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Objective To investigate the impact of phellodendrine on treating bacterial vaginosis (BV) mice. Methods BV model mice were induced by vaginal injecting with EPEC and MRSA. After the mice in control and model groups being administrated with phellodendrine (10, 20, and 40 mg/kg) for 7 d, the mice were sacrificed under deep anesthesia. The mice vaginal pathology changes were observed using HE staining method. IL-1β, IL-6, IL-8, and TNF-α secretion in vagina and PGE2, PGF2α levels in uterus were detected using ELISA assay. Uterine MMP-9, TLR4, and NF-κB expression levels were measured using western blotting. Results Vaginal pathology changes were improved by phellodendrine. Specifically, phellodendrine could reduce IL-1β, IL-6, IL-8, and TNF-α secretion in vagina, it could also reduce PGE2 and PGF2α content, and inhibit MMP-9, TLR4, and NF-κB expression levels in uterus tissue. Conclusion Phellodendrine could effectively treat BV and reduce the rise of pre-term birth by simultaneously inhibiting endovaginal inflammatory response and regulating intrauterine TLR4/NF-κB signal pathway in BV mice.
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OBJECTIVE:To establish a method for simultaneous determination of sodium danshensu,protocatechuic aldehyde,salvianolic acid B and phellodendrine hydrochloride in Shenbai shuxin granules.METHODS:HPLC method was adopted.The separation was performed on Waters sunfire-C18 column with mobile phase consisted of acetonitrile-0.05 ％ trifluoroacetic acid (gradient elution) at the flow rate of 1.0 mL/min.The detection wavelength was set at 288 nm,and the column temperature was 30 ℃.The sample size was 5 μL.RESULTS:The linear ranges of sodium danshensu,protocatechuic aldehyde,salvianolic acid B and phellodendrine hydrochloride were 0.040 01-1.600 46 μg(r=0.999 9),0.013 84-0.553 7 μg(r=0.999 9),0.049 32-1.972 94 μg(r=0.999 6),0.014 46-0.578 6 μg(r=0.999 8).The limits of quantitation were 7.68,2.66,4.74,1.38 ng,and the limits of detection were 1.92,0.66,2.36,0.69 ng,respectively.RSDs of precision,stability and reproducibility tests were all lower than 2％.The recoveries were 98.846％-100.762％ (RSD=0.77％,n=6),96.632％-99.463％ (RSD=0.98％,n=6),98.541％-100.432％ (RSD=0.82 ％,n =6),98.607 ％-101.521％ (RSD =1.11％,n =6),respectively.CONCLUSIONS:The method is simple,accurate and precise.It can be used for 4 components in Shenbai shuxin granules.
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OBJECTIVE: To develop an ultra-high performance liquid chromatography(UHPLC) method for simultaneous determination of berberine hydrochloride, phellodendrine hydrochloride, palmatine hydrochloride, magnoflorine, and rhizoma atractylodis in Ermiao pills. METHODS: The UHPLC analyses were performed on a Waters BEH C18 column (2.1 mm×100 mm, 1.7 μm)eluted with acetonitrile and 0.1% phosphoric acid(each 100 mL containing 0.1 g sodium dodecyl sulfate). The flow rate was 0.3 mL·min-1, the detection wavelength was set at 280 and 340 nm, and the column temperature was set at 30℃. RESULTS: The calibration curves of the five compositions had good linear relationship in the ranges of the tested concentrations. The precision, stability and repeatability complied with the requirements of methodology. The recoveries were between 96.8%-99.3% respectively. The RSDs were below 2.8%. CONCLUSION: The developed method is simple, sensitive, rapid and accurate. This work provides helpful information for the comprehensive quality evaluation of Ermiao pills.
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Objective To optimize the particle size of Huoxue powder by contents comparison of emodin,phellodendrine,berberine and jatrorrhizine before and after permeabilized skin absorption of Huoxue powder in different particle size of 0.150,0.075,0.048,0.038 mm.Methods The contents of emodin,phellodendrine,berberine and jatrorrhizine in transparent absorbent fluid of Huoxue power in different particle size within 24 hours were measured by ultra performance liquid chromatography tandem mass spectrometry,and optimize its particle size by contents comparison of the effective components.Results The contents of the active components in Huoxue power with the particle size of 0.075 mm were high before and after percutaneous absorption.Conclusion Particle size of 0.075 mm is best for Huoxue powder.
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Objective To establish the method of thin layer identification of Phellodendri Chinensis Cortex and solid phase extraction - high performance liquid chromatography (SPE-HPLC) determination of the content of phellodendrine in Leshu Lotion. Methods Thin layer chromatography (TLC) was used for qualitative identification of Phellodendri Chinensis Cortex in Leshu Lotion and the Bond Elutplexa PCX strong cation-exchange solid phase was used for extraction small column purification. Platisil-NH2 column was as chromatographic column; column temperature was 30 ℃; triethylamine pH 3.82 (phosphoric acid), tetrahydrofuran, acetonitrile was 18:12:70, isocratic elution; flow rate was 1.0 mL/min; detection wavelength was 286 nm. Results TLC could obviously identify active ingredients of phellodendrine in Phellodendri Chinensis Cortex. The phellodendrine had good linear relationship between concentration and peak area within the scope of 1.22–12.2 μg (r=0.9999); the average recovery rate of phellodendrine was 101.18%, RSD was 1.71%. Conclusion This method is simple to operate, with accuracy and repeatability, and can be used for quality control of Leshu Lotion.
