Identification of the absorptive constituents and their metabolites in vivo of Puerariae Lobatae Radix decoction orally administered in WZS-miniature pigs by HPLC-ESI-Q-TOFMS.

In this study, the technique of high-performance liquid chromatography coupled with electrospray ionization quadrupole time-of-flight mass spectrometry (HPLC-ESI-Q-TOFMS) was used to analyze and identify the absorptive constituents and their metabolites in drug-containing urine of Wuzhishan (WZS)-miniature pigs administered with Puerariae Lobatae Radix (PLR) decoction. With the accurate mass measurements (<5 ppm) and effective MS(2) fragment ions, 96 compounds, including eight original constituents and 88 metabolites, were identified from the drug-containing urine. Among these, 64 metabolites were new ones and their structures can be categorized into five types: isoflavones, puerols, O-desmethylangolensins, equols and isoflavanones. In particular, puerol-type constituents in PLR were first proved to be absorptive in vivo. Meanwhile, the metabolic pathways of PLR in vivo were investigated. On the basis of relative content of the identified compounds, 13 major metabolites accounting for approximately 50% of the contents, as well as their corresponding 12 prototype compounds, were determined as the major original absorptive constituents and metabolites of PLR in vivo. The HPLC-ESI-Q-TOFMS technique proved to be powerful for characterizing the chemical constituents from the complicated traditional Chinese medicine matrices in this research.

[Determination of dehydroabietic acid and abietic acid in aqueous alkali extract of Liquidambaris Resina by HPLC].

OBJECTIVE: To develop an HPLC method for content determination of dehydroabietic acid and abietic acid in aqueous alkali extract of Liquidambaris Resina. METHOD: The determination was carried out on a DIONEX C18 column (4.6 mm x 250 mm, 5 microm) eluted with acetonitrile and water containing 0.1% acetic acid. The flow rate was 1 mL x min(-1), and the detected wavelength was set at 210, 240 nm. RESULT: The peak areas and the sample quantity of the two components had good linear relationship in the range of 0.4-3.4 microg for dehydroabietic acid, and 0.6-4.8 microg for abietic acid. The average recoveries were 99.53%, 101.9%, respectively. CONCLUSION: The method was proved to be simple, accurate and used for the quality evaluation of Liquidambaris Resina.

[Effect of aloe polysaccharides pretreatment on the cerebral inflammatory response and lipid peroxidation in severe hemorrhagic shock rats first entering high altitude].

OBJECTIVE: To investigate the effect of aloe polysaccharides pretreatment on the cerebral inflammatory response and lipid peroxidation in severe hemorrhagic shock rats first entering high altitude. METHODS: Forty healthy male SD rats weighing 250-300 g were randomly divided into 5 groups (n = 8 each): sham group, shock group, AP group was further divided into 3 subgroups (AP1 0.75 mg/kg; AP2 1.50 mg/kg; AP3 3.00 mg/kg). The different doses AP were given iv respectively at 30 min before hemorrhagic shock. The mean blood pressure (MAP) was maintained at (35 ± 5) mmHg (1 mmHg = 0.133 kPa) for 60 minutes. The animals were killed at 2 hours after resuscitation. Blood samples were obtained from femoral artery for detecting tumor necrosis factor α (TNF-α), IL-6 and IL-10 concentrations; the frontal and parietal lobes brain and the hippocampus were separated from brain tissues on the ice for detecting superoxide dismutase (SOD) activity and myeloperoxidase (MPO) activity, malondialdehyde (MDA) concentration, brain Wet-dry weight ratio (W/D). RESULTS: Compared with sham group, hemorrhagic shock significantly increased serum TNF-α ((76 ± 11) ng/L), IL-6 ((1303 ± 141) ng/L) and IL-10 concentrations ((95 ± 14) ng/L), MPO activity ((20.72 ± 2.28)×10(-2) U/g) and MDA concentration ((80 ± 13) nmol/mgprot) in the brain tissue and brain W/D (6.21 ± 0.18) (t = 6.928 - 14.565, P < 0.05), while SOD activity ((56 ± 11) U/mgprot) decreased significantly (t = -5.374, P < 0.05). There were no significant difference between shock and AP1 groups. AP2 group significantly inhibited hemorrhagic shock-induced increase serum TNF-α ((54 ± 12) ng/L), IL-6 ((846 ± 78) ng/L) and IL-10 concentrations ((66 ± 11) ng/L), MPO activity ((13.13 ± 1.23)×10(-2) U/g) and MDA concentration ((56 ± 9) nmol/mgprot) in the brain tissue and brain W/D (5.71 ± 0.18) (t = -6.905 - -3.357, P < 0.05), while SOD activity ((86 ± 12) U/mgprot) increased significantly compared to shock group (t = 4.240, P < 0.05). There were no significant difference between AP2 and AP3 groups. CONCLUSION: AP pretreatment can attenuate the cerebral ischemia and reperfusion injury in severe traumatic-hemorrhagic rats first entering high altitude through inhibiting systemic inflammatory response and leukocyte aggregation and lipid peroxidation in the brain.

