Inhibition of intracellular proton-sensitive Ca2+-permeable TRPV3 channels protects against ischemic brain injury

Ischemic brain stroke is pathologically characterized by tissue acidosis, sustained calcium entry and progressive cell death. Previous studies focusing on antagonizing N-methyl-d-aspartate (NMDA) receptors have failed to translate any clinical benefits, suggesting a non-NMDA mechanism involved in the sustained injury after stroke. Here, we report that inhibition of intracellular proton-sensitive Ca2+-permeable transient receptor potential vanilloid 3 (TRPV3) channel protects against cerebral ischemia/reperfusion (I/R) injury. TRPV3 expression is upregulated in mice subjected to cerebral I/R injury. Silencing of TRPV3 reduces intrinsic neuronal excitability, excitatory synaptic transmissions, and also attenuates cerebral I/R injury in mouse model of transient middle cerebral artery occlusion (tMCAO). Conversely, overexpressing or re-expressing TRPV3 increases neuronal excitability, excitatory synaptic transmissions and aggravates cerebral I/R injury. Furthermore, specific inhibition of TRPV3 by natural forsythoside B decreases neural excitability and attenuates cerebral I/R injury. Taken together, our findings for the first time reveal a causative role of neuronal TRPV3 channel in progressive cell death after stroke, and blocking overactive TRPV3 channel may provide therapeutic potential for ischemic brain injury.

Chemical components from Lianhua Qingwen Capsule (III) / 中草药

Objective: To study the chemical constituents of Lianhua Qingwen Capsule and illuminate its substance foundation. Methods: The compounds were isolated and purified by LPLC and preparative HPLC from the 30% ethanol fraction of Lianhua Qingwen Capsule macroporous resin column chromatography. Their chemical structures were elucidated by the spectral analyses. Results: 18 compounds were isolated and identified as forsythoside A (1), forsythoside I (2), forsythoside H (3), lugrandoside (4), isolugrandoside (5), ferruginoside A (6), lianqiaoxinoside C (7), calceolarioside C (8), forsythoside E (9), ferruginoside B (10), D-amygdaloside (11), L-amygdaloside (12), sambunigrin (13), cornoside (14), 4-hydroxy-4-methylenecarbomethoxy-cyclohexa- 2,5-dienone (15), liriodendrin (16), liquiritigenin-7-O-β-D-glucopyranoside (17), and 3,4-dihydroxy benzaldehyde (18). Conclusion: Compounds 2-8, 10, and 13-18 are isolated from Lianhua Qingwen Capsule for the first time, and compounds 4-6, 10, 15, and 16 are isolated from single herb in Lianhua Qingwen Capsule compound for the first time. The spectral data in DMSO-d6 solution of compound 8 are reported firstly with 2D NMR spectral data. The above results show the high polar chemical constitutions of Lianhua Qingwen Capsule, which provides more chemical information of Lianhua Qingwen Capsule.

Spectrum-effect relationship of wind-dispersing and exterior syndrome-relieving of Shufeng Jiedu Capsules / 中草药

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.

Spectrum-effect relationship of anti-inflammatory activity of Shufeng Jiedu Capsule based on neural network analysis / 中草药

Objective: To establish the spectrum-effect relationship network model of Shufeng Jiedu Capsule (SJC) for anti- inflammatory activity and explore potential effective compounds of its anti-inflammatory effects. Methods: On the basis of the establishment of fingerprint analysis method for SJC, the uniform design method was used to divide eight herbs in SJC into groups of different proportions to determine their inhibition ratios of TNF-α and IL-6 released by LPS-induced RAW264.7 cells. Pharmacodynamic data and chemical information of MS fingerprints of each group were analyzed with the BP artificial neural network to get the anti-inflammatory effect of each peak to establish the chromatography-efficacy relationship. Results: According to the optimal neural network model, 14 characteristic peaks were calculated and found to be significantly correlated with the anti- inflammatory activity of SJC. Conclusion: Through the spectrum-effect study, it is speculated that the anti-inflammatory components of SJC may be the 14 compounds including rhein, emodin, verbenalin, verbascoside, pinoresinol-β-D-glucoside, forsythoside A, polydatin, etc., which provides a reference for the quality control and determination of effective components.

