Treatment of wet process hardboard plant VOC emissions by a pilot scale biological system

A pilot scale biological treatment system for air emissions was installed and tested at a forest products plant in western Oregon, USA, which collected and treated gaseous emissions from the hardboard steam press vents on the top of the plant building. This system was installed mainly to demonstrate the effectiveness of biological treatment technologies in removing volatile organic compounds (VOC) and hazardous air pollutants (HAP) from the wet-process hardboard press emissions, and to test the efficiency of the system on fine particles and condensable organics with the presence of a pre-treatment wet dust collector. The bio-oxidation system was comprised of a particle pre-treatment unit Type W Rotoclone (wet hydrocyclone), a biotrickling filter and a biofilter with airflow capacity of up to 4.72 m³/s. This unit operated at approximately 0.71 m³/s, which is the optimal flow required for the Rotoclone's throughput, and provided an EBCT (empty bed contact time) of 45 s. Analysis of total VOC measurements from the system indicated removals down to less than 5 ppm in the effluent emissions. Evaluations of opacity reductions also met project objectives with routine outlet measurements of 0–5%, which are in compliance with state regulatory guidelines. Emissions air samples were collected at different locations in the biological system for GC–MS analysis and characterization to identify specific VOCs and their removals.     

Analysis of cyclosporine A and its metabolites in rat urine and feces by liquid chromatography–tandem mass spectrometry

A sensitive and specific liquid chromatography–tandem mass spectrometry (LC–MS/MS) method was developed and validated for the quantification of cyclosporine A (CyA) and the identification of its metabolites in rat urine and feces. The analytes were extracted from waste samples via liquid–liquid extraction. A Turboionspray source was used as a detector. It was operated in a positive ion mode with transitions of m/z 1225 → m/z 1112 for CyA and in a selected multiple reactions monitoring (MRM) mode with transitions of m/z 1239 → m/z 1099 for the internal standard (cyclosporine D, CyD). Linear calibration curves were obtained for CyA concentration ranges of 12.5–250 ng mL^?1 in urine and 2.5–375 ng mg^?1 in feces. The intra- and inter-day precision values (relative standard deviation) obtained were less than 8%, and the accuracy was within ±15% for each of the analytes. Extraction recoveries of CyA and CyD were both over 80%. The identification of the metabolites and elucidation of their structure were performed on the basis of their retention times and mass spectrometry fragmentation behaviors. A total of seven metabolites in rat feces were identified as dimethyl CyA, hydroxy CyA, and dihydroxy CyA after the oral administration of cyclosporine A-Eudragit^® S100 nanoparticles (CyA-NP). Six of these metabolites were also detected in rat urine. A possible metabolic pathway was also proposed. The newly developed method was proven to be sensitive, simple, reproducible, and suitable for the rapid determination of CyA. It was successfully employed to study the excretion of CyA in rats and could be used to better understand the in vivo metabolism of CyA-NP, a potentially effective nanoparticle system.     

Sensitivity of Norway spruce physiology and terpenoid emission dynamics to elevated ozone and elevated temperature under open-field exposure

Volatile organic compounds (VOCs) emitted from vegetation to the atmosphere contribute to global climate change, but climate change factors also affect VOC emission from vegetation. Soil-grown Norway spruce seedlings were exposed to elevated ozone (1.4 × ambient ozone concentration) and elevated temperature (ambient + 1.3 °C) alone and in combination as well as to ambient ozone and temperature treatments under open-field conditions. VOC emissions (mainly terpenoids), genes involved in early steps of plastidial monoterpene and isoprene synthesis, photosynthetic parameters and growth were measured. In July, when daytime elevated ozone concentrations had been over 40 ppb, ozone doubled the total terpenoid emissions by increasing the emissions of many monoterpenes and sesquiterpenes. Elevated temperature changed the terpenoid profile by increasing the emissions of oxygenated monoterpenes, but did not influence total emissions. Terpenoid emission profiles also differed between elevated ozone alone and elevated ozone in combination with elevated temperature. In August, when daytime elevated ozone concentrations had been ca. 30 ppb, significant treatment effects were not found. Ozone and temperature reduced the expression of DXS2B (1-deoxy-d-xylulose 5-phosphate synthase type II), and ozone that of DXR (1-deoxy-d-xylulose 5-phosphate reductoisomerase) in August. Elevated temperature reduced the stem diameter growth, net photosynthesis and stomatal conductance, but elevated ozone did not have effects on these parameters. Results suggest that elevated temperature may not modify the ozone responses, or vice versa, in terms of gas exchange, growth or total terpenoid emission rates of young Norway spruces in a near-future climate. However, observed changes in terpenoid emission profiles may be important in the future climate, as reactivity in the troposphere differs between individual terpenoids.     

