For a negative control, homogenates were pre-incubated for 10 min at 37 °C with commercial inhibitors to caspase-1 (Ac-WEHD-CHO,

1 μM) or caspase-3 (Ac-DEVD-CHO, 1 μM), followed by the addition of the respective substrate. Activity was measured continuously over 90 min on a GENius Tecan Austria G.M.B.H. Spectrofluorimeter, ABT-199 using λex = 360 nm and λem = 465 nm. The peptide hydrolysis reaction velocities were expressed as units of fluorescence per min (RFU/min). Variance analysis (Two-way ANOVA) and Bonferroni post hoc were used to compare the estimative of neuronal cell numbers, including the right and left hemispheres. Data are presented as mean ± S.E. and differences were considered significant when p ≤ 0.05. One-way ANOVA followed by Tukey’s test was used to compare the activity of the different VX-770 mouse caspases. Data are presented as mean ± S.D. and differences were considered significant when p ≤ 0.05. Freeman-Halton extension for Fisher’s exact test (table 2X4) was used to compare the survival rates in different experimental

groups. All procedures were approved by the Local Ethics Committee (CEP. 1913/06) and are in accordance with the National Institutes of Health Guide for the Care and Use of Laboratory Animals. Every effort was taken to minimize the number of animal used and distress of the animals. A significant reduction of hippocampal neurons (CA1 and hilus) was observed in the group Pilo + Saline, when compared to the control group Saline + Saline (Table 1, Fig. isothipendyl 1). SE-induced neuronal loss in CA1 was completely prevented in rats treated with pyruvate plus oxaloacetate (Group Pilo + Pyr + Oxa). Treatment with pyruvate or oxaloacetate alone did not prevent neuronal loss in CA1. On the other hand, SE-induced neuronal loss in the hilus was prevented only in

rats that received pyruvate alone (Group Pilo + Pyr). Seven days after pilocarpine-induced SE, a significant increase in the caspase-1 and caspase-3 activity was observed in all experimental groups when compared to controls (p < 0.001) ( Table 1). Treatment with Oxa and Pyr + Oxa to rats presenting SE, reduced significantly the caspase-1 activation in the hippocampus whereas have no effect on caspase-3. The administration of pyruvate or oxaloacetate did not change seizure semiology and severity during SE in experimental rats. Mortality during SE was 34% in the group Pilo + Saline, 29% in the group Pilo + Pyruvate, 7% in the group Pilo + Oxa and 25% in the group Pilo + Pyr + Oxa. Fisher’s exact test did not show significant differences amongst groups (P = 0.38). In humans, several brain insults are characterized by excessive Glu brain levels. These include acute disorders such as stroke, traumatic brain injury, bacterial meningitis and prolonged seizures (Castillo et al., 1996, Spranger et al., 1996, Zauner et al., 1996, Men et al., 2000 and Ma et al.

MF: Declares no potential conflict of interest. MCJM is a Wellcome Trust Senior Research Fellow, and acknowledges the Wellcome Trust for research Funding. “
“To date, more than 150 human papillomavirus (HPV) types have been completely sequenced (Fig. 1), along with over 60 animal papillomaviruses (PV) (see Papillomavirus KPT-330 purchase Episteme (PaVE); http://pave.niaid.nih.gov/#home) and [1]). The presence of PVs in mammals, as well as in various diverse hosts, including birds, turtles and snakes, suggests that they may be ubiquitously present amongst

present day amniotes (i.e., mammals, birds and reptiles) [2]. Papillomavirus types found in humans are divided into five genera based on DNA sequence analysis, with the different types having different life-cycle characteristics and disease associations [1], [3], [4] and [5] (Fig. 1). In recent years, it has become clear that many HPV types, including the majority of those contained within the Beta and Gamma genera, cause only asymptomatic infections in immunocompetent individuals and can be detected in skin swabs, and for some Gamma types, also in mucosal rinses [6], [7], [8] and [9]. selleck inhibitor Such viruses are well adapted to their host, and can in most instances complete their life-cycle and be maintained in the population without causing any apparent disease [5] and [10]. Such characteristics suggest that the PV-host

interactions are very old, and that over time, this has lead to a balance between viral replication and immune tolerance [11]. Indeed, the evolutionary origins

