis Rhizoma was simulated by texture analyzer （hand pinch method）， and the softening degree of the finished products was determined after optimizing the relevant parameters. ResultLF-NMR/MRI showed that the water could penetrate through the core and distribute evenly in Corydalis Rhizoma softened by Zhangbang method. The water first entered into the medicinal material from the epidermis and stem marks in the soaking stage as the form of free water， and then penetrated into the inner core to achieve redistribution in the moistening stage. Under stereoscopic microscope， it was observed that Corydalis Rhizoma softened by the Zhangbang method could be sliced well， but the core bursting slices were easy to appear if the softening time was not enough， and the softening of samples was caused by the keratine-like powder after absorbing water. HPLC measurement showed that the loss of tetrahydropalmatine in the softening method was small， its content decreased about 5% in the soaking process， and its content was almost unchanged during the moistening process. The softening degree of Corydalis Rhizoma could be quantified by the texture analyzer， and the optimum parameters were 2 mm·s-1 of speed before test， test speed and speed after test， 20 g of the trigger force， 20% of compression degree. The compressive force of the qualified softened Corydalis Rhizoma was 12.75-15.69 N with the relative standard deviation （RSD） of 6.8%. ConclusionModern scientific methods and techniques can characterize the scientificity and rationality of the traditional processing methods， and confirm that the Zhangbang softening method has the advantages of high efficiency， convenience and small loss of index components. The texture analyzer can simulate the softening endpoint judgment method （hand pinch method）， and realize the goal from subjective experience judgment to objective technology quantification， which has a good demonstration role for the modern inheritance of traditional processing technology.

Scientific Characterization of Traditional Softening Method of Corydalis Rhizoma / 中国实验方剂学杂志

ObjectiveModern scientific methods and techniques were used to scientifically characterize the traditional softening process of Corydalis Rhizoma， so as to clarify the scientificity and rationality of the traditional process， and provide reference for inheriting the processing methods and experience of traditional Chinese medicine. MethodLow-field nuclear magnetic resonance imaging （LF-NMR/MRI） was used to characterize the water types and distribution in the softening process of Corydalis Rhizoma. Samples during the softening process was cut into thick slices and its section was observed by stereoscopic microscope. High performance liquid chromatography （HPLC） was employed to determine the content change of tetrahydropalmatine during the softening process with the mobile phase of methanol-0.1% phosphoric acid solution （60∶40， triethylamine regulated to pH 6.5） and detection wavelength at 280 nm. The determination method of softening endpoint of Corydalis Rhizoma was simulated by texture analyzer （hand pinch method）， and the softening degree of the finished products was determined after optimizing the relevant parameters. ResultLF-NMR/MRI showed that the water could penetrate through the core and distribute evenly in Corydalis Rhizoma softened by Zhangbang method. The water first entered into the medicinal material from the epidermis and stem marks in the soaking stage as the form of free water， and then penetrated into the inner core to achieve redistribution in the moistening stage. Under stereoscopic microscope， it was observed that Corydalis Rhizoma softened by the Zhangbang method could be sliced well， but the core bursting slices were easy to appear if the softening time was not enough， and the softening of samples was caused by the keratine-like powder after absorbing water. HPLC measurement showed that the loss of tetrahydropalmatine in the softening method was small， its content decreased about 5% in the soaking process， and its content was almost unchanged during the moistening process. The softening degree of Corydalis Rhizoma could be quantified by the texture analyzer， and the optimum parameters were 2 mm·s-1 of speed before test， test speed and speed after test， 20 g of the trigger force， 20% of compression degree. The compressive force of the qualified softened Corydalis Rhizoma was 12.75-15.69 N with the relative standard deviation （RSD） of 6.8%. ConclusionModern scientific methods and techniques can characterize the scientificity and rationality of the traditional processing methods， and confirm that the Zhangbang softening method has the advantages of high efficiency， convenience and small loss of index components. The texture analyzer can simulate the softening endpoint judgment method （hand pinch method）， and realize the goal from subjective experience judgment to objective technology quantification， which has a good demonstration role for the modern inheritance of traditional processing technology.

