Optimization of extraction process of Sarcandra glabra by combination of entropy weight method with Box-Behnken response surface method / 中国药房

OBJECTIVE To optimize the extraction process of Sarcandra glabra. METHODS The contents of rosmarinic acid and isofraxidin in S. glabra were determined by HPLC； ultrasonic time， ultrasonic temperature， solid-liquid ratio （mL/g） and methanol volume fraction were investigated by single factor test. Based on the results of single factor test， experimental scheme was designed by Box-Behnken response surface method， and the entropy weight method was used to assign the weight of each index and calculate the comprehensive score. Taking the comprehensive score as the evaluation index， the extraction process of S. glabra was optimized， and then optimized extraction process was verified. RESULTS The optimal extraction technology of S. glabra included ultrasonic time of 40 min， ultrasonic temperature of 45 ℃， liquid-solid ratio of 50∶1， methanol volume fraction of 70%. The results of 3 times of verification experiment showed that average comprehensive score was 0.988 6， and the RSD was 0.50%. The deviation between the actual value and the predicted value （0.985 1） of each comprehensive score was within ±1%. CONCLUSIONS The optimized extraction method is stable， feasible and repeatable， which can provide reference for extraction of S. glabra.

Quality standard study of Sarcandrae Herba Dispensing Granules based on specific chromatogram and multiple ingredients determination / 中草药

Objective: To establish HPLC and multiple components determination method of Sarcandrae Herba Dispensing Granules (SHDG), in order to compare the difference of the quality in various SHDG samples and provide an effective method to ensure the quality of SHDG. Methods: HPLC-UV method was used to establish the characteristic chromatogram of SHDG, and acetonitrile-0.2% formic acid solution was used as the mobile phase with the gradient elution. The common peaks were identified by comparison with the reference standards and HPLC-Q/TOF. At the same time, the method for simultaneous determination of chlorogenic acid, isofraxidin, and rosmarinic acid was established with the same approach. Chemometrics software Chempattern was employed to analyze the data. Through cluster analysis and principal component analysis, different preparations from different manufacturers and different batches from the same manufacturer were classified, and the main components causing the differences were clarified. Results: The SHDG fingerprint was established to confirm and identify seven characteristic peaks, namely, neochlorgenic acid, chlorogenic acid, cryptochlorogenic acid, caffeic acid, isofraxidin, rosmarinic acid-4-O-β-D-glucoside, and rosmarinic acid. The main chromatographic peaks of SHDG can be completely separated within 55 min. There was a good linear relationship between the peak area and concentration of chlorogenic acid, isofraxidin, and rosmarinic acid. The average recovery rates were 98.92% (RSD 1.54%), 98.20% (RSD 1.12%), and 99.58% (RSD 1.12%), respectively. The chlorogenic acid content of 18 batches of samples was 0.33-1.39 mg/g, the content of isocyanidine was 1.31-2.74 mg/g, and the content of rosmarinic acid was 1.11-4.54 mg/g. The similarity between 18 batches of samples and the common mode was 0.688-0.992. There were certain differences in the content of chlorogenic acid, isofraxidin, rosmarinic acid in the SHDG of various batches and manufacturers. Conclusion: The proposed specific HPLC characteristic chromatogram and quantitation method of three components for SHDG offered more comprehensive reference for quality control of the crude drug.

In vivo metabolism of three coumarins from Chimonanthi Radix based on UHPLC-QTOF-MS/MS / 中国中药杂志

Scopolin (SC-1), scopoletin (SC-2) and isofraxidin (IS-1) are the main active constituents in Chimonanthi Radix. However, the in vivo metabolism of SC-1, SC-2 and IS-1 have not been comprehensively clarified. In this study, the in vivo metabolic profiles of these three coumarins in the rat plasma, urine and feces were analyzed. Ultra-high performance liquid chromatography-quadrupole time of flight mass spectrometry (UHPLC-QTOF-MS/MS) method was applied to characterize the prototypes and metabolites of SC-1, SC-2 and IS-1 in rat feces, urine, and plasma after intravenous administration. A total of 11 metabolites of the three parent compounds were tentatively identified. The main metabolic pathways were analyzed by identification of metabolites, and it was found that these three coumarins underwent multiple in vivo metabolic reactions including glucuronidation, sulfonation, isomerism and reduction. In this study, the analysis of metabolites of three coumarins basically demonstrated their in vivo metabolic process, providing basis for the further pharmacokinetics and pharmacological evaluations of SC-1, SC-2 and IS-1.

