[Overview of metalomics research and its application prospect in mineral Chinese medicine research].

Metallomics is a frontier interdisciplinary subject at its vigorous development stage. Its goal is to systematically study the content, distribution, chemical species, structural characteristics and functions of metal elements in biological system. It is also a comprehensive discipline to study the existing state and function of free or complex metal elements in life. Metallomics is an ideal tool to study the biological behavior of inorganic elements, which can be used to solve many problems in the research of mineral Chinese medicine(MCM). It provides a strong theoretical basis and technical support for the research of MCM. Its theory and methods provide re-ference and enlightenment for the in-depth study of MCM, and also provide new ideas and open up new ways for the research of MCM. The application of metallomics theory and methods in the research of MCM is of great significance to reveal the material basis and mec-hanism of MCM, promote the process of basic research on MCM, fully exploit and utilize medicinal mineral resources and carry forward the traditional MCM treasure in China. In this paper, we introduced the concept, academic development, research content and research methods of metallomics, and discussed the application prospects of metallomics in the analysis of inorganic element composition characteristics and quality control, material basis and mechanism of MCM, so as to provide reference for further researches on MCM.

[Mineral composition analysis and hemostatic effect of limonitum with different mineral genesis].

Models were established in mice with warfarin sodium method, and their bleeding time and hemostasis time were measured by tail cutting method and slide method respectively. Rats were administered for 15 consecutive days to measure their recalcification time, plasma viscosity, platelet adhesion rate, platelet aggregation rate and other blood indexes. As compared with the blank group, the bleeding time was prolonged in model groupn(P<0.05). As compared with the model group, the results showed that the positive vitamin K, the leaching type water decoction and the sediment type decoction could significantly shorten the bleeding time (P<0.01); positive vitamin K significantly (P<0.01) shortened clotting time, and the leaching type water decoction, the sediment type water decoction and the sediment type powder could also shorten the clotting time (P<0.05). As compared with blank group, low dose, medium dose of leaching type water decoction, medium dose of powder, high dose of sediment type decoction and low dose of drug residues could reduce plasma viscosity (P<0.05), and high dose of leaching powder and low dose of water decoction could significantly reduce (P<0.01) plasma viscosity. As compared with blank group, Limonitum leaching type decoction high dose group could significantly reduce the platelet adhesion rate (P<0.05), while sediment type water decoction could significantly increase the platelet adhesion rate (P<0.05); the high dose of leaching type water decoction, high dose of drug residues, low dose of leaching type powder and low dose of drug residues could decrease the platelet aggregation rate (P<0.05), while high dose of leaching type water decoction and high dose of the powder could increase the platelet aggregation rate (P<0.05). Analysis of mineral compositions was conducted by polarized light microscopy and X-ray diffraction (XRD). The results of the both methods showed that Limonitum mineral compositions contained goethite, quartz, and kaolinite, and sedimentary type also contained illite and albite. Sediment type of Limonitum showed better hemostatic effect, which may be related to the high content of goethite and illite.

[Application of Thermogravimetric-Differential Scanning Calorimetry(TG-DSC)in Quality Control of Limonitum].

Objective: To control the quality of Limonitum by investigating the thermoanalysis curves. Methods: Analysis Limonitum samples from different origins by Thermogravimetric-Differential Scanning Calorimetry( TG-DSC),and the processed samples and fake samples were analyzed to compare the difference of them at the same time. Results: Thermal analysis curves showed that most of Limonitum samples had three weight loss steps in 30 ~ 1 000 ℃,and the process of dehydration weight loss of goethite was obviously in about309 ℃. There was a positive correlation between the weight loss rate of the second step and the content of iron. Conclusion: The Thermal analysis method can provide reference to the identification and quality control of Limonitum.

[Intervention Study of Mineral Chinese Medicine Chloriti Lapis in PTZ-kindled Epileptic Rat].