Tetrahydropalmatine alleviated diabetic neuropathic pain by inhibiting activation of microglia via p38 MAPK signaling pathway / 中国中药杂志

Neuropathic pain is one of the common complications of diabetes. Tetrahydropalmatine(THP) is a main active component of Corydalis Rhizoma with excellent anti-inflammatory and pain-alleviating properties. This study aims to investigate the therapeutic effect of THP on diabetic neuropathic pain(DNP) and the underlying mechanism. High-fat and high-sugar diet(4 weeks) and streptozotocin(STZ, 35 mg·kg~(-1), single intraperitoneal injection) were employed to induce type-2 DNP in rats. Moreover, lipopolysaccharide(LPS) was used to induce the activation of BV2 microglia in vitro to establish an inflammatory cellular model. Fasting blood glucose(FBG) was measured by a blood glucose meter. Mechanical withdrawal threshold(MWT) was assessed with von Frey filaments, and thermal withdrawal latency(TWL) with hot plate apparatus. The protein expression levels of OX42, inducible nitric oxide synthase(iNOS), CD206, p38, and p-p38 were determined by Western blot, the fluorescence expression levels of OX42 and p-p38 in the dorsal horn of the rat spinal cord by immunofluorescence, the mRNA content of p38 and OX42 in rat spinal cord tissue by qRT-PCR, and levels of nitric oxide(NO), interleukin-1β(IL-1β), interleukin-6(IL-6), tumor necrosis factor-α(TNF-α), interleukin-10(IL-10), and serum fasting insulin(FINS) by enzyme-linked immunosorbent assay(ELISA). RESULTS:: showed that the mo-del group demonstrated significant decrease in MWT and TWL, with pain symptoms. THP significantly improved the MWT and TWL of DNP rats, inhibited the activation of microglia and p38 MAPK signaling pathway in rat spinal cord, and ameliorated its inflammatory response. Meanwhile, THP promoted the change of LPS-induced BV2 microglia from the pro-inflammatory M1 phenotype to the anti-inflammatory M2 phenotype, suppressed the activation of the p38 MAPK signaling pathway, decreased the expression levels of inflammatory factors NO, IL-1β, IL-6, and TNF-α, and increased the expression level of anti-inflammatory factor IL-10. The findings suggested that THP can significantly ameliorate the pain symptoms of DNP rats possibly by inhibiting the inflammatory response caused by M1 polarization of microglia via the p38 MAPK pathway.

Comparison of the sedative and anti-anxiety effects of levo-tetrahydropalmatine and diazepam on rats with conditioned fear / 中华行为医学与脑科学杂志

Objective:To compare the sedative and anti-anxiety effects of levo-tetrahydropalmatine (L-THP) and diazepam on conditioned fear model rats.Methods:According to the random number table method, 32 adult male rats were divided into blank group, model group, diazepam group and L-THP group(with 8 rats in each group). The conditioned fear model was reproduced by the plantar electric shock method. Four days after the modeling, the rats in diazepam group and L-THP group were given diazepam (3.6 mg/kg) and L-THP (25 mg/kg) were respectively gavaged once a day for 10 days, the rats in blank group and model group were given the same volume of saline. After the administration, the elevated plus maze test and the open field test were used to measure the anxiety behavior of the rats, and the sleep energy monitoring system was used to detect changes in sleep and energy-related indicators. SPSS 23.0 and Graphpad Prism 7.0 softwares were used for data analysis, multiple samples between groups were compared by one-way ANOVA, and LSD test was used for pairwise comparison.Results:The results of the elevated plus maze experiment showed that compared with the model group, the percentage of open-arm entry times ((11.27±8.78)%, (30.11±14.59)%, P<0.05) and the percentage of open-arm residence time ((1.94±1.48)%, (17.53±8.21)%, P<0.05) in diazepam group were all significantly increased. Compared with the model group, the open arm residence time, the percentage of open arm residence time and the percentage of open arm entry times in L-THP group showed an upward trend, but there was no statistical significance (all P>0.05). The results of the open field experiment showed that compared with the model group, the time of entering the central grid ((2.99±1.83) s, (6.94±3.52) s, P<0.05) and the time of entering the peripheral field ((297.01±1.83) s, (293.30±3.52) s, P<0.05) in diazepam group both increased. Compared with the model group, there was no significant difference in the changes of various indexes in L-THP group (all P>0.05). The results of locomotor activities showed that the autonomic activity times of model group in nighttime was significantly lower than that of blank group((758.79±375.37)times/h, (1 101.93±525.96)times/h, P<0.05). Compared with the model group, the number of autonomous activities of rats in L-THP group in daytime ((820.57±364.60) times/min, (502.40±228.54)times/min, P<0.05) decreased, and the number of autonomous activities in the nighttime ((758.79±375.37) times/min, (1 146.85±309.69)times/min, P<0.05) increased, but there was no significant change in the number of autonomous activities in the whole day. Correlation analysis of energy metabolism related indexes and sleep time of rats in each group were analyzed. The experimental results showed that the daytime sleep time were negatively correlated with heat value ( r=-0.335, P<0.05), and the night sleep time was positively correlated with daytime heat value ( r=0.352, P<0.05). Conclusion:L-tetrahydropalmatine has no significant anti-anxiety effect in the concentration range used in this study, but its sedative and improving sleep activity rhythm are better than diazepam.

