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Aim To study the antiviral anrl antipyretic mechanism of Phillvrin in vitro and in vivo.Methods By respiratory virus infection cell model, SI index and antiviral activity of forsythia glycosides virus activity in vitro were detected.A mouse model of influenza virus infection was established, and hemagglutination titer, lung index, lung histopathology pathology were detec¬ted.Hemagglutination titer, lung index, lung histopa¬thology pathology were observed and in vivo anti-influ¬enza virus and pneumonia effects were investigated.Dry yeast induced rat fever model was established, temperature and plasma and hypothalamus thermoregu¬lation and inflammation of the related factors were test¬ed , and its antipyretic mechanism was investigated.By AutoDock Vina software for molecular docking, the docking results were plotted with PyMol software.Re¬sults Phillyrin had certain inhibitory effects on H3N2, RSV, E71 , ADV-3, HSV-1 and HSV-2 (SI >2).Phillyrin could reduce hemagglutination titer of infected lung tissue, decrease lung index, and alleviate lung lesions, especially interstitial pneumonia.Phill- vrin could also significantly reduce the body tempera¬ture of rats with fever, and its antipyretic mechanism might he related to the decrease of PGE2 and IL-ip levels in plasma and hypothalamus of rats.Molecular docking results showed that the binding energies of Phillyrin with a-MSH, IL-lp, PEG2, A VP, cAMP and other proteins were all less than - 5 kcal • mol 1.Conclusions Phillyrin has obvious antiviral, antipy-retic and improvement of pulmonary inflammatory le¬sions, and it is speculated that it can play an anti-in¬fluenza effect through "treating both symptoms and root causes".
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Objective To establish a HPLC fingerprints of Biyuanjing capsules. Methods The column was Agilent SB-C18(4.6mm×250 mm, 5 µm). The mobile phase was acetonitrile-water with gradient elution at a flow rate of 1.0 ml/min. The detection wavelength was 210 nm. The detection time was 80 min. Results The HPLC fingerprints of Biyuanjing capsules were established. Twenty common peaks were confirmed, of which, 15 peaks were belonging to each crude drug and 5 peaks were identified as chemical components. The overall similarity of the fingerprints of 10 batches of samples was above 90% comparing with the control. Conclusion This method can be used for the quality control of Biyuanjing capsules.
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Objective: To explore the potential effective compounds of ‘wind-dispersing and exterior syndrome-relieving’ of Shufeng Jiedu Capsule (SJC). Methods: The fingerprints of 22 samples with different proportions of SJC were established by LC-MS. The stimulation rates of 22 samples to the M3 receptor were measured by Chinese hamster ovarian cancer cells with high expression of M3 acetylcholine receptor which related to ‘wind-dispersing and exterior syndrome-relieving’. Finally, the spectrum-effect relationship of relative peak areas of characteristic peaks and stimulation rates was analyzed by BP-neural network. Results: According to the established optimal neural network model, 16 characteristic peaks were calculated and found to be significantly correlated with the activity of SJC on the M3 receptor stimulation. Conclusion: Through the spectrum-effect study, it is speculated that the effective components of SJC on ‘wind-dispersing and exterior syndrome-relieving’ may be the 16 compounds including dihydrodylate glycoside, emodin, rhein, phillyrin, forsythoside E, saikosaponin d, polydatin, verbascoside, etc, which provides a basis for the determination of quality markers.