Principal Components Analysis in Determination of Seven Chemical Constituents in Different Parts and Collecting Time of Sarcandra glabra by HPLC / 中国药学杂志

OBJECTIVE: To develop an HPLC method for simultaneous determination of seven active constituents including neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, quercetin-3-O-β-D-glucuronide, isofraxidin, kaempferol-3-O-β-D-glucuronide and rosmarinic acid in different parts and various collecting time of Sarcandra glabra. METHODS: The DIKMA Diamonsil C18 (4.6 mm × 250 mm, 5 μm) column was used. acetonitrile(A) -0.2% phosphoric acid(B) was used as the mobile phase with gradient elution(0 - 10 min, 5% A→15% A; 11 - 18 min, 15% A→25% A; 18 - 25 min, 25% A; 25 - 40 min, 25% A→40% A), at a flow rate of 1.0 mL•min-1. The detection wavelength was set at 344 nm and the column temperature was maintained at 35 ℃. RESULTS: The calibration curves of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, quercetin-3-O-β-D-glucuronide, isofraxidin, kaempferol-3-O-β-D-glucuronide and rosmarinic acid were in good linearity over 9.32 - 466.00, 11.25 - 562.50, 10.94 - 547.00, 8.68 - 434.00, 10.48 - 524.00, 9.66 - 483.00 and 10.86 - 543. 00 μg•mL-1 (r = 0.999 6 - 0.999 9), respectively. The average recoveries (n = 6) were in the range of 99.0% - 101.5% and RSD were in the range of 0.85% - 1.8%. The optimal parts and collecting time of Sarcandra glabra are aerial parts and from September to November according to the comprehensive evaluation. CONCLUSION: The method is precise and reliable, can be used to determin the content of seven chemical constituents in Sarcandra glabra and provide a reference for collecting in cultivate area.

Determination of Three Active Components in Shuanghua Caoshanhu Hanpian by HPLC / 医药导报

Objective To establish a HPLC method for determination of three active components in Shuanghua Caoshanhu Hanpian.Methods Agilent TC-C18 (4.6 mm × 250 mm,5 μm) column was used.The mobile phase was acetonitrile-water containing 0.2％ phosphoric acid with gradient elution mode at a flow rate of 1.0 mL· min-1,the column temperature was 30 ℃,the detection wavelength was 327 mn (0-10 min) for chlorogenic acid,342 nm (10-35 min) for isofraxidin and msmarinic acid.Results Chlorogenic acid had a good linearity in the range of 0.052-0.939 μg (r =0.999 8,n =6),the average recovery was 99.23％ with RSD of 1.4％ (n =9).Isofraxidin had a good linearity in the range of 0.004-0.077 μg (r =0.999 8,n =6),the average recovery was 100.33 ％ with RSD of 1.8 ％ (n =9).Rosmarinic acid had a good linearity in the range of 0.012-0.213 μg (r =0.999 9,n =6),the average recovery was 99.42％ with RSD of 1.4％ (n =9).Conclusion The method is simple,accurate and reliable.It can be used to determine chlorogenic acid,isofraxidin and rosmarinic acid in Shuanghua Caoshanhu hanpian at the same chromatogram condition.

