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Table of Content

    15 June 2000, Volume 9 Issue 2
    Contents
    Contents list
    Journal of Chinese Pharmaceutical Sciences
    2000, 9(2):  1-01. 
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    Full Papers
    RAPD Differentiation of Five Medicinal Dysosma Species
    Fu Rongchao, Shaw Pangchui, Wang Jun, But Paulhay, Shi Dawen, Sun Yongru*
    2000, 9(2):  57-60. 
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    Traditional methods of authentication are not always reliable in differentiation of Dysosma species. We explored the potentiality of random amplified polymorphic DNA (RAPD) technique for the differentiation of the medicinal plants. Among the forty-two primers screened, twelve were able to generate reproducible bands that distinguished the seven samples of five Dysosma species, D. versipellis (Hance) M. Cheng from 3 localities, D.majorense (Gagnep.) Ying, D. pleiantha (Hance) Woodson, D. furfuracea S.Y Bao, and D. veitchii (Hemsl. et Wils.) Fu. Characteristic RAPD marker bands unique to each of the seven Dysosma samples were identified. RAPD polymorphism was lower among the intra-specific samples of D. versipellis, but higher in inter-specific samples. Our results demonstrate that RAPD marker technique can effectively authenticate these seven Dysosma samples.
    Authentication of Bungarus parvus and Its Adulterants by DNA Molecular Method Using Diagnostic Primer
    Wang Yiquan, Zhou Kaiya, Xu Luoshan, Xu Guojun
    2000, 9(2):  61-66. 
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    In order to develop a more applicable DNA molecular marker identification method for the authentication of Bungarus parvus and its adulterants, DNA templates were extracted from Bungarus parvus, adulterants of the crude drugs and their original. animals. Cyt b gene fragment was amplified from all these templates respectively using universal primers. PCR products were purified and sequenced directly. After sequence alignment and comparing the sequence of Bungarus parvus to its adulterants, we find that Cyt b gene is a good molecular marker for authentication of the crude drugs since interspecific differentiation is more evident than that of intraspecies in DNA sequence of the gene. Based on the sequence data of Cyt b gene fragment, a pair of highly specialized primers, used as diagnostic primers, was designed for the PCR identification of the crude drugs. Using diagnostic primers for the PCR identification, Bungarus parvus samples could be absolutely distinguished from all its adulterers when annealing temperature is at 60 ºC~65 ºC, and no incorrect or missing discrimination was found under these PCR conditions. Mixed powder of the qualified crude drug and its adulterants could also be detected by diagnostic PCR. The results indicate that diagnostic PCR using the primers designed in the present study is a simple, effective and applicable method for the authentication of Bungarus parvus, and this method might also be a new way for examining the compositions of Chinese patent medicine.
    Study on Mineral Chinese Medicine YuYuLiang
    Li Gang, Jin Tongshun, Xu Qunwei
    2000, 9(2):  67-70. 
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    Studies were made on mineral Chinese medicine YuYuLiang with X-ray powder diffraction and ICP-AES. The analysis results are given on qualitative, quantitative, crystal size and trace elements, which indicate that YuYuLiang is mainly made up of goethite, hematite, quartz and clay minerals. The clay and its content, the crystal size and trace elements related to human health were found to vary with the origin of the medicine. The results may provide basis for quality evaluation of the medicine.
    Triterpenoids from Tripterygium wilfordii
    Li Chunyu, Feng Huijin, Li Yuanchao*
    2000, 9(2):  71-72. 
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    A triterpenoid named 3β, 22α, 30-trihydroxy-urs-12-en (1) was isolated from the ethanol extract of Tripterygium wilfordii together with four known compounds. The structure of (1), which was for the first time isolated from nature, was established by spectral analysis and by comparison with that of the relative compound.
    A Novel Platelet-Activating Factor Antagonist Isolated from a Chinese Herbal Drug Viscum coloratum
    Guan Zengwei, Liu Xuehui, Liu Haixin, Cui Yuxin*
    2000, 9(2):  73-76. 
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    A novel natural platelet-activating factor (PAF) antagonist was isolated from a Chinese medicinal. herb Viscum coloratum and its structure was identified as ho moeriodictyol-7-O-β-D-glycoside, which existed in ethyl acetate extract fraction. At the final concentration of 10-5 mol·L-1, it could significantly inhibit human and rabbit platelet aggregation induced by PAF, with IC50 being 8.0×10-7 mol·L-1, but had no inhibitory effect on platelet aggregation induced by ADP.
    Synthesis and Antitumor Activity of Antineoplaston A10 Analogs
    Song Lin, Xie Yuli, Xie Yuyuan*
    2000, 9(2):  77-79. 