of PVs can be traced to the origin of the amniotes themselves (approximately 350 million years ago [12], [13] and [14]), with many evolutionary mechanisms contributing Metalloexopeptidase to their current diversity, including host/virus co-evolution, recombination, host-switching and the possible extinction of the PV lineage in some hosts [15]. In humans, the PV types that cause visible papillomas are generally of most concern for the individual, especially when they occur at oral or genital sites and are persistent. Approximately one-third of individuals who present for treatment with genital warts will still have their lesions 3 months later, with recurrence after treatment being a significant problem [16]. The low-risk Alpha types that cause these lesions (typically the Alpha 10 species [e.g., HPV6 and 11]; Fig. 1) are also implicated in the development of respiratory papillomatosis (RRP) [17]. Although rare, juvenile RRP (which affects around 4 per 100,000 children [18], [19] and [20]) is a serious condition that can only be managed by repeated surgery, and can progress to cancer in a small percentage (approximately 5%) of persistently infected individuals where the infection spreads to the lung [20] and [21]. The various types of epithelial disease that HPVs cause (i.e.

This study was carried out in the Cardiothoracic Surgical Unit, Auckland City Hospital, a tertiary click here referral hospital in New Zealand. One control group participant inadvertently received physiotherapy intervention as per the experimental group until discharge

from hospital. Another control group participant required physiotherapy input for a postoperative neurological complication, including transfer to a stroke rehabilitation unit, however as the neurological problem was cerebellar, this did not include specific shoulder and thoracic cage exercises. There were no reports of additional shoulder and thoracic cage exercises implemented during the inpatient phase for experimental group participants beyond those in the protocol. Two participants from each group reported that they had independently sought

treatment for problems related to their shoulder on the operated side following discharge from hospital. Data from all these participants have been analysed using intention-totreat principles. Experimental group interventions were provided see more as scheduled on 81% of occasions during the inpatient phase of the trial. For the experimental group, the median (range) number of physiotherapy treatment sessions received was 6 (1 to 18) and the median (range) total physiotherapy time per participant in 15-minute units of service was 12 (2 to 47) units. For the 76 randomised participants, data on pain, shoulder function and quality of life were obtained 83% of the time. Missing data most frequently resulted from nonreturned or incomplete questionnaires. For the subgroup of 47 participants who were scheduled to participate in measures of range of motion and strength, data were obtained 82% of the time. Missing data most often resulted from unwillingness or inability to attend for measurement. Exercise diaries were completed by only 8 (19%) of the 42 experimental group participants, so data from the diaries have not been reported. The physiotherapists who acted as independent assessors were asked to report any episodes of unblinding to group allocation. Five reports of inadvertent unblinding were received from the 122 follow-up assessment occasions (4%):

2 of these episodes occurred at the time of discharge, and 3 episodes occurred at the 3 months ADP ribosylation factor postoperative follow-up. When unblinding occurred, an alternative blinded assessor performed the outcome measures on all subsequent occasions. Group data at baseline and follow-up are shown in Table 2 for pain and range of motion and in Table 3 for muscle strength, shoulder function and quality of life. Individual data for all outcomes are provided in Table 4 (see eAddenda for Table 4). The experimental group had significantly less shoulder pain at discharge than the control group, by 1.3 units (95% CI 0.3 to 2.2). The experimental group also had significantly less total pain than the control group at discharge, by 2.2 units (95% CI 0.2 to 4.3).

The Committee also established a sub-committee for the investigation of vaccine-related injuries, which was separated from the KACIP

and became the Advisory Committee on Vaccine Injury Compensation in 2003. Committee members are appointed to 2-year terms that all begin at the same time, and thus a new committee is formed every 2 years. However certain officials, who serve as a result of their position within the government will remain on the Committee for as long as they remain in their position (see next section). Despite this intention, the duration of the current – seventh – committee, which was formed in October 2007, has been extended www.selleckchem.com/products/fg-4592.html to a third year, because of the many issues it has been dealing with that still need to be resolved. This is the first time that the Committee’s term has been extended and the terms will go back to 2 years

in 2010. Among the items on the agenda of the current committee have been: a review of national immunization strategies; the control of measles; how to control a hepatitis A outbreak; the control of varicella and mumps; whether to change the strain of Bacillus Calmette–Guérin (BCG) vaccine and route of administration (from intradermal to transdermal); and the issue of subsidizing the cost of Expanded Program of Immunization (EPI) vaccines provided through the private sector, through which the majority of immunizations in Korea are given. Based on a recommendation by the KACIP, the Government has decided to partially Ku-0059436 solubility dmso subsidize the