Chemical composition identification and mechanism study of Fufang Shangtong Capsule based on UHPLC-Q-Orbitrap HRMS and network pharmacology / 中草药

Objective: To systematically study the main chemical components of Fufang Shangtong Capsule and explore the main mechanism of its effect, and provide some reference for the research of its pharmacodynamic substance. Methods: In this study, UHPLC-Q-Orbitrap HRMS was used to comprehensively analyze the main chemical components of Fufang Shangtong Capsule. The chromatographic column was Waters Acquity UPLC® BEH C18 chromatographic column (50 mm ×2.1 mm, 1.7 μm) and the mobile phase was acetonitrile (A)-0.1% formic acid water (B). According to the MS/MS spectrometry information of compounds, and the comparison with standards or references, the chemical information of drugs can be quickly and accurately identified. On this basis, the network pharmacology method was used to analyze the chemical composition target of the drug, enrich its function, preliminarily select the main effective substances of the drug, and simultaneously explore its mechanism of action. Results: A total of 36 chemicals were identified in this study from Fufang Shangtong Capsule. The target function of enrichment analysis showed that the drug mainly played its therapeutic effect on regulating vascular endothelial, vascular smooth muscle pain, affecting platelet function, promoting energy supply, reducing inflammation and relieving pain, so as to exert its efficacy in promoting blood circulation and removing stasis, invigorating qi and relieving pain. Conclusion: In this study, UHPLC-Q-Orbitrap HRMS combined with network pharmacology was used, wich provided scientific theoretical basis and important reference for the identification of effective ingredients, screening of quality markers and the study of potential mechanism of action of Fufang Shangtong Capsule.

Diversity and community structure of endophytic fungi in Corydalis yanhusuo tuber and their correlations with tetrahydropalmatine content / 中草药

Objective: To analyze the diversity and community structure of endophytic fungi in Corydali yanhusuo tuber and their correlations with tetrahydropalmatine content. Methods The endophytic fungi in C. yanhusuo tuber from eight localities (Pan’an of Zhejiang Province, Dongyang of Zhejiang Province, Jinyun of Zhejiang Province, Sanhe of Shanxi Province, Dongjiaying of Shanxi Province, Kaixian of Chongqing, Xuancheng of Anhui Province and Xinyang of Henan Province) were isolated and identified according to the morphology and ITS sequences, so as to analyze the community structure of endophytic fungi; HPLC was used to determine tetrahydropalmatine content, and Excel and SPSS were used to analyze their correlations and establish multiple linear regression equations at different levels of dominant genera and species. Results: A total of 1 742 endophytic fungi were isolated and divided into 19 taxa (14 were identified to species level and five to genera level), belonging to 3 phyla, 5 classes, 10 orders, 14 families and 15 genera; The tetrahydropalmatine content in C. yanhusuo tuber from eight localities was higher than the standard of Chinese Pharmacopoeia (2015 edition); The richness (S) and diversity index (H’) of endophytic fungi in C. yanhusuo tuber and tetrahydropalmatine content in Zhejiang Province were extremely significant or significantly higher than those in the other localities. The diversity index (H’) was significantly positive correlated with the tetrahydropalmatine content; There was the largest positive correlation between the tetrahydropalmatine content and T83 (Trichoderma sp.) (correlation coefficient: 0.793). Conclusion: There are abundant endophytic fungi resources in C. yanhusuo; T83 as a dominant endophytic fungus probably related to the accumulation of tetrahydropalmatine.

