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Objective: To establish an HPLC fingerprint of Cnidii Fructus formula granule analysis method for simultaneous determination of six main coumarin components, including osthol, xanthotoxin, xanthotol, bergapten, imperatorin and isopimpinellin, in order to provide reference for the study of the material basis of Cnidii Fructus formula granule. Methods: The method was performed by high performance liquid chromatography with a Waters XBridge C18 (250 mm × 4.6 mm, 5 μm) column and methanol (A)-0.1% acetic acid (B) as the mobile phase for gradient elution. The flow rate was 0.5 mL/min, the injection volume was 10 μL and the column temperature was 40 ℃. The detection wavelength was set at 320 nm. The chromatographic fingerprint evaluation system published by the State Pharmacopoeia Commission (2012 Edition) was used to establish the fingerprint of Cnidii Fructus formula granule, and the content of six main coumarin components was simultaneously determined. Results: The research on the 18 batches of Cnidii Fructus formula granule showed that the fingerprint similarity was greater than 0.992 and 19 common peaks were calibrated with satisfied peak resolution. The content determination results showed that the content of both xanthotoxin and osthol were the main coumarin components in Cnidii Fructus formula granule. According to the methodological investigation, the precision RSD values were all less than 1.6%. The sample was stable within 48 h and this method had good repeatability. The average recovery rates of xanthotol, xanthotoxin, imperatorin, isopimpinellin, bergapten and osthol were 100.69%, 101.03%, 99.48%, 100.88%, 101.27% and 100.35%, respectively. All of these coumarin components’ RSD were less than 2.5%. The six components showed a good linear relationship within a certain concentration range. The results of the content determination of xanthotol, xanthotoxin, isopimpinellin, bergapten, imperatorin and osthol respectively were 8.01-8.29, 2.37-2.63, 4.30-4.61, 4.04-4.40, 3.45-3.90 and 6.02-6.80 mg/g among the 18 batches of the Cnidii Fructus formula granule. Conclusion: The fingerprint method and the determination method of six main coumarin components in the Cnidii Fructus formula granule established in this study are simple, stable, accurate and reliable. This method can be used for the quality control of the Cnidii Fructus formula granule.
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Objective::To investigate the effect of bergapten on the apoptosis of HepG2 and Hep3B cells through phosphatidylinositol 3 kinase (PI3K)/protein kinase B(Akt) pathway. Method::Bergamot (5, 50, 200 μmol·L-1) groups and blank group were set up. The effect of bergapten at different concentrations on proliferation of HepG2 and Hep3B cells for 24, 48 h were detected by thiazolyl blue(MTT) assay. Apoptosis was detected by Annexin V-FITC/propidium iodide double staining. Quantitative real-time fluorescence reverse transcriptional polymerase chain reaction (Real-time PCR) and Western blot assay were used to detect relevant mRNA and proteins expressions. The clone formation rate and the effect of HepG2 and Hep3B cells in each group were evaluated by plate cell clone formation. Result::MTT assay showed that bergapten could significantly inhibit the proliferation activity of HepG2 and Hep3B cells in a time-dependent manner. Flow cytometry analysis showed that bergapten in 200 μmol·L-1 concentration groups had significant pro-apoptotic effect on HepG2 and Hep3B cells after 48 h (P<0.05). Western blot results showed that bergamolactone could up-regulate the protein expressions of Caspase-3, Caspase-8 (P<0.05), and down-regulate protein expressions of B-lymphocytoma-2 (Bcl-2), PI3K (P<0.05). Real-time PCR results showed that mRNA expressions of PI3K and Akt were decreased(P<0.05). The results of plate cell clone formation experiment showed that with the increase of the concentration of bergamolide, the cell clone formation rate of each group showed a decreasing trend, particularly in 200 μmol·L-1 concentration group (P<0.05). Conclusion::Bergapten can inhibit the proliferation of HepG2 and Hep3B cells, which may be induced through the PI3K/Akt signaling pathway.
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Despite the wide ethnomedicinal applications of Ficus exasperata, little is known about the active principles responsible for the observed biological effects, thus limiting opportunities for further therapeutic applications. The bioassay guided chemical investigation of F. exasperataroot bark resulted in the isolationof a furocoumarin (D-1) shown to be partly responsible for the acclaimed anti-diabetic effect of the plant.
