The 1RT and 2RT groups participated in a progressive high intensi

The 1RT and 2RT groups participated in a progressive high intensity protocol using a weights machine and free weights for resistance with a training regimen of 2 sets of 6 to 8 repetitions for arm and leg exercises. The BAT group’s

selleck chemical program consisted of exercises for stretching, range of motion, pelvic floor and balance, and relaxation techniques. Outcome measures: The primary outcome was change in the executive cognitive function of selective attention and conflict resolution as measured by the Stroop test at 6 and 12 months. The Stroop test assesses the time taken to name words of colours typed in incongruent ink colours. Secondary outcome measures were cognitive functions of set shifting and working memory, whole-brain volume, and functional measures of gait speed and muscular performance. Results: 135 participants (87%) completed the study and were included in the analysis. At 6 months there was no between-group difference but at 12 months, task performance in the Stroop test had improved by –2.9 s in the 2RT group compared to BAT (95% CI –12.2 to –0.8) and –4.3 s in the 1RT compared to BAT (95% CI –13.8 to –2.5) representing improvement of 11% and 13% in 2RT and 1RT groups, respectively, and deterioration of 0.5% in the BAT group. Peak quadriceps muscle power increased by 13% in the 2RT group, but decreased by 8% in 1RT

and 16% in the BAT group. There was a small but significant reduction in whole brain volume in 1RT and 2RT compared with BAT. The groups did not differ significantly on the remaining secondary outcomes. Conclusion: Twelve months of once or twice-weekly resistance training can improve almost Afatinib concentration the cognitive functioning of older women living in the community. This randomised controlled trial (RCT) contributes to the growing body of literature showing that physical activity can improve cognitive function in cognitively healthy

older adults (Angevaren et al 2008). Liu-Ambrose and colleagues demonstrated that only one 60-minute session of supervised progressive resistance training per week for 12 months improved participants’ selective attention and conflict resolution in comparison to a twice weekly balance and tone training control group. This improvement was greater in the once weekly resistance training group than in the twice weekly group. However, the authors did not offer any explanations for this dose effect. The authors conclude that the positive cognitive effect may be selective for executive functions since other secondary cognitive outcomes did not improve, however the battery of cognitive tests used was small. Furthermore the authors reported that the improvement in executive functions was significantly associated with increased gait speed. This important finding adds further weight to the relevance of gait speed for cognitive function and survival (Soumaré et al 2009, Hardy et al 2007).

1) In many comparisons, the difference between LAIV and placebo

1). In many comparisons, the difference between LAIV and placebo recipients was statistically significant. In study 3, responses were observed after a single dose but the differences compared to placebo recipients were more apparent after receipt of 2 doses of vaccine. Among subjects receiving only 1 dose of vaccine in year 1, a

greater difference versus placebo was observed at GW786034 molecular weight the second versus first sample collection (approximately 2 months versus 1 month postvaccination). When the percentage of subjects with a ≥4-fold increase was evaluated, a similar pattern was observed, although response rates were lower. For LAIV and placebo recipients respectively, response rates were 26–39% versus 12–30% for A/H1N1, 33–48% versus 20–27% for A/H3N2, and 46–59% versus 14–38% for B. When subjects were stratified by baseline

serostatus, similar IgA responses were observed among seronegative and seropositive subjects. Postvaccination GMFRs for strain-specific IgA ratios among LAIV recipients after 2 doses of vaccine in year 1 ranged from 1.4 to 6.2, compared to 0.5–2.0 among placebo recipients (Table 1). In year 2, GMFRs ranged from 1.2 to 4.6 among LAIV recipients and 0.8–2.2 among placebo recipients (Table 1). Postvaccination GMFRs in absolute strain-specific IgA, uncorrected for total IgA, trended higher than postvaccination Selleck Adriamycin GMFRs in strain-specific IgA ratios. Among LAIV and placebo recipients, total IgA increased from prevaccination to postvaccination by 1.0- to 2.4-fold in year 1 and 0.7- to 1.2-fold in year 2 (Table 2). Year 1 of study 3 was responsible for the greatest observed responses for LAIV and placebo recipients and 4 of the 5 statistically significant GMFRs. Because of the observed increases in total IgA from prevaccination to postvaccination in both placebo and vaccine recipients in year 1 of study 3, subject-level data by site were reviewed. In study 3, but not in studies 1 and 2, the total IgA content in year 1 prevaccination samples was lower among the initial subjects enrolled