Simultaneous Determination of 7 Components in Niuhuang Qingwei Pills by HPLC-wavelength Switching Method / 中国药房

OBJECTIVE： To develop a method for simultaneous determination of 7 components of Niuhuang qingwei pills as chlorogenic acid， geniposide， forsythoside A， narirutin， baicalin， ammonium glycyrrhetate， chrysophanol. METHODS： HPLC-wavelength switching method was adopted. The determination was performed on Agilent ZORBAX SB-C18 column with mobile phase consisted of acetonitrile （A）-0.1％ phosphoric acid （B） gradient elution at the flow rate of 1.0 mL/min. The detection wavelengths were set at 348 nm （chlorogenic acid）， 238 nm （geniposide）， 330 nm （forsythoside A）， 280 nm （narirutin and baicalin）， 237 nm （ammonium glycyrrhetate）， 254 nm （chrysophanol）. The column temperature was 30 ℃， and sample size was 10 μL. RESULTS： The linear ranges of chlorogenic acid， geniposide， forsythoside A， narirutin， baicalin， ammonium glycyrrhetate， chrysophanol were 0.011 67-0.233 4 μg （r＝0.999 4）， 0.042 91-0.858 1 μg （r＝0.999 4）， 0.125 0-2.500 μg （r＝0.999 9）， 0.118 0- 2.360 μg （r＝0.999 9）， 0.119 6-2.392 μg （r＝0.999 7）， 0.030 57-0.611 4 μg （r＝0.999 6）， 0.006 201-0.124 0 μg（r＝0.999 4）， respectively； the limits of quantitation were 1.167， 0.858， 1.250， 1.180， 1.196， 0.611， 0.620 μg/mL， respectively； RSDs of precision tests were 0.98％， 1.04％， 0.59％， 1.50％， 0.83％， 1.24％ and 1.32％ （n＝6）， respectively. RSDs of stability tests were 1.21％， 0.97％， 1.42％， 0.71％， 0.98％， 1.87％ and 1.63％ （n＝6， 12 h）， respectively. Average recoveries were 98.32％， 98.11％， 98.81％， 98.50％， 98.30％， 98.16％ and 97.83％， and the RSDs were 1.37％， 1.41％， 0.64％， 1.01％， 1.18％， 1.16％ and 1.16％ （n＝6）， respectively. CONCLUSIONS： Established method is easy and reproducible. It can be used for the quality control of Niuhuang qingwei pills.

Simultaneous determination of Forsythoside A and other four components in Xiao'er-Ganmao-Keli from multivendor by HPLC / 国际中医中药杂志

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.

Optimization of method for determination of Forsythia Fructus and discussion on the establishment of quality control standard for green (old) Forsythia Fructus / 中国中药杂志

In order to obtain the optimum method for content determination of Forsythia Fructus (FF), a variety methods for the sample preparation of FF were evaluated by the content determination methods of Chinese Pharmacopoeia. And an optimum method was screened and as follows: 30 times with 70% ethanol solution in ultrasonic extractor for half an hour. The method can achieve the best effect of simultaneously extracting forsythoside A and forsythin. Then, a HPLC method for simultaneous determination of forsythoside A and forsythin was established by methodology. The HPLC chromatographic conditions: the mobile phase consisted of acetonitrile (A)-0.4% acetic acid solution (B) with gradient elution [0-33 min，15%A，33-43 min，15%-25%A，43-60 min，25% A] was at the flow rate of 1 mL·min⁻¹, the column temperature was 25 °C, and the detection wavelength was 330 and 277 nm. Moreover, the contents of forsythoside A and forsythin for 10 Green Forsythia Fructus (GF) and 5 Old Forsythia Fructus (OF) were determined by this method and Chinese Pharmacopoeia. The result not only displayed that the established method is effective, rapid, and simple, but also showed that the contents of forsythoside A and forsythin for GF and OF were significantly different. Which implied that the forsythoside A and forsythin limit standard for GF and OF should be controled by different values. This studies provide an important basis for the establishment of the content determination of FF and the quality control standard for GF and OF.

Comparison of Effects of Different Drying Methods on Contents of Effective Components of Forsythiae Fructus Leaves Tea / 中国中医药信息杂志

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.

Simultaneous quantification of eight constituents in the seed of Forsythia suspense (Thunb.) Vahl by HPLC-MS/MS method / 中草药

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.