[Analysis of plasma metabonomics of mini-swines with qi deficiency and blood stasis syndrome due to chronic myocardial ischemia].

OBJECTIVE: To study the changes of plasma metabolites of mini-swines with qi deficiency and blood stasis syndrome due to chronic myocardial ischemia and to explore the relationship between plasma metabonomics and qi deficiency and blood stasis syndrome. METHODS: Twenty-two mini-swines were used in this study and were divided into sham-operation group (n=10) and model group (n=12). Ameroid constrictors were placed around the left anterior descending coronary artery of mini-swines in model group to induce chronic myocardial ischemia. Twelve weeks after Ameroid placement, physical signs, coronary angiography and echocardiography were used to evaluate qi deficiency and blood stasis syndrome; gas chromatography-mass spectrometry (GC-MS) and data mining method were used to analyze plasma metabolites of the mini-swines. RESULTS: Twelve weeks after operation, mini-swines in model group were confirmed with qi deficiency and blood stasis syndrome. Principal component analysis (PCA) found that plasma GC-MS spectra of the two groups were significantly different. Compared with sham-operation group, endogenous metabolites in plasma were changed in model group. The levels of lipid metabolites such as glycerol, acetic acid and tricosadiynoic acid, myo-inositol, as well as amino acid metabolite proline were raised, while concentrations of glucose (eg. d-glucose, and a-D-glucoside), amino acid (eg. alanine, phenylalanine, and urea) and lipid metabolites (eg. hexadecanoic acid, propionic acid, butanoic acid, and octadecanoic acid) were declined. CONCLUSION: Significant differences in metabolic spectrum exist in mini-swines with qi deficiency and blood stasis syndrome due to chronic myocardial ischemia and normal mini-swines. Disorders in glucose, amino acid and lipid metabolism result in a deterioration of coronary artery disease; citric acid, glucose, octadecanoic acid, hexadecanoic acid, glycerol and acetic acid are contributory to separation of the healthy from qi deficiency and blood stasis syndrome. These metabolites pattern may be used in clinical diagnosis and treatment of coronary artery disease, and also can provide a new approach to objective research in traditional Chinese medicine syndrome.

[Effect of fluid resuscitation for hemorrhagic shock on oxygenation of subjects in early period of first visit of high altitude area, an experimentation on dogs].

