Pharmacokinetic research strategies of compatibilities and synergistic effects of classical Danshen herb pairs based on pharmacokinetics of "Danshen-Bingpian" and "Danshen-Honghua" / 中国中药杂志

Herb pairs are usual clinical compatibility forms and one of compound prescription sources in Chinese medicine. Pharmacokinetic research in vivo is one of the important items in elucidating the mechanism for synergistic and attenuated mechanisms of herb pairs. The paper comprehensively summarized and systemized the pharmacokinetic researches of marker-ingredients about Danshen-Honghua and Danshen-Bingpian in order to elucidate the rationality and scientificity of herb pairs and provide some feasible suggestions on the pharmacokinetics of drugs in the future. In view of complicated system of Traditional Chinese medicines and a chemical system that is not separated from its natural state, comparative pharmacokinetic researches on marker-ingredients from the herb pairs are reasonable to elucidate the synergistic and attenuated mechanisms of monarch-subjects compatible herbs and monarch-guide compatible herbs. Such pharmacokinetic research can better explain the mechanism of drug compatibility, while the pharmacokinetic researches based on the monomer chemical compositions and marker-ingredients that have been separated from complex chemical environment of traditional Chinese Medicine are still unreasonable and should be discussed deeply.

Analysis of aristolochic acids, aristololactams and their analogues using liquid chromatography tandem mass spectrometry / 中国天然药物

More than 80 aristolochic acids (AAs) and aristololactams (ALs) have been found in plants of the Aristolochiaceae family, but relatively few have been fully studied. The present study aimed at developing and validating a liquid chromatography tandem mass spectrometry (LC/MS(n)) for the analysis of these compounds. We characterized the fragmentation behaviors of 31 AAs, ALs, and their analogues via high performance liquid chromatography coupled with electrospray ionization mass spectrometry. We summarized their fragmentation rules and used these rules to identify the constituents contained in Aristolochia contorta, Ar. debilis, Ar. manshurensis, Ar. fangchi, Ar. cinnabarina, and Ar. mollissima. The AAs and ALs showed very different MS behaviors. In MS(1) of AAs, the characteristic pseudomolecular ions were [M + NH4](+), [M + H](+), and [M + H - H2O](+). However, only [M + H](+) was found in the MS(1) of ALs, which was simpler than that of AAs. Distinct MS(n)fragmentation patterns were found for AAs and ALs, showing the same skeleton among the different substituent groups. The distribution of the 31 constituents in the 6 species of Aristolochia genus was reported for the first time. 25 Analogues of AAs and ALs were detected in this genus. A hierarchical schemes and a calculating formula of the molecular formula of these nitrophenanthrene carboxylic acids and their lactams were proposed. In conclusion, this method could be applied to identification of similar unknown constituents in other plants.

Study on compatibility of Salviae Miltiorrhizae Radix et Rhizoma and Chuanxiong Rhizoma based on pharmacokinetics of effective components salvianolic acid B and ferulic acid in rat plasma / 中国中药杂志

A study was made on the pharmacokinetic regularity of effective components salvianolic acid B and ferulic acid in Salviae Miltiorrhizae Radix et Rhizoma (SMRR) and Chuanxiong Rhizoma(CR) in rats, so as to discuss the compatibility mechanism of Salviae Miltiorrhizae Radix et Rhizoma and Chuanxiong Rhizoma. Rats were randomly divided into three groups and intravenously injected with 50 mg x kg(-1) salvianolic acid B for the single SMRR extracts group, 0.5 mg x kg(-1) ferulic acid for the single CR extracts group and 50 mg x kg(-1) salvianolic acid B + 0.5 mg x kg(-1) ferulic acid for the SMRR and CR combination group. The blood samples were collected at different time points and purified by liquid-liquid extraction with ethyl acetate. With chloramphenicol as internal standard (IS), UPLC was adopted to determine concentrations of salvianolic acid B and ferulic acid. The pharmacokinetic parameters of salvianolic acid B and ferulic acid were calculated with WinNonlin 6.2 software and analyzed by SPSS 19.0 statistical software. The UPLC analysis method was adopted to determine salvianolic acid B and ferulic acid in rat plasma, including linear equation, stability, repeatability, precision and recovery. The established sample processing and analysis methods were stable and reliable, with significant differences in major pharmacokinetic parameters, e.g., area under the curve (AUC), mean residence time (MRT) and terminal half-life (t(1/2)). According to the experimental results, the combined application of SMRR and CR can significantly impact the pharmacokinetic process of their effective components in rats and promote the wide distribution, shorten the action time and prolong the in vivo action time of salvianolic acid B and increase the blood drug concentration and accelerate the clearance of ferulic acid in vivo.