Objective: To study the intervention effect of Chloriti Lapis in PTZ-kindled epileptic rat. Methods: Rats were kindled by pentylenetetrazol( PTZ),and successful kindled model were administered with drugs, then taken out the hippocampus of the brain. HE staining method was used to observe lesion in hippocampus, immunohistochemical method was used to test protein expression of nNOS, xanthine oxidase method was used to measure the activity of T-SOD, thiobarbituric acid method was used to measure the content of MDA,and phosphorus determination method was used to detect the activities of Na+,K+-ATPase and Ca2 +,Mg2 +-ATPase. Results: Each group of Chloriti Lapis( powder group, dregs group and decoction group) decreased the lesion grade, MDA content,nNOS protein expression, while increased the T-SOD activities, Na+,K+-ATPase and Ca2 +,Mg2 +-ATPase activities in the hippocampu of rats. Conclusion: Chloriti Lapis have antiepileptic effects, the mechanism may be related to increasing brain antioxidant activities, eliminating free radicals, protecting membrane function, maintaining dynamic balance of ion concentration in the braiofn rat, inhibiting brain abnormalities discharge, and ultimately achieve the goal of epilepsy treatment.

Application of Fourier Transform Infrared Spectra (FTIR) Fingerprint in the Quality Control of Mineral Chinese Medicine Limonitum.

In the present paper, the fingerprint of Limonitum (a mineral Chinese medicine) by FTIR was established, and the spectrograms among crude samples, processed one and the adulterant sample were compared. Eighteen batches of Limonitum samples from different production areas were analyzed and the angle cosine value of transmittance (%) of common peaks was calculated to get the similarity of the FTIR fingerprints. The result showed that the similarities and the coefficients of the samples were all more than 0.90. The processed samples revealed significant differences compared with the crude one. This study analyzed the composition characteristics of Limonitum in FTIR fingerprint, and it was simple and fast to distinguish the crude, processed and the counterfeit samples. The FTIR fingerprints provide a new method for evaluating the quality of Limonitum.

Simultaneous determination of eleven major flavonoids in the pollen of Typha angustifolia by HPLC-PDA-MS.

INTRODUCTION: The pollen of Typha angustifolia L. has been used for the treatment of dysmenorrhea, stranguria and metrorrhagia. Flavonoids are major active compounds in this pollen and their quantification is important for its quality control. OBJECTIVE: To establish an HPLC-PDA-MS method for simultaneous determination of the 11 majority flavonoids in the pollen of T. angustifolia. METHODOLOGY: The optimal condition of separation was achieved on a reversed-phase C18 column with gradient elution of acetonitrile and 0.05% formic acid (v/v) at a flow-rate of 0.8 mL/min; the column temperature was set at 35 °C. RESULTS: All calibration curves showed good linear regression (r² > 0.9992). The method provided good accuracy, precision, recovery and sensitivity for the quantification of the 11 compounds analysed. CONCLUSION: The HPLC method established is appropriate for the quality assurance of the pollen of T. angustifolia.

[Study on water-soluble iron, heavy metals and harmful elements of Magnetitum].

OBJECTIVE: To measure the contents of the water-soluble iron, five heavy metals and harmful elements in Magnetiturn and provide a basis for the quality control and safety evaluation of Magnetitum. METHOD: Iron (Fe), lead (Pb), cadmium (Cd) and copper (Cu) were determined by atomic absorption spectrometry (AAS); arsenic (As) and mercury (Hg) were determined by atomic fluorescence spectrometry (AFS). RESULT: The mean content of element iron is 764.30 mg x kg(-1). The contents of five water-soluble heavy metals and harmful elements in Magnetitum were within the safety range. The recovery of the standard addition was in the range of 93.7% - 110.6%, and the RSD was less than 5.0%. CONCLUSION: Analyzing the water-soluble iron, heavy metals and harmful elements in Magnetitum is effective to the quality control and the safety evaluation of magnetitum.

[Study on the fingerprints of magnetitum by FTIR].