OBJECTIVE: To investigate the effect of fluid resuscitation on oxygenation of subjects with hemorrhagic shock in early period of first visit to area of high altitude, an experiment in dogs was performed. METHODS: A model of serious hemorrhagic shock was reproduced by exsanguination resulting in a lowering of mean arterial pressure to (35+/-5) mm Hg (1 mm Hg=0.133 kPa) maintain for 1 hour. Thirteen mongrel dogs were carried to an area of 3,780 metres above sea level from an area of 1,510 metres, and they were randomly divided into three groups, namely lactated Ringer's (LR) group, 6% hydroxyethyl starch (HES) group, and control group. The dogs in LR group were infused intravenously LR in 1.5 times the volume of blood loss; those in 6% HES group were given HES in equal volume. No fluid infusion was given in the control group. After 1 hour of resuscitation, LR was intravenously given at 5 ml.kg(-1).h(-1) in all groups as maintenance dose. The changes in oxygenation were observed. RESULTS: All animals in control group were dead after 2 hours. One hour after establishment of shock, the oxygen consumption (VO(2)), oxygen delivery (DO(2)), oxygen extraction ratio (O(2)ER), arterial oxygen saturation (SaO(2)) in two resuscitation groups were significantly lower than those before shock, but venous oxygen saturation (Sv(2)) and alveolar-arterial oxygen difference (A-aDO(2)) were significantly higher (all P<0.05). In LR group, the oxygenation parameters including VO(2), DO(2), O(2)ER, SaO(2) after 2 hours of resuscitation were significantly higher than those 1 hour after shock, while A-aDO(2) was significantly lower (all P<0.05); and in HES group, VO(2), DO(2), O(2)ER were significantly higher than those 1 hour after shock, while SvO(2) was significantly lower (all P<0.05). CONCLUSION: All dogs with serious hemorrhagic shock would die of hemorrhagic shock in the early period of entering a high altitude area if fluid resuscitation is denied. Two hours after infusion of LR in 1.5 times of quantity of blood loss, oxygenation can be restored to expected normal values. Infusion of 6% HES with an equal volume of blood loss, oxygenation dose not reach expected level 2 hours after resuscitation.

[Determination of lead, cadmium, mercury, chromium and arsenic in acrylonitrile-butadiene-styrene copolymer using microwave digestion-ICP-MS].

A method was studied for the analysis of Cr, As, Cd, Hg and Pb in acrylonitrile-butadiene-styrene copolymer by using ICP-MS. The instrument parameters were optimized and the introduction system was developed systematically. The sample is decomposed by microwave digestion. The digestion condition was optimized concerning digestion system, proportion of acids and digestion procedure, which affords reference for the preparation of the same kinds of polymer samples. The detection limits of the method for all sample elements were 0.7-6.5 ng x g(-1), the recoveries were 89.8%-110.8%, and the RSDs were 2.8%-11.3%. The analytical method presented was characterized with good precision and accuracy, simplicity, rapidness, low limits of detection and no matrix matching requirements.

[Determination of se in blood and tissue of animal samples by normal ICP-MS and ICP-MS with octupole reaction system].

The Se in the blood and tissue of animal samples was determined by ICP-MS instrument. The method was not only proved simple and rapid, but also showed satisfying precision and low detection limit. The optimum conditions of ICP-MS for the method were investigated. Results by the digestion method, the isotopes selection, and the difference between the normal ICP-MS without ORS (Octopole Reaction System) and the ICP-MS with ORS were compared, which afford reference to the determination of the same kinds of sample. The standard materials were determined to proved the reliability of the normal ICP-MS and the ORS-ICP-MS methods. The ORS-ICP-MS can effectively remove the interference based on polyatomic ions, so the the detection limits were lowed, and the accuracy was improved. The detection limits of the method for normal ICP-MS is 0.024 ng x g(-1) and that for ORS-ICP-MS is 0.004 6 ng x g(-1), the RSDs are between 1.8% and 5.5%, and the recoveries of the method are in the range of 90.8%-107.2%.

[Application of high performance liquid chromatography and hyphenated techniques in analysis of arsenic species].

Arsenic is a very important nonmetal element in the environment and is vitally related to human lives. It is well known that the biological availability and toxicological effects of arsenic compounds depend upon their chemical forms, so it is necessary to detect the species of arsenic in some environmental samples, food, medicine, and body fluids. High performance liquid chromatography (HPLC) is widely used in the research on arsenic species because of its high separation capability. The detection techniques used for arsenic species analysis have been improved rapidly with the gradually increasing demand for sensitivity and accuracy. Various hyphenated techniques have also been developed. Inductively coupled plasma mass spectrometry has many advantages, such as high sensitivity, simultaneous multi-element detection, wide linear range and so on. Therefore, high performance liquid chromatography-inductively coupled plasma mass spectrometry is widely used in the analysis of arsenic species.

