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To study the chemical constituents and their biological activities in the rhizomes of Curcuma phaeocaulis, silica gel column chromatography, reverse medium pressure liquid chromatography, preparative thin layer chromatography, and semi-preparative high performance liquid chromatography were used for isolation and purification and modern spectroscopic methods were used to determine the structure of the isolated compound. Moreover, the effect of the compound on the proliferation of HUVECs was determined by the MTT assay. A new elemane-type sesquiterpenoid glycoside was isolated from the n-butanol soluble fraction of 95% ethanolic extract of the rhizomes of Curcuma phaeocaulis. Its structure was identified as (1Z)-2-hydroxy-curzerenone 2-O-β-D-glucoside. It showed no inhibitory effect on the proliferation of HUVECs.
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Objective:To establish the grading standard of <italic>Curcuma phaeocaulis</italic> seed rhizoma，and explore the effect of seed rhizoma classification on the growth，yield and quality of <italic>C. phaeocaulis</italic>. Method:The purity，diameter，weight，length，germination rate and water content of <italic>C. phaeocaulis</italic> seed rhizoma samples were determined. The grading index was determined by partial correlation analysis，variance analysis and cluster analysis. According to actual conditions of production，the classification standard of<italic> C. phaeocaulis</italic> seed rhizoma was established. Then the field experiments were carried out with different grades of seed rhizoma for treatment， so as to analyze the effect of seed rhizoma grade on plant growth，yield and quality of medicinal materials. Result:With seed rhizoma diameter and seed rhizoma weight as classification indexes，<italic>C. phaeocaulis</italic> seed rhizoma were divided into three grades. The field experiments showed that with the growth and development of <italic>C. phaeocaulis</italic>，there were significant differences in plant height and leaf length among different grades of seed rhizoma during the root tuber expansion period （<italic>P</italic><0.05）. The yield of Curcumae Radix and Curcumae Rhizoma produced by different grades of seed rhizoma was in the order of first-class ginger>second-class ginger>third-class ginger>substandard seed rhizoma. According to the analysis of the proportion of the commodity grade of medicinal materials，the higher the grade of seed rhizoma was，the higher the proportion of the first-grade Curcumae Radix occupied. According to the analysis of the quality of medicinal materials，there was no significant difference in the content of extract，volatile oil and germanone in Curcumae Radix and Curcumae Rhizoma produced by different grades of seed rhizoma. Conclusion:The grading standard of seed rhizoma was established based on the systematic study on the classification method of <italic>C. phaeocaulis</italic> seed rhizoma. According to the field experiment， grading planting of seed rhizoma can promote plant growth and development，increase the yield of medicinal materials，and improve the proportion of first-grade medicinal materials， with no impact on the internal quality of medicinal materials. Therefore， classification of <italic>C. phaeocaulis</italic> seed rhizoma is scientific and necessary，and can lay the foundation for the standardized cultivation of <italic>C. phaeocaulis</italic>.
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Objective: To study the chemical constituents and the cytotoxic activity of curcumins from the rhizome of Curcuma phaeocaulis. Methods: The 95% ethanol extract from the rhizome of C. phaeocaulis was extracted with petroleum ether, ethyl acetate, and n-butanol. The separation and purification of ethyl acetate fraction was carried out by silica gel column, sephadex LH-20 column, reversed-phase medium pressure chromatography, preparative thin-layer chromatography, and semi-preparative high performance liquid chromatography. The structures of the isolated components were identified by modern spectroscopy techniques. The isolated compounds were screened for cytotoxic activity by MTT assay. Results: Four curcuminoids were isolated from the ethyl acetate extract of the rhizome of C. phaeocaulis, and identified as 1,7-bis (4-hydroxyphenyl)-1E,6E-heptadien-3-one (1), 1,7-bis (4-hydroxyphenyl)-1,4,6-heptatrien-3-one (2), 1,7-bis (4-hydroxyphenyl)-4E,6E-heptadien-3-one (3), and (1R,5S,6S)-1,5-epoxy-6- hydroxy-1,7-bis (3-methoxy-4-hydroxy-phenyl)-heptane (4). MTT experiments showed that compounds 1-3 inhibited HGC-27 cells proliferation, and only compound 2 inhibited MDA-MB-231 cells proliferation. Compounds 2 and 3 also showed strong toxic effects on human normal liver cells. Conclusion: Four curcuminoids were isolated from C. phaeocaulis. Compound 1 was a new compound named curcumin P. Compounds 1-3 had a certain inhibitory effect on the proliferation of HGC-27 cells. Notably, compound 1 selectively inhibited the proliferation of HGC-27 cells and showed no obvious toxic effects on L-02 cells.
