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In this study, the marker compounds of Curcumae Rhizoma (CR) were simultaneously quantified by high-performance liquid chromatography equipped with a photodiode array detector and the anti-inflammatory effects of CR extract and marker compounds in human benign prostatic hyperplasia epithelial-1 (BPH-1) cell lines were investigated. The marker components (4S,5S)-(+)-germacrone-4,5-epoxide, furanodienone, and germacrone, were separated on Gemini C₁₈ columns (250 mm × 4.6 mm, 5 µm) at 40 ℃ by using a gradient of two mobile phases eluting at 1.0 mL/min. Prostaglandin E₂ (PGE₂) levels in Human BPH-1 cells were determined with an ELISA kit. The coefficients of determination in a calibration curve of each analyte were all 0.9997. The limits of detection and quantification of the three compounds were 0.10 – 0.32 µg/mL and 0.30 – 0.98 µg/mL, respectively. The content of three compounds, (4S,5S)-(+)-germacrone-4,5-epoxide, furanodienone, and germacrone, in the CR sample were found to be 5.79 – 5.92 mg/g, 4.72 – 4.86 mg/g, and 1.06 – 1.09 mg/g, respectively. Regarding pharmacological activity against benign prostatic hyperplasia, CR and its components significantly suppressed PGE₂ levels of BPH-1 cells. The established analysis method will help to improve quality assessment of CR samples and related products. In addition, CR and its components exhibit anti-inflammatory activity in BPH-1 cells, suggesting the inhibitory efficacy of these compounds against the pathogenesis of BPH.
Subject(s)
Humans , Calibration , Cell Line , Chromatography, Liquid , Curcuma , Enzyme-Linked Immunosorbent Assay , Limit of Detection , Methods , Prostatic Hyperplasia , RhizomeABSTRACT
Objective:To analyze and compare volatile components in raw and fried products of Curcuma wenyujin radix, and find out the material basis for the differences in efficacy between them,in order to provide powerful support and reference for the rational use of C. wenyujin radix of different processed products and establish a more perfect quality evaluation system. Method:HS-SPME combined with GC-MS was used to analyze and compare volatile components and relative percentage in different processed products of C. wenyujin radix. Result:Totally 42 peaks were detected from the raw product of C. wenyujin radix,and 23 components were identified. Totally 59 peaks were detected from vinegar-baked C. wenyujin radix,and 34 components were identified. Totally 64 peaks were detected from wine-baked C. wenyujin radix,and 40 components were identified. Compared with C. wenyujin radix,22 kinds of new ingredients were produced. Compared with C. wenyujin radix,29 kinds of new ingredients were produced in wine-baked C. wenyujin radix. The composition and content of volatile components in different processed products of C. wenyujin radix were changed. Conclusion:The headspace solid phase micro-extraction method is simple in operation and fast in extraction speed,with no use of organic solvent, no pollutant in the environment and a low operation cost. The method using headspace solid phase micro-extraction combined with gas chromatography-mass spectrometry has a high stability and reliability. It is suitable for rapid analysis of volatile components between C. wenyujin radix and its different processed products. It can provide a scientific basis for the quality evaluation of C. wenyujin radix and its different processed products,and a scientific reference for the rational use of C. wenyujin radix resources.
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OBJECTIVE： To establish the method for simultaneous determination of curdione， curcumenol， germacrone， furanodiene and β-elemene in Curcuma wenyujin， and to optimize its processing method in production place. METHODS： HPLC method was adopted. The determination was performed on ODS SP C18 column with mobile phase consisted of acetonitrile-0.05％ phosphoric acid solution （gradient elution） at the flow rate of 1.0 mL/min. The detection wavelength was set at 214 nm， and column temperature was 30 ℃. The sample size was 10 μL. 18 batches of sample medicinal materials were prepared by steaming-drying （10， 20， 30， 40 min）， or cooking-drying （drying temperature 50， 60， 70 ℃）， and then the contents of 5 components were determined. Relative relational degree （ri） obtained by grey relational analysis method as indexes was used to optimize the processing methods of C. wenyujin in production place using. RESULTS： The linear range of curdione， curcumenol， germacrone， furanodiene and β-elemene were 25.56-409.0， 2.05-32.8， 6.36-102.0， 9.14-146.0， 8.62-138.0 μg/mL （r≥0.999 2）， respectively. RSDs of precision， stability and reproducibility tests were all lower than 2.0％ （n＝6）. Quantitative limits were 0.34， 0.62， 0.11， 0.14 and 1.20 μg/mL， and detection limits were 0.10， 0.19， 0.03， 0.04 and 0.42 μg/mL， respectively. The average recoveries were 101.3％， 98.7％， 99.0％， 99.6％， 96.4％， respectively （RSD＜2.8％， n＝6）. The results of grey relational analysis showed that the relative relational degree （ri） from the processing technology of “boil for 10 min-drying at 50 ℃” was greatest， which had relatively small comprehensive effect on the contents of curdione， curcumenol， germacrone， furanodiene and β-elemene respectively. CONCLUSIONS： Established HPLC method can be used for content determination of 5 components in C. wenyujin. The results of optimization can provide reference for the selection of processing method of C. wenyujin in production place.