[Investigating the distribution of 11 kinds of trace elements in root-like and rootstalk-like dried medicinal herbs by using microwave digestion-ICP-MS].

A method was studied for the analysis of 11 kinds of trace elements(Cr, Mn, Ni, Co, Cu, Zn, As, Se, Mo, Cd and Pb) in root-like and rootstalk-like dried medicinal herbs by microwave digestion-ICP-MS. A digestion method was developed for trace elements analysis for two types of herbs. The Microwave digestion condition was optimized in terms of digestion system, proportion of acids, and digestion procedure, which affords reference to the preparation of the same kinds of dried medicinal herbs. The detection limits of the method for all sample elements were in the range of 0. 001-0. 260 microg x g(-1), the RSDs were between 0. 4%-3. 1% and the recoveries were between 90% and 110%.

[Study on the method for the determination of soil available B by ICP-MS].

A method was established for the determination of soil available B by inductively coupled plasma mass spectrometry (ICP-MS). Compared to the traditional method for B analysis, the recommended method does not need filtration, so it escapes the contamination. The method not only proved simple and rapid, but also showed satisfying precision and accuracy. The detection limit of this method is 0.009 ng x g(-1). The relative standard deviation is 2.66%, and the recovery is 93.0%-102.0%.

[Determination of As and Se in animal and plant samples by the normal ICP-MS with and without octupole reaction system and the ICP-MS with ORS].

Scme certified reference materials (CRMs) were digested and the elemental concentrations were determined by ICP-MS instrument, and the results include not only the trace key elements such as As, Pb, Hg and Cd but also the major components such as Ca, Fe, K, Na etc. The digestion method, the recoveries, and the detection limits are discussed to piove the reliability of ICP-MS and EPA200.8 method for the determination of the elements in fcxxl and plant samples. The ICP-MS instrument with ORS (octupole reaction system) was applied to the determination of As, Se etc, and the results are compared with those from the normal ICP-MS without ORS. The removal of ArCl and ArAr interference is discussed, and the advantage of ORS is proved by measuring the As and Se in some CRMs.

[In situ thin layer chromatography-fourier transform-surface-enhanced Raman spectrum study on ingredients of berberine].

Surface Enhanced Raman Scattering (SERS) combined with Thin Layer Chromatography (TLC) has been used for studying characteristic spectrum of molecules in situ in micrograms samples. There are very few report for applying the SERS-TCL method in the study of the effective ingredients of Chinese traditional herbs. Coptis Chinensis France is an often-used clinic Chinese traditional medicine. Its main effective components include berberine and so on, which have antibiotic very wide and also have curative effect on improving the functions of heart vascular cycles. Therefore the concentrations of berberine are very important for the quality control of the medicine. In this work, the ethanol extract of Coptis Chinensis France was first separated by TLC, the SERS was then measure directly after dropping silver gel on the separated spots. The method can be used for the finger print analysis of the berberine. 3 microL of alcohol extract of Coptis Chinensis France with total alkaloids concentration of 1.0 mg.mL was placed on silicon GF254 TLC plate. The sample was separated by developing solvent of n bulanol-Acitic acid-H2O (7:2:1 V/V). The positions of berberine in the sample were confirmed by the standard alkaloid solutions. The Rf values for berberine are 0.29. The silver gel was used as surface enhanced substrate and placed on the separated berberine spots. FT-SERS was measured directly by a Nicolet FT-Raman 910 spectrometer. Berberine belong to isoquinoline alkaloids. His structure can be found in reference. The date of spectrum of berberine can be seen that the band at 1,396 cm-1 due to Ar-OCH3 deformation vibrations was greatly enhanced, indicating that the molecule was absorbed on silver gel strongly through lone-pair electron in Ar-OCH3. The ring stretching mode occurring around 1,548 cm-1 represents isoquinoline ring in the molecule. The band at 727 cm-1 due to CH (ring) deformation vibrations was also enhanced.