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Objective: To qualitatively and quantitatively analyze the terpene-conjugated curcuminoid compound- Bisabolocurcumin ether (BCE) in Curcuma species. Methods: UPLC-Q-TOF-MS/MS was used to establish a qualitative analysis of BCE, and its distribution in Curcuma species was studied. UPLC was used to establish quantitative analysis of BCE and three main curcuminoids in three Curcuma species (Rhizoma Curcumae Longae, HuangsiYujin, Curcuma phaeocaulis), in order to study the difference of BCE content in different Curcuma species and its content difference with other curcuminoids. Results: In the qualitative analysis, by analyzing the primary, secondary pyrolysis fragments and the cracking law of BCE, it was found that only Jianghuang and HuangsiYujin contained it, but BCE could not be detected in the PengEzhu. In the quantitative analysis, BCE and the three main curcuminoids had good linear relationship in their linear range (r ≥ 0.999 3); The average recovery rates were in range of 99.407%-113.631%, and RSD < 4%. It was found that the content of three curcuminoids in Jianghuang were significantly higher than the other two herbs, and the content of BCE in the three herbs was very different. The BCE content in Jianghuang was four times than that of HuangsiYujin. Conclusion: The qualitative and quantitative analysis methods established in this study can provide reference for the discovery and analysis of more novel BCE structure’s compounds, and provide a basis for improving the quality standards of Curcuma species.
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Both raw and vinegar products of the rhizome of Curcuma phaeocaulis are common drugs for promoting blood circulation and removing blood stasis in traditional Chinese medicine,which could be reflected in the inhibition of tail thrombosis in mice. As the traditional processing theory instructs,vinegar tastes sour and bitter,but can activate blood circulation and remove stasis after being infiltrated into the rhizome of C. phaeocaulis as an excipient. In this study,under the help of the ultrafast liquid chromatography-quadrupole time-offlight mass spectrometry( UFLC-Q-TOF-MS),the spectrum-effect relationship between the inhibition of tail thrombosis in mice and the rhizome of C. phaeocaulis both before and after the vinegar processing,were established to explore the functional changes of blood circulation and stasis after vinegar process. Based on the peak area from the fingerprint of UFLC-Q-TOF-MS of the alcohol extracts from the raw and vinegar-processed rhizome of C. phaeocaulis and their efficacy for inhibiting tail thrombosis,the correlation between the chromatography of UFLC-Q-TOF-MS and the inhibition of tail thrombosis in mice were analyzed by orthogonal partial least squares discriminant analysis( OPLS-DA) method. The results,produced by Simca-P software,showed that effective components consisted of eight peaks 16,24( aromadendrene oxide),3,11,22( dehydro-α-curcumene),19[( R)-(-)-α-curcumene],23 and 10 from the fingerprint,making great contribution to distinguish C. phaeocaulis raw products and the corresponding vinegar processed products. Therefore,from the perspective of inhibiting the formation of tail thrombosis in mice,the marker components could be found through the spectrum-effect relationship to distinguish C.phaeocaulis raw and vinegar products. This study provided new basis to explain the difference between the raw and the processed products of traditional Chinese medicine in the functional change of promoting blood circulation and removing blood stasis.