Simultaneous determination of puerarin,daidzein, baicalin,wogonoside and liquiritin of GegenQinlian decoction in rat plasma by ultra-performance liquid chromatography–mass spectrometry

A specific ultra-performance liquid chromatography–mass spectrometry (UPLC–MS) method was developed for the simultaneous determination of puerarin, daidzein, baicalin, wogonoside and liquiritin in rat plasma. Chromatographic separation was performed on a C₁₈ column packed with 1.7 μm particles by a linear gradient elution. The analytes and carbamazepine (internal standard, I.S.) were monitored in a selected-ion reaction (SIR) mode with a positive electrospray ionization (ESI) interface by the following ions: m/z 417.2 for puerarin, m/z 255.2 for daidzein, m/z 271.0 for baicalin, m/z 461.0 for wogonoside, m/z 441.0 for liquiritin and m/z 237.2 for carbamazepine (I.S.), respectively. The calibration curves of these analytes were linear over the concentration ranges from 0.00254–1.02 μg mL^?1 to 0.0102–10.2 μg mL^?1. Within-batch and between-batch precisions (RSD%) were all within 15% and accuracy (RE%) ranged from ?10% to 10%. The extraction recoveries were on average 79.8% for puerarin, 90.8% for daidzein, 74.4% for baicalin, 70.2% for wogonoside and 84.7% for liquiritin. The validated method was successfully applied to investigate the pharmacokinetics of five bioactive compounds of GegenQinlian decoction (GQD) in rats.     

Determination of picamilon concentration in human plasma by liquid chromatography–tandem mass spectrometry

A rapid liquid chromatography–tandem mass spectrometry (LC–MS/MS) method was developed and validated for the determination of picamilon concentration in human plasma. Picamilon was extracted from human plasma by protein precipitation. High performance liquid chromatography separation was performed on a Venusil ASB C₁₈ column with a mobile phase consisting of methanol ?10 mM ammonium acetate–formic acid (55:45:01, v/v/v) at a flow rate of 0.65 ml/min. Acquisition of mass spectrometric data was performed in selected reaction monitoring mode, using the transitions of m/z 209.0 → m/z (78.0 + 106.0) for picamilon and m/z 152.0 → m/z (93.0 + 110.0) for paracetamol (internal standard). The method was linear in the concentration range of 1.00–5000 ng/ml for the analyte. The lower limit of quantification was 1.00 ng/ml. The intra- and inter-assay precision were below 13.5%, and the accuracy was between 99.6% and 101.6%. The method was successfully applied to characterize the pharmacokinetic profiles of picamilon in healthy volunteers. This validated LC–MS/MS method was selective and rapid, and is suitable for the pharmacokinetic study of picamilon in humans.     

Simultaneous determination of tolbutamide,omeprazole, midazolam and dextromethorphan in human plasma by LC–MS/MS—A high throughput approach to evaluate drug–drug interactions