[Pollution characteristics and source identification of atmospheric particulate matters n-alkanes in Baoding City].

Organic matter has been a very important component in the ambient particulate matter of big cities in China. In order to investigate the mass concentrations and sources of the organic matter which are adsorbed in ambient particulate matters in the industry cities of Hebei province, aerosol samples were collected with Anderson sampler during Sep. 2009 to Aug. 2010, in Baoding city, Hebei province. The concentration of n-alkanes was determined via GC-MS. About 66.7% of the daily average concentrations of fine particulate matters were higher than the 24-hour average threshold value of class II standard of the ambient air quality standard (GB 3095-2012, 75 microg.m-3). About 96% of the daily average concentrations of inhalable particles were higher than 150 microg.m-3)(GB 3095-2012). The total concentration of n-alkanes was in the range of 111.23-979.81 ng.m-3 with an average of 264.2 ng.m-3. The n-alkene homologues from C14 to C32 were detected with different peak carbon numbers ranging from 20 to 27 in different seasons. In summer, the peak carbon number was 27, while it was C20, C21 or C22 in winter and spring. The CPI values were 0. 97, 1. 24, 0. 92 and 0. 86 in spring,summer, autumn and winter, respectively, with an average of 1.01. These results indicated that the incomplete combustion of fossil fuel and vehicle emissions was the main resource of n-alkanes in winter and spring, and the high plant waxes were playing a major role in summer and autumn. The primary sources were anthropogenic activities all year round.

[Analysis on water-soluble inorganic ions in the atmospheric aerosol of Xinglong].

Size-segregated aerosol samples were collected using Andersen cascade sampler from Sep. 2009 to Aug. 2010 in Xinglong, a regional atmospheric background station. The water-soluble inorganic ions were analyzed by IC. The result showed that the annual concentrations of the total water-soluble inorganic ions were (89.66 +/- 47.66), (54.44 +/- 34.08) and (44.39 +/- 29.95) microg x m(-3) in TSP, PM2.1 and PM1.1 respectively. SO4(2-), NO3(-), Ca(2) and NH4(+) were the dominant contributors of water-soluble inorganic ions. The total water-soluble inorganic ions in PM2.1 accounted for 61% of TSP. The total water-soluble inorganic ions in PM1.1 accounted for 50% and 82% in TSP and PM2.1, respectively. The seasonal variations of the total water-soluble inorganic ion concentration in TSP, PM2.1 and PM1.1 were the same, following the order of summer > autumn > springs > winter. The mean molar ratio of NH4(+) to SO4(2-) was larger than 2, indicating that NH4(+) was not completely neutralized by SO4(2-). NH4(+) -SO4(2-) and NH4(+) -NO3(-) concentrations were strongly correlated (R2 0.96 and 0.87), indicating that NH4(+) was mainly present as (NH4) 2SO4 and NH4NO3.

The antidepressant effect of secoisolariciresinol, a lignan-type phytoestrogen constituent of flaxseed, on ovariectomized mice.

Secoisolariciresinol (SECO) is a natural lignan-type phytoestrogen constituent mainly found in flaxseed. It can be metabolized in vivo to mammalian lignans of enterodiol and enterolactone, which have been proven to be effective in relieving menopausal syndrome. Depression is one of the most common symptoms of menopausal syndrome, and is currently treated with estrogen replacement and antidepressant therapy. However, due to the serious side-effects of such agents, there are urgent needs for safer and more tolerable treatments. In this paper, using two classical depression models, the forced swimming test and the tail suspension test, we report the antidepressant effect of SECO on ovariectomized (OVX) mice by intragastric administration for 14 consecutive days at doses of 5, 10 and 20 mg/kg. The results showed that SECO (10 mg/kg) treatment could significantly reduce the duration of immobility of OVX mice in these two models compared with the control group (OVX mice + vehicle), which was similar to the positive control imipramine. In addition, SECO treatment could substantially increase brain monoamine (norepinephrine and dopamine) levels in OVX mice. The present studies showed that SECO can reverse depressive-like behavior and exhibit monoamine-enhancing effects.