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Animals , Mice , Acetic Acid , Chromatography, High Pressure Liquid , Curcuma , Chemistry , Drugs, Chinese Herbal , Chemistry , Pharmacology , Mass Spectrometry , Rhizome , Chemistry , Thrombosis , Drug TherapyABSTRACT
Objective: To Compare the difference of the intestinal absorption kinetics of volatile oil of Sichuan genuine herb Curcuma phaeocaulis before and after vinegar processing. Methods: Rat model of systemic intestinal perfusion were established and were divided into four groups including volatile oil of C. phaeocaulis group, volatile oil of C. phaeocaulis processed with vinegar group, volatile oil of C. phaeocaulis added with curcuminoids group and volatile oil of C. phaeocaulis processed with vinegar added with curcuminoids group to investigate the absorption of effective constituents of germacrone and furanodiene in the intestinal segements, and calculate the absorption rate constant (Ka), absorption half-life (t1/2), and absorption rate per unit time (P). Results: The results showed that the absorption degree and Ka value of volatile oil of C. phaeocaulis after vinegar processing was less than that before vinegar processing. The degree and Ka value of volatile oil after addition of curcuminoids was less than that before compatibility. Conclusion: C. phaeocaulis after processing with vinegar could alleviate drug property through delaying the absorption of volatile oil to stabilize the blood concentration, with view to meeting the need of clinical safety ad effectivity.
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OBJECTIVE:To develop the determination method for plasma concentration of effective components in essential oil from Curcuma phaeocaulis,and to study its integrated pharmacokinetics. METHODS:Sixteen rats were given the extract of essential oil from C. phaeocaulis 1.0 g/kg(by crude drug)intragastrically;blood samples 300-400 μL from orbit were collected 0, 0.17,0.5,1,2,2.5,3,4,6,8,10,12,24 h after medication. The plasma concentration of α-pinene,1,8-cineole,borneol, β-elemene,curcumol,germacrone and curdione in rats were determined by GC-MS. The determination was performed on DB-5 capillary column,using helium as carrier gas,at the flow rate of 1.2 mL/min. The injector temperature was 270 ℃,by temperature programming,and split ratio was 20∶1. The sample size was 1 μ L. The ion source was electrospray ion source. The selective reaction monitoring mode was used for the positive ion scanning in the range of m/z 20-500. Pharmacokinetic parameters of above effective components were calculated by using DAS 2.0 software. The weight coefficients were customized according to the proportion of AUC0-∞in the sum of AUC0-∞. The integrated pharmacokinetic parameters of multiple effective components in essential oil from C. phaeocaulis were calculated. RESULTS:The linear range of α-pinene,1,8-cineole,borneol,β-elemene, curcumol, germacrone, and curdione were 2.71-173.54, 7.76-496.88, 3.37-215.72, 21.68-1 387.50, 40.21-2 573.44, 24.84-3 179.69,47.78-3 057.81 ng/mL,respectively (r＞0.99). The lower limits of quantitation were 2.71,7.76, 3.37,21.68,40.21,24.84,47.78 ng/mL,respectively. The precision,accuracy and matrix effects were in line with related requirements of quantitative analysis of biological samples. The pharmacokinetic parameters of α-pinene,1,8-cineole,borneol, β-elemene,curcumol,germacrone,and curdione were as follows that cmaxwere (34.72 ± 9.97),(99.86 ± 5.54),(16.10 ± 3.37), (248.98±86.19),(673.75±104.15),(2 353.64±637.83),(2 420.04±708.51)ng/mL;tmaxwere(2.33±0.29),(0.67±0.29), (1.33±0.58),(1.83±0.76),(0.83±0.29),(0.89±0.18),(1.17±0.76)h;t1/2were(8.64±1.46),(8.98±1.63),(12.43± 2.88),(19.86 ± 4.05),(15.63 ± 5.50),(14.17 ± 4.13),(7.14 ± 0.67)h;AUC0-twere (189.78 ± 89.10),(454.74 ± 82.43), (100.55±8.27),(1 067.37±216.55),(3 154.16±405.94),(16 501.24±663.88),(12 524.92±3 222.10)ng·h/mL;AUC0-∞were(229.57±93.50),(524.32±81.67),(146.28±10.74),(2 092.70±416.18),(5 388.65±661.86),(28 198.87±4 102.62), (14 139.35 ± 3 109.19)ng·h/mL,respectively. After integration,the pharmacokinetic parameters were as follows that cmaxwas 1 880.94 ng/mL; tmaxwas 0.50 h; t1/2was 11.22 h;AUC0-twas 13 050.89 ng·h/mL;AUC0- ∞was 19 015.21 ng·h/mL. CONCLUSIONS: The method can be used for the detection of plasma concentration of effective components in rats;pharmacokinetic parameters of essential oil from C. phaeocaulis after integration are greatly different from single effective component,which can provide reference for characterization of its overall pharmacokinetics.