Percutaneous Absorption in Vitro of Jiegugao Blended and Pasted by Commonly Used ;Ointment Matrices in Tujia Minority / 中国中医药信息杂志

Objective To establish a method to simultaneously determine syringing and isofraxidin by HPLC;To investigate the features of percutaneous absorption in vitro of Jiegugao blended and pasted by white vinegar, honey and vaseline; To discuss the mechanism of commonly used ointment matrices in Tujia Minority. Methods Rat abdomen skin in vitro was as transdermal barrier;the modified Franz diffusion pool was used to simulate human skin medication; the content of syringin and isoprofen was determined by HPLC; the percutaneous absorption equation was established and the related parameters, such as cumulative permeation rate and permeation rate, were calculated. Results When using Syncronis C18 (250 mm × 4.6 mm, 5 μm) as chromatographic column, acetonitrile-0.1%phosphoric acid as mobile phase, 1.0 mL/min as perfusion speed and 265 nm as determine wavelength, regression equation of syringingwas A=10 686.454 6C+1565.778 8 (r=1.000 0), regression equation of isofraxidin was A=12 297.305 4C-5913.729 9 (r=0.999 9). Cumulative permeation quantity of syringing in Jiegugao blended and pasted by white vinegar, honey, vaseline and blank were 7.549 2, 4.580 3, 3.890 8 and 5.378 4 μg?cm-2?h-1 respectively and permeation rate were 25.66%, 16.11%, 13.73% and 18.78%. Meanwhile, cumulative permeation quantity of isofraxidin were 2.536 9, 1.941 8, 1.178 2 and 2.293 6 μg?cm-2?h-1 respectively and permeation rate were 47.04%, 35.06%, 22.11%and 41.11%. Conclusion Using white vinegar as the ointment matrix can promote the percutaneous absorption of effective composition in Jiegugao blended. However, it will retard the percutaneous absorption of effective composition in Jiegugao when using honey and vaseline as the ointment matrices, but honey and vaseline can be used as a slow-release matrix.

Phenolic constituents from fruit of Gardenia jasminosides var. radicans / 中草药

Objective To investigate the phenolic constituents in the fruit of Gardenia jasminosides var. radicans. Methods The phenolic constituents were isolated and identified by chromatography on silica gel, Toyopearl HW-40, Sephadex LH-20, MCI gel CHP-20, ODS, and RP-HPLC. Their structures were elucidated on the basis of physicochemical properties and spectral analyses. Results Seventeen compounds were isolated from 50% aqueous acetone extract from the fruit of Gardenia jasminosides var. radicans and identified as 3,3',5-trimethoxy-9,9'-epoxylignane-4,4'-diol (1), 3,4-divanillyltetrahydrofuran (2), marphenol C (3), sinapaldehyde (4), hydroxycinnamic acid (5), isoferulic acid (6), caffeic acid (7), isofraxidin (8), isoscopoletin (9), 6-methoxy-7-hydroxycoumarin (10), 6,7-dihydroxycoumarin (11), syringic acid (12), 3-hydroxy-4-methoxybenzoic acid (13), 3,4-dihydroxybenzoic acid (14), p-hydroxybenzoate (15), 4,5-dihydroxy-3-methoxy-benzoic acid (16), and gallic acid (17). Conclusion All 17 compounds are isolated from this plant for the first time.

High performance liquid chromatography for determining the isofraxidin content in Feitai Capsule / 医学研究生学报

Objective Feitai Capsule,a compound of traditional Chinese herbal medicine,has been screened and refined repeatedly for many years and shown to have a good anti-tumor effect.Strict quality control and further screening of the efficacious components of the compound are of great clinical significance.The purpose of this study was to establish the methods for determining the isofraxidin content in Feitai Capsule.Methods We determined the content of isofraxidin in Feitai Capsule by high performance liquid chromatography (HPLC),using the chromatographic column Hypersil ODS-C18 (4.6 mm?150 mm,5 ?m),with the mobile phase as acetonitrile 0.2% phosphoric acid solution (21∶79),the flow rate of 1.0 ml/min,the detective wavelength of 344 nm and the column temperature at 30℃.Results Isofraxidin showed a good linearity,within the range of 2.00-10.80 ?g/ml (y=69 427x+15961,r = 0.999 9),with the average recovery of 97.89% and RSD of 1.64% (n = 6).Conclusion HPLC,accurate and reproducible,is suitable for the determination of the isofraxidin content in Feitai Capsule.