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    Analogs of Antineoplaston A10 were synthesized. Eight new Schiff bases were confirmed by elemental analysis and 1H NMR. Crystal of compound 7a was examined by X-ray in order to study the similarity of these molecules with Antineoplaston A10. Antitumor assay shows that they can inhibit P388 and A-549 at 10-4 mol·L-1.
    Determination of Magnolol and Honokiol in Cortex Magnoliae Officinalis by Capillary Zone Electrophoresis
    Li Yan, Long Hong, Liu Huwei*
    2000, 9(2):  80-83. 
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    Two Pharmacologically active phenols (magnolol and honokiol) isolated from the Chinese herbal drug, Cortex Magnoliae Officinalis, were separated by capillary zone electrophoresis. After a series of optimization, baseline separation of magnolol and honokiol was obtained within 12 min by using the buffer system consisting of 0.03 mol·L-1 borate and 0.01 mol·L-1 phosphate, containing 20% methanol, at pH 10.80, and by DAD detection at 294 nm. With this simple and rapid method, the contents of two phenols in Cortex Magnoliae Officinalis were determined simultaneously by external standard method.
    Determination of Adenosine and Related Substances by RP-HPLC and Studies on Their Retention Behavior
    Li Huiyi, Zhou Tieling, Wang Jing, Xu Kangsen
    2000, 9(2):  84-87. 
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    A rapid and simple reversed-phase HPLC(RP-HPLC) method for the determination of adenosine and its related substances was developed. On an ODS column (5μm, 150mm×4.6mm), adenosine and its related substances (adenine, guanosine, cytidine and uridine) were separated by using 0.01 mol·L-1 KH2PO4 (pH 3.7) -CH3OH (94:6) as mobile phase. The flow rate was 1.0 mL·min-1, p-aminobenzoic acid was used as the internal standard and UV detection was performed at 260 um. The calibration curve showed good linearity over the range of 0.01007-0.2014 μg, r = 0.9999 and the detection limit was 0.2 ng (S/N ≥ 3). The recovery was 100.l% and the precision of the method was 0.5% (n = 8).
    The method is accurate, sensitive and reproducible. It has been used for the quality control of adenosine and its preparations and also for monitoring the purification and refinement process of adenosine.
    The relationship between retention behavior of the analytes and properties of mobile phase was also studied and discussed in this paper. The results can help to predict the chromatographic retention behavior and understand the retention mechanism better.
    Microdialysis Sampling for Transdermal Delivery Study of Ondansetron HCl in Rats
    Ding Pingtian, Xu Hui, Zheng Junmin
    2000, 9(2):  88-91. 
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    The characterization of microdialysis probe was performed in vitro and in vivo. No net flux method and retrodialysis method were used to find the recovery in vitro and in vivo respectively. On the basis of the probe characterization, transdermal delivery of ondansetron HCI (ON) solution in propylene glycol (PG) with a widely used enhancer oleic acid (OA) was carried out in rats. Dialysate samples collected from the probe were analyzed by HPLC. There was no significant difference between the in vitro recovery (35.46±4.2%) and the in vivo recovery (32.53±1.8%) (P>0.1). The ON concentration in both dialysate and dermis reached a plateau at 1.5 h by coapplication of 5% OA in solution of ON in PG, while at about 3.5 h by co-application of 2% OA. And the plateau ON concentration in case of 5% OA was about two fold of that of 2% OA. Oleic acid at concentrations of 2% and 5% (w/v) increased the delivery rates from 0.001 to 0.030 and 0.058 μg·h-1 respectively. OA proved to be an effective enhancer for transdermal delivery of ON in rats.
    Study on Pharmacokinetics and Pharmacodynamics of High Dose Epirubicin in Cancer Patients
    Dong Mei, Feng Fengyi*, Fu Qiang, Zhu Zhu
    2000, 9(2):  92-95. 
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    Objective: This study was designed to characterize the pharmacokinetics and pharmacodynamics of high dose epirubicin in cancer patients. Methods: Eleven patients with malignant tumors were administered with a dose of 100 mg.m-2 epirubicin. The concentration of epirubicin was determined by high-performance liquid chromatographic (HPLC) assay. The modelling data were performed with a compartment pharmacokinetic modelling program (PCNONLIN). Hematological toxicity was used as the pharmacodynamic index. The relationships among the pharmacokinetics and pharmacodynamics and other factors affecting dose modulation were assessed. Results: The pharmacokinetics of high dose epirubicin was best described by a typical three-compartment model. It showed wide interindividual variation. There was no correlation between its pharmacokinetics and pharmacodynamics. Age was closely correlated with epirubicin clearance. Conclusions: There was no difference in the pharmacokinetic parameters between high dose and low dose. Total clearance appeared to decrease with age, which indicates the necessity of reducing dose for the elderly patients. The tolerance was good for patients receiving a dose of 100 mg.m-2 epirubicin.