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OBJECTIVE:To study the decoction rate changes of 4 active ingredients in the volatile oil of Curcuma phaeocaulis before and after combined with Sparganium stoloniferum,and provide reference for showing the compatibility mechanism of C. pha-eocaulis and S. stoloniferum. METHODS:HPLC was used to simultaneously determine the contents of 4 active ingredients(curdi-one,curcumol,germacrone and β-elemene) in the volatile oil of C. phaeocaulis after C. phaeocaulis single decoction and com-bined decoction with S. stoloniferum. And the fried rates of each ingredient were compared. RESULTS:Compared with C. phaeo-caulis single decoction,there was significant difference in the fried amounts of curdione,curcumol,germacrone in the volatile oil after combined decoction(P0.05). CONCLUSIONS:The com-bined decoction of C. phaeocaulis and S. stoloniferum can effectively improve the dissolution of curdione,curcumol,germacrone in the volatile oil. It may be the reason for the enhancement of efficacy after compatibility.
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Objective: To determine the dynamic variation of pH value and seven components including bisdemethoxycurcumin, demethoxycurcumin, curcumin, curdione, curcumol, germacrone and β-elemene during Curcuma Phaeocaulis' processing, in order to study its curcuminoids and volatile oils components in the process of rice vinegar production affected by external factors such as water, heat, and acid. Methods: C. phaeocaulis were processed by using distilled water, 9° rice vinegar, and 9% acetic acid aqueous. The pH value of the decoction at a specific time point in 10 batches was detected. The data were analyzed by SPSS software. The content of seven components in the samples of the raw crude group, the blank group, the vinegar group, and the reference group were determined by HPLC method, and the change of the content was analyzed. Results: Compared with the blank group, the vinegar group and reference group showed low pH value, and there was no significant difference; Compared with the blank group and reference group, the contents of three curcuminoids in the vinegar group increased [the mass fraction of bisdemethoxycurcumin is (0.002 320 ± 0.000 344) mg/g, the mass fraction of demethoxycurcumin is (0.059 65 ± 0.015 64) mg/g, the mass fraction of curcumin is (0.272 5 ± 0.125 2) mg/g], and the contents of volatile oils decreased in different degrees. Conclusion: During the processing progress, the main active constituents of pharmaceutic adjuvant 9° rice vinegar might be acetic acid; The acetic acid and other organic principles of rice vinegar might protect the chemical constituents in C. phaeocaulis by regulating pH value; The toxicity degree of C. phaeocaulis before and after processing with vinegar was regulated by different protecting degrees of different component. It not only reflects the scientificity of 9° rice vinegar as a pharmaceutic adjuvant, but also offers a new method to study the material basis of C. phaeocaulis processing with rice vinegar.