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Objective: To analyze the quality of Angelicae Dahuricae Radix (ADR) from national drug evaluation sampling, continuously improve and revise the standard determination method, and ensure the safety of public drug use, six index components of ADR medicinal material and decoction pieces were determined by combining HPLC fingerprint chromatography and quantitative analysis of multi-components by single marker (QAMS). Methods: The determination and exploratory study were carried out according to the National Standard. The establishment of steamed Angelicae dahurica HPLC fingerprint, method for multiple assessment of six index components to conduct a methodological survey determination of multi-index component content of 20 batches of Angelicae dahurica tablets, the final determination of the quality of Angelicae dahurica. Result:s A total of 239 batches of ADR were received from 189 enterprises, in which 225 batches were qualified and 14 batches were unqualified. The qualified rate was 94%. A method that determined content of six kinds of coumarin in ADR decoction pieces using imperatorin as control was established, which could solve the problem of difficulty in obtaining control and consuming long determination time. It would better indicate the drug quality. Conclusio:n The method we had established could be used in qualitative and quantitative analysis of oxypeucedanin hydrate, byakangelicin, oxypeucedanin, berapten, imperatorin and isoimperatorin in ADR decoction pieces. Through this random inspection, it is found that there were still some quality problems in ADR decoction pieces. The quality supervision of traditional Chinese medicine should be strengthened further.
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ABSTRACT A validation study of a reverse-phase high-performance liquid chromatographic assay for the quantification of two furanocoumarins (psoralen and bergapten) in soft extract obtained from Brosimum gaudichaudii Trécul, Moraceae, roots was conducted. The developed method was sensitive, rapid, reproducible, easy and precise, and showed linearity (r > 0.99) in the range of 10-64 µg/ml for psoralen, and 9-56 µg/ml for bergapten. It also showed a good efficiency for the photodegradation analysis of psoralen and bergapten in the soft extract. The photostability results showed that the Higuchi model presented the best fitting to the obtained data. Both chemical markers showed stability over 2.6 days, suggesting potential applications of the extract in obtaining intermediate products from this plant material. Furanocoumarins take around 30 min to be activated by UV light, reaching the maximum biological potential. Thus, the results obtained to the Higuchi model, corresponding to 2.6 days of stability, shows feasibility with future applications of these chemical markers.
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Objectives: To investigate the effects of bergapten of Angelicae Dahuricae Radix on the transport of vincristine and its possible mechanism. Methods: The apparent permeability coefficient (Papp) for the transport of vincristine through the membrane of MDCK-MDR1 cells was used as an indicator of the effect of bergapten on vincristine transport. Molecular docking was employed to predict the binding force between bergapten and P-glycoprotein (P-gp). The effects of bergapten on P-gp function and P-gp ATPase activity were determined by rhodamine 123 (Rho123) accumulation and activity analysis, respectively. 1,6-Diphenyl-1,3,5-hexatriene (DPH) was used to study the effects of bergapten on membrane fluidity, and Western blotting and quantitative real-time PCR assays were performed to analyze the effect of bergapten on the protein and mRNA expression of P-gp, respectively. These experiments clarified the effects of bergapten on the transport of vincristine and allowed exploration of the possible mechanism underlying the effects of bergapten. Results: The results showed that bergapten could inhibit the transport of vincristine in MDCK-MDR1 cells, and the binding force between bergapten and P-gp was weaker. Bergapten could reduce the accumulation of Rh123 in MDCK-MDR1 cells, increase the membrane fluidity, and upregulate P-gp protein and mRNA expression but it had no effect on P-gp ATPase activity. Conclusions: Overall, we concluded that the possible mechanism through which bergapten inhibits vincristine transport was related to the bergapten-mediated upregulation of P-gp protein and mRNA expression, membrane fluidity or P-gp enzyme activity.
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Objective To study the chemical constituents in 95% ethanol aqueous extract of Trigonostemon lutescens. Methods The open silica gel, MCI, Sephadex LH-20 column chromatography, and the semi-preparative HPLC were used to isolate and purify the chemical constituents from the EtOAc fraction of T. lutescens. The structures of the isolates were elucidated by their physiochemical properties, NMR, and MS spectroscopic data, as well as comparison with literature data. Results Eleven compounds were isolated from the EtOAc fraction of the 95% aqueous EtOH extract of T. lutescens, and their structures were identified as seven coumarins, auraptenol (1), meranzin hydrate (2), xanthotoxin (3), bergapten (4), isoimpinellin (5), alloisoimperatorin (6), and isodemethylfuropinarine (7); Two phenylalanine glycosides, 3,4-dihydroxy allylbenzene-4-O-β-D-glucopyranoside (8) and 1-O-β-D-glucopyranosyl-4- allylbenzene (9); One phenylethanol, 1-(2-ethylphenyl)-1,2-ethanediol (10); One alkaloid, 8-hydroxy-3-methoxy-5H-pyrido [2,1-c] pyrazin-5-one (11). Conclusion All these compounds are isolated from the genus Trigonostemon for the first time.