    Delivery of Folio Acid Conjugated Liposomes Into Cultured HeLa Cells
    Li Hong, Liu Min, Jia Yongfeng, Xie Chengying, Lu Weiyue*
    2000, 9(2):  96-99. 
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    Effect of folio acid on delivery of liposomes was examined using cultured HeLa cells. Folateliposomes encapsulating calcein and liposomes were cultured with HeLa cells respectively. Liposomes taken up bycells were measured by a fluorescence spectrophotometer. The cell fluorescence increased slowly at the 4 h timepoint, and the saturated folate-liposome concentration was 0.45 mg·mL-1. The binding of folate-liposomes to HeLacells could be competitively inhibited by excess free folate. Enzyme phospholipase D (PLD) or phosphatidylinositolspecific phospholipase C (PIPLC) influenced liposome uptake via folate receptor. Folate targeting improved the specificity of liposomes for cells which overexpressed the folate receptors.
    Determination of Levofloxacin in Patients' Plasma and Cerebrospinal Fluid(CSF) by HPLC and its Pharmacokinetics
    Zhang Ruhong, Lei Jiachuan, Li Rongling, Luo Shunde, He Wen, Cai Hongsheng, Zhou Yan'an
    2000, 9(2):  100-103. 
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    The blood and cerebrospinal fluid concentrations of levofloxacin in patients undergoing neurosurgical operations were determined by RP-HPLC and its pharmacokinetics was studied. C18H37 column was eluted with the mobile phase of mmol·L-1 KH2PO4-10 mmol·L-1(C4H9)4NBr-CH3CN (45:45: 10 v/v, pH 3.0) and the ultraviolet absorbance was monitored at 295nm. Ciprofloxacin was used as internal standard. The mean extractionrecoveries were 74.76% in plasma and 82.43% in CSF, with the lowest detectable limits of 10 μg·L-1 and 6 μg·L-1 respectively. The intra-day and inter-day RSD were less than 5%. A single oral dose of 300 mg·LVFX tablets was taken by 10 patients undergoing neurosurgical operations. The pharmacokinetic parameters in plasma and in CSF could be described by one compartment open model. The pharmacokinetic parameters were: plasma Ke 0.125±0.042 h-1, T1/2 6.054±1.680 h, Tpeak l.049±0.288 h, Cmax 3.670±0.416 mg.L-1, AUC 33.427±7.316 mg·h·L-1, Cls 9.463±2.531 L·h-1, Vd 77.486±7.393 L, CSF Ke 0.109±0.041 h-1, T1/2 6.946±1.883 h, Tpeak 3.555±1.235 h, Cmax 1.682±0.249 mg·L-1, AUC 23.697±5.615 mg·h·L-1, Cls 13.702±5.110 L·h-1, Vd 126.606±13.198 L.

    The Kinetics of Nimodipine Release from Swellable Hydrophilic Matrix
    Zhao Ganlin, Shen Xiaobin
    2000, 9(2):  104-107. 
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    Nimodipine (NMDP) sustained release formulations were designed with hydroxypropyl methylcellulose (HPMC) as swellable polymer and with solid dispersion of the drug in polyvinyl pyrrolidone (K30), a water-soluble system, and compressed into tablets by direct compression. Drug release kinetics from tablets with different compositions was investigated in vitro. Results showed that dissolution process of NMDP from porous hydrophilic matrix conformed to zero-order release kinetics. The addinon of microcrystalline cellulose to the porous swellable release system increased the NMDP release rate constant. The presence of hydroXyethylcellulose (HEC) in the system, which had a lower viscosity, increased the drug release rate constant. When tablets were prepared with HPMC 40 mesh granulation, the release rate of the drug was sustained. These show that the dosage form may be formulated as a drug-polymer system with constant release at a desired rate. The release properties of the sustained release tablets made by pure drug instead of drug solid dispersion were also investigated. In the present study, the optimal formulations that constantly released the drug during 12 h were selected for in vivo analysis.
    Study on the Gene Transcription Level of Hippocampus NGF in C57 Mouse Development
    Liu Xiaojun, Wu Wutong*, Zhang Linyuan, Gao Xiangdong
    2000, 9(2):  108-111. 
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    β-Nerve growth factor (β NGF) mRNA levels in hippocampa from C57BL/6J mice of different ages were compared by semi-quantitative RT-PCR method, taking β-actin gene as an internal control. The levels of 6-and 12-month-old C57 mice were much lower compared with that of the l-month-old mouse. However, there was no significant difference between these two groups. This result suggests that gene transcription level of hippocampus β NGF in a C57, mouse descends evidently as it grows up. Base T at Site 294 of β NGF cDNA was substituted by C in the C57, mouse, as revealed by DNA sequencing for the first time. Nevertheless, this polymorphism of nucleotide did not make any difference in amino acid composition.