Rapid establishment of Q-Marker database for Qingreling Granules with UPLC coupled with hybrid quadrupole-orbitrap mass spectrometry / 中草药

Objective: To establish the Q-Marker database for Chinese medicine Qingreling Granules (QG) based on the concept of Q-Marker by the method of ultra high performance liquid chromatography coupled with hybrid quadrupole-orbitrap mass spectrometry (UPLC-quadrupole-orbitrap MS) to recognize and identify the chemical constituents, and study the quality of QG. Methods: The 50% methanol extract was separated on an Acquity UPLC BEH C18 column (100 mm × 2.1 mm, 1.7 μm), and eluted with a gradient of methanol-water containing 0.1% formic acid. Constituents of QG were identified by HRMS in the negative ion mode using both full scan and two-stage threshold-triggered mass modes. Eleven Q-Marker were docked by Autodock vina 1.1.2 software, using H5N1 avian influenza virus as the Neuraminidase receptor, and the active site of target protein were determined by the Auto Dock 1.5.6 program. Results: A total of 39 compounds were identified, including 25 flavonoids, 6 phenylethanoid glycosides, 4 organic acids, 2 triterpenoids, and 2 lignan ingredients. Totally 15 components were specific and 32 chemical compositions were reported for the first time in QG by the method of UPLC-quadrupole-orbitrap MS. Baicalin, wogonoside, forsythoside A, liquiritin, forsythoside E, forsythoside B, isoliquiritin, baicalein, phillyrin, wogonin, and liquirtigenin could be used to establish the Q-Marker database. Conclusion: To establish Q-Marker database for Chinese medicine QG and identify the compounds in QG by the method of UPLC-quadrupole-orbitrap MS.

Determination of chemical constituents and quantitative fingerprints of Xiao’er Xiaoji Zhike Oral Liquid by HPLC-ESI-Q-TOF/MS / 中草药

Objective To establish the HPLC multi-index quantitative fingerprints of Xiao’er Xiaoji Zhike Oral Liquid (XXZOL), and to provide methods and technical support for improving its quality standard. Methods Using an Agilent Zorbax SB-C18 (250 mm × 4.6 mm, 5.0 μm) column, acetonitrile-0.4% glacial acetic acid mobile phase system, gradient elution, flow rate of 0.6 mL/min, column temperature 25 ℃, detection wavelength 254 nm; The HPLC-ESI-Q-TOF/MS technique was used to identify the chemical constituents (peak 1-synephrine, peak 13-chlorogenic acid, peak 16-quercetin-3-O-β-D-glucose-7-O-β-D-gentiobioside, peak 22-hyperoside, peak 24-forsythoside A, peak 26-naringin, peak 27-hesperidin, peak 29-neohesperidin, peak 33-phillyrin, peak 37-3,29-dibenzoyl rarounitriol) in the XXZOL, and the content of the main components was determined by HPLC-DAD. The HPLC fingerprint was established on the basis of multi-batch analysis. Results The fingerprints of the 10 batches of samples were established, and the similarities of them were above 0.9, which were 0.999 3, 0.999 6, 0.999 2, 0.999 2, 0.999 4, 0.999 3, 0.999 4, 0.999 5, 0.999 1, and 0.999 7, respectively. XXZOL has 38 common peaks, 32 of them was identified, six of them was unknown; the contents of the 10 chemical constituents were 2 542.42, 158.61, 147.02, 34.35, 432.42, 1 813.01, 1 339.70, 2 580.71, 175.32, and 1 615.17 μg/mL. Conclusion The establishment of multi-index analysis by HPLC and fingerprints of XXZOL provides a scientific basis for improving its quality standards.