Effect of precipitation inhibitors on pH-induced supersaturated phase behavior of dl-tetrahydropalmatine / 中草药

Objective: To investigate the effect of three types of precipitation inhibitors (PPI) HPMC K4M, HPMC AS MG and Soluplus on the pH-induced supersaturated phase behavior of dl-tetrahydropalmatine (dl-THP) at oral clinical doses. Methods: dl-THP pH solubility phase diagram and desaturation curve during pH-shift were drawn, and the solubility phase diagram was used to support dl-THP phase behavior. Area under the concentration-time curve and supersaturation ratio were used to analyze the effect of PPI on the phase behavior of dl-THP; Polarized light microscope and differential scanning calorimetry were used to analyze the precipitation properties. Results: Under the clinical dosage, the maximum supersaturation of dl-THP during the pH-shift was 3.93, and the supersaturation was lost over time; HPMC K4M, HPMC AS MG, and Soluplus could all maintain the supersaturation within 180 minutes during the pH-shift dissolution. HPMC K4M, HPMC AS MG, and Soluplus maintained supersaturation levels of 1.19, 1.89 and 1.36 respectively at a concentration of 5%, 1.30, 2.35 and 1.86 at a concentration of 20%, and 1.30, 2.60 and 2.07 at a concentration of 50%. Polarized light microscopy and differential scanning calorimetry results showed that crystalline precipitation occurred. Conclusion: All precipitation inhibitors can improve the pH-induced supersaturated phase behavior of tetrahydropalmatine, and this improvement behavior varies with the type and concentration of precipitation inhibitors. HPMC AS MG has the best effect.

Preparation of dl-tetrahydropalmatine ethosomes and their ex vivo transdermal permeation properties / 中草药

Objective To prepare dl-tetrahydropalmatine (dl-THP) ethosomes (ETS) and elucidate their transdermal absorption properties. Methods Dl-tetrahydropalmatine ethosomes (dl-THP ETS) were prepared by ethanol injection combined with pH-gradient active drug-loading method. Their physicochemical properties including elasticity, vesicle size, morphology and entrapment efficiency were characterized. Franz diffusion cells were used to investigate the ex vivo skin permeation characteristics of the formulation with liposomes (LPS) and tinctures being used as reference preparations. Results According to a preferred formulation of dl-THP ETS [dl-THP 100 mg, vitamin E 1.3 mg, soybean lecithin 1 200 mg, cholesterol 120 mg, absolute ethanol 9 mL and citrate buffered saline (pH 3.0) 21 mL, 0.1 mol/L NaOH solution suitable quantity (to adjust the pH value to 5.5) ], the obtained dl-THP ETS had an elasticity index of (20.1 ± 1.1) mL, an average size of (85.8 ± 0.9) nm with a polydispersity index of (0.082 ± 0.003) and an entrapment efficiency of (81.7 ± 3.2)%. The cumulative permeated drug quantity per unit area (Qn) of dl-THP ETS in 24 h was (2 306.4 ± 592.3) μg/cm2 with no significant difference compared with the Qn of the LPS [(2 434.2 ± 564.4) μg/cm2] (P > 0.05) and about 4 times of that of the tincture [(633.1 ± 218.0) μg/cm2] (P < 0.05). And the averages of RSD of the Qn at each time point were (28.37 ± 10.9)% and (62.83 ± 44.1)% for the ETS and LPS, respectively, indicating that the Qn fluctuation among samples of the ETS was smaller than that of the LPS (P < 0.05). Average correlation coefficients of (0.968 ± 0.033) and (0.882 ± 0.078) (P < 0.05) were obtained for the ETS and LPS respectively when their 24 h permeation curves were fitted to linear relationship, indicating the permeation of the former was closer to zero-order kinetics than that of the latter. Conclusion The dl-THP ETS have a high elasticity, a suitable size, a high entrapment efficiency, and enhanced and stable percutaneous absorption in line with zero-order kinetics.