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Objective To establish a method for the simultaneous determination of Forsythoside A, Phillyrin, (R,S)-Epigoitrin, Chlorgenic Acid and Isochlorogenic Acid A by HPLC, and test 16 batches of samples from 14 manufacturers. Methods The test was performed on Kinetex EVO C18 column (150 mm × 4.6 mm, 5 μm) with the column temperature at 35 ℃ . The gradient elution was adopted with the mobile phase of acetonitrile and 0.3% phosphoric acid aqucous at a flow rate of 1.0 ml/min. The detection wavelength of (R,S)-Epigoitrin and Phillyrin were set as 236 nm, the detection wavelength of Forsythoside A, Cholorogenic Acid and Isochlorogenic Acid A were set as 327 nm. Results The good linear relationships were displayed within the linear range of 0.050 45-2.018 00 μg for Forsythoside A (r=0.999 9), 0.018 21-0.728 40 μg for Phillyrin (r=0.999 9), 0.010 16-0.406 40 μg for (R,S)-Epigoitrin (r=0.999 9), 0.006 60-0.263 90 μg for Cholorogenic Acid (r=0.999 9) and 0.0040 44~0.161 76 μg for Isochlorogenic Acid A ( r=0.999 5). The RSDs of reproducibility and stability tests were lower than 2%; recoveries were 97.01%, 98.28%, 99.35% and 96.21%, RSD were 3.19%, 1.19%, 0.81%, 2.88% and 2.96%. The content ranges of Forsythoside A, Phillyrin, (R,S)-Epigoitrin, Chlorgenic Acid and Isochlorogenic Acid A from 16 batches of samples from 14 manufacturers were 0.057 43-1.508 71 mg/g, 0.017 72-0.350 15 mg/g, 0.005 68-0.177 13 mg/g, 0.007 53-0.226 33 mg/g and 0.00308-0.11908 mg/g. Conclusions The established method is simple and accurate, and has a good repeatability. It can be used for the quality analysis of Forsythoside A, Phillyrin, (R,S)-Epigoitrin, Chlorgenic Acid and Isochlorogenic Acid A. The content of the tested chemical components from 16 batches of samples from 14 manufacturers have significant differences which indicate that a reinforcement of the quality control is needed.
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OBJECTIVE:To establish a method for the content determination of chlorogenic acid,rutin,phillyrin,quercetin and glycyrrhizic acid in Sangju ganmao granules. METHODS:HPLC method was adopted. The determination was performed on WondasilTM-C18 column with mobile phase consisted of acetonitrile-0.4% phosphoric acid solution(gradient elution)at the flow rate of 0.8 mL/min. The column temperature was set at 30 ℃,and the detection wavelength was set at 254 nm. The sample size was 5 μL. RESULTS:The linear range of chlorogenic acid,rutin,phillyrin,quercetin and glycyrrhizic acid were 0.014-0.560 μg (r＝0.999 2),0.022-0.880 μg(r＝0.999 6),0.010-0.400 μg(r＝0.999 9),0.016-0.640 μg(r＝0.999 9),0.011-0.440 μg(r＝0.999 1), respectively. The limits of quantitation were 0.752,0.647,0.483,0.372,0.242 mg/L;the limits of detection were 0.253,0.185, 0.157,0.162,0.098 mg/L. RSDs of precision,stability and reproducibility tests were all lower than 2.0%. The average recoveries were 97.4%-100.9%(RSD＝1.4%,n＝6),95.7%-99.0%(RSD＝1.1%,n＝6),95.1%-98.1%(RSD＝1.3%,n＝6),96.4%-99.8%(RSD＝1.4%,n＝6),97.8%-100.4%(RSD＝1.0%,n＝6),respectively. CONCLUSIONS:The method is simple,precise,stable and reproducible,and can be used for simultaneous determination of chlorogenic acid,rutin,phillyrin,quercetin and glycyrrhizic acid in Sangju ganmao granules.
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Objective Influenza A (InfA) is an acute respiratory infectious disease persistently threatening human health and social stability. The high mutation of the InfA virus makes very significant the development of anti-influenza drugs. Traditional Chinese herbal drugs have shown distinct advantages in the prevention and management of InfA. This study aimed to investigate the effect of baicalin combined with phillyrin on the gene expression of InfA virus nucleoprotein (NP).Methods The nucleoprotein eukaryotic expression vector of InfA virus was constructed by transient transfection of InfA virus NP eukaryotic recombinant plasmid pcDNA3.1 (+)/NP into hela cells. Six groups were designed for the experiment: hela cells, liposome, recombinant plasmid (transfected pcDNA3.1(+)/NP), baicalin, phillyrin, baicalin (15.625 μg/mL-1) + phillyrin (12.5 μg/mL-1), each treated with respective agents after transfection. After 48 hours of culture, the number of the copied InfA virus NPs in the hela cells was measured by RT-qPCR and the effect of baicalin combined with phillyrin on the gene expression of InfA virus NP in the target cells evaluated by the statistical method.Results RT-qPCR showed that the gene expression of InfA virus NP was significantly lower in the baicalin + phillyrin than in the recombinant plasmid group (t=6.966, P1) and a synergistic effect of medium- and high-dose baicalin + medium- and high-dose phillyrin (CI<1).Conclusion Baicalin combined with phillyrin can decrease the gene expression of InfA virus NP and has a synergistic effect within a certain dose range.