Simultaneous Determination of Eight Active Components in Liuwei Wuling Tablet Using HPLC / 中草药·英文版

Objective To develop an effective method for simultaneous determination of eight major components in Liuwei Wuling Tablet (LWT). Methods Reversed phase HPLC method was used with variously improved conditions. Results Salidroside, forsythoside A, specnuezhenide, phillyrin, schisandrol A, schizandrol A, schizandrin A, and schizandrin B in LWT were successfully separated on a Kromasil 100-5-C18 reverse phase column (250 mm × 4.6 mm, 5 μm) using acetonitrile-0.1% phosphoric acid (gradient elution) as mobile phase. The detection wavelength was 275 nm with flow rate of 1.0 mL/min, and the column temperature was maintained at 35 oC. The recovery rate of the method was within the range of 95.13%–104.56% and the precision (RSD) was less than 3% for all eight analytes. All the compounds showed good linearities (r2 > 0.9980) in a relatively wide concentration range. Conclusion Simultaneous quantification of the multiple components by HPLC would be a better strategy for the quality evaluation of LWT.

Phenylethanoid glycosides from fruits of Forsythia Suspensa / 中草药

Objective: To study the chemical constituents from the fruits of Forsythia suspensa. Methods: Compounds were isolated by combination of various chromatographic techniques including column chromatography over macroporous resin, silica gel, Sephadex LH-20, and reversed-phase HPLC. Their structures were elucidated by physicochemical property and spectroscopic analysis. Results: Two compounds were isolated from the 50% ethanol extract of the fruits of F. suspensa and identified as 7R-suspensaside methyl ether (1) and 7S-suspensaside methyl ether (2). Conclusion: Compound 1 is a new compound named forsythoside K.

Optimization of Ultrahigh Pressure Extraction of Forsythoside from Forsythia Suspensa by Box-Behnken Experimental Design / 中国药师

Objective: To optimize the ultrahigh pressure extraction ( UPE) process for forsythoside from Forsythia suspensa by Box-Behnken experimental design. Methods:On the basis of single factor screening, a three-factor and three-level Box-Behnken ex-perimental design was employed with liquid/material ratio( X1 ) , extraction pressure ( X2 ) and extraction time ( X3 ) as the independent variables. The response variable was the extraction yield of forsythoside. Results:The optimal UPE conditions for forsythoside were as follows:the ratio of solvent to material (ml·g-1) was 70, the extraction pressure was 151 MPa, and the extraction time was 114 s. With the optimal extraction process, the extraction yield of forsythoside was 13. 15 mg·g-1 . Conclusion:As a novel extraction tech-nology for Chinese herbal medicines, UPE procedure has higher extraction yield, lower extraction temperature, shorter extraction time and less power consumption, which provides a brand-new method for the extraction of forsythoside from Forsythia suspensa.

HPLC specific chromatogram of Lamiophlomis Herba and its counterfeit and determination of four effective components / 中国中药杂志

This study is to establish the HPLC specific chromatogram and determine four main effective components of Lamiophlomis Herba and its counterfeit.Chlorogenic acid, forsythoside B, acteoside and luteoloside were reference substance.HPLC analysis was performed on a Waters XSelect C₁₈ column (4.6 mm×250 mm,5 μm).The mobile phase was acetonitrile-0.5% phosphoric acid solution (18∶82) with isocratic elution.The flow rate was 1.0 mL•min⁻¹, the detection wavelength was 332 nm and the column temperature was 30 ℃.Chemometrics software Chempattern was employed to analyze the research data.HPLC specific chromatogram of Lamiophlomis Herba from different samples were of high similarity, but the similarity of the HPLC specific chromatogram of its counterfeit were less than 0.65.Both of cluster and principal component analysis can distinguish certified products and adulterants.The HPLC specific chromatogram and contents of four effective components can be used for the quality control of Lamiophlomis Herba and its preparations.It provided scientific basis to standardize the use of the crude drug.

Fingerprint Chromatogram of Forsythia Suspensa Dispensing Granule by HPLC / 医药导报

Objective To establish the fingerprint of Forsythia suspensa dispensing granule by HPLC. Methods The total of 10 samples of Forsythia suspensa were analyzed by HPLC on a Waters SunFire C18 column(4. 6 mm×250 mm,5 μm) with mobile phase of A:acetonitrile and mobile phase B:0. 4% acetic acid solution as gradient elution, with flow rate at 1. 0 mL·min-1 , wavelength at 277 nm and column temperature at 30 ℃. Results There were 13 characteristic common peaks in the Forsythia suspensa dispensing granule, in which two peaks identified as forsythoside A and forsythin, the similarities were more than 0. 999. Conclusion The HPLC fingerprint of Forsythia suspensa dispensing granule is stable, and is easy to manipulate, which can provide more information for the quality control of Forsythia suspensa dispensing granule.