HPLC fingerprint and multicomponents determination of Corydalis Decumbentis Rhizoma / 中草药

Objective: To establish the HPLC fingerprint and multicomponents determination of Corydalis Decumbentis Rhizoma, and provide a scientific basis for the improvement of its quality standards. Methods The separation was performed on a chromatographic Diamonsil C18 column (250 mm × 4.6 mm, 5 μm), with acetonitrile-0.2% acetic acid (adjusting pH to 6.0 with triethylamine) as the mobile phase for gradient elution. Volume flow rate was 1.0 mL/min. Column temperature was 35 ℃. Injection was 10 μL and the detection wavelength was 280 nm. The fingerprints of 15 batches of Corydalis Decumbentis Rhizoma were established and evaluated by the similarity evaluation system of TCM (version 2012A), which were divided into two categories by clustering analysis and principal component analysis. Meanwhile, the content of protopine, palmatine, bicuculline and tetrahydropalmatine was determinated. Results:The fingerprint of Corydalis Decumbentis Rhizoma was established. There were 10 common peaks in the fingerprint. Protopine, palmatine, bicuculline and dl-tetrahydropalmatine were separated with good linearity relationships (r > 0.999 9). The average recovery rates of the investigated compounds were 97.12%, 100.09%, 98.53%, and 99.71%, respectively. Conclusion:HPLC fingerprint combined with multicomponents determination can provide the reference for the quality evaluation of Corydalis Decumbentis Rhizoma.

Analysis of Correlation Between Color and Intrinsic Quality of Vinegar-processed Corydalis Rhizoma Decoction Pieces / 中国实验方剂学杂志

Objective: Based on the color space technology of CIE-LAB,the color of vinegar-processed Corydalis Rhizoma decoction pieces was digitized,combining with the contents of 10 major alkaloids in the decoction pieces,to discuss the correlation between the color and contents of main ingredients of vinegar-processed Corydalis Rhizoma decoction pieces,and investigate the intrinsic quality difference in the decoction pieces with different color. Method: The precision colorimeter was used to determine the color parameters of vinegar-processed Corydalis Rhizoma decoction pieces;HPLC was employed to determine contents of main chemical components in the decoction pieces,which was performed on Agilent ZORBAX SB-C18 column(4.6 mm×250 mm,5 μm) with mobile phase of acetonitrile(A)-0.1% potassium dihydrogen phosphate aqueous solution(B) for gradient elution(0-10 min,5%-22%A;10-30 min,22%-25%A;30-50 min,25%-60%A;50-70 min,60%-95%A),detection wavelength of 280 nm,column temperature at 30℃ and flow rate of 1.0 mL·min-1. Result: The quality of vinegar-processed Corydalis Rhizoma decoction pieces with different color was in line with the requirement of the 2015 edition of Chinese Pharmacopoeia,but there were differences in the intrinsic quality between the decoction pieces.The total content of chemical components in the samples showed a positive correlation with the a*(green-red axis) and total chromatic aberration value(ΔE) in the CIE-LAB color space, and it was significantly negative correlated with L*(lightness) and b*(blue-yellow axis).In the 10 tested components,except for D-tetrahydrojatrorrhizine and tetrahydrocoptisine,contents of protopine and other 6 components were positively correlated with color,and only the content of corydaline was negatively correlated with color. Conclusion: Color analysis technology can objectively quantify the color of the decoction pieces,and can achieve a quick evaluation of quality of the decoction pieces by analyzing correlation between the color and the contents of main active ingredients.