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OBJECTIVE:To establish the quality standard of Huoxuesan. METHODS:Microscopic identification method was adopted for the qualitative identification of Rhizoma pinelliae,Eupolyphaga seu,Phellodendron amurense and Fructus kochiae in the preparation. HPLC was adopted for the contents determination of phellodendrine,polydatin,jatrorrhizine,berberine and emo-din:the column was ZORBAX Eclipse Plus C18 with methanol-0.1% phosphoric acid(gradient elution)at a flow rate of 0.4 ml/min,the detection wavelength was 240 nm,column temperature was 30 ℃,and the injection volume was 5 μl. RESULTS:Micro-scopic identification map of R. pinelliae,E. seu,P. amurense and F. kochiae was clear. The linear range was 0.159-5.073 μg(r=0.997 4)for phellodendrine、0.149-4.761 μg(r=0.999 9)for polydatin、0.239-7.649 μg(r=0.995 5)for jatrorrhizine、0.165-5.268 μg (r=0.997 2)for berberine、0.012-0.382 μg(r=0.999 9)for emodin;RSDs of precision,stability and reproducibility tests were low-er than 3.0%;recoveries were 98.9%-104.1%(RSD=1.9%,n=6),96.1%-101.7%(RSD=2.5%,n=6),99.6%-105.1%(RSD=2.6%,n=6),90.3%-98.2%(RSD=2.9%,n=6)and 92.9%-96.4%(RSD=2.0%,n=6)respectively. CONCLUSIONS:The es-tablished standard can be used for the quality control of Huoxuesan.
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Objective To establish quality standards forAnshu Liquid.Methods Phellodendri Chinensis Cortex and Kochiae Fructus were qualitatively identified by TLC method. The contents of berberine hydrochloride and phellodendrine hydrochloride in the preparation were determined by HPLC method. The chromatographic column was ZORBAX SB-C18 (4.6 mm×250 mm, 5μm); the mobile phase was acetonitrile-0.1% phosphoric acid (0.5% diethylamine) gradient elution; the flow rate was 1.0 mL/min; the detection wavelength was 284 nm.Results Berberine hydrochloride and phellodendrine hydrochloride in Phellodendri Chinensis Cortex and saponin Ic in Kochiae Fructus were detected by TLC method. The linear relationship of berberine hydrochloride in the preparation was Y=23.109X?30.548,r2=0.999 8, RSD=2.97%, showing that the linear range of berberine hydrochloride was 0.050 5– 2.525 0 μg. The linear relationship of phellodendrine hydrochloride in the preparation wasY=10.038X?2.446 6, r2=0.999 9, RSD=1.24%, showing that the linear range of phellodendrine hydrochloride was 0.050 0–2.500 0 μg. Conclusion The method is accurate, rapid, stable and reliable, with good reproducibility, which can be used for the quality control and evaluation ofAnshu Liquid.
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Objective To establish fingerprint of analgesic ingredients in Cangbaiqutong capsules by HPLC, and control different production batches of Cangbaiqutong capsules’ quality. Methods Waters XSELECT CSH-C18 (4.6 mmxl50 mm, 5.0μm) chromatographic column was used and eluted with acetonitrile -0.1% phosphoric acid solution gradiently in the flow rate of 1.0 ml/ min. The detection wavelength was 284 nm with column temperature of 30°C. rrith phellodendrine chloride, gentiopicroside, paeoniflorin, tetrandrine and berberine hydrochloride as the object references, ten batches of Cangbaiqutong capsules were tested and analyzed by similarity comparison. Results fingerprint spectrum in Cangbaiqutong capsules had 20 common peaks in total, and characteristic spectra of Cortex Phellodendri, Gentiana, Radix Paeoniae Rubra and Radix Stephaniae Tetrandrae were found, while similarity of HPLC fingerprint was more than 0.9 among those batches of samples. Conclusion Using HPLC fingerprint can be used to evaluate the Cangbaiqutong capsules% quality in the round, which could improve the quality control standard in productive process of Cangbaiqutong capsules.