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AIM To establish an HPLC method for the simultaneous content determination of four constituents in Xiaojie'an Capsules (Forsythiae Fructus,Leonuri Herba,Spatholobi Caulis,etc.).METHODS The analysis of chloroform extract of this drug was carried out on a 30 ℃ thermostatic Diamond C1scolumn(250 mm ×4.6 mm,5 μm),with the mobile phase comprising of acetonitrile (A)-0.05 mol/L monosodium phosphate (B) flowing at 1.0 mL/min in a gradient elution manner,and the detection wavelength was set at 278 nm.RESULTS Berberine hydrochloride,palmatine chloride,phillyrin and rutin showed good linear relationships within the ranges of 0.033 7-0.337 2 μg (r =0.999 1),0.054 8-0.548 3 μg (r =0.999 0),0.025 9-0.258 8 μg (r=0.999 2) and 0.008 4-0.084 2 μg (r =0.999 6),whose average recoveries were 98.8％ (RSD =1.3％),99.8％ (RSD =0.7％),98.8％ (RSD =1.3％) and 96.8％ (RSD =1.0％),respectively.CONCLUSION This sensitive and accurate method can be used for the quality control of Xiaojie'an Capsules.
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Objective: To simultaneously detect five components (phillyrin, chlorogenic acid, chlorpheniramine maleate, vitamin C and acetaminophen) in compound paracetamol vitamin C Yinqiao tablets.Methods: Using a Waters Xselect C18 column(250 mm×4.6 mm, 5 μm), the mobile phase consisted of acetonitril-0.1 mol·L-1 NaH2PO4(pH 3.0) with gradient elution at a flow rate of 1.0 ml·min-1.The detection wavelength was 230 nm and the column temperature was 35℃.Results: The linear range of vitamin C, acetaminophen, chlorogenic acid, chlorphenamine maleate and phillyrin was 0.500-200.200, 0.499-199.600, 0.501-200.100, 0.502-200.900 and 0.488-195.400 μg·ml-1(r＞0.999), respectively.The average recovery was 99.3%(RSD=1.5%), 97.8%(RSD=1.7%), 98.5%(RSD=1.1%), 97.1%(RSD=1.7%)and 99.8%(RSD=1.2%)(n=9), respectively.Conclusion: The simple and sensitive method can be applied in the quality control of compound paracetamol vitamin C Yinqiao tablets.
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Objective: To develop an HPLC-MS/MS method for the simultaneous determination of forsythoside A, phillyrin, forsythiaside, rutin, quercetin, isoquercitrin, ferulic acid, and hesperidin in the seed of Forsythia suspense (Thunb.) Vahl. Methods: Chromatographic separation was performed on a Diamonsil C18 column (150 mm × 4.6 mm, 5 μm) with linear gradient elution of methanol and 0.1% formic acid at a flow rate of 800 μL/min, and the injection volume was 10 μL. Multiple-reaction monitoring (MRM) was employed with switching electrospray ion source polarity in negative mode. The ion spray voltage was set at-4 500 V and the turbo spray temperature was maintained at 650℃. Results: All calibration curves showed good linearity within the test ranges. The average recoveries of the compounds ranged from 98.1% to 101.8%. The precisions (RSD) for the investigated components were less than 0.84%. The contents of forsythoside A, phillyrin, forsythiaside and rutin are higher than quercetin, isoquercitrin, ferulic acid and hesperidin. And the average contents of forsythoside A, phillyrin, forsythiaside and rutin are 1 609.78, 80.08, 86.64 and 373.86 μg/g, respectively. Conclusion: An efficient, rapid, and sensitive method is first established for the qualitation and quantification of eight major components in the seed of Forsythia suspense (Thunb.) Vahl. The satisfactory results demonstrate that the method could be applied as a reliable quality control method for the seed of Forsythia suspense (Thunb.) Vahl.