[Chemical characteristics and sources of trace metals in precipitation collected from a typical industrial city in Northern China].

To investigate the chemical characteristics and possible sources of trace metals in precipitation of Tangshan, a typical industrial city in Northern China, precipitation samples were collected using an automated wet-only sampler for one year from December 2009 to November 2010 and subjected to chemical analysis using ICP-MS. The results showed that crustal elements (Ca, Mg, Na, K, Fe and Al) accounted for 97.72% of the total concentrations while heavy metals (Zn, Mn, Pb, Ba, Sb, Cu, Ni, As, V, Co and Se) only contributed 2.25% of the total. Zinc was the most abundant heavy metal and calcium had the highest concentration among the crustal elements, with volume-weighted mean concentrations of 88.7 microg x L(-1) and 4.5 mg x L(-1), respectively. Seasonal variations of trace metals were significant, with higher values observed in winter and spring than in summer and autumn. The calculation of crustal enrichment factors with Al as the reference element indicated that Tl, As, Ag, Zn, Pb, Cd, Se and Sb were mainly derived from anthropogenic sources and the enrichment factor values of those elements ranged from 10(2) to 10(5). In addition, the concentrations of heavy metals such as Pb, Cu and Ni in precipitation of Tangshan city were higher than those reported for Mt. Tai. The results of principle component analysis demonstrated that steel smelting emissions, fugitive dusts and coal combustion were probably the major contributors of trace metals in the precipitation of Tangshan City.

[Size distribution and characterization of EC and OC in aerosols during the Olympics of Beijing, China].

Samples of airborne PM with different size were collected by cascade impactor (Andersen) in Beijing during the Beijing Olympics. Organic carbon (OC) and elemental carbon (EC) in particles were determined by DRI Model 2001 A t carbon analyzer of USA. PM (56%), OC (55%) and EC (73%) were associated with the fine particle below 2.1 microm. OC and EC account for 25% and 5% of particle below 2.1 microm, respectively, which indicates that carbonaceous aerosols are key components for controlling fine particles pollution in Beijing. It is found that Beijing auto measures were effective in abatement of carbonaceous aerosols. The ratios of OC/EC indicated that the main origins of carbonaceous aerosols in fine particle were automobile exhaust fumes and coal combustion and in coarse particle were biomass burning and cooking. The average percentage of estimated secondary OC (OC(sec)) in the total OC (OC(tot)) in TSP was 74%. The results of correlation analysis show that the correlation coefficient between the OC and secondary water soluble ions (SO4(2-), NO3-, NH4+, NO2-) were very high in PM2.1 (R2 = 0.88), while the correlation coefficient was very faint in particles greater than 2.1 microm (R2 = 0.21).

Simultaneous analysis of multiple bioactive constituents in Rheum tanguticum Maxim. ex Balf. by high-performance liquid chromatography coupled to tandem mass spectrometry.

A simple, rapid method was developed using on-line high-performance liquid chromatography (HPLC)-diode array detection (DAD)/electrospray ionization mass spectrometry (ESI-MS) to simultaneously analyze multiple bioactive constituents in the extract of Rheum tanguticum Maxim. ex Balf. (Dahuang), a traditional Chinese medicine (TCM). Many bioactive constituents gave prominent [M-H]- ions in the negative ion ESI mass spectra. Among them, 41 different constituents including 16 anthraquinone derivatives, 7 phenylbutanone glucopyranosides, 4 stilbenes and 14 tannins were unambiguously identified or tentatively characterized based on their retention times, UV spectra and mass spectra in comparison with the data from standards or references. Meanwhile, some principles of fragmentation behavior for different types of constituents were proposed, which could contribute to the elucidation of these constituents in Rheum tanguticum.