cost of all EPI vaccines administered at private health facilities that agree to participate in this program, starting in 2009 (with parents now paying 70% instead of 100% of the vaccine cost). The KACIP consists Ketanserin of a Chairperson and specialists in internal medicine, paediatrics, obstetrics, microbiology, preventive medicine and nursing. The Committee also includes a representative from a consumer group, the Director of Disease Prevention at the Korea Centers for Disease Control and Prevention (KCDC), and the Director of Biologics at the Korea Food and Drug Administration (KFDA). Apart from the two government officials mentioned above, all other members usually come from the affiliated organizations shown in Fig. 1, which each nominate one member. The total number of Committee members is usually around 15. The Secretariat of the Committee is within the KCDC, which funds, organizes and prepares for the meetings, and at whose headquarters the meetings are held. The Chairperson rotates every term (i.e., 2 years) and can be selected from any field or affiliated organization. Over the years, Committee members have made recommendations to include more female members, representatives from civil society, and people from rural areas, though to date there are no minimum requirements or quotas for representation of these groups.

First, numerous studies have shown that despite a lack of sarcolemma depolarisation or crossbridge cycling, a stretched muscle cell can not be considered metabolically dormant. In 1932, Feng (1932) showed that a passively stretched in vitro muscle was metabolically active. He found that passively stretched muscles exhibited increased heat production and oxygen consumption. Later research corroborated these findings; Clinch (1968) reported increased heat production, while Whalen and colleagues (1962) and Barnes (1987) added reports of increased oxygen consumption. In other related studies, passive stretch increased carbon dioxide production

( Eddy and Downs, 1921), increased glycogen breakdown RAD001 ( Barnes and Worrell, 1985), increased lactic acid production ( Barnes, 1987), and decreased phosphocreatine concentrations ( Barnes, 1987). Since increased metabolic activity is related to increased activation of the adenosine monophosphate kinase (AMPK) facilitated glucose transporter (GLUT 4) activation pathway ( Dohm, 2002), it is possible that the increased metabolic activity accompanying passive muscle stretching could have activated the incorporation of GLUT 4 into the stretched muscles. Other research also points to the possibility

of stretching increasing GLUT4 incorporation. For instance, protein kinase B activity Proteasome inhibitor partially controls GLUT 4 incorporation and activation, and Sakamoto and colleagues (2003) found that protein kinase B was stimulated by passively stretching isolated muscles for ten minutes. Second, mitogenactivated protein kinase activity stimulates muscle cell glucose uptake (Ho et al 2004), and the activity of mitogenactivated protein kinases directly reflects the magnitude of the mechanical stress (ie, actively or passively generated

Isotretinoin tension) applied to the muscle (Martineau and Gardiner, 2001). Third, exercise-induced increases in nitric oxide result in increased glucose transport (Roberts et al 1997), and nitric oxide released from excised soleus muscles can be increased 20% by a single two-minute passive stretch (Tidball et al 1998). Finally, ischaemia can increase GLUT 4 translocation to the sarcolemma as well as increasing glucose uptake (Sun et al 1994, Young et al 1997), and passive stretching has the potential to cause ischaemia (Poole et al 1997, Wines and Kirkebo 1976). Wisnes and Kirkebo (1976) found an increased resistance to blood flow during passive stretching. In addition, Poole and colleagues (1997) reported that muscle stretching reduces bulk blood delivery, alters capillary flow dynamics, and impairs blood tissue oxygen exchange. Regardless of the responsible mechanisms, it is clear that passive static stretching had a significant positive effect on blood glucose levels.

L’HTPPNN a été décrite après l’utilisation des IRS par la femme enceinte et reste une forme d’HTP grave selleck chemicals avec une mortalité élevée. Dans la classification des HTP Dana Point (2008), l’HTPPNN faisait partie du groupe 1, mais dans la nouvelle classification elle a été encadrée dans un groupe 1”, car elle présente plus de différences que de similitudes avec les HTAP du groupe 1 [1] and [21]. C’est probablement la forme la plus fréquente d’HTP. À part les valvulopathies, le mécanisme le plus

souvent retrouvé consiste en une dysfonction diastolique du ventricule gauche qui va entraîner une élévation des pressions de remplissage de celui-ci, une augmentation de la pression auriculaire gauche et, par conséquence, une augmentation passive de la pression artérielle pulmonaire [31]. Sur le plan hémodynamique, ces patients avec une HTP post-capillaire « pure » ont une PCP > 15 mmHg et un gradient de pression diastolique (GPD = PAPd-PCP) < 7 mmHg. L’objectif pour ces patients est l’optimisation de leur traitement cardiologique en essayant