Effects of Jiawei Foshou San capsule on rat hepatic cytochrome P450 enzymes invitro and invivo / 药学学报

This study aimed to evaluate the effects of Jiawei Foshou San capsule (JWFSSC) on CYP1A2, CYP2C6, CYP2D2, CYP2E1 and CYP3A1/2 enzyme activities in rat liver microsomes in vitro and in vivo, and to provide pharmacokinetic data for its combined use with other medicines. After incubating liver microsomes with a cocktail of probe drugs, the metabolites were quantitated with LC-MS/MS to assess the CYP enzyme activity. The hepatic pathological changes were evaluated by histology after hematoxylin and eosin (HE) staining. With the dose range up to 3 200 mg·L-1, the IC50 of JWFSSC for CYP2D2, CYP2E1 and CYP3A1/2 in vitro was 229.3 mg·L-1, 361.9 mg·L-1 and 274.6 mg·L-1 respectively. Compared with the vehicle control group, the enzyme activities of CYP1A2, CYP2C6 and CYP3A1/2 showed a significant increase in animals given JWFSSC 180 mg·kg-1·d-1 (P<0.01). Based on histology, several pathological changes were observed in JWFSSC groups: there was less inflammatory infiltration compared to the tetrahydropalmatine (THP) group. These results of inhibition in vitro and induction in vivo suggest a strengthened efficacy and a prolonged effective time of drugs metabolized by CYP2D2 and CYP2E1 enzymes when combined with JWFSSC in use. The dosage of parent drugs should be appropriately reduced when used in combination with JWFSSC. However, if a drug is metabolized by CYP1A2 and CYP2C6 when used in combination with JWFSSC, the effect of the drug is likely reduced and the dosage should be increased appropriately. In addition, the combination of ferulic acid (FA), ligustrazine (LZ) and THP can significantly reduce the toxicity of THP in rat livers. In this study, the program of animal testing had been approved by Committee on the management and usage of experimental animal in the College of Pharmaceutical Sciences, Southwest University.

Study on mechanism of enhanced HaCaT cellular uptake of tetrahydropalmatine by asarum essential oil and sinapine in Sanfu Patch / 中草药

Objective To study the mechanism of enhanced HaCaT cellular uptake of tetrahydropalmatine (THP) by asarum essential oil (AEO) and sinapine thiocyanate (SPT) in Sanfu Patch. Methods Effect of SPT, AEO, and THP on cell viability of HaCaT were determined by MTT. HaCaT cellular uptake of THP and the enhancing effects of AEO and SPT on THP uptake were visualized with confocal laser scanning microscope (CLSM) based on the green autofluorescence of THP, and the THP uptake content by HaCaT was further determined with HPLC. Moreover, HaCaT cells were labeled with diphenylhexatriene (DPH). After the labeled cells were treated with AEO, SPT, and THP, respectively, the cellular membrane fluidity was determined with fluorescence polarization technology. Results THP fluorescence intensity within HaCaT cells was significantly increased when THP was co-delivered with AEO or SPT respectively, and the THP content with each group within the cells was also significantly higher than that of THP delivered alone. In addition, AEO was superior to SPT in enhancing THP uptake by HaCaT cells. The fluorescence polarization and membrane micro-viscosity of HaCaT cells were significantly decreased after AEO treatment, which indicated that membrane fluidity was increased by the treatment with AEO. However, SPT or THP did not present the character of increasing the membrane fluidity.Conclusion HaCaT cellular uptake of THP can be enhanced by AEO and SPT of Sanfu Patch, in which AEO enhances the cellular uptake of THP through increasing the cellular membrane fluidity, while the mechanism of SPT in enhancing THP cellular uptake remains further clarification.

Study on biopharmaceutical properties of active fraction from Xiangfu Siwu Decoction by composite phospholipid liposome-based artificial skin membrane / 中草药