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OBJECTIVE Bergapten (BG), is a furanocoumarin derived from herbal and citrus extracts can act as antioxidant and selective anticancer agents.The current study aimed to investigate whether bergapten would attenuate immunosenescence and to exploreits immunomodulatory effects on immune responses in D-galactose-induced aging BALB/c mice.METHODS Firstly,mice were given D-galactose(180 mg·kg-1)subcutaneous injections for 30 d.To evaluate the establishment of the aging-related effect in mice, serum samples of BALB/c mice were collected from tail vein. Aging BALB/c mice were freely divided into three groups: negative control group received 1% Tween 80 solution only, named D-gal group. Positive groups were received BG administration at the dose of 20 and 100 mg·kg-1, named D-gal+BG(20)group and D-gal+BG(100)group,respectively.Effects of bergapten on T lympho-cyte proliferation and flow cytometry were assessed by using the splenic cell suspension. Enzyme linked immunospot kits were used to quantitatively determine interferon-γ(IFN-γ)and interleukin-4(IL-4) levels of the isolated serum. Immunophenotype was determined by using mixture of antibodies includ-ing anti-CD3,anti-CD4,and anti-CD8.RESULTS Bergapten(20 mg·kg-1)therapy can modulate immu-nity against viral epidemics and attenuate aging-induced immune deficiency(P<0.01),which was correlat-ed with the decline in the activation of the Th and Tc responses in D-galactose induced aging BALB/c mice.According to the in vivo results,bergapten exposure up-regulated the secretion of IFN-γ and IL-4 in T-helper 1(Th1)and T helper 2(Th2)cells(P<0.05,P<0.01).Additionally,BG(20 mg·kg-1)restored antigen-specific CD4+and CD8+T cells in aging models (P<0.05, P<0.01), which may help to curing chronic infections. CONCLUSION The beneficial effect of bergapten in D-galactose induced aging BALB/c mice may be due to the Th and Tc responses activation.
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OBJECTIVE:To develop a method for simultaneous determination of psoralen,isopsoralen,bergapten,imperato-rin,trimethylpsorale,neobavaisoflavone and bavachin in Qing'e pills. METHODS:HPLC method was adopted. The separation was performed on Kinetex-C18 column with mobile phase consisted of methanol-0.1% glacial acetic (gradient elution) at the flow rate of 1.0 mL/min. The detection wavelength was set at 248 nm. The column temperature was 30 ℃. The sample size was 20 μL. RESULTS:The linear ranges were 1.012-101.2 μg/mL for psoralen(r=0.9999),1.007-100.7 μg/mL for isopsoralen(r=0.9997), 1.010-101.0μg/mL for bergapten(r=0.9999),1.021-102.1μg/mL for imperatorin(r=0.9999),1.002-100.2μg/mL for trimethyl-psorale(r=0.9996),1.008-100.8 μg/mL for neobavaisoflavone(r=0.9999),1.025-102.5 μg/mL for bavachin(r=0.9998),re-spectively. The limits of quantitation were 0.15,0.15,0.30,0.30,0.15,0.30,0.30 μg/mL,and the limits of detection were 0.05, 0.05,0.10,0.10,0.05,0.10,0.10 μg/mL,respectively. RSDs of precision,stability and reproducibility tests were lower than 2.0%. The recoveries were 95.3%-99.6%(RSD=1.9%,n=6),96.1%-99.3%(RSD=1.2%,n=6),95.2%-98.4%(RSD=1.4%,n=6),95.4%-99.2%(RSD=1.5%,n=6),96.1%-99.3%(RSD=1.5%,n=6),95.6%-98.9%(RSD=1.4%,n=6), 95.2%-99.6%(RSD=1.6%,n=6),respectively. CONCLUSIONS:The method is simple and accurate,can be used for simultane-ous determination of 7 kinds of components in Qing'e pills.