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To analyse the quality of three processed products of Rhizome of Curcuma wenyujin by establishing an ultra performance liquid chromatography (UPLC) method for simultaneously determining five sesquiterpene components in three processed products of rhizome of C. wenyujin and establishing UPLC fingerprints. Component determination was achieved on Waters ACQUITY UPLC BEH C₁₈ column（2.1 mm×50 mm, 1.7 μm), with acetonitrile-water as mobile phase for gradient elution. The flow rate was 0.3 mL·min⁻¹; column temperature was 30 °C; the detection wavelength was set at 214 nm and injection volume was 1 μL. The similarity was analyzed with "Similarity Evaluation System for Chromatographic Fingerprint of Chinese Materia Medica (2012.130723)", and hydrophobic cluster analysis (HCA), principal components analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA) were conducted by using simca-p14.1 software to investigate the differences in components among these three kinds of processed products. The curzerene, curdione, curcumol, germacrone, furanodiene and β-elemene showed good linearity relationship with chromatographic peak area within the ranges of 10.8-320（=0.999 9）, 10.36-259（=0.998 1）, 10.54-263.5（=0.999 3）, 30.2-755（=0.999 6）and 34.38-862（=0.999 9）mg·L⁻¹, respectively; their average recoveries were 98.75%, 98.69%, 98.63%, 99.76% and 99.57% respectively, with RSD of 2.67%, 1.47%, 1.29%, 2.54% and 0.87% respectively. The similarity of 30 batches of samples was larger than 0.9, indicating good consistency of the samples. The samples can be clearly classified into three categories for HCA, PCA and OPLS-DA pattern recognition, the differential chromatographic peak among three processed products was found respectively. The results showed that the pharmacology basis had changed obviously after processing of Rhizome of C. wenyujin, so it can provide the scientific basis for rational clinical application and establishing quality standards of three processed products of Rhizome of curcuma wenyujin.
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Objective To establish the qualitative and quantitative analytic method for quality control of Wujin capsules.Methods Curcuma wenyujin Y.H.Chen et C.Ling and Crataegus pinnatifida Bge.in Wujin capsules were identified by TLC method.The content of tenacissoside H was determined by HPLC.Chromatographic separation was achieved at Waters Xbridge C18 column (100 mm× 3.0 mm, 3.5 μm) with an evaporative light scattering detector (ELSD).Acetonitrile and H2O were used as the mobile phase for isocratic elution.Results The qualitative identification by TLC was specific.The well separated clear spots were exhibited on each thin layer plate.Tenacissoside H had a linear equation in the range of 11.60-580.00 μg/ml(r=0.999 0).The average recovery was 101.54％.Conclusion The TLC identification of Curcuma wenyujin Y.H.Chen et C.Ling and Crataegus pinnatifida Bge.was simple and well established.The HPLC-ELSD method was specific,sensitive and reliable.Those methods can be used for the quality control of Wujin capsules.
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OBJECTIVE: To establish a method for the simultaneous determination of curcumin and germacrone in Curcuma wenyujin by ultrasound-assisted ionic liquid-reversed phase liquid chromatography. METHODS: An Agilent ZORBAX Eclipse Plus C18 column (4. 6 mm × 250 mm, 5 μm) was used. Gradient elution was conducted with the mobile phase consisting of acetonitrile and 0.1% phosphoric acid at the flow rate of 1.0 mL·min-1. The detection wavelength was set at 265 nm, the flow rate was set at 1.0 mL·min-1, and the column temperature was maintained at 30 ℃. RESULTS: The result showed that curcumin and germacrone had the highest extraction yield when the [BMIM]PF6 methanol solution concentration of 0.6 mol·L-1 was used as the extraction solvent and the solid-liquid ratio was 1:20. Under the optimized condition, good linearity of curcumin was shown for curcumin in the range of 0.041 2-2.06 μg (r=0.999 9)and the average recovery was 98.31%. The good linearity of germacrone was shown for germacrone in the range of 0.010 04-0.502 μg (r=0.999 8)and the average recovery was 97.62%. CONCLUSION: This method is convenient, environmentally friendlygreen and highly reproducible. It is beneficial important to study the extraction method of the active gradients of the traditional Chinese medicine Curcuma wenyujin active gradient and its quality control of Curcuma wenyujin.
