1.Determination of Paeoniflorin in Gujing Pill by HPLC
Traditional Chinese Drug Research & Clinical Pharmacology 1993;0(01):-
Objective To establish a method for the content determination of paeoniflorin in Gujing Pill. Methods Orthogonal test was used to optimize the condition of sample extraction and paeoniflorin content was detected by HPLC. HPLC was performed with Merck- LiChrospher RP- C18 column(4.6? 250 mm,5 ? m); methanol- water (25 ∶ 75) served as mobile phase and the detection wavelength was at 230 nm, the flow rate being 0.8 mL/min. Results Paeoniflorin has a good linearity within 0.27~ 1.35? g(r=0.9998) and the average recovery was 97.08 % with RSD 2.02 % (n=10). Conclusion This method is simple, sensitive, specific and can be used for the quality control of Gujing Pill.
2.Orthogonal Test for Optimization of Rhubarbs Extraction Process and Pharmacodynamic Studies the Extracts
Traditional Chinese Drug Research & Clinical Pharmacology 1993;0(03):-
Objective To optimize the solvent extraction of Rhubarbs. Methods With different concentrations of alcohol as the solvent, the amount of solvent, the extracting time and the extracting frequencies were assayed by orthogonal test to detect their influences on the extraction rate of rhubarb anthraquinone. And the extracts were analyzed and their pharmacodymamic effects were studied. Results Indicated by the content of total anthraquinones, the best conditions was: reflux extraction with 5- fold 50 % alcohol for 3 times, 0.5 h per time. The purgative activity of the extracts was stronger than that in the positive control group. Conclusion Optimization of the extracting conditions can promote the purgative activity of the extracts.
3.Study on the Effects and Mechanism of Celastrol on Proliferation and Apoptosis of Human Hepatoma HepG2 Cells in vitro
China Pharmacy 2017;28(10):1342-1345
OBJECTIVE:To study the effects of celastrol on the proliferation and apoptosis of human hepatoma HepG2 cells, and investigate its mechanism. METHODS:CCK-8 method was used to determine the cell activity 24,48,72 h after treated by 2, 5,10 μmol/L celastrol,and the proliferation inhibition rate and half inhibitory concentration(IC50)were calculated;flow cytome-try was conducted to detect the cell apoptosis rate and cycle change 24 h after treated by 2,5,10μmol/L celastrol,and the DMSO was used as negative control;rhodamine 123 staining method was used to determine the mitochondrial membrane potential 48 h af-ter treated by 2,5,10 μmol/L celastrol,and the DMSO was used as negative control;Western blot was adopted to detect the pro-apoptotic related genes Bax and B lymphoma 2(Bcl-2)protein expressions 0,12,24,36 h after treated by 5 μmol/L celastrol. RESULTS:2,5,10 μmol/L celastrol can inhibit cell proliferation,IC50 was 5.834 μmol/L. 2,5,10 μmol/L celastrol can induce apoptosis;5,10 μmol/L celastrol can block cell in G0/G1,S phases,compared with negative control group,with significant differ-ences(P<0.05 or P<0.01),and the above effects all showing certain concentration-dependent manner. 5 μmol/L celastrol can in-crease Bax protein expression and decrease Bcl-2 protein expression after cultured for 12,24,36 h,showing certain time-depen-dent manner;compared with 0 h,there was significant difference(P<0.05 or P<0.01). CONCLUSIONS:Celastrol can obvious-ly inhibit the proliferation of human hepatoma HepG2 cells and induce their apoptosis,and the mechanism may be related with strengthening mitochondrial permeability and promoting the release of apoptosis-inducing factor.
4.Fingerprint analysis of Cortex Phellodendri by RP-HPLC
Chenchen ZHU ; Jianxia MO ; Chaozhan LIN
Traditional Chinese Drug Research & Clinical Pharmacology 2000;0(05):-
Objective To establish a new method for the identification of Cortex Phellodendri. Methods A HPLC fingerprint at 220 nm has been established for identification of Cortex Phellodendri, using a Merck-Lichrospher RP-C_(18) column(4.6?250mm, 5?m) and ACN(A) with the buffer of 0.3% H_3PO_4-NH(CH_2CH_3)_2 (B) in gradient as the mobile phase. The results were compared with the chromatograms of three species of Rbizoma Coptidis under the same conditions. Results HPLC fingerprint of Cortex Phellodendri at 220nm consists of 14 specific peaks. The steady appearance of the peaks and their relative contents were considered as important signs for the identification of this crude drug. The chromatograms of Rhizoma Coptidis were obviously different from the fingerprint of Cortex Phellodendri. Conclusion This method has been proven to be feasible for the identification of Cortex Phellodendri.
5.Determination of Berberine and Phellodendrine in Cortex Phellodendri by HPLC
Chenchen ZHU ; Chaozhan LIN ; Jianxia MO
Traditional Chinese Drug Research & Clinical Pharmacology 1993;0(04):-
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.
6.Research on HPLC Fingerprint of Salt- prepared Cortex Phellodendri
Quan XIA ; Chenchen ZHU ; Suiqing MI
Traditional Chinese Drug Research & Clinical Pharmacology 1993;0(04):-
Objective To establish a HPLC fingerprint analysis method of salt-prepared Cortex Phellodendri. Methods C18 column was used, with gradient elution by the mobile phase consisted of acetonitrile-0.1 % phosphoric acid (contained 0.2 % triethylamine). The detection wavelength was at 230 nm, and the flow rate was at 0.8 mL/min. Results Seventeen characteristic peaks were selected and the fingerprints on HPLC was set up. Conclusion HPLC fingerprint method is reproducible, accurate and stable, and can be used for the quality control of salt-prepared Cortex Phellodendri.