Drug–drug interactions involving cytochrome P450 (CYP450s) are an important factor for evaluation of a new chemical entity (NCE) in drug development. To evaluate the potential inhibitory effects of a NCE on the pharmacokinetics of a cocktail of representative probes of CYP enzymes (midazolam for CYP3A4, tolbutamide for CYP2C9, omeprazole for CYP2C19 and dextromethorphan for CYP2D6) and the safety and tolerability of the NCE in the presence of probe substrates, a high throughput liquid chromatography/tandem mass spectrometry (LC–MS/MS) method was developed and validated for the simultaneous determination of tolbutamide, omeprazole, midazolam and dextromethorphan in human plasma using tolbutamide-d₉, midazolam-d₄, (±)-omeprazole-d₃, and dextromethorphan-d₃ as the internal standards (ISs). Human plasma samples of 50 μL were extracted by a simple protein-precipitation procedure and analyzed using a high performance liquid chromatography electrospray tandem mass spectrometer system. Reversed-phase HPLC separation was achieved with a Hypersil GOLD AQ column (50 mm × 4.6 mm, 5 μm). MS/MS detection was set at mass transitions of 271 → 172 m/z for tolbutamide, 346 → 198 m/z for omeprazole, 326 → 291 m/z for midazolam, 272 → 171 m/z for dextromethorphan, 280 → 172 m/z for tolbutamide-d₉ (IS), 349 → 198 m/z for (±)-omeprazole-d₃ (IS), 330 → 295 m/z for midazolam-d₄ (IS), and 275 → 171 m/z for dextromethorphan-d₃ (IS) in positive mode. The high throughput LC–MS/MS method was validated for accuracy, precision, sensitivity, stability, recovery, matrix effects, and calibration range. Acceptable intra-run and inter-run assay precision (<10%) and accuracy (<10%) were achieved over a linear range of 50–50,000 ng/mL for tolbutamide, 1–1000 ng/mL for omeprazole, 0.1–100 ng/mL for midazolam and 0.05–50 ng/mL for dextromethorphan in human plasma. Method robustness was demonstrated by the 100% pass rate of 24 incurred sample analysis runs and all of the 50 clinical study samples used for incurred sample reproducibility (ISR) test having met the acceptance criterion (%Diff within 20%). The overall ISR results for all compounds showed that over 95% of the samples had a %Diff of less than 10%. The method is simple, rapid and rugged, and has been applied successfully to sample analysis in support of a drug–drug interaction study.     

Simple and selective method for the determination of various tyrosine kinase inhibitors used in the clinical setting by liquid chromatography tandem mass spectrometry

A fast, sensitive, universal and accurate method for the determination of four different tyrosine kinase inhibitors from biological material was developed using LC–MS/MS techniques. Utilizing a simple protein precipitation with acetonitrile a 20 μl sample volume of biological matrixes can be extracted at 4 °C with minimal effort. After centrifugation the sample extract is introduced directly onto the LC–MS/MS system without further clean-up and assayed across a linear range of 1–4000 ng/ml. Chromatography was performed using a Dionex Ultimate 3000 with a Phenomenex prodigy ODS3 (2.0 mm × 100 mm, 3 μm) column and eluted at 200 μl/min with a tertiary mobile phase consisting of 20 mM ammonium acetate:acetonitrile:methanol (2.5:6.7:8.3%). Injection volume varied from 0.1 μl to 1 μl depending on the concentration of the drug observed. Samples were observed to be stable for a maximum of 48 h after extraction when kept at 4 °C. Detection was performed using a turbo-spray ionization source and mass spectrometric positive multi-reaction-monitoring-mode (+MRM) for Gefitinib (447.1 m/z; 127.9 m/z), Erlotinib (393.9 m/z; 278.2 m/z), Sunitinib (399.1 m/z; 283.1 m/z) and Sorafenib (465.0 m/z; 251.9 m/z) at an ion voltage of +3500 V. The accuracy, precision and limit-of-quantification (LOQ) from cell culture medium were as follows: Gefitinib: 100.2 ± 3.8%, 11.2 nM; Erlotinib: 101.6 ± 3.7%, 12.7 nM; Sunitinib: 100.8 ± 4.3%, 12.6 nM; Sorafenib: 93.9 ± 3.0%, 10.8 nM, respectively. This was reproducible for plasma, whole blood, and serum. The method was observed to be linear between the LOQ and 4000 ng/ml for each analyte. Effectiveness of the method is illustrated with the analysis of samples from a cellular accumulation investigation and from determination of steady state concentrations in clinically treated patients.     