de corriger leurs facteurs de risques. En fonction des résultats du cathétérisme cardiaque droit, les patients ayant un GPD > 7 mmHg ont une HTP post-capillaire avec une composante pré-capillaire et les traitements spécifiques de l’HTAP ont été déjà testés dans de petites Androgen Receptor antagonist études sans résultats concluants jusqu’à présent [1], [31] and [32]. Les mécanismes impliqués dans cette forme d’HTP sont soit une hypoxie alvéolaire, conséquence d’un apport insuffisant en oxygène, soit une vasoconstriction hypoxique, mécanisme réflexe dans les maladies respiratoires chroniques obstructives ou restrictives. En fonction de l’hémodynamique artérielle pulmonaire, on distingue trois groupes de patients avec des maladies respiratoires chroniques : (i) patients sans HTP (PAPm < 25 mmHg), (ii) patients avec une HTP (PAPm > 25 mmHg), et (iii) patients avec

une HTP sévère (PAPm > 35 mmHg ou PAPm > 25 mmHg et Index Cardiac < 2 l/min/m2) [33]. Concernant l’HTP, l’utilisation des termes « proportionnée » ou « disproportionnée » n’est pas recommandée. Les patients ayant une HTP sévère sont peu nombreux et nécessitent une évaluation exhaustive sur le plan fonctionnel, respiratoire, gazométrique et de l’imagerie thoracique below dans des centres experts. Jusqu’à présent, il existe peu de données concernant l’utilisation des traitements spécifiques de l’HTAP pour ces patients et, par conséquence, leur utilisation doit être limitée à des situations particulières. L’hypertension pulmonaire post-embolique (HTPPE) est liée à la persistance et l’organisation fibreuse des caillots après une ou plusieurs embolies pulmonaires aiguës. Cette forme d’HTP est de plus en plus diagnostiquée et est potentiellement curable en cas d’obstruction vasculaire proximale accessible à une thrombo-endartérectomie [34]. Tous ces patients doivent être évalués sur le plan hémodynamique et de l’imagerie, dans des centres experts, afin de pouvoir décider de l’opérabilité.

In addition to physiological repercussions, social defeat and VBS stress engender behavioral disturbances that are strikingly isomorphic to symptoms of clinical depression. After exposure to social defeat using the resident-intruder paradigm, rats that adopted a passive coping response (SL rats) in the face of repeated brief exposure to social stress exhibited enhanced susceptibility to displaying depressive-like behaviors, as indicated by increased immobility in the Porsolt forced swim MEK activation test (Wood et al., 2010), and decreased sucrose preference as well as increased social anxiety (unpublished findings), while

the LL phenotype remained generally resistant to these changes. The impact of coping strategies and dominance/submissive roles on stress-related pathology has also MI-773 manufacturer been demonstrated following social stress in tree shrews. In nature, when tree shrews fight, the subordinated animal must leave the territory. However seminal studies by Von Holst (1972) forced the subjugated animal to be in constant visual and olfactory contact with the victor. Under these conditions, the subordinate animal spent the majority

of the day lying motionless in the corner of the cage and many of them eventually died. In a more recent, related model of social stress in tree shrews, subordinate animals exhibit reductions in general motor activity, grooming, and food and water intake (Kramer et al., 1999). Similarly, subordinate rats in the VBS also demonstrate reduced food intake and exaggerated weight loss, decreased sexual and social behaviors, and altered sleep cycles (Blanchard and Blanchard,

1989). Behavioral disturbances and dysfunction within the Cediranib (AZD2171) HPA axis are reported as persistent outcomes and mimic maladaptive changes seen in people with psychiatric diseases (Wood et al., 2010, Bhatnagar and Vining, 2003, Buwalda et al., 1999 and Stefanski, 1998). These studies emphasize how a variation in coping response influences the pathogenic potential of social stress. Gender differences in both prevalence and symptomatology of affective disorders are well-established (Garber, 2006). Women display up to two-fold higher rates of depression, anxiety and seasonal affective disorders than men (Kessler et al., 1994). Higher suicide rates are found in men while increased numbers of suicide attempts are found in women (Hawton, 2000). Depressed women are also more likely to display atypical symptoms than men, including weight gain, increased appetite and increased sleep (Rappaport et al., 1995). Considerable sex differences exist in the social relationships of adolescent humans (described more below) and adult humans. Current theory posits that adult females exhibit affiliative behavior (a “tend and befriend” response) whereas males exhibit more of a fight or flight response to stress (Rose and Rudolph, 2006).