at sites and higher among subjects enrolled subsequently; because linear regression analysis controlling for site showed that total IgA content in prevaccination samples increased significantly over calendar time in study 3 (P = 0.002). Across studies, data for both HAI and IgA responses following receipt of 2 doses was available for 392 LAIV recipients and 213 placebo recipients in year 1. Four-fold increases in HAI antibody titer for A/H1N1 were observed for 61% of LAIV recipients compared to 13% of placebo recipients (P < 0.001); for A/H3N2 and B, responses were 74% versus 16% (P < 0.001) and 76% versus 12% (P < 0.001) for LAIV versus placebo recipients, respectively. Among LAIV recipients, IgA responses were more frequently seen among subjects with an HAI response. Across studies, IgA responses to A/H1N1 were observed among 48% of subjects with a 4-fold HAI response, compared to 33% of those without a 4-fold HAI response (P < 0.001).

In this study factorial design based on the response surface meth

In this study factorial design based on the response surface method was adopted to optimize effective factors for the release of the drug from the microspheres. Analysis of variance (ANOVA) and all statistical analysis were also performed using the software. Calculation of the effects was performed. The significant effects would constitute the model. The F-value was then calculated by comparing the treatment variance with

the error variance. The multiple correlation co-efficient was calculated which is a measure of the amount of variation about the mean, which is explained by the model. The main effects and interactions are plotted and results interpreted. All assumptions underlying the ANOVA are checked. For statistical purposes, the assumption is selleck chemical made that residuals are normally distributed check details and independent with constant variance. Eudragit microspheres of tinidazole were successfully prepared by emulsion solvent evaporation technique. The results shown in Table 3 indicates that optimum concentration of surfactant (1% w/v) and stirring speed (2500 rpm) showed higher percent of entrapment

efficiency while change in stirring speed up to optimum range and change the surfactant concentration up to optimum range change the percent entrapment efficiency (Table 4). Also the percentage yield of microspheres of all formulations was found in the range of 68.6–77.5 %. The microspheres were characterized for particle size analysis within range of 585.6 μm–986 μm (Table 4). The FTIR spectra of

pure drug, Eudragit and tinidazole microspheres were shown in (Fig. 1). It shows that no incompatibility reactions took place between drug and excipients. The value of angle of repose of formulation within the range of 17°.97′ ± 0.51–26°.22′ ± 0.22 indicating Dichloromethane dehalogenase good flow properties for the microspheres. The bulk density values ranged between 0.148 ± 0.001 and 0.278 ± 0.004 gm/cm3. The tapped density values ranged between 0.206 ± 0.002 and 0.401 ± 0.03 (gm/cm). The Carr’s index values ranged between 17.55 ± 3.0 % and 42.80 ± 1.2% and Hausner’s ratio values ranged between 1.2140 ± 0.04 to 1.7148 ± 0.08 which can described by Table 5. The in vitro release study was carried out by buffer change method to mimic the GIT environment. Drug release for the initial 2 h i.e. in 0.1 N HCL, the drug release was found to be low in all cases. Then drug release is found 92.74% at the end of 8 h in pH 7.4 phosphate buffer, shown in Fig. 2. The produced microspheres were spherical, non aggregated with rough and porous surface, as shown in scanning electron micrographs (Fig. 3). The surface of microspheres was rough due to arising as a trace of solvent evaporation during the process. ANOVA results indicated that concentration of surfactant and stirring speed showed individual effect on % drug release. There is no significant interaction between surfactant and stirring speed.

05) with range of motion at six months ( Table 3) However, only

05) with range of motion at six months ( Table 3). However, only 1% to 17% of the variation in range of motion was explained by these predictors. Multivariate analysis: As several of the candidate predictors were highly correlated with each other, only five of the candidate

predictors (age, pre-morbid function, strength, spasticity, and pain) were entered into the multivariate analysis ( Table 4). Muscle strength was the only predictor selected in more than 80% of bootstrap samples. Even when all five predictors were forced into the model, they only explained 6% to 20% of variation in contracture development (adjusted r2 of full model for elbow extension = 0.19, wrist extension = 0.20, ankle dorsiflexion = 0.06). This study provides the first robust estimates of the incidence of contractures in a representative sample of patients presenting to hospital with stroke. The data indicate that contractures Endocrinology antagonist are common; half the cohort (52%) developed at least one contracture. Contractures are most common at the shoulder and hip, and more common in those with moderate to severe strokes (NIHSS > 5). The data do not provide any further guidance on which patients 5-FU nmr are most susceptible to contractures. It is widely believed that factors such as strength, pain, spasticity, and severity