Thirteen different magnetitum samples were analyzed by FTIR, and the FTIR fingerprint was set up with them. Processed samples and the crudes were compared with the fingerprint. It was found that all the similarities of the samples are more than 0.97. The similarities and the correlation coefficients of the processed samples are decreased. The FTIR fingerprint could be used for evaluating the quality of magnetitum for commodities.

[Analysis on competitive interaction between arenobufagin and verapamil hydrochloride with serum albumin].

OBJECTIVE: The present study was performed to investigate competitive interaction between arenobufagin and verapamil hydrochloride with serum albumin. METHODS: Equilibrium dialysis and high-performance liquid chromatography were used to analyze the binding rates of the two medicines to serum protein. The interactions based on bovine serum albumin (BSA) and human serum albumin (HSA) were investigated by using spectrofluorimetry. The interaction mode of arenobufagin and verapamil hydrochloride binding to serum proteins was simulated by molecular docking. RESULTS: The rate of arenobufagin (0.1µg/mL) binding to bovine serum was (61.1±0.2)%. Verapamil hydrochloride (0.025 to 0.1µg/mL) significantly reduced the bovine serum binding rate of arenobufagin, from (60.2±3.7)% to (36.9±3.4)%. However, arenobufagin at the tested doses had no marked effects on the binding rate of verapamil hydrochloride. Furthermore, the verapamil hydrochloride had an active effect on the arenobufagin-induced fluorescence quenching of BSA and HSA. The molecular docking results showed that verapamil hydrochloride and arenobufagin binded to HSA at site I. Molecular interaction energy of verapamil hydrochloride binding to site I was stronger than that of arenobufagin. CONCLUSION: Verapamil hydrochloride reduces the binding of arenobufagin to bovine serum. The mechanism may be a competitive interaction of arenobufagin and verapamil hydrochloride at site I on HSA.

[X-ray diffraction Fourier fingerprint of mineral Chinese medicine Chloriti Lapis].

The technology of powder X-ray diffraction (XRD) was used for analysis Chloriti Lapis and the XRD Fourier fingerprints were established. The dates were analyzed by fuzzy cluster and fingerprint similarity evaluation software to compare the similarity of samples. XRD fingerprint with 10 common peaks of 14 batches of Chloriti Lapis were established. The average, median coefficients of crystal lattice spacing d (A), peak position 2 theta, relative intensity value I/I0 (%) were all more than 0.95. And similarity( angle cosine value) were all more than 0. 97. There were small number samples differed from others. And obvious differences between the pre-and post-processing samples. This paper shows the powder XRD Fourier fingerprint can be used for appraisal and study of the Chloriti Lapis.

[Bio-activity evaluation of Qinlian Siwu decoction on inhibiting mice uterine contraction in vitro and its components analysis].

OBJECTIVE: To evaluate the bio-activity of Qinlian Siwu decoction on in vitro uterus contraction model and exploit the relationship between chemical components and the bio-activity. METHOD: The samples were prepared by macroporous adsorptive resins. The in vitro uterus contraction model was adopted to appraise the bio-activities of Qinlian Siwu decoction and its different separated fractions. HPLC-DAD- ESI -MS method was applied to analyze and identify the components in the fraction QL-3. RESULT: It was found that five active fractions (QL-1, QL-3, QL-5, QL-7 and QL-11) were separated from Qinlian Siwu decoction, mainly contributed to the observed antagonismto the contraction of the mouse uterus. 28 compounds in the fraction QL-3 were identified as malic acid, gallic acid, catalpol, protocatechuic acid, aucubin, chuanxiongzine hydrochloridum, vanillic acid, caffeic acid, paeoniflorin, berberastine, albiflorin, tetrahydropalmatine, coptisine, jatrorrhizine, leonuride, worenine, ferulic acid, palmatine, berberine, scutellarin, baicalin-7-0-glucoside, baicalin, rehmannioside C, wogonoside, chrysin-7-glucuronide, ttetuin, baicalein, wogonin and oroxylin-A. CONCLUSION: In vitro inhibiting the contraction of the isolated mouse uterine of Qinlian Siwu decoction was mainly attributed to the fraction QL-1 and QL-3. The active fractions (QL-5, QL-7 and QL-11) were obtained from QL-3 on the macroporous adsorptive resins by the gradient elution using ethanol.