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Objective: To control the quality of Fici Hirtae Radix by establishing quality standards. Methods: The origin mistakes of Fici Hirtae Radix in Chinese Pharmacopoeia (1977 edition) were modified by searching and mining plant morphology characters, distribution of origin, historical evolution, clinical efficacy and modern research, medicinal standards in different regions on Ficus hirta Vahl and Ficus simplicissima. Morphological identification, microscopical characteristics based on transverse section and powder, and thin layer chromatography were adopted to identify commercial medicinal materials of Fici Hirtae Radix from different sources. The content of psoralen and bergapten was determined by HPLC. Moisture content, total ash, acid-insoluble ash, and ethanol-soluble extractives were determined by methods described in appendices of Chinese Pharmacopoeia (2010 edition). Results: The origin of Fici Hirtae Radix was not F. simplicissima Lour., but F. hirta Vahl. This study collected 20 Fici Hirtae Radix commercial medicines mixed 40% non-medicinal parts samples. TLC data, morphological characteristics and microscopic characteristics, both showed obvious distinction. The concentration of psoralen and bergapten showed the content of psoralen range from 0.00848% to 0.150%, and the content of bergapten range from 0.00587% to 0.026 3%, and the contents of two index components of Fici Hirtae Radix were significantly higher than the non-medicinal parts samples. The moisture content ranged from 4.73% to 8.27%, total ash ranged for 2.58% to 4.37%, acid insoluble ash ranged for 0.054% to 0.710%, ethanol-soluble extracts ranged for 6.23% to 8.34%. Conclusion: This paper revised the origin error of Fici Hirtae Radix of Chinese Pharmacopoeia (1977 edition) that its specification medicinal part should be the roots of F. hirta, and established quality standards of Fici Hirtae Radix, and these methods could comprehensively and effectively control the quality of commercial medicinal materials of Fici Hirtae Radix. This study could provide evidence for rational, safe, and effective clinical application of Fici Hirtae Radix.
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Objective: To study the effects of coumarins in Angelicae Dahuricae Radix (imperatorin, isoimperatorin, bergapten, and oxypeucedanin) on the activities of adenosine triphosphate (ATPase) in P-glycoprotein (P-gp) in vitro and to investigate the P-gp mediated meschanism in transport of drug. Methods: The ATPase activity assay was used to identify whether the coumarins in Angelicae Dahuricae Radix were P-gp substrates or not. Results: The average fluorescent values (ΔRLUTC) of imperation and oxypeucedanin at each concentration were greater than average basic fluorescence value (ΔRLUbasal). The average fluorescent values (ΔRLUTC) in medium and low concentration of isoimperatorin and bergapten groups were lower than average basic fluorescence value (ΔRLUbasal). Conclusion: Imperation and oxypeucedanun are non-concentration dependent P-gp ATP inhibitors. Isoimperatorin and bergamot lactones could inhibit the ATPase activity at low concentration and have the inhibitory effect on P-gp. At high concentration, the activity of ATPase can be induced so as to induce P-gp.
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Objective:To develop an HPLC method for the simultaneous determination of four active ingredients ( psoralen, ber-gapten, desmethylwedelolactone and wedelolactone) in Xichuan pills. Methods: A Hypersil C18 column was used for the chromato-graphic separation of the four analytes. The mobile phase consisting of methanol-acetonitrile (2∶ 1) and 0. 5% glacial acetic acid solu-tion was used for gradient elution. The flow rate was 1. 1 ml·min-1 . The column temperature was 25℃. Psoralen and bergapten were detected at 222 nm, and desmethylwedelolactone and wedelolactone were detected at 351 nm. Results:There was good linear relation-ship between the concentration of psoralen, bergapten, desmethylwedelolactone and wedelolactone and peak area within the concentra-tion range of 5.380-107.600 μg·ml-1(r =0.999 5),6.870-137.400 μg·ml-1(r =0.999 7),4.580-91.600 μg·ml-1(r =0. 999 2) and 7. 300-146. 000 μg·ml-1(r=0. 999 9),respectively. The average recovery(RSD) was 96. 82%(0. 77%), 98. 81%(1. 40%),99. 06%(1. 20%) and 98. 39% (0. 80%), respectively (n=6). Conclusion:The developed method is accurate, highly sensitive and well reproducible, which can be used for the determination of the four active ingredients in Xichuan pills.