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Objective To compare the contents of total flavonoids, forsythoside A and phillyrin in Forsythiae Fructus leaves tea under different drying conditions; To determine the best drying method for Forsythiae Fructus leaves tea. Methods With the sodium nitrite-aluminum trichloride-sodium hydroxide solution as color reagent, total flavonoids in forsythiae Fructus leaves tea were determined by UV spectrophotometry at the wavelength of 500 nm. HPLC was used to determine the contents of forsythiaside A and phillyrin. The analysis was performed on a Diamosil C18 (2) column (4.6 mm × 250 mm, 5 μm); the mobile phase was composed of acetonitrile and 0.4% acetic acid with gradient elution; the detection wavelength was set at 277 nm; the flow rate was 1.0 mL/min at column temperature of 30 ℃. Results The content of total flavonoids of Forsythiae Fructus leaves tea under different drying conditions was basically the same; the sequence of the contents of forsythoside A and Forsythin of Forsythiae Fructus leaves tea under different drying conditions was: ventilated drying > vacuum drying > blast drying. Conclusion Different drying conditions have no effect on the contents of total flavonoids in Forsythiae Fructus leaves tea, but have obvious effects on the contents of forsythiaside A and phillyrin. Ventilation shade is better than blast drying and vacuum drying for preverence of forsythiaside A and forsythin.
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Objective:To study the effect of phillyrin on immune function in mice treated with cyclophosphamide.Methods: Mice model of immunosuppression was induced by intraperitoneal injection of cyclophosphamide,the mice in the experimental group received phillyrin with 50,100 and 150 mg/kg doses by gavage.While model group and control group received normal salin,all treatments continued 7 d.The thymus and spleen organ coefficients was measured by conventional methods.The phagocytosis of peritoneal macrophages was measured by FITC microspheres.The levels of IL-2,IL-4 and cAMP in peripheral blood were detected by ELISA.The effects of different doses of phillyrin on the above indexes were analyzed in immunosuppressed mice.Results: A mice model of immunosuppression was established after 7 d,the results showed that the immune organ index,phagocytic capacity of macrophages and the level of immune factors in serum could be improved,and the content of cAMP in peripheral blood lymphocytes was decreased in different degree by phillyrin.Conclusion: Phillyrin has the function of immune regulation,which can resist the immunosuppression of cyclophosphamide.
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OBJECTIVE:To develop a method for simultaneous determination of harpagide,harpagoside,chlorogenic acid, caffeic acid and phillyrin in Xiaoer qingyan granules. METHODS:HPLC method was adopted. The determination was performed on Zorbax SB-C18 column with mobile phase consisted of acetonitrile-0.1% phosphoric acid solution (gradient elution) at a flow rate of 1.0 mL/min. The detection wavelength was set at 210 nm (0-13 min,harpagide),327 nm (13-25 min,chlorogenic acid and caffeic acid),277 nm(25-29 min,phillyrin),210 nm(29-40 min,harpagosid);the column temperature was 30℃,and sam-ple size was 10 μL. RESULTS:The linear ranges of harpagide,harpagosid,chlorogenic acid,caffeic acid and phillyrin were 8.400-168.0 ng(r=0.9996),11.30-226.0 ng(r=0.9998),128.8-257.6 ng(r=0.9993),8.110-162.2 ng(r=0.9996),29.69-593.8 ng(r=0.9994),respectively. LOQs of the 5 components were 33.39,451.2,515.2,324.5,1188 ng/mL;LODs were 8.348, 112.8,128.8,81.12,297.0 ng/mL,respectively;RSDs of precision,stability and reproducibility tests were all lower than 2.0%. The recoveries were 96.39%-98.64%(RSD=0.83%,n=6),96.60%-98.89%(RSD=0.89%,n=6),96.28%-99.22%(RSD=1.25%,n=6),96.49%-99.54%(RSD=1.16%,n=6),96.26%-99.70%(RSD=1.30%,n=6),respectively. CONCLUSIONS:The method is simple,accurate and suitable for simultaneous determination of 5 components in Xiaoer qingyan granules
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OBJECTIVE:To establish the quality standard for ShutonganⅠgranule.METHODS:TLC was conducted for the qual-itative identification of Citrus aurantium and Forsythia suspensa. HPLC was conducted for the content determination of naringin in the preparation;the column was Agilent C18 with mobile phase of acetonitrile-water(20:80,V/V)at a flow rate of 1.0 ml/min,the detection wavelength was 283 nm,the column temperature was 30 ℃,and the injection volume was 10 μl. HPLC was also used for the content determination of phillyrin;the column was Agilent TC-C18 with mobile phase of acetonitrile- water(23:77,V/V)at a flow rate of 1.0 ml/min,the detection wavelength was 277 nm,the column temperature was 30 ℃,and the injection volume was 10 μl. RESULTS:The C. aurantium and F. suspensa of TLC showed clear spots and good separation. The linear range was 57.843-289.214 μg/ml for naringin(r=0.999 9)and 37.434-187.169 μg/ml for phillyrin(r=0.999 6);RSDs of precision,stability and reproducibility tests were less than 2%;recoveries were 97.20%-102.04%(RSD=1.7%,n=9)and 97.14%-102.27%(RSD=2.0%,n=9),respectively. CONCLUSIONS:The standard can be used for the quality control of ShutonganⅠgranule.