Objective: To evaluate the feasibility of the application of composite phospholipid liposome-based artificial skin membrane (CPLASM) for measuring biopharmaceutical properties of active fraction extracted from Xiangfu Siwu Decoction (XSD) via transdermal administration. Methods: The HPLC method was established for the determination of active ingredients (ferulic acid, tetrahydrocolumbamine, tetrahydropalmatine, senkyunolide I, and senkyunolide H) in active fraction of XSD. The oil/water partition coefficient values of these active ingredients were measured. The CPLASM was prepared to determine the in vitro permeability parameters of active fraction of XSD. The obtained parameters were further compared with those obtained by porcine ear skin. Results: The oil-water partition coefficients of ferulic acid, tetrahydrocolumbamine, tetrahydropalmatine, senkyunolide I, and senkyunolide H in active fraction of XSD (pH 5.5) were measured to be 1.220 ± 0.280, 0.670 ± 0.085, 0.920 ± 0.110, 1.040 ± 0.092, 1.030 ± 0.093 (n = 3), respectively. The permeability parameters of the active ingredients through porcine ear skin and CPLASM indicated a significant correlation (r > 0.9). Compared with the classical oil-water partition coefficients, the 6 h- cumulative permeation ratio of five active ingredients through CPLASM can be characterized as effectively predicting permeability parameters through porcine ear skin. Conclusion: The biopharmaceutical properties of active fractions from Chinese material medica can be effectively characterized by the application of CPLASM for the determination of in vitro permeability parameters of active ingredients.

Quaily Analysis of Weiling Granules / 中国药师

Objective: To analyze 167 batches of Weiling granules according to the quality standard and exploratory research, and evaluate the overall quality and standard condition of the preparation. Methods: A TLC method was established for the identification of Atractylodis macrocephalae Rhizoma;an HPLC method was established for the fingerprint and the content determination of paeoniflorin, tetrahydropalmatine and ammonium glycyrrhizinate. An Agilent Poroshell120,SB-C18analytical column (100 mm×4. 6 mm, 2. 7 μm) was employed with gradient elution of acetonitrile-0. 1% phosphoric acid as the mobile phase at the flow rate of 1. 8 ml·min-1, and the sample size was 3 μl. Acid base titration method was used for measuring acid-neutralizing capacity. Results: No interference from the negative controls was shown to the TLC identification of Atractylodis macrocephalae Rhizoma. The fingerprint exhibited better separation of each peak. The precision, reproducibility and stability of the method were good,and the RSDs of the relative retention time and rela-tive peak area were less than 3. 0% . The linear range of paeoniflorin, tetrahydropalmatine and ammonium glycyrrhizinate was 0. 057-0. 568 μg(r=0. 999 9), 0. 035-0. 353 μg(r=0. 999 9)and 4. 244×10 -3-42. 44×10 -3μg(r=0. 999 9), respectively, and the av-eragerecoverywas99.3%(RSD=1.0%,n=6),100.0%(RSD=0.8%,n=6) and99.8%(RSD=1.2%,n=6),respectively. The average recovery of acid-neutralizing capacity was 99. 5% (RSD=0. 5% ,n=6). Conclusion: Exploratory research increases the specificity, controllability and safety of the standards, which provides reference for the further drug standards revision and the drug quality control.

Effect of 60Co Irradiation on 3 Active Ingredients in Compound Hemostatic Capsules / 中国药师

Objective: To establish an HPLC method for the simultaneous determination of 3 active components in compound he-mostatic capsules and investigate the content changes before and after 60Co-γ ray irradiation. Methods: An Agilent-C18column (250 mm×4. 6 mm, 5 μm) was adopted and the flow rate was 1. 0 ml·min-1. The mobile phase consisted of acetonitrile-0. 5% phosphor-ic acid solution (triethylamine was used to adjust pH to 6) with gradient elution, the detection wavelength was 254 nm (0-35 min) and 281 nm (35-55 min), and the column temperature was 35℃. The compound hemostatic capsules were irradiated at 5, 8 and 10 kGy, respectively, and the contents of the 3 active components in compound hemostatic capsules were compared before and after the radia-tion,and the t-test was applied to investigate the significance. Results: Typhaneoside,isorhamnetin-3-O-neohesperidin and tetrahydro-palmatine respectively within the concentration range of 0. 018-0. 11 mg·ml-1,0. 020-0. 120 mg·ml-1and 0. 011-0. 066 mg·ml-1 showed a good linear relationship with the peak area. The average recovery was 97. 7% , 98. 7% and 99. 3% with the RSD of 0. 9% , 0. 9% and 0. 5% , respectively (n=6). After the irradiation, the contents of the three active components in compound hemostatic capsules all showed changes. Under the dose of 8 kGy, the content changes of typhaneoside and isorhamnetin-3-O- neohesperidin showed no statistical significance (P>0. 05),and when the dose increased to 10 kGy,the content of tetrahydropalmatine exhibited sig-nificant difference (P<0. 05). Conclusion: The established determination method for compound hemostatic capsule shows such ad-vantages as high recovery, good repeatability, simple operation and promising separation, which can be used as a quality control meth-od for compound hemostatic capsules. The sterilization effect of 60Co-γ ray irradiation on compound hemostatic capsules is significant without notable changes in the active components. When the irradiation dose is equal to or below 5 kGy,the change of each component is not obvious,which can provide reference for the radiation sterilization of compound hemostatic capsules.