Simultaneous Determination of Imperatorin, Phillyrin and Forsythoside A in Kanggan Jiedu Granules by UPLC / 中国药师

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.

Simultaneous determination of three phenolic glycosides in Callicarpa nudiflora by UHPLC-MS methods and analysis of their pharmacokinetics in plasma of rats / 中草药

Objective: An UHPLC-MS/MS method was developed for the simultaneous determination of acteoside, isoacteoside, and forsythoside B in plasma of rats and the pharmacokinetic parameters for three phenolic glycosides were calculated as well. Methods: Samples of plasma of rats were obtained at different time after rats were administrated with Callicarpa nudiflora extract (5 g/kg). After the addition of acidification (hydrochloric acid, 0.25 mol/L) and deproteinization by acetonitrile, plasma samples were separated on a Phenomenex® Kinetex C18 column (50 mm × 2.1 mm, 1.7 μm) with gradient elution using acetonitrile-0.005% formic acid as mobile phase. Mass spectrometric detection was carried out by multiple reaction monitoring (MRM) using electrospray ionization in negative ion mode. Results: A good linearity of acteoside, isoacteoside, and forsythoside B was shown in the ranges of 7.77 - 3 880.00 ng/mL (r2 = 0.995 5), 5.04 - 2 520.00 ng/mL (r2 = 0.994 9), and 1.78 - 890.00 ng/mL (r2 = 0.995 1), respectively. The mean extraction recoveries of analytes were in the range of 75.2% - 89.9%, and the intra- and inter-day RSD values were less than 8.8%. The tmax of acteoside, isoacteoside, and forsythoside B was about 30 min, AUC0~t were (93 881.65 ± 18 326.65), (29 204.97 ± 8 499.88), and (15 027.05 ± 3763.82) ng∙min/mL, Cmax were (2 179.00 ± 355.60), (737.57 ± 210.31), and (227.30 ± 48.38) ng/mL, t1/2z were (235.41 ± 117.90), (151.56 ± 49.23), and (161.68 ± 63.92) min, respectively. Conclusion: The method is proved to be simple, rapid, and specific, and to be suitable for the simultaneous determination of acteoside, isoacteoside, and forsythoside B in plasma of rats and the pharmacokinetic study.

Content determination of Forsythoside A in Yinqiaosan granules by HPLC / 药学实践杂志

Objective To establish a method for determination of the content of Forsythoside A in Yinqiaosan granules . Methods HPLC method was adopted to carry on the determination .Welch Ultimate XB‐C18 column (250 mm × 4 .6 mm ,5μm) was used with the mobile phase of methanol‐0 .2% Glacial acetic acid solution by gradient elution . The flow rate was 1 .0 ml/min ,the detection wavelength was 330 nm ,and the column temperature was 20 ℃ .Results Forsythoside A showed a good linear relationship at the range from 0 .080 3~3 .211 μg (r=1 .000) .The average recovery rates were 103 .78% ,and RSD=0 .82% .Conclusion The method was convenient and accurate .Meanwhile ,it also has good specificity and reproducibili‐ty .It can be used effectively for the quality control of Yinqiaosan granules .

Comparison of Contents of Forsythoside A and Forsythin in Fructus Forsythia and Its Dispensing Granule / 中国中医药信息杂志

Objective To compare the contents of forsythoside A and forsythin in Fructus Forsythia and its dispensing granule. Methods HPLC-gradient elution method was used with SunFire C18 column (4.6 mm×250 mm, 5μm), mobile phase A of acetonitrile and B of acetic acid, flow rate of 1.0 mL/min, detection wavelength of 277 nm, and column temperature at 30 ℃. HPLC was used to determine the contents of forsythoside A and forsythin in Fructus Forsythia and its dispensing granule, and compare the difference between the two contents. Results The content of forsythoside A in dispensing granule was less than that of raw material of Fructus Forsythia, and the concentration of the major components in the commercial Lianqiao Granule were not equivalent to that in the decoction of Fructus Forsythia. The content of forsythin in dispensing granule was equivalent with that of raw material of Fructus Forsythia. Conclusion The original formula granule production process needs to be improved, and the standardized criteria for the quality control and reasonable quality standard of granule should be established.