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Objective: To optimize the extraction process of Xiaoer Wenre Oral Liquid (XWOL). Methods: Through the analgesic, anti-inflammatory and antipyretic pharmacological experiments, the best extraction process route was primarily optimized, then taking the volatile oil amount as index, single factor test was adopted to investigate the effects of the soaking time and extracting time on extraction process of volatile oil; With contents of phillyrin, baicalin, and overall desirability as comprehensive evaluation indexes, central composite design-response surface method was adopted to predict the water extraction process. Results: Adding 10 times water, extracting for 6 h, collecting volatile oil and water extract, then the dregs and other medicines were extracted twice with 13 times water, for 80 min each time. Conclusion: The optimized extraction process is reasonable and feasible, and it can be used in a large scale industrial production.
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OBJECTIVE:To establish a method for determining the plasma concentration of paeoniflorin and phillyrin and phar-macokinetic study before and after intragastric administration of Qianliean granules. METHODS:LC-MS/MS method was adopted. The column was Waters C18 with mobile phase consisted of acetonitrile(A)-2 mmol/L ammonium acetate(containing 0.05% formic acid)(B)(0-9 min:15%A→50%A;9-11 min:50%A→90%A;11-17 min:90%A;17-19 min:90%A→15%A;19-20 min:15%A),at the flow rate of 0.6 ml/min;column temperature was 35 ℃ and the volume was 20 μl;quantitative ions were paeoniflorin m/z 525.2 → m/z 449.0,phillyrin m/z 552.3 → m/z 355.3. 7 SD male rats were docked to collect blood 0.5 ml from angular vein 0.25,0.5,0.75,1,1.5,2,3,4,6,8,10,12,24 h after administration Qianliean granule solution 1 g(medicinal materials)/kg to determine the blood concentration of drugs. DAS 2.1.1 software was employed to calculate pharmacokinetic parameters. RE-SULTS:The linear range of paeoniflorin and phillyri were 5.0-2500.0 μg/L(r=0.9979)and 2.0-2000.0 μg/L(r=0.9982),re-spectively;RSD of precision test was less than 5.5%(n=5);the method recovery were 96.0%-104.0% and 92.0%-107.0%,the extration recovery were 71.4%-83.5% and 81.5%-92.3% and RSD of stability test was less than 5.0%(n=3). The pharmacokinet-ic parameters of paeoniflorin and phillyrin were as follows as t1/2 of (2.206 ± 0.631) and (1.355 ± 0.317) h;cmax of (1504.069 ± 620.885) and (79.043 ± 15.568)μg/L;tmax of (1.000 ± 0.250) and (1.214 ± 0.267) h;AUC0-24 h of (4897.645 ± 2207.577) and (263.475±54.795)μg·h/L;CL of(5.025±2.773)and(76.253±13.986)L/(h·kg). CONCLUSIONS:The method is highly sensi-tive,exclusive,simple,accurate and reliable,and can be applied to study the pharmacokinetic characteristics of paeoniflorin and phillyrin in rats in vivo.