Self-microemulsifying drug delivery system of Acanthopanax senticosus total saponin / 中草药

Objective To develop self-microemulsifying drug delivery system(SMEDDS) of Acanthopanax senticosus total saponins(ASTS) and the determination method.Methods The equilibrium solubility of ASTS in different compositions of oils,emulsifier and assistant emulsifier was investigated.The self-microemulsion formula was optimized by constructing the pseudo-ternary phase diagrams of blank SMEDDS and the studying the self-microemulsifying efficiency and the stability of drug-loaded SMEDDS.Taking isofraxidin,an effective component of A.senticosus,as the content determination index,the content was determined by HPLC.Results The optimal self-microemulsion formula was composed of Caf,propanediol,and ethyl linolenate.The ratio of them was 16∶4∶5.The average particle size was 40 nm.The average content of isofraxidin was 92.6 ?g/mL.Conclusion The acquired microemulsion with small particle size is stable.The content determination method of taking isofraxidin as the quality control index is accurate and reliable.

Application of flocculation and membrane separation in purification of Caoshanhu extract / 中草药

Objective To investigate the different pretreatments of Caoshanhu extract and their effects on the downstream process,membrane separation.Methods Based on the index related to isofraxidin,the different pretreatment methods of Caoshanhu extract were evaluated and their effects on membrane separation downstream were analyzed by the comparison of different pretreatments and further orthogonal test of ZTC_(1+1) Ⅲ,chitosan,and(KAl(SO_4)_2?12H_2O).Results After the pretreatment of microfiltration,both the best membrane flux and the highest percentage of isofraxidin filtered through the membrane were obtained and the time used was the least,and the ratio of isofraxidin to insoluble substances was tiptop in all pretreatment methods in comparison to other methods,such as ZTC_(1+1) Ⅲ,chitosan,and(KAl(SO_4)_2?12H_2O) methods.Conclusion Microfiltration is a better pretreatment method in preparation of Caoshanhu extract.

Determination of Isofraxidin in Pientzehuang Oral Tablet by RP-HPLC / 中药新药与临床药理

Objective :To establish a RP-HPLC method for determination of isofraxidin in Pientzehuang oral tablet. Methods: RP-HPLC analysis was carried out on Nava-Pak C18 column and with acetonitrile: 0.1%phosphoric acid (15∶85) as a mobile phase. Results: The linear range was 12.9～90.3 ?g/mL(r=0.9999,n=7).The average recovery rate was 102.65 %and RSD=0.71 %.The intra-day and inter-day RSD was less than 3 %.Conclusion :This method is simple, rapid and accurate and suitable for the quality control of the preparations of Chinese herbal medicine containing isofraxidin.

Study on microwave-assisted extraction process for active ingredients in Radix acanthopanacis senticosi / 中成药

Objective: To study the microwave assisted extraction process for saponins and isofraxidin in Radix Acanthopanacis Senticosi as a marker. Methods: Using the uniform design, the optimization process of microwave assisted extraction for saponins and isofraxidin in Radix Acanthopanacis Senticosi was gained by continuous microwave radiation. Results: Some parameters such as microwave power, radiation time, solvent concentraction, solvent consumption, pulverized degree and dipping time had interactions on the extraction of active constituents in Radix Acanthopanacis Senticosi. Conclusion: The optimization process was obtained which could be described as following: 510W as the microwave power, 30 min as the radiation time, 85% ethanol as the solvent, 1∶5 as the proportion of raw material to solvent, 40 mesh as the grinding degree and 0.5 hr as the dipping time. Meanwhile, the model developed were proved to be reasonable by the pilot experiment.

Determination of isofraxidin in Xiaoerkaiwei Oral Liquid by HPLC / 中成药

Objective: To establish a method for determination of isofraxidin in Xiaoerkaiwei Oral Liquid (Radix Acanthopanacis Senticosi, Endothelium Corneum Gigeriae Galli, Semen Alpiniae Katsumadai, Semen Pharbitidis, etc.) by RP HPLC. Methods: The HPLC system consisted of Symmetry C 18 column(3.9m?150mm, 5?m), methanol water (0.1% formic acid) (30∶70) mixture as a mobile phase, detection wavelength at 342nm, 0.8mL?min -1 of flow rate and column temperature at 30℃.Results: The calibration curves of the isofraxidin were linear ( r =0.9993). The precision were perfect ( RSD =1.87). The samples were stable in 6 h. The average recovery of Xiaoerkaiwei Oral Liquid was 102.4% with RSD of 1.76%.Conclusion: This method is simple, quick and specific, suitable for the quality control.