Two new cerebrosides from the pollen of Typha angustifolia.

Two new cerebrosides, 1-O-(beta-d-glucopyranosyloxy)-(2S,3S,4R,8Z)-2-[(2'R)-2'-hydroxytricosanoylamino]-8-nonadecene-3,4-diol (1) and 1-O-(beta-D-glucopyranosyloxy)-(2S,3R,4E,8Z)-2-[(2'R)-2'-hydroxynonadecanoylamino]-4,13-nonadecene-3-diol (2), were isolated from the pollen of Typha angustifolia. Their structures were elucidated by chemical and spectral means. This is the first report on the occurrence of cerebroside in Typha (Typhaceae). Compounds 1 and 2 exhibited effect on the proliferation of cultured vascular smooth muscle cell (VSMCs) induced by fatal bovine serum (FBS).

Separation, characterization and dose-effect relationship of the PPARgamma-activating bio-active constituents in the Chinese herb formulation 'San-Ao decoction'.

San-ao decoction (SAD), comprising Herba Ephedrae, Radix et Rhizoma Glycyrrhizae and Seneb Armeniacae Amarum, is one of the most popular traditional Chinese medicine (TCM) formulae for asthma. Peroxisome proliferator-activated receptors (PPARs) areey regulators of lipid and glucose metabolism and have become important therapeutic targets for various deseases, PPARgamma activation might exhibit anti-inflammatory properties in different chronic inflammatory processes. The EtOAc fraction of SAD showed a significant effect on PPARgamma activation. A simple and rapid method has been established for separation and characterization of the main compounds in the PPARgamma-activating fraction of SAD by ultra-fast HPLC coupled with quadropole time-of-flight mass pectrometry (UPLC-Q-TOF/MS). A total of 10 compounds were identified in the activating fraction of SAD, including amygdalin (1), liquiritin (2), 6'-acetyliquiritin (3), liquiritigenin (4), isoliquiritigenin (5), formononetin (6), licoisoflavanone (7), glycycoumarin (8), glycyrol (9) and uercetin (10). The results also characterized formononetin as a predominant component in this fraction. The dose-effect relationship comparison study of formononetin and the EtOAc fraction of SAD by adding formononetin was performed, the results suggested that formononetin was the major component of the EtOAc fraction of SAD responsible for activating PPARgamma, and the method will possibly be applied to study the complex biological active constituents of other TCMs.

[Comparative analysis of volatile oils of Wuao decoction and its major constituing herbs by GC-MS].

OBJECTIVE: To evaluate the correlativity between volatile components of Wuao decoction and its major constituting herbs. METHOD: The chemical compositions of essential oil, obtained by hydrodistillation from Wu-ao Decoction and its major constituting herbs (Herba Ephedrae, Semen Armeniacae Amarum, Radix Platycodi, Herba Schizonepetae), were analyzed by GC-MS. RESULT: The volatile components of Wu-ao Decoction were mostly derived from Herba Ephedrae, Radix Platycodi and Herba Sehizonepetae. CONCLUSION: The method of GC-MS can be used to investigate the volatile component changes in traditional Chinese medicine formulae.

[Study on the SFE condition for curcumin in Curcuma longa].

OBJECTIVE: To optimize the conditions of supercritical fluid extraction (SFE) for curcumin in Curcuma longa. METHOD: Optimum extraction conditions were studied by orthogonal tests. The extracts were analyzed by HPLC. RESULT: The optimal extraction conditions were pressure 25 MPa, temperature 55 degrees C, static time 4 h, dynamic time 5 h, flow rate of CO2 3.5 L x min(-1), co-solvent ethanol 30% (mL x g(-1)). CONCLUSION: It is feasible to extract curcumin by SFE.