of stroke help predict contractures yet in our models none of these factors explain more than 20% of variation in range of motion at six months. Few cohort studies have investigated the incidence of contractures after stroke (Fergusson et al 2007). Current estimates of the incidence proportion of contractures vary from 23% to 60% in the year after stroke (Pinedo and de la Villa 2001, Sackley et al 2008). Direct comparisons of our estimates to these studies are difficult due to the

difference in characteristics of cohorts and lack of detailed information regarding measurement and definitions of contractures. However, our estimates broadly align with those of earlier studies. Our estimates may have been higher if we had measured incidence of contractures at one year rather than six months after stroke. It is not clear why we were not better able to predict those susceptible to contractures. The predictors were chosen because they are believed to be associated with the development of contractures. Interestingly, even spasticity, the which is widely believed to predict contractures (Ada et al 2006), was not a good predictor (it was selected in only 25% to 48% of bootstrap samples). This was despite the high incidence of spasticity at baseline (25 elbows, 11 wrists, 21 ankles). Pain was arguably a better predictor than spasticity (selected in a greater number of bootstrap samples than spasticity) even though few joints were painful (4 elbows, 2 wrists, 6 ankles). It is also possible that our failure to predict contractures could have been due to errors associated with the measurement of either predictors or outcomes (contractures).

The saponins could be responsible for the observed antidiabetic,

The saponins could be responsible for the observed antidiabetic, lipid and cholesterol lowering activities. 11 From the results obtained correlation among antiradical and α-amylase inhibitory potential was established. It could be concluded that the

aqueous and ethyl acetate fractions possess significant antiradical property and inhibitory potential on α-amylase. All authors have none to declare. The authors are thankful to Prof. Ashok Kumar, Vice-Chancellor, C.S.J.M. University, Kanpur for providing the necessary facilities at University Institute of Pharmacy. “
“Hyperlipidemia is the major cause of atherosclerosis and atherosclerosis-associated conditions, such as coronary heart disease (CHD), ischemic cerebro-vascular disease, and peripheral vascular disease. Although the incidence of these atherosclerosis-related events Selleck BI6727 has declined in the United States, these conditions still account for the majority of morbidity and mortality among middle-aged and older adults.

The incidence and absolute number of annual events will likely increase over the next decades because of the epidemic of obesity and the aging of the U.S. population. Therefore, there is a great need for methods for treatment of lipid disorders, especially those which predispose a patient to cardiovascular problems such as myocardial infarction, angina conditions, stroke, coronary artery buy INCB024360 disease, etc.1 and 2 Fluvastatin sodium (FVS) is the first fully synthetic HMG-CoA reductase inhibitor approved for clinical lipid lowering therapy. FVS is subjected to extensive first pass metabolism in the liver and the plasma half-life of the drug is approximately 3 h with 40%–60% bioavailability. The physicochemical characteristics of drug like low molecular mass (411.46 g/mol) and log Po/w (3.24) favors molding of it in transdermal drug delivery system.3 Through literature review, it was revealed that so far no one

has attempted transdermal delivery or novel drug delivery of fluvastatin sodium. In the present research work, transdermal matrix patch was fabricated with use of FDA approved commercial acrylate-co-polymer based pressure sensitive adhesives. Resminostat Effect of different permeation enhancers, Eudragit polymer and matrix fillers were investigated.4 Fluvastatin sodium was a gift sample from Biocon Limited, India. Durotak 87-9301 (DT 9301) & Durotak 87-900A (DT 900A) were obtained from Henkel Ltd. (Salisbury NC, USA). Transcutol P (TC) was obtained from Colorcon Asia, Mumbai, India. Isopropyl myristate (IPM) and Oleyl alcohol (OLA) were purchased from S D Fine-Chem Limited, Mumbai. Oleic acid (OA), Propylene glycol (PG), Colloidal silicone dioxide (CSD) and Eudragit RL 100 (E RL 100) were obtained from Loba Chem pvt ltd, Mumbai, India.