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Objective:To establish a method for determining the content of imperatorin,phillyrin and forsythoside A in Kanggan Jiedu granules by UPLC. Methods:An Agilent Eclipse Plus C18 column(100 mm × 2. 1 mm,1. 8 μm)was used,the mobile phase was acetonitrile-0. 2% acetic acid with gradient elution,the flow rate was 0. 2 ml· min-1 and the column temperature was 30℃. The de-tection wavelength was set at 335 nm in the first 8 min for forsythoside A, and then changed to 277 nm for phillyrin between 8 and 10 min followed by 300 nm for imperatorin between 10 and 15 min. Results:The linear range of imperatorin was 0. 374-3. 366 μg·ml-1 (r=0. 999 6)and the average recovery was 99. 12%(RSD=1. 07%),that of phillyrin was 0. 568-5. 112 μg·ml-1(r=0. 999 4)and the average recovery was 100. 39%(RSD=0.93%,n=9),and that of forsythoside A was 0.738-6.642μg·ml-1(r=0.999 7)and the average recovery was 99. 78%(RSD=1. 14%,n=9). Conclusion:The method is rapid,simple,accurate and specific,which can be used for the determination of imperatorin,phillyrin and forsythoside A in Kanggan Jiedu granules.
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OBJECTIVE:To research the mechanism of in vitro permeation of phillyrin through blood-brain barrier. METH-ODS:After Madin-Darby canine kidney epithelial cells transfected with colorectal cancer MDR1 gene (MDCK-MDR1) were cul-tured with phillyrin solution of 0(negative control),10,25,50,75 and 100μg/ml for 24 h,cell viability was determined by resa-zurin method and cell survival rate was calculated. After MDCK-MDR1 cells were cultured with phillyrin solution of 10,25,50, 75 and 100 μg/ml for 10 min,the content of phillyrin in the cells was determined,and concentration-uptake rate curve was drawn. Following 3 h culture of MDCK-MDR1 cells with phillyrin solution of 0 (negative control),50 and 100 μg/ml,the structure of cell tight junction protein was observed under the inverted microscope. RESULTS:Compared to the negative control group,after 24 h cell culture with phillyrin solution of 10-100 μg/ml,no obvious change in cell survival rate occurred. MDCK-MDR1 cells cultured with the phillyrin at a mass concentration of 10-100 μg/ml demonstrated a nonlinear relationship with concentration of phillyrin and a gradual saturation trend. After the cells were cultured with phillyrin of 50 and 100 μg/ml for 3 h,cell tight junction protein was intact. CONCLUSIONS:The absorption of phillyrin through the simulated blood-brain barrier may be in the form of passive transportation combined with active transportation,the concertration has effect on cell tight junction protein.
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Phillyrin is an important lignans compound from Forsythia. It is not only found in the raw medicinal material with high content, but also shows a variety of pharmacological activities. In the paper, the recent advances in extraction methods, pharmacology-toxicology, pharmacokinetics and the other studies of phillyrin were reviewed in order to provide reference and suggestions for the fur-ther research.
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Objective:To establish an HPLC method for the simultaneous determination of hesperidin and phillyrin in Siji Gamao capsules. Methods:HPLC was applied with the chromatographic conditions as follows:the chromatographic column was Agilent-XDB C18 (250 mm × 4. 6 mm,5μm) at 30℃, the mobile phase was methanol-0. 1% phosphoric acid solution with gradient elution, the flow rate was 1. 0 ml·min-1 , the detection wavelength was 277nm and the injection volume was 5μl. Results:The linear range of hesperi-din and phillyrin was 12.000-60.000 μg·ml-1(r=0.999 6) and 18.000-90.000 μg·ml-1(r=0.999 3),respectively; the average recovery was 96.61%(RSD=1.40%) and 97.66%(RSD=1.27%), respectively. Conclusion: The method is simple, accurate and repeatable, which can be used in the quality control of Siji Ganmao capsules.
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This study was aimed to establish a HPCE method for the content determination of hesperidin and phillyrin in Bao-He-W an (BHW). Fused silica capillary (75 cm í 50 μm) was employed and 30 mmol·L-1 borax so-lution (8% acetonitrile, pH9.64) was served as the running buffer. Other conditions were as follows: electrokinetic injection was 50 kPa í 20 s; analytical voltage was 20 kV; temperature was 20℃; and detection wavelength was 254 nm. The silica capillary was flushed with 0.1 mol·L-1 sodium hydrate and the running buffer for 10 min before each injection, respectively. The results showed that the linearity of hesperidin was in the range of 0.10~2.40 mg·mL-1 (r=0.999 4), the average recovery was 99.85% and RSD=2.34%. The phillyrin was in the range of 0.07~0.84 mg·mL-1 (r=0.999 2), the average recovery was 99.16% and RSD=2.78%. It was concluded that the method was rapid and sensitive. It can be used for the quality control of content determination of hesperidin and phillyrin in BHW.