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OBJECTIVE:To study the preventive and therapeutic effect and its mechanism of medicine pair decoction liquid of Curcuma phaeocaulis-Sparganium stoloniferum on rats with uterine myoma. METHODS:Rats were randomly divided into normal group,model group,positive group(mifepristone,2.25 mg/kg)and medicine pair decoction liquid group(C. phaeocaulis-S. stolon-iferum decoction liquid,6.0 g/kg),10 in each group. Except for the normal group,rats in other groups were injected Estradiol ben-zoate injection (0.5 mg/kg) intramuscularly every Monday,Wednesday and Friday,for 12 weeks. From the 13th week,rats in modeling group were added Progesterone injection(5 mg/kg)intramuscularly as well as relevant medicines intragastrically,once a day,until the 16th week. After administration,uterine coefficient of rats was detected. HE staining was used to observe the patho-logical changes of uterus and determine the thickness of smooth muscle;immunohistochemical staining was adopted to detect the transforming growth factor β3 (TGF-β3),matrix metalloproteinase 11 (MMP-11) protein expressions in uterus tissue of rats. RE-SULTS:Compared with normal group,uterine coefficient was increased in model group,pathological changes were obvious in uterus,thickness of smooth muscle was increased,TGF-β3 and MMP-11 protein expressions were enhanced(P<0.05 or P<0.01). Compared with model group,above-mentioned changes were improved significantly in positive group and medicine pair decoction liquid groups(P<0.05 or P<0.01). CONCLUSIONS:Medicine pair decoction liquid of C. phaeocaulis-S. stoloniferum shows cer-tain preventive and therapeutic effect on rats with uterine myoma. The mechanism may be associated with downregulating the TGF-β3,MMP-11 protein expressions in uterus tissue.
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Chinese herbs are mostly used to make decoction, which would form precipitation after standing for cooling and abandoned by patients. Processing with vinegar can change the property of the herbal pieces, such as the transfer rate of heavy metal into decoction. To analyze the transfer rate change of heavy metal in the decoction and precipitation of Curcuma phaeocaulis before and after processing with vinegar, inductively coupled plasma mass spectrometry (ICP-MS) was used to establish the determination method on five heavy metals in C. phaeocaulis, including Copper (Cu), arsenic (As), chromium (Cd), lead (Pb), mercury (Hg), using microwave to digest the samples, indium (In) as the internal standard, and national level standard material tea leaves GBW10016 (GSB-7 tea) as the quality control standard material. Then, the content of five heavy metals in the herbal pieces, decoction and five heavy metals of 6 representative batches of C. phaeocaulis and their vinegar-processing products was determinated. After computation, the transfer rates of heavy metals in the decoction and precipitation of C. phaeocaulis Val. before and after the processing with vinegar were obtained. The results showed that, after the processing with vinegar, total transfer rate of Pb and Hg was decreased significantly; total transfer rate of Cd and Cu was slightly decreased; total transfer rate of As was slightly increased, however heavy metals in all the precipitation were decreased. The results indicated that processing with vinegar had certain influence on heavy metal transfer rate, with certain synergistic and attenuated effect.
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Objective To study pharmacokinetics of curcumin in Curcuma phaeocaulis in rats in vivo.Methods HPLC method was used to determine the curcumin in rat plasma.The conditions were column: Lichrospher-5-C_(18)(250 mm?4.6 mm, 5 ?m); column temperature: 25 ℃;mobile phase: CH_3CN-5% HAc water solution(45∶55);flow: 1 mL/min;detection wavelength: 420 nm.Results The calibration curve was liner(r=0.999 5) at the range of 6.5—104 ?g/mL.The average recovery was 98.5%.RSD was 2.41%(n=5).The pharmacokinetic parameters of curcumin were as follows: k_a was 0.53/h,k_e was 0.10/h,t_(1/2ka) was 1.32 h,t_(1/2k) was 6.89 h,t_(peak) was 3.89 h,C_(max) was 93.15 ng/mL,AUC was(1 369.38) ng/mL.Conclusion This method is stable,simple,and reliable,which can be applied for the determination of curcumin in plasma and pharmacokinetic studies.