Subjects were randomized

Subjects were randomized click here (1:1:1:1:1:1) to receive control vaccine at M0,1,6 or one of 5 different formulations/dose schedules of tetravalent vaccine: (i) one formulation with the same concentration of HPV L1 VLPs (20 μg each) and adjuvant system (AS04) as the control vaccine; (ii) two formulations with new adjuvant systems (AS01 and AS02) and containing half the amount of HPV-33 and -58 L1 VLPs (10 μg each) while maintaining the same amount of HPV-16 and -18 L1 VLPs (20 μg each); (iii) finally the AS01 formulation was also tested

using two different 2-dose schedules: classic 2-dose (M0,6) or accelerated 2-dose (M0,3). Subjects were followed for 6 months after the last vaccine dose. The trial was open with regard to dose schedule (2-dose or 3-dose) and was observer-blind within the 3-dose groups. Syringes were prepared and administered by qualified medical personnel not otherwise involved in the conduct of the study or in the assessment of symptoms. For both trials the randomization list was generated at

GlaxoSmithKline Biologicals SA using a standard Statistical Analysis System program; a randomization blocking scheme was used to ensure that balance was maintained. Vaccine allocation at all sites was performed using a central randomization call-in system on Internet. Trials were see more approved by the appropriate Independent Ethics Committee for each center and carried out in accordance with The Code of Ethics of the World Medical Association (Declaration of Helsinki). Written informed consent was obtained from subjects prior to the performance of any study-specific procedures, after the nature and consequences of the trial had been fully explained. Healthy women aged 18–25 years at the time of first vaccination who had had no more than 6 lifetime sexual partners were eligible for each trial. Subjects of childbearing potential had to have used adequate

contraception for 30 days prior isothipendyl to vaccination, have a negative pregnancy test, and continue contraceptive precautions for 2 months after completion of the vaccination series. Other standard eligibility criteria are detailed in the registry. All vaccines were developed and manufactured by GlaxoSmithKline Biologicals SA. The AS04 adjuvant system contains 3-O-desacyl-4’-monophosphoryl lipid A (MPL; 50 µg) adsorbed on aluminum salt (500 µg Al3+). AS04-adjuvanted vaccines were provided as a liquid suspension in individual pre-filled syringes for single use (0.5 mL). AS01E is an adjuvant system containing 25 μg MPL, 25 μg Quillaja saponaria Molina fraction 21 (QS21) and liposome. AS02W is an adjuvant system containing 25 μg MPL and 25 μg QS21 in an oil-in-water emulsion. For AS01 and AS02 vaccines, the HPV L1 VLPs were provided as a lyophilized pellet which was reconstituted with 0.5 mL adjuvant immediately prior to administration. All vaccines were administered (0.

The reaction was detected with a secondary antibody HRP conjugate

The reaction was detected with a secondary antibody HRP conjugated anti-human IgG (Chemicon, Australia) and enzyme substrate solution, TMB (3,3′,5,5′-tetramethylbenzidine, KPL, USA) followed by a 1 M H3PO4 stop solution. The absorbance (OD) was measured at 450 nm (reference filter 630 nm) on a Bio-Tek Elx808 (Bio-Tek Instruments, USA). OD was converted to antibody concentrations (μg/ml) using KCJunior software (Bio-Tek Instruments, USA). Sample dilutions were analyzed in duplicate and three controls (low, medium and high) were included on each plate to assess assay performance and inter-assay

variation. Results from Ipatasertib cell line an inter-laboratory comparison between Wyeth Vaccines and the KTL Finland laboratory demonstrated a good correlation in measurement of serotype-specific antibody concentrations [28]. Laboratory staff members were

blinded to the group allocation of each serum sample. Cleaned data were exported to Stata version 9.0 (Stata Corporation, College Station, Texas) for analysis. Serotype-specific CH5424802 ic50 antibody concentrations by ELISA were log transformed (to base e) to calculate GMC. Comparisons of pre- and post-mPPS GMC and between group comparisons were performed using a paired t-test and two sample t-test, respectively. Simple and multi-variable regression analyses were undertaken to adjust for both the pre-mPPS log antibody concentration for all 23 serotypes, and the number of PCV doses either administered for all seven PCV serotypes. A p-value of <0.05 was considered statistically significant. The primary endpoint was serotype-specific