For AHSV serotypes 1, 3, 7, 8 and 9, open reading frames based on amino acid sequences of VP2 proteins (GenBank accession number: CAP04841; U01832; AAN74570; ABI96883, respectively), were designed for optimized expression in insect cells

(Gene Art, Regensburg, Germany). VP2 genes were amplified by PCR with specific primers containing BamHI or SmaI site for cloning purposes into the transfer vector pAcYM1 [27]. Recombinant vectors pAcYM1 with VP2 genes were purified and co-transfected into Sf9 cells with linearized baculovirus DNA (strain BAC10:KO1629), using Cellfectin® II Reagent (Invitrogen) according to the manufacturer’s instruction. On day six after transfection, 200 μl of the supernatants were transferred to fresh Sf9 cells in 12-wells plates. After learn more the first passage,

supernatants were transferred to fresh Sf9 cells every 3–5 days until virus infection was confirmed by light microscopy. The virus titer was measured by standard plaque assay using Sf21 cells. Recombinant http://www.selleckchem.com/products/s-gsk1349572.html baculoviruses expressing AHSV VP2 were used to infect Sf9 cells with a multiplicity of infection (moi) of 5. Infected cells were incubated at 28 °C for 72 h. Then, infected cells were harvested by centrifugation, washed with phosphate buffered saline (PBS) and pelleted by centrifugation. Cell pellets were suspended in 25 mM sodium bicarbonate (NaHCO3, pH 8.39) at 1.0 × 107 cells/ml. Cells were disrupted by dounce homogenization and after centrifugation at 6000 rpm for 3 min, supernatants containing soluble VP2 protein were collected. To examine the amount of VP2 proteins, soluble VP2 were mixed with equal volumes of SDS-PAGE sample buffer (10 mM Tris-HCl, pH 6.8, 2% (w/v) SDS, 2% β-mercaptoethanol,

20% glycerol, 0.05% bromophenol blue). After heating at 95 °C for 1 min, the samples were analyzed by SDS-PAGE with BSA as concentration standard and protein molecular weight standard (Page Ruler, SM0671, Fermentas). Concentrations of all samples were adjusted to 100 μg of VP2 per ml by 25 mM sodium bicarbonate and stored at −80 ° C until use. All experiments with live animals were performed under the guidelines of the European to Community (86/609) and were approved by the Committee on the Ethics of Animal Experiments of the Central Veterinary Institute (Permit numbers: 2011-042 and 2011-170). Adult female guinea pigs were purchased from a registered breeding farm for guinea pigs and were randomly divided into groups of six animals. Nine groups were immunized with VP2 protein from each AHSV serotype, two groups were immunized with cocktails of different combinations of VP2 proteins (one consisting of serotypes 1, 3, 7, 8 and other, serotypes of 2, 4, 5, 6, 9, respectively) and one group was immunized with phosphate buffered saline (PBS). Shortly before immunization, recombinant VP2 proteins or PBS in 1.5 ml were warmed to 37 °C and mixed with an equal volume of Montanide 206VG (Seppic) by vortexing.

15 according to Eq. (A.6). The log Ppara, log Pfilter, and log PABL were added as fixed contributions, as log P0 http://www.selleckchem.com/products/byl719.html and log Puptake were refined ( Appendix A.5) for the non-inhibitor and added-inhibitor (50 μM PSC833) sets. Both the intrinsic and the uptake permeability values appeared to be affected by efflux ( Table 3). The two sets were

then combined, with the repeated refinement yielding log P0 = −5.28 ± 0.04, log Puptake = −5.73 (kept fixed), and log Pefflux = −5.80 ± 0.04 for the non-inhibitor set and log Pefflux < −8 for the +50 μM PSC833 set. This suggested that efflux was essentially suppressed by the inhibitor. With the log Pefflux of −5.80, it was possible to rationalize the extent to which the individual-set refined log Puptake and BMS-354825 mw log P0 in the two sets were different. Fig. 4c and d shows colchicine and digoxin with added efflux inhibitor (checkered circle) and no-inhibitor (black circles). The addition of inhibitors increases the apparent permeability by nearly the same amount in both drugs, consistent with the suppression of efflux

transporter. To assess the ability to predict in vivo BBB permeability of a compound from permeability data measured using the PBEC model, P0 (in vitro) derived from our PBEC model permeability data was plotted against P0in situ (in vivo) derived from in situ brain perfusion data in rodents ( Fig. 5). Published data from other in vitro porcine BBB models were also included in the linear regression analysis. The r2 value Chlormezanone of 0.61 shows a good correlation for the pooled data. The in vitro blood–brain barrier