The application of HPLC-MS/MS for comprehensively monitor the feeding analysis of Yuanhu analgesic capsules / 国际中医中药杂志

Objective The HPLC-MS/MS is used to comprehensively monitor the feeding conditions of raw materials in Yuanhu analgesic capsules, and the content of the index components can be detected at the same time. Methods The Inertsil ODS-3 (4.6 mm×250 mm, 5 μm) was used with the column temperature 40 ℃, Flow phase: 0.1% formic acid-acetonitrile; the gradient elution program, active ingredients were separated by HPLC, and the Electrospray Ionization Mass (ESI) source was applied and operated in the negative ion mode, and reactions ion monitoring mode (MRM) for quantitative analysis were selected. Results Through analysis and contrast of the medicinal materials, reference substance of primary mass spectrogram showed the same characteristic peak, and the proprietary Chinese medicine can be judged by prescription feeding process. The tetrahydroxene and imperatorin had a good linear relationship in 5.08×10-5-30.45×10-5μg (r=0.999 4), 5.02×10-5-30.09×10-5μg (r=0.999 2). Precision test were 0.99% and 1.14%, the recovery rate were 97.02%-99.66%, 97.62%-99.94%. Conclusions The method is simple, accurate and reliable, high sensitive and fast. It is suitable to monitor the feeding condition and quality of yuanhu analgesic capsules.

Simultaneous Determination of Paeoniflorin, Tetrahydropalmatine and Osthole in Gujinwan Capsules by HPLC / 中国药师

Objective:To develop an HPLC method for the content determination of paeoniflorin, tetrahydropalmatine and osthole in Gujinwan capsules. Methods:The determination was performed on an Eclipse XDB-C18 column (250 mm × 4. 6 mm,5 μm) with the mobile phase consisting of 0. 1％ phosphoric acid (adjusting pH to 6. 6 with triethylamine)-acetonitrile with gradient elution. The flow rate was 1. 0 ml·min-1 . The column temperature was 30 ℃. The detection wavelengths were set at 230 nm and 280 nm. Re-sults:The calibration curves were found to be linear within the range of 9. 56-33. 65 μg· ml-1 for paeoniflorin(r=0. 9998), 3. 65-12. 86 μg· ml-1for tetrahydropalmatine(r=0. 9999) and 5. 81-20. 45 μg· ml-1for osthole(r=1. 0000). The average recovery of paeoniflorin, tetrahydropalmatine and osthole was 98. 8％(RSD=1. 1％), 98. 4％(RSD=0. 8％) and 99. 1％(RSD=1. 4％)(n=6),respectively. Conclusion:The method is simple, accurate and reproducible, which can be used for the quality control of Gujinwan capsules.

Optimization of microwave processing technology for vinegar Corydalis Rhizoma by response surface methodology / 中草药

Objective To optimize the microwave processing technology of vinegar Corydalis Rhizoma. Methods On the basis of single factor test, the index was evaluated by total rating of content of tetrahydropalmatine, protopine and the total alkaloid, which were determined by HPLC and UV spectrophotometry method, four factors (fire, stuffy time, processing time and dosage of vinegar) were studied by response surface, the microwave processing technology of vinegar Corydalis Rhizoma was optimized by response surface methodology. Results The optimal parameters of microwave processing technology were as follows: microwave power of 70%, stuffy time of 1.5 h, microwave time of 2.6 min, vinegar dosage of 27.5%, the content of tetrahydropalmatine, protopine, and total alkaloid were 0.112 4%, 0.041 8%, and 0.85%. Conclusion Microwave processing can be used as a processing method to enrich the traditional processing technology.