Aphid antixenosis in cotton is activated by the natural plant defence elicitor cis-jasmone

Upon insect herbivory, plants can release blends of volatile organic compounds (VOCs) that modify herbivore and natural enemy behaviour. We have shown recently that cotton, Gossypium hirsutum, emits a blend of defence VOCs that repels the cotton aphid, Aphis gossypii, upon herbivory by this notorious crop pest, including (Z)-3-hexenyl acetate, (E)-4,8-dimethyl-1,3,7-nonatriene (DMNT), methyl salicylate and (E,E)-4,8,12-trimethyl-1,3,7,11-tridecatetraene (TMTT). In this study, we investigated changes in the defence VOC profile of G. hirsutum induced by the naturally-occurring plant elicitor cis-jasmone (CJ) and whether these changes modify the behaviour of A. gossypii. In four-arm olfactometer assays, VOCs from untreated plants were significantly attractive (P < 0.05), whilst VOCs from CJ-treated plants were significantly repellent (P < 0.05). The VOCs induced by CJ appeared to comprise (Z)-3-hexenyl acetate, DMNT, methyl salicylate and TMTT. In quantitative VOC collection studies, sustained release of DMNT and TMTT was observed in CJ-treated plants over a period of five days, with levels becoming statistically significantly higher than for control treated plants on the fifth day in most cases. Despite earlier indications, no statistically significant differences were observed in levels of (Z)-3-hexenyl acetate or methyl salicylate between CJ and control treatments on any day. Furthermore, DMNT and TMTT emissions from CJ-treated plants were further enhanced by subsequent addition of A. gossypii. CJ treatment induced statistically significantly higher DMNT and TMTT expression levels as early as day three, when A. gossypii was present. The results in this study show that CJ can induce the production of A. gossypii-induced VOCs from G. hirsutum, with potential for deployment in novel crop protection strategies.     

Enantiomeric determination of tramadol and O-desmethyltramadol in human plasma by fast liquid chromatographic technique coupled with mass spectrometric detection

A rapid and sensitive method using liquid chromatography–tandem mass spectrometry (LC–MS/MS) for enantiomeric determination of tramadol and its primary phase metabolite O-desmethyltramadol in human plasma has been developed. Tramadol hydrochloride – ¹³C, d_3, was used as an isotopic labeled internal standard for quantification. The method involves a simple solid phase extraction. The analytes and internal standard were separated on Lux Cellulose-2 packed with cellulose tris(3-chloro-4-methylphenylcarbamate) using isocratic elution with hexane/isopropanol/diethylamine (90:10:0.1, v/v/v) at a flow rate of 1.3 mL/min. The APCI positive ionization mass spectrometry was used with multiple reaction monitoring of the transitions at m/z 264.2 → 58.2 for tramadol, m/z 250.1 → 58.2 for O-desmethyltramadol and m/z 268.2 → 58.2 for internal standard. Linearity was achieved between 1–800 ng/mL and 1–400 ng/mL (R² ≥ 0.999) for each enantiomer of tramadol and O-desmethyltramadol, respectively. Intra-day accuracies ranged among 98.2–102.8%, 97.1–109.1% and 97.4–102.9% at the lower, intermediate, and high concentration for all analytes, respectively. Inter-day accuracies ranged among 95.5–104.1%, 99.2–104.7%, and 94.2–105.6% at the lower, intermediate, and high concentration for all analytes, respectively. This assay was successfully used to determine the concentration of enantiomers of tramadol and O-desmethyltramadol in a pharmacogenetic study.     

Stable isotope liquid chromatography-tandem mass spectrometry assay for fatty acid amide hydrolase activity

Fatty acid amide hydrolase (FAAH) is the main enzyme responsible for the hydrolysis of the endocannabinoid anandamide (arachidonoyl ethanolamide, AEA) to arachidonic acid (AA) and ethanolamine (EA). Published FAAH activity assays mostly employ radiolabeled anandamide or synthetic fluorogenic substrates. We report a stable isotope liquid chromatography–tandem mass spectrometry (LC–MS/MS) assay for specific, sensitive, and high-throughput capable FAAH activity measurements. The assay uses AEA labeled with deuterium on the EA moiety (d₄-AEA) as substrate and measures the specific reaction product tetradeutero-EA (d₄-EA) and the internal standard ¹³C₂-EA. Selected reaction monitoring of m/z 66 → m/z 48 (d₄-EA) and m/z 64 → m/z 46 (¹³C₂-EA) in the positive electrospray ionization mode after liquid chromatographic separation on a HILIC (hydrophilic interaction liquid chromatography) column is performed. The assay was developed and thoroughly validated using recombinant human FAAH (rhFAAH) and then was applied to human blood and dog liver samples. rhFAAH-catalyzed d₄-AEA hydrolysis obeyed Michaelis–Menten kinetics (K_M = 12.3 μM, V_max = 27.6 nmol/min mg). Oleoyl oxazolopyridine (oloxa) was a potent, partial noncompetitive inhibitor of rhFAAH (IC₅₀ = 24.3 nM). Substrate specificity of other fatty acid ethanolamides decreased with decreasing length, number of double bonds, and lipophilicity of the fatty acid skeleton. In human whole blood, we detected FAAH activity that was inhibited by oloxa.     