7.Research on HPLC Fingerprint of Salt-prepared Cortex Phellodendri
Quai XIA ; Chenchen ZHU ; Suiqing MI
Traditional Chinese Drug Research & Clinical Pharmacology 2009;20(4):335-337
Objective To establish a HPLC fingerprint analysis method of salt-prepared Cortex Phellodendri. Methods C18 column was used, with gradient elution by the mobile phase consisted of acetonitrile-0. 1% phosphoric acid (contained 0. 2 % triethylamine). The detection wavelength was at 230 nm, and the flow rate was at 0. 8 mL/min. Results Seventeen char-acteristic peaks were selected and the fingerprints on HPLC was set up. Conclusion HPLC fingerprint method is repro-ducible, accurate and stable, and can be used for the quality control of salt-prepared Cortex PheUodendri.
8.Determination of Chimonin and Forsythiaside in Kangbingdu Oral Liquid by RP-HPLC
Fangmei LI ; Zhaozhan LIN ; Songfeng ZHEN ; Chenchen ZHU
Traditional Chinese Drug Research & Clinical Pharmacology 1993;0(01):-
Objective To establish a method for the determination of chimoni and forsythiaside in Kangbingdu Oral Liquid(KOL) and to develop a new quality standard for KOL. Methods A RP-HPLC method was adopted for the determination of chimonin and forsythiaside of KOL. The chromatographic conditions were as follows:Kromasil RP-C18(250 mm? 4.6 mm,5 ? m) column was used,a mixture of acetonitrile -0.1 % acetic acid(15 ∶ 85,v/v) served as mobile phase,the flow rate was 0.80 mL? min-1,and detective wavelengths of time process were 0→ 25 min:258 nm,and 25→ 30 min:277 nm. Theoretical plate number of chimoni was over 6000 and that of forsythiaside was over 5000. Results The chimonin showed a good linearity in the range of 48.8ng~ 1525 ng(r=0.999 4),and the linear range of forsythiaside was in the ranged of 56ng~ 1750 ng(r=0.999 5).The mean recovery of chimonin and forsythiaside was 97.65 % and 99.21 % respectively. The content of chimonin in the 10 batches of samples was in the range of 14.40~ 20.41 ? g? mL-1,and that of forsythiaside was in the range of 28.70~ 36.01 ? g? mL-1. Conclusion The developed method has been proven to be simple,stable and reproducible,and can be applied for quality control of the Kangbingdu Oral Liquid.
9.Quality Analysis of Herba Ephedra
Chaozhan LIN ; Chenchen ZHU ; Jinyan YANG ; Kang CHEN
Traditional Chinese Drug Research & Clinical Pharmacology 1993;0(01):-
Objective To supply evidence for quality control of Herba Ephedra by determining content limit in different batches of medicinal material of Herba Ephedra.Methods A RP-HPLC method was used for the determination of ephedrine and pseudo-ephedrine. The chromatographic conditions were: Kromasil RP-C18(250?4.6 mm,5 ?m) column, a mixture of 0.3 %phosphoric acid-methanol (10∶90) as mobile phase and 213nm as detected wavelengths.Results The contents of the above two alkaloids showed great difference among the 14 patches of tested samples. The content of ephedrine ranged 0.361 %~1.538 %and that of pseudo-ephedrine 0.332~2.087 %. The suggested content limitation of ephedrine should be 1.082 %, and that of psedoephedrine be 1.008 %in the crude Herba Ephedra.Conclusion The established method has been proven to be simple, stable and repeatable, and can be applied for quality control of the crude crude Herba Ephedra and compound prescriptions containing crude Herba Ephedra.
10.Triterpenes from Callicarpa integerrima Champ.
Chenchen ZHU ; Li GAO ; Zhongxiang ZHAO ; Chaozhan LIN
Acta Pharmaceutica Sinica 2012;47(1):77-83
A new triterpenoid saponin and fourteen known triterpenoids were isolated from the methanol extract of the stems and leaves of Callicarpa integerrima Champ, which is used in Chinese folk medicine for stopping bleeding, expelling the wind, dissipating stagnation, and treating scrofula, by using various chromatographies, such as silica gel, Sephadex LH-20 and RP-C18 column chromatography. Their structures were identified as a new compound 2alpha, 3beta, 19alpha, 23-tetrahydroxy-olean-12-en-28-oic acid-28-O-beta-D-glucopyranosyl-(1 --> 4)-beta-D-glucopyranoside (1), together with fourteen known compounds: oleanolic acid (2), 3-acetyl oleanolic acid (3), 3beta-O-acetyl ursolic acid (4), 2alpha-hydroxy-ursolic acid (5), 2alpha, 3beta, 19alpha, 23-tetrahydroxy-urs-12-en-28-oic acid (6), alpha-amyrin-3-O-beta-D-glucopyranoside (7), pomolic acid (8), betulinic acid (9), ursolic acid (10), 2alpha, 3beta, 19alpha, 23-tetrahydroxy-olean-12-en-28-oic acid (arjungenin) (11), 2alpha-hydroxy-oleanolic acid (12), hederagenin (13), 2alpha, 19alpha-dihydroxy-ursolic acid (14) and pruvuloside A (15), by the spectroscopic techniques of NMR, HMBC, IR and MS, separately. All these compounds were obtained from this plant for the first time, and compounds 3, 4 and 15 were isolated from genus Callicarpa L. for the first time.