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Objective: To establish the RP-HPLC method for the simultaneous determination of phellodendrine, rutin, baicalin, baicalein, berberine hydrochloride, quercetin, and atractylodin in Longsha Granules. Methods: The seven components were measured on a C18 column (250 mm × 4.6 mm, 5 μm) of Kromasil with a gradient elution of acetonitrile -0.02% phosphoric acidsolution at the wavelengths of 205 nm (phellodendrine, rutin, baicalin, baicalein, and quercetin) and 340 nm (berberine hydrochloride and atractylodin), a flow rate of 1.0 mL/min, and the column temperature of 30 ℃. Results: Phellodendrine, rutin, baicalin, baicalein, berberine hydrochloride, quercetin, and atractylodin showed a good correlation in the ranges of 0.011-0.168 mg/mL (r = 0.9999), 0.010-0.160 mg/mL (r = 0.9999), 0.053-0.842 mg/mL (r = 0.9999), 0.014-0.228 mg/mL (r = 0.9998), 0.016-0.266 mg/mL (r = 0.9998), 0.009-0.144 mg/mL (r = 0.9999), and 0.002-0.024 mg/mL (r = 0.9997). The average recovery rates were 99.83%, 100.09%, 99.98%, 99.54%, 100.14%, 99.67%, and 100.06%. The RSD were 1.9%, 2.6%, 3.7%, 2.4%, 2.1%, 1.8%, and 3.2%. Conclusion: The method is simple, accurate, and excellent repetition, and can be used to control the quality of Longsha Granules.
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Objective To establish fingerprint of analgesic ingredients in Cangbaiqutong capsules by HPLC, and control different production batches of Cangbaiqutong capsules′ quality. Methods Waters XSELECT CSH-C18 (4.6 mm×150 mm, 5.0μm) chromatographic column was used and eluted with acetonitrile -0.1% phosphoric acid solution gradiently in the flow rate of 1.0 ml/min. The detection wavelength was 284 nm with column temperature of 30℃. rrith phellodendrine chloride, gentiopicroside, paeoniflorin, tetrandrine and berberine hydrochloride as the object references, ten batches of Cangbaiqutong capsules were tested and analyzed by similarity comparison. Results fingerprint spectrum in Cangbaiqutong capsules had 20 common peaks in total, and characteristic spectra of Cortex Phellodendri, Gentiana, Radix Paeoniae Rubra and Radix Stephaniae Tetrandrae were found, while similarity of HPLC fingerprint was more than 0.9 among those batches of samples. Conclusion Using HPLC fingerprint can be used to evaluate the Cangbaiqutong capsules′quality in the round, which could improve the quality control standard in productive process of Cangbaiqutong capsules.
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Objective: To analyze the content variation of phellodendrine and berberine from Phellodendron chinense in different years, different seasons, and different parts, and to provide the basis for increasing the planting benefit of Chinese herbal medicine. Methods: High performance liquid chromatography (HPLC) analysis method was used to determine the contents. Results: The contents of phellodendrine and berberine from different parts of P. chinense showed as the following order: root barks > barks > roots. The contents of the two kinds of alkaloids changed each year. In first three years, the content change trend was obvious, and the content in barks was close to 5% in the third year. The young P. chinense had development value. After the third year, the contents of the two kinds of alkaloids changed slowly. The contents of phellodendrine and berberine in October were higher. Conclusion: The metabolic accumulation rules of berberine and phellodendrine are summarized, in order to provide the basis for the sustainable utilization of P. chinensis resources.
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Objective To establish a method for the determination of berberine hydrochloride and phellodendrine in Lishukang Capsules by HPLC. Methods Waters Symmetry C18 column (4.6 mm× 250 mm, 5μm) was used, the mobile phase was acetonitrile-0.1%phosphoric acid (0.1 g sodidum dodecyl sulfonate in 100 mL water), with the flow rate of 1 mL/min by gradient elution. The detection wavelength was set at 284 nm. Results Berberine hydrochloride showed good linear relationship (r 2=0.999 1) in the range of 403.4-10 084.7 μg, the average recovery rate was 99.8%(RSD=1.7%). Phellodendrine showed good linear relationship (r 2=0.999 2) in the range of 169.137 6-4228.44 μg, the average recovery rate was 95.5%(RSD=3.2%). Conclusion The method is specific, accurate and convenient to be used for the quality control of Lishukang Capsules.
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Objective To establish a method for determination of Berberine and Phellodendrine in Cortex Phellodendri(CP).Methods Reverse-phase HPLC method was used for determination of Berberine and Phellodendrine in different batches of CP.The chromatographic conditions were:Merck-lichrospher RP-C18 column(4.6?250 mm,5 ?m),flow rate being 0.8 mL/min,column temperature at 25 ℃,detection wavelength at 284nm for Phellodendrine and 245nm for Berberine.Results The content of Berberine was 4 times and that of Phellodendrine in Chuan CP 2～3 times as much as that of Guan CP.Conclusion The method is proved to be feasible for quality assessment of Cortex Phellodendri.Limited contents of Berberine and Phellodendrine should be laid out for the great difference in Chuan CP and Guan CP.
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Objective: To determine phellodendrine in Cortex Phellodendri by HPLC. Methods : HPLC condition consists of C 18 column (Phenomenex, 150mm, 5?), actonitrile:250mL of 0.05mol?L -1 phosphoric acid +0.4mL of diethylamine (9∶91, v/v) as mobile phase, detection wavelength at 284nm. Results : The averagy recovery of phellofendrine was 101.43% ( RSD =1.56%, n =5) and the linear range of phellodrine was 0.4368～2.1840?g??L -1 , r =0.9998. Conclusion : The method is simple, accurate and reproducible, and can be used for the determination of phellodendrine in Cordex Phellodendri.