Simultaneous quantification of rosiglitazone and its two major metabolites,N-desmethyl and p-hydroxy rosiglitazone in human plasma by liquid chromatography/tandem mass spectrometry: Application to a pharmacokinetic study

We present a simple, rapid, and sensitive liquid chromatography (LC)–tandem mass spectrometry (MS/MS) method for the simultaneous quantification of rosiglitazone and its two major metabolites via CYP2C8/9, N-desmethyl and p-hydroxy rosiglitazone, in human plasma. The procedure was developed and validated using rosiglitazone-d₃ as the internal standard. Plasma samples (0.1 ml) were prepared using a simple deproteinization procedure with 0.2 ml of acetonitrile containing 40 ng/ml of rosiglitazone-d₃. Chromatographic separation was carried out on a Luna C₁₈ column (100 mm × 2.0 mm, 3-μm particle size) using an isocratic mobile phase consisting of a 60:40 (v/v) mixture of acetonitrile and 0.1% formic acid_(aq). Each sample was run at 0.2 ml/min for a total run time of 2.5 min per sample. Detection and quantification were performed using a mass spectrometer in selected reaction-monitoring mode with positive electrospray ionization at m/z 358.1 → 135.1 for rosiglitazone, m/z 344.2 → 121.1 for N-desmethyl rosiglitazone, m/z 374.1 → 151.1 for p-hydroxy rosiglitazone, and m/z 361.1 → 138.1 for rosiglitazone-d₃. The linear ranges of concentration for rosiglitazone, N-desmethyl rosiglitazone, and p-hydroxy rosiglitazone were 1–500, 1–150, and 1–25 ng/ml, respectively, with a lower limit of quantification of 1 ng/ml for all analytes. The coefficient of variation for assay precision was less than 14.4%, and the accuracy was 93.3–112.3%. No relevant cross-talk and matrix effect were observed. This method was successfully applied to a pharmacokinetic study after oral administration of a 4-mg rosiglitazone tablet to healthy male Korean volunteers.     

Interplay between thermal and immune ecology: effect of environmental temperature on insect immune response and energetic costs after an immune challenge

Although the study of thermoregulation in insects has shown that infected animals tend to prefer higher temperatures than healthy individuals, the immune response and energetic consequences of this preference remain unknown. We examined the effect of environmental temperature and the energetic costs associated to the activation of the immune response of Tenebrio molitor larvae following a lipopolysaccharide (LPS) challenge. We measured the effect of temperature on immune parameters including phenoloxidase (PO) activity and antibacterial responses. Further as proximal and distal costs of the immune response we determined the standard metabolic rate (SMR) and the loss of body mass (m_b), respectively. Immune response was stronger at 30 °C than was at 10 or 20 °C. While SMR at 10 and 20 °C did not differ between immune treatments, at 30 °C SMR of LPS-treated larvae was almost 25–60% higher than SMR of PBS-treated and naïve larvae. In addition, the loss in m_b was 1.9 and 4.2 times higher in LPS-treated larvae than in PBS-treated and naïve controls. The immune responses exhibited a positive correlation with temperature and both, SMR and m_b change, were sensitive to environmental temperature. These data suggest a significant effect of environmental temperature on the immune response and on the energetic costs of immunity.     