(BBB) model from primary porcine brain endothelial cells (PBEC) which shows a restrictive paracellular pathway was used for permeability studies of small drug-like compounds of different chemistry: acid, bases, neutrals and zwitterions. Assay at multiple pH was conducted for the ionizable compounds propranolol, acetylsalicylic acid, naloxone and vinblastine to plot permeability vs. pH. The pCEL-X software (Section 2.5 and Appendix A) was used for detailed permeability data analysis, including aqueous boundary layer (ABL) correction. The ABL was found to restrict propranolol permeability, which was also limited by low pore density of the Transwell®-Clear polyester filter membrane. The intrinsic transcellular permeability P0 showed good correlation with in situ data, indicating the predictive power of the in vitro model. Stirring helps to diminish the ABL thickness, but it cannot reduce it entirely. This is because the aqueous medium adjacent to the membrane surface is less mobile due to hydrogen bonds formed at the interface (Loftsson and Brewster, 2008). Hence, even vigorous stirring is unable to remove the ABL totally. Furthermore, excessive stirring is undesirable, since it can compromize tight junction integrity (cf., Zhang et al., 2006: 600 RPM). Application of the pKaFLUX method for ABL correction using pCEL-X proved useful particularly for ionizable compounds.

Few analytical methods have been reported for the verification of steroidal hormone drugs, especially for those with similar selleck chemical properties. In this paper, our aim was to develop a set of simple High-performance liquid chromatographic (HPLC) with evaporative light scattering detection12, 13, 14 and 15 (ELSD) and with dual

ESI ionization mass spectrometry (LCMS) methods are presented to distinguish and qualitatively analyze used to identify of Dexamethasone, Testosterone and Estrone (E1) in the combination form. Pure standards of Dexamethasone, Testosterone and Estrone (E1) were obtained from the Sigma–Aldrich, India. Organic solvents for chromatography were purchased in LCMS grade, ACS grade Acetonitrile was purchased from Honeywell-Burdick & Jackson (USA), water was obtained from ultra-purified from Elix Advantage 5 system equipped with Milli-Q Biocel (Millipore), all the chemicals used were of analytical reagent grade, and the solvents were of ACS. The purity of each reference standard was determined by HPLC PDA, ELSD detectors and dual ESI (LCMS). All solvents and samples were filtered through MILLEX FG (Millipore),

13 mm, 0.2 μM, fluoropore, non-sterile membrane sample filter paper before injecting into system. The analyses were performed using an Agilent 1200 Series HPLC system, equipped with a binary pump, an auto-sampler, a column oven, PDA detector and Selleck LDN 193189 a mass hunter software version B.02.01 (B2116.20) Ketanserin (Agilent Technologies, USA). Agilent 1260 Infinity Evaporative Light Scattering Detector (ELSD) instrument, operated by the Agilent 35900E multichannel interface which converts analog signal to digital (A/D) (Agilent Technologies, USA), was connected to the liquid chromatography for detection of steroids. The separation was carried out on a reverse phase Shodex C18, 3 μm, 4.6 × 100 mm at ambient temperature. The isocratic elution mode with a mobile phases

Acetonitrile and 0.1% formic acid in water and eluted by the following program at the flow 1 mL/min, runtime 6 min. The drift tube temperature for ELSD was set at 50 °C and the nitrogen flow rate was 53 psi. Agilent 6520 Quadrupole time-of-flight (Q-TOF) mass spectrometer. Coupled to an Agilent 1200 series HPLC system (Agilent Technologies, USA) is equipped with binary pump, auto sampler, thermostatted column compartment, variable wavelength detector, auto sampler thermostatted (G 1330B). The Agilent Q-TOF (6520) mass spectrometer is equipped with dual electrospray ionization (ESI) ion source, and the HPLC conditions were identical to those used for HPLC–ELSD analyses mentioned above. Mass spectra were acquired in positive mode with scan range from m/z 100 to 500 Da. The conditions of dual ESI source were as followed: drying gas (N2) flow rate, 30.