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Objective: To research on the most appropriate SRAP-PCR amplication system of Curcuma wenyujin. Methods: The concentration of Mg2+, dNTP, template DNA, primer, and Taq polymerase was optimized by amplifying technology and method of PCR based on isolating genomic DNA from C. wenyujin. Results: The best amplification system for C. wenyujin was as follows: total reaction volume of 25 μL, including Mg2+ (25 mmol/L) 2.0 μL, Taq polymerase(5 U/μL)0.4 μL, random primer (20 μmol/L) 2 μL, template DNA (5 ng/μL) 2.0 μL, dNTP (25 mmol/L) 2.0 μL, 10 × PCR buffer 2.5 μL. Conclusion: The stripes of amplification products are clearness, high brightness, and good reproducibility. It indicates that the reaction system and reaction procedures which are confirmed in this experiment are applied to the SRAP molecular markers in C. wenyujin, based for the genetic diversity of C. wenyujin.
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The aim of the study was to investigate the sesquiterpene constituents from the rhizomes of Curcuma wenyujin Y．H．Chen et C．Ling．The isolation and purification of the constituents from the 50％ EtOH extracts of the rhizomes were performed with repeated column chromatography over sillica gel and macroporous resin．Eight sesquiterpenes were obtained and identified as wenyujinlactone A(1),neolitamone A(2),zedoarondiol(3),isozedoarondiol(4),aerugidiol(5),curcumol (6),curdione(7) and (1R,10R)-epoxy-(-)-1,10-dihydrocurdine (8) by means of spectral analysis．Among them,compound 1 was found to be a new eudesmane sesquiterpene lactone,whilst compounds 2-5 were obtained from this plant for the first time．
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[Objective]To discuss the analgesic effects of Radix Curcuma extracts on mice.[Method]The writhing tests were adopted to observe the mice writhing times.[Results]The Radix Curcuma extracts middle-dose group can obviously inhibit the mice average writhing times,with statistically significant differences in comparison to contral group,P
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Objective To establish the chromatographic fingerprint of Curcuma wenyujin by HPLC.Methods The chromatographic fingerprint of C.wenyujin was established by HPLC with Shimadzu VP-ODS C18 column(250 mm?4.6 mm,5 ?m),acetonitrile-water(0.1 phosphoric acid) gradient elution,the flow rate of 1.0 mL/min,column temperatures of 30 ℃,and detective wavelength at 210 nm.ResultsTaking germacrone as the reference peak,14 common-peaks were selected as the fingerprint peaks.The similarities among samples of C.wenyujin among 19 approved samples and their standard chromatographic fingerprints were calculated with a software "Similarity Evaluation System for Chromatographic Fingerprint of Chinese Materia Medica" published by GPC(Version 2004A) and cluster analysis.ConclusionThis method with better accuracy and reproducibility could be used in the quality control of C.wenyujin.
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Object To develop a new method for the determination of curcumol in essential oil from rhizoma Curcumae L.. Methods The contents of curcumol were determined by high performance capillary gas chromatography with sequential increase of temperature on a HEWLETT PACKARD 5890A gas chromatograph. Results The method can be used to determine curcumol with accuracy at a recovery of 101.4% and RSD of 0.40%. Conclusion The present study provided a satisfactory method for the determination of curcumol, and it was found that its contents in four different species (C. wenyujin, C. longa, C. aeruginose, and C. kwangsiensis) were markedly different.
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Objective:To study optimum inclusion process conditions for volatile oil from Curcuma wenyujin. Methods: The study was carried out with orthogonal design. The process conditions were studied by determining the ultizision ratio of volatile oil from Curcuma wenyujin , the oil bearing rate and extract ratio of inclusion compound. Results: The optimum preparation conditions for inclusion were established as∶oil∶? CD was 1∶9, The inclusion temperature and time were at 40 ?C and for 1.5h , The ultizision ratio of oil was 86.5% . Conclusions: The rate is high and the method is applicable.
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AIM: To find the optimum extraction of volatile oil. METHOD: To extract the volatile oil by steam distillation or supercritical fluid extraction (SFE). RESULT: (1) The optimum extraction of volatile oil extracting by steam distillation is: Ligustricum Chuanxiong Hort.:Soak time 12 hrs, thin powder, extract time 3hrs, and to add water with 1∶10; Alpinia Officinarum Hance: Soak time 0 hr, thick powder, extract time 3 hrs, and to add water with 1∶10; Curcuma Wenyujin Y.H. chen C. Ling: Soak time 24hrs, mild powder, extract time 1.5hrs, and to add water with 1∶5; Notopterygium incisum Ting: Soak time 0 hr, thick powder, extract time 3 hrs, and to add water with 1∶10. (2) The best condition of volatile oil extracting by SFE is: Ligustricum Chuanxiong Hort.: 60℃, thin powder, press 25MPa, extract time 1.5hrs; Alpinia Officinarum Hance: 60℃, mild powder, press 25MPa, extract time 0.5hr; Curcuma Wenyujin Y.H. chen C. Ling: 60℃, thin powder, press 20MPa, extract time 1 hr; Notopterygium incisum Ting: 60℃, thin powder, press 25MPa, extract time 1hr. CONCLUSION: Supercritical fluid extraction (SFE) is superior to steam distillation.