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Objective: To investigate the influence of furan coumarins from Angelicae Dahuricae Radix on intestinal absorption of puerarin, paeoniflorin, and vincristine, and to explore the influence law of furan coumarins on the intestinal absorption of the drugs with different structures. Methods: The apparent permeability coefficient (Papp) of drugs across the Caco-2 cell monolayers as evaluated index and Caco-2 cell model was used to study the absorption effects of furan coumarins (imperatorin, isoimperatorin, bergapten, and oxypeucedanin) on the transport of puerarin, paeoniflorin, and vincristine across Caco-2 cell monolayers. Results: There are different effects of the four coumarins on puerarin, paeoniflorin, and vincristine. For imperatorin, isoimperatorin, and oxypeucedanin could improve the transport of puerarin on Caco-2 cell significantly (P < 0.05, 0.01), while bergapten has no effect on the transport of puerarin across Caco-2 cell monolayers. For imperatorin and bergapten have no effect on the transport of paeoniflorin: However isoimperatorin and oxypeucedanin could significantly inhibit the transport of paeoniflorin across Caco-2 cell monolayers (P < 0.05, 0.01). Iperatorin, isoimperatorin, and oxypeucedanin could improve the transport of vincristine across Caco-2 cell monolayers significantly (P < 0.05, 0.01), but bergapten could significantly restrain the transport of vincristine across Caco-2 cell monolayers (P < 0.05). Conclusion: Different furan coumarins have different effects on the intestinal transport of the same medicine. Although the structures all are the furan type, the influence shows the opposite effect on drug transport. The same furan coumarin combined with different drugs have the different influences on the intestinal transport of compatibility drugs.
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Objective: To study the chemical constituents from the root bark of Changium smyrnioides. Methods: Compounds were isolated by various kinds of column chromatographies on silica gel, Sephadex LH-20, and recycling preparative HPLC from the ethanol extract in the root bark of C. smyrnioides, and their structures were elucidated by the physicochemical characteristics and spectral analyses. Results: Fifteen chemical constituents were obtained and identified as imperatorin (1), phellopterin (2), xanthotoxol (3), 5-hydroxy-8-methoxy-psoralen (4), vanillic acid (5), alloimperatorin (6), psoralen (7), bergapten (8), 8-O-β-D-glucopyranosyl- 5-methoxylpsoralen (9), isopimpinellin (10), caffeic acid (11), aurantiamide acetate (12), vaginatin (13), β-sitosterol (14), and succinic acid (15). Conclusion: Compounds 6-13 are isolated from the plants in Changium Wolff for the first time.
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OBJECTIVE: To investigate the blocking effect of bergapten on cell cycle in nasopharyngeal carcinoma (NPC) cells. METHODS: The inhibition was tested by CCK-8 assay after the NPC cells (CNE-2 and HONE-1) being exposed to bergapten for 48 h. The bergapten concentrations were as follows: 1, 10 and 100 μmol·L-1. The cell cycle was assayed by PI staining and following flow cytometry analysis, and the CDK4, CDK6, CDK2, Cyclin D1, Cyclin E2, β-catenin, GSK-30 and p-GSK-3p (S9) protein levels were detected by Western blotting. RESULTS: Bergapten was able to significantly inhibit CNE-2 and HONE-1 NPC cancer cell proliferations, and the effect was dose-(CNE-2: r=0.926, P<0.05; HONE-1: r=0.959, P<0.05) and time-dependent (CNE-2: r=0.991, P<0.05; HONE-1: r=0.963, P<0.05). Drug treatment induced a block in the G0/G1 phase and decreased protein levels of CDK4, CDK6, CDK2, Cyclin D1, Cyclin E2 and β-catenin, but the protein levels of GSK-3β and the phosphorylation of p-GSK-3β (S9) didn't change. CONCLUSION: Bergapten can block cells cycle and inhibit NPC cells' proliferations, and those effects might be related to suppressing of cells cycle-related protein and interfering with the wnt/p-catenin signal pathway.