Extremely low fine root biomass in Larix sibirica forests at the southern drought limit of the boreal forest

Mongolia's Larix sibirica forests at the southern fringe of the Eurosiberian boreal forest belt are exposed not only to very low winter temperatures, but also to frequent summer droughts. It is not completely known how Siberian larch adapts to these stressors. We examined whether (i) these forests differ in their fine root bio- and necromass from more humid boreal forests further in the North and (ii) inter-annual fluctuations in fine root biomass are related to tree vitality. In two exceptionally dry summers, we found only 4–5 g DM m^?2 of fine root biomass (in 0–20 cm depth), which is far less than typical conifer fine root biomass figures from boreal forests (c. 200–400 g m^?2) and the lowest forest fine root biomass reported worldwide; in a moist summer, fine root biomass was 20 fold higher. In contrast to fine root biomass, both necromass and non-tree root mass were high in all three years. From the large increase of fine root biomass in the moist summer and the generally high root necromass, we conclude that drought-induced fine root dieback was the likely cause of the very small amount of live root mass in the dry summers. Larch fine roots seem to be more drought-sensitive than shoots, since marked needle loss did not occur under the extreme conditions.     

Performance of tiloronoxim and tilorone determination in human blood by HPLC–MS/MS: Method validation,uncertainty assessment and its application to a pharmacokinetic study

A highly sensitive and selective HPLC–MS/MS method is presented for the quantitative determination of tiloronoxim and its metabolite tilorone in human blood. An aliquot of 200 μl human blood was extracted with a mixture of chloroform/ethyl ether (1/2, v/v), using metoprolol as the internal standard (the IS). Separation was achieved on an Xterra MS C18 column (50 mm × 2.1 mm, 5 μm) with a gradient mobile phase of methanol/water containing 15 mM ammonium bicarbonate (pH 10.5). Detection was performed using positive MRM mode on a TurboIonSpray source. The mass transitions monitored were m/z 426.3 → 100.0, m/z 411.3 → 100.0 and m/z 268.3 → 116.1 for tiloronoxim, tilorone and the IS, respectively. The method was fully validated using total error theory, which is based on β-expectation tolerance intervals and include trueness and intermediate precision. The method was found to be accurate over a concentration range of 1–100 ng/ml for both compounds. The measurement uncertainty based on β-expectation tolerance intervals was assessed at each concentration level of the validation standards. This method was successively applied to a pharmacokinetic study of tiloronoxim in healthy volunteers.     

Quantitative analysis of RU38486 (mifepristone) by HPLC triple quadrupole mass spectrometry

A sensitive liquid chromatography–mass spectrometric method was validated for the quantification of RU38486 (mifepristone) in human and murine plasma. The analyte and internal standard (alfaxolone) were extracted by liquid–liquid extraction with diethyl ether, resolved on a C18 column using gradient elution with methanol and ammonium acetate and detected after positive electrospray ionization (m/z 430 → 372; m/z 333 → 297, respectively). Quantification was linear over the range 0.5–500 ng (r² > 0.997), precise and accurate (intra-assay RSD ≤ 7.2%, RME ≤ 8.2%; inter-assay RSD ≤ 15.7% RME ≤ 10.2%). The limit of quantification (LOQ) was 50 pg injected on column, permitting reproducible analysis of RU38486 in small volumes of plasma.     

Determination of metoclopramide in human plasma by LC–ESI-MS and its application to bioequivalance studies

An LC–MS method for the determination of metoclopramide in human plasma was developed and validated. Sample preparation involved extraction with ethyl acetate. Chromatographic separation was performed on a Thermo Hypersil-Hypurity C₁₈ (150 mm × 2.1 mm, 5 μm) with the mobile phase consisting of 40 mM ammonium acetate–methanol–acetonitrile. A single-quadrupole mass spectrometer with an electrospray interface was operated in the selected-ion monitoring mode to detect the [M+H]⁺ ions at m/z 300 for metoclopramide and at m/z 384 for the internal standard (prazosin). The method was validated over 0.78–50.00 ng mL^?1 for metoclopramide. The recovery was 67.8–83.1%, and the limit of quantitation (LOQ) detection was 0.78 ng mL^?1 for metoclopramide. The intra- and inter-day precision of the method at three concentrations was 5.0–13.6% with accuracy of 99.2–104.0%. Stability of compounds was established in a battery of stability studies. The method was successfully applied to bioequivalence studies of metoclopramide hydrochloride tablets to obtain the pharmacokinetic parameters.     