GMC response to mPPS at 18 months of age in children who had received the 12 months 23vPPS compared to children who had not received the 23vPPS. We defined hyporesponsiveness to a particular serotype as a significantly lower GMC observed post-mPPS, in the 12 month 23vPPS group compared to the no 12 month 23vPPS group, controlling for pre-mPPS antibody levels, using multivariable regression analysis. To prevent an inflated type 1 error due to multiple comparisons, and obtain a single p-value for the null hypothesis of mPPS having no impact on the antibody response to any of the 23 serotypes, a joint test of all the regression coefficients from the aforementioned multivariable regression analysis was performed [29]. The study was approved by the Fiji National Research Ethics Review Committee and the University of Melbourne Human Research Ethics Committee. There were 552 children enrolled in the study (Fig. 1) which represent a consent rate of 30.5%. There were 90 (16.3%) withdrawals and no child was withdrawn due to an adverse event resulting from administration of any of the vaccines. Characteristics and the number of children randomized to the eight groups are shown in Table 1. Following the 12 month 23vPPS, there were significantly higher GMC (each p < 0.001) for all PCV serotypes.

Deficits in spontaneous spatial recognition and working memory pe

Deficits in spontaneous spatial recognition and working memory performance have been reported (Vallee et al., 1999). Additionally, PNS offspring have been shown to have impaired prepulse-inhibition responses and increased locomotor activity after amphetamine administration, Selleck SCH772984 both of these phenotypes have been associated with development of a schizophrenia-like phenotype (Koenig et al., 2005). There is a large body of literature on the effects of PNS on

stress responsivity and hypothalamus-pituitary-adrenal (HPA)-axis functioning. Exposure to prenatal stress has been shown to alter corticosterone levels throughout the circadian cycle; in adult male rats increased corticosterone levels have been found at the end of the light phase, a time

period where typically the highest corticosterone levels are observed (Koehl et al., 1999). Consistent with heightened corticosterone levels, hypertrophy of the adrenals has been reported (Lemaire et al., 2000). Furthermore, several studies showed increased glucocorticoid levels and associated decreased negative feedback of the HPA-axis after acute stress (Koehl et al., 1999, Henry et al., 1994, Barbazanges et al., 1996 and Maccari et al., 1995). At the level of the brain, alterations in the glucocorticoid system have been shown; the binding capacity of both the mineralocorticoid receptor and the glucocorticoid receptor were decreased in PNS offspring (Koehl et al., 1999 and Maccari et al., 1995). In addition to effects CYTH4 on stress-related traits, prenatal stress has also been reported to this website affect the metabolic phenotype of the offspring. Lesage and colleagues showed that chronic restraint stress during the last week of pregnancy induced hyperphagia and impaired glucose tolerance in adult male offspring (Lesage et al., 2004).

Similar to the human studies, PNS offspring had lower birth weights than control, which may have contributed to their metabolic phenotype later in life. Metabolic syndrome-predisposing effects of PNS in rats were confirmed in a study that used a variable stress paradigm during the last week of pregnancy and in this study differences in birth weight were not found. Tamashiro and colleagues showed that offspring of prenatally stressed dams were also impaired in an oral glucose tolerance test. However, these differences were only apparent in PNS rats that were weaned onto a high fat diet (Tamashiro et al., 2009). Stress exposure earlier during pregnancy seems to have some contrasting effects, offspring of mice exposed to stress during the first week of pregnancy were shown to gain less weight on a high fat diet, whereas they were hyperphagic on a standard chow diet (Pankevich et al., 2009). This suggests that the timing of the stress is an important variable in the metabolic risk associated with prenatal stress exposure.

These antibodies also detected bands of the predicted size for VP

These antibodies also detected bands of the predicted size for VP2 (∼110 kDa), VP5 (∼60 kDa) and VP7 (∼38 kDa) in BTV-4(SPA2003/01) infected cell lysates by western-blotting (Fig. 1e, f, g). In contrast to expressed proteins that had been ‘CAPS-denatured’, antisera against the soluble amino terminal domain of VP2 contained NAbs with titres of 1.505–1.602 (Table 1), giving ≥50% plaque reduction. Lower titres of neutralising antibodies (0.301–0.477, P < 0.05) were found in antisera against the carboxy-terminal domain. Sera from mice immunised

with: VP2D1 + VP2D2; VP2D1 + VP2D2 + VP