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Objective To screen the active component from Fructus Citri Sarcodactylis which have aortic vascular relaxant effect. Methods The active components from Fructus Citri Sarcodactylis were screened by combining vascular smooth muscle/cell membrane chromatography (VSM/CMC) and liquid chromatography/mass spectrometry (LC/MS) with pharmacological assay in vitro. Results Bergapten was the active component of Fructus Citri Sarcodactylis that functions in vascular. The pharmacological experiment showed that bergapten had vasodilating effect. The capacity factor of this compound on the VSM/CMC model was found to be significantly correlated with its pharmacological effect. Conclusion Association between compound and membrane protein (receptor) can be reflected by using the VSM/CMC model. VSM/CMC-offline-LC/MS technology can be used to quickly screening new active components from natural medicine.
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Objective: To develop an HPLC-DAD method for the simultaneous determination and comparative analysis on the contents of prim-O-glucosylcimifugin, 4′-O-β-D-glucosyl-5-O-methylvisamminol, cimifugin, sec-O-glucosylhamaudol, psoralen, imperatorin, bergapten, and xanthotoxin in the roots of Saposhnikovia divaricata from Longxi areas. Methods: The extracts were obtained by methanol reflux method and the contents of the eight compounds were determined by HPLC-DAD. The mobile phase, consisting of acetonitrile-water, was programmed for a gradient elution. The flow rate was 1.0 mL/min, the detection wavelength was 254 nm, and the column temperature was 25°C. Results: Excellent linearity with correlation coefficents (r) of 0.9992-0.9999 was obtained. The average recoveries of the eight compounds were 96.2%-104.1% and all RSD values were less than 3%. The contents of the four coumarins in the roots of S. divaricata were much less than those four chromones. The contents of the four coumarins in the roots of Carum carvi and Peucedahum ledebourielloides were more than those in S. divaricta, while no chromones were detected except sec-O-glucosylhamaudol in P. ledebourielloides. Conclusion: The method appears to be simple, accurate, and well reproducible, which could be used for the simultaneous determination of the above-mentioned eight compounds in S. divaricata. According to the above analysis, C. carvi and P. ledebourielloides could not be used as the succedaneum of S. divaricata on the basis of the eight compound contents.
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ObjectiveTo identify and analyze the volatile constituents in the leaves and fruits ofFicus carica.MethodsGas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS) were used.ResultsThe major components detected in volatile oil of the leaves were psoralen (10.12％),β-damascenone (10.17％),benzyl alcohol (4.56％),behenic acid (4.79％),and bergapten (1.99％),etc.The major components detected in volatile oil of the fruits were furfural (10.55％),5-methyl-2-furaldehyde (10.1％),and benzeneacetaldehyde (6.59％),etc.Conclusion A total of 121 volatile constituents are identified in the leaves and 108 in the fruits ofF.carica,among which 103 constituents are identified for the first time in the leaves and 100 in the fruits.Eighteen volatile constituents are identified in both leaves and fruits.
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Objective To establish a HPLC method for the determination of psoralen and bergapten in Ficus hirta Vahl.Methods A Merck-lichrospher C18(4 mm?250 mm,5 ?m)column was adopted.The mobile phase was acetonitrile-water(35∶65) with the flow rate of 1.0 mL/min.The column temperature was 35 ℃,and the detection wavelength was set at 222 nm.Results Psoralen's linearity was obtained in the range of 0.12 ?g～1.20 ?g(r=0.999 7),and bergapten's linearity in the range of 0.03 ?g～0.30 ?g(r=0.999 5).Psoralen's average recovery was 100.9 %with RSD 2.9 %,and that of bergapten was 99.6 %with RSD 1.9 %.Conclusion This is the first report of simultaneous determination of psoralen and bergapten in Ficus hirta Vahl.by HPLC,and the results showed the method is accurate,reproducible,and can serve as quality evaluation method for Ficus hirta Vahl.
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Objective To determine the appropriate collecting -time of Exocarpium citri grandis,the chemical compo-sition in Exocarpium citri grandis in different fruit -ages was compare d and analyzed.Methods Contents of naringein and bergapten were compared by TLC an d the total dihydroflavones in Exocarpium citri grandis was determined by spec-trophotometmic analysis with detection wavelength at 384nm.Results With the increase of fruit -age,the c ontents of naringein and total dihydroflavones in Exocarpium citri grandis decreased and the content of bergapt en increased.Con-clusion Fruit -age has an obviously effect on the chemical composition in Exocarpium citri grandis and collecting -time can influence the quality of medicin e materials.Further studies of Exocarpium citri grandis should be combined pharma-codynamics for fixing collecting -t ime.