Influence of slope on root system anchorage of Pinus yunnanensis

In southwestern China, Yunnan pines (Pinus yunnanensis) have been extensively cultivated on barren hills for reforestation and ecological engineering. The objective of this work is to study the influences of slope gradient on anchorage of root systems of P. yunnanensis. Pulling experiments were carried out at low (5–6°), moderate (25–26°) and high gradient (42–43°) by using selected 15-year-old P. yunnanensis. The results showed that the anchorage resistances induced by the slope gradient were significantly (P < 0.01) different from each other and followed in the order of high slope > moderate slope > low slope. Anchorage strength of upslope-grown roots increased with size and length of first-order lateral roots as well as the number of second-order lateral roots. Contributions of upslope-grown roots for preventing plants from overturning varied considerably between those from different slopes. The contribution of roots growing on upslope side on high slope to the anchorage of root system reached 50%, whereas those on moderate and low slopes were about 44% and 37%, respectively. It was concluded that the anchorage resistance was closely related to slope gradient and root distribution, while upslope-grown roots were positively related to anchorage resistance.     

Determination of eptifibatide concentration in human plasma utilizing the liquid chromatography–tandem mass spectrometry method

A sensitive and selective liquid chromatography–tandem mass spectrometry (LC–MS/MS) method was developed to determine the concentration of eptifibatide in human plasma. Following protein precipitation, the analyte was separated on a reversed-phase C₁₈ column. Acetonitrile:5 mM ammonium acetate:acetic acid (30:70:0.1, v/v/v) was used at a flow-rate of 0.5 mL/min with the isocratic mobile phase. An API 4000 tandem mass spectrometer equipped with a Turbo IonSpray ionization source was used as the detector and was operated in the positive ion mode. “Truncated” multiple reaction monitoring using the transition of m/z 832.6 → m/z 832.6 and m/z 931.3 → m/z 931.3 was performed to quantify eptifibatide and the internal standard (EPM-05), respectively. The method had a lower limit of quantification of 4.61 ng/mL for eptifibatide. The calibration curve was demonstrated to be linear over the concentration range of 4.61 ? 2770 ng/mL. The intra- and inter-day precisions were less than 10.5% for each QC level, and the inter-day relative errors were 2.0%, 5.6%, and 2.8% for 9.22, 184, and 2490 ng/mL, respectively. The validated method was successfully applied to the quantification of eptifibatide concentration in human plasma after intravenous (i.v.) administration of a 270-μg/kg bolus of eptifibatide and i.v. administration of eptifibatide at a constant rate of infusion of 2 μg/(kg min) for 18 h in order to evaluate the pharmacokinetics.     

Short-time response in root morphology of Vallisneria natans to sediment type and water-column nutrient

Rooted submerged macrophytes can absorb significant amounts of nutrients from both sediment and water. We investigated root morphology of Vallisneria natans in mesocosm plastic bins, in response to three types of sediment (sandy loam, clay, and a 50:50 (v/v) mixture of the two sediments) and two levels of water-column nutrient (well water and nutrient medium). Compared to the plants grown in the clay or mixed sediments, root diameter decreased (0.39–0.41 versus 0.36–0.37 mm) but total root length per plant increased (0.87–1.27 versus 1.14–1.62 m) when grown in sandy loam. Increase of nutrient availability in water column led to decreased specific root length (306–339 versus 258–281 m g⁻¹). However, both sediment type and water-column nutrient had no impacts on root number (ranged from 19 to 24 number of roots per plant). Root weight ratio, root:leaf mass ratio and root:leaf length ratio generally decreased with enhanced nutrient availability in sediment or water. Plant growth was affected by sediment type alone (P < 0.05), rather than water-column nutrient (P > 0.05). However, plant N and P contents were significantly impacted by both sediment type (P ≤ 0.001) and water-column nutrient (P < 0.05). Increase of nutrient availability in sediment or water led to increased plant N (ranged from 2.47 to 4.77 mg g⁻¹) and P concentrations (ranged from 42.8 to 62.0 mg g⁻¹). These results indicate that considerable variation in root morphology of V. natans exists in response to the fertility of the sediment it is rooted in.