Study on Determination of Total Isofraxidin in Ciwujia Tablets by HPLC / 中成药

Objective: To establish the determination method of total isofraxidin in Ciwujia Tablets. Methods: In this method phenomenex C 18 column was used, acetonitrile -0.05mol?L -1 potassium dihydrogen phosphate solution (15∶85) was used as a mobile phase. The detection wavelength was at 344nm.Results: The recovery of the added sample was 102.8% and RSD was 2.2%. Conclusion: This method is simple and the result is reliable.

Determination of isofraxidin and rosmarinic acid in Zhongjiefeng Tablet by HPLC / 中成药

AIM:To determine the contents of isofraxidin and rosmarinic acid in Zhongjiefeng Tablet(Sarcandra glabra(Thunb)). METHODS:HPLC was performed on Diamonsil C18 column(250 mm ? 4. 6 mm,5 ?m),the mobile phase consisted of acetonitrile-0. 1% phosphoric acid solution(80 ∶ 20) at a flow rate of 1. 0 mL/min. The column temperature was set at 35 ℃,UV detection wavelength was at 342 nm. RESULTS:Contents of isofraxidin and rosmarinic acid detecded showed good linear relation(R2 =0. 999 99,0. 999 92,respectively) and the average recoveries(n =6) of 98. 7% (RSD =2. 22% )and 96. 9% (RSD =2. 76% ). CONCLUSION:The method is fea-sible,accurate and reliable,thereby available for quality control.

Simultaneous determination of isofraxidin and schisandrin in Ciwujia Naoling (Oral) Liquid by HPLC / 中成药

AIM: To establish a method of simultaneously determining effective components in Ciwujia Naoling Oral Liquid by RP-HPLC. METHODS: Stationary phase: Alltech C_(18) column,the mobile phase was methanol(A)-0.4% HAC-water(B) with linear gradient elution.Detection was performed at wavelength of 344 nm,254 nm respectively for isofraxidin and schisandrin. RESULTS: The two effective components could be detected by this method,and their contents could be determined.A good linearity of isofraxidin and schisandrin was in the range of 0.88-17.6 ?g/mL,0.76-15.2 ?g/mL respectively.The average recoveries were 100.64%,100.33% respectively,RSD were 1.07%,0.65% respectively. CONCLUSION: The method is simple,feasible and reproducible and can be used for the quality control of Ciwujia Naoling Oral Liquid.

Simultaneous determination of isofraxidin and formononetin in Aidi Freezing Dried Powder for Injection by HPLC / 中成药

AIM: To establish a method for determining isofraxidin and formononetin in Aidi Freezing Dried Powder for Injection(Radix et Rhizoma Ginseng,Radix Astragali,Radix et Rhizoma seu Caulis Acanthopanacis senticosi,etc.) by HPLC. METHODS: Chromatographic assay was performed on Hypersil C_(18) column(200 mm?4.6 mm,5 ?m).The mobile phase consisted of acetonitrile(A) and water(B).Gradient program was adopted as follows:0-15 min:16% A,15-45 min:40% A.The flow rate was 1.0 mL/min and the detection wavelength was 344 nm and 254 nm. RESULTS: Isofraxidin had a good linearity(r=(0.999 7)) in the range of 36.80-184.0 ?g/mL.The average recovery was 97.2%,and RSD=2.1%(n=9).Formononetin had a good linearity(r=0.999 7) in the range of 36.40-182.0 ?g/mL.The average recovery was 98.8%,and RSD=2.1%(n=9). CONCLUSION: The above method is simple,sensitive and accurate.It can be used for quality control of Aidi Freezing Dried Powder for Injection.