HPLC-UV-ELSD characteristic figure and chemical pattern recognition of Panacis Quinquefolii Radix / 药学学报

The paper is to report the establishment of a method of characteristic figure analysis for the quality control of Panacis Quinquefolii Radix. Application of HPLC-UV-ELSD techniques was connected in series and applied. The separation was carried out on the Agilent Extend-C18 (250 mm x 4.6 mm, 5 microm) column. The mobile phase consisted of water and acetonitrile with gradient elution. The flow rate was 1.0 mL x min(-1) and the wavelength of measurement was 203 nm. The temperature of drift tube was maintained at 106.5 degrees C and the flow rate of air was set at 2.9 L x min(-1). Twenty batches of the Panacis Quinquefolii Radix were determined. Hierarchical cluster analysis (HCA) and principal component analysis (PCA) were applied to study on the HPLC characteristic figure and chemical pattern recognition. The HPLC-UV and HPLC-ELSD characteristic figure of Panacis Quinquefolii Radix was developed, the ginsenosides Rg1, Re, Rb1, Rc, Rb2, Rb3, Rd and the pseudoginsenoside F11 were identified. This method is accurate and reliable, and it can be used to control the quality of the Panacis Quinquefolii Radix.

UPLC fingerprint for quality assessment of ginsenosides of ginseng radix et rhizoma / 药学学报

This paper is aimed to establish the method of fingerprint analysis of chemical constituents by reversed-phase ultra-performance liquid chromatography (UPLC) for the quality control of the roots and rhizomes of Panax ginseng (Ginseng Radix et Rhizoma). The method was performed on a ACQUITY UPLC BEH C18 (50 mm x 2.1 mm ID, 1.7 microm) with a mixed mobile phase of water and acetonitrile in a gradient mode. The flow rate was 0.3 mL x min(-1) and the wavelength of measurement was 203 nm. Eleven batches of the Ginseng Radix et Rhizoma were determined. The UPLC chromatographic fingerprints of chemical constituents were established from the eleven batches of the Ginseng Radix et Rhizoma and showed fifteen characteristic common peaks, among which fifteen peaks were recognized and nine compounds (ginsenosides Rg1, Re, Rf, Rg2, Rb1, Rc, Rb2, Rb3 and Rd) were determined by comparison with chromatographic behaviors and UV spectra of the authentic compounds. The eleven batches of samples were classified as two clusters by hierarchical clustering analysis (HCA) and principle component analysis (PCA), and six samples were confirmed to establish the mutual model. The quality was assessed by similarity evaluation system for chromatographic fingerprint of TCM (2004B Version). The convenient and high specific method can be used to identify and evaluate the quality of the Ginseng Radix et Rhizoma.

Affection of different preparations on the content of toxic constituents in traditional Chinese medicines, examplified with Caulis Aristolochiae Manshuriensis / 中国中药杂志

<p><b>OBJECTIVE</b>Taking Caulis Aristolochiae Manshuriensis (Guanmutong in Chinese, derived from the stem of Aristolochia manshuriensis) as an example, to study the affection of different preparations on the content of toxic constituents in traditional Chinese medicines.</p><p><b>METHOD</b>The separation was performed on a zorbax SB-C18 column with mobile phase of acetonitrile-3.7 mmol x L(-1) phosphoric acid buffer, detected at 260 nm.</p><p><b>RESULT</b>The extraction percentage of aristolochic acids I, II and IV a in water extraction (1 h x 2) of Guanmutong were 53.4%, 75.5% and 61.9%, respectively; the remaining quantity of aristolochic acids I, II and IVa in the dregs of the decoction were 22.3%, 15.7% and 30.3%, respectively; Aristolochic acid I was still main substance among these aristolohic acids in the decoction of Guanmutong.</p><p><b>CONCLUSION</b>The content of toxic constituents of the traditional Chinese medicines varies evidently with different preparations of Guanmutong. So the preparation methods of traditional Chinese medicines should be suitably selected according to characteristics of the toxic constituents so as to lessen the body damages of human.</p>

Injury in renal proximal tubular epithelial cells induced by aristololactam I / 中国中药杂志

<p><b>OBJECTIVE</b>To study whether aristololactam I (AL-I) induces injury in human renal proximal tubular epithelial cells.</p><p><b>METHOD</b>Cultured human renal proximal tubular epithelial cell line HK-2 was used as the subject. Aristolochic Acid I (AA-I) was used as a positive control. Cell toxicity of AL-I was detected by LDH releasing rate. Cell apoptosis was evaluated by cellular morphology, DNA content and expression of cell membrane phosphatidylserine (PS). The secretion level of fibronectin (FN) and TGF-beta1 in HK-2 cells were assayed by ELISA.</p><p><b>RESULT</b>AL-I had a direct toxicity on HK-2 in a dose dependent manner from 2.5 mg x mL(-1) to 20 mg x mL(-1); In these range of concentration, AL-I could induce cell apoptosis which was detectable by measurements of morphology, DNA content and expression of PS. AL-I could stimulate the secretion of FN and TGF-beta1. The potency of AL-I cell toxicity was higher than AA-I at the same concentration. The effects of AL-I on apoptosis, secretion of FN and TGF-beta1 were all weaker than AA-I.</p><p><b>CONCLUSION</b>AL-I as one metabolite of AA-I in vivo induces direct injury in renal proximal tubular cells. Its effects are similar to those of AA-I. AL-I may be one of toxic metabolites in Chinese herbs containing AA which participate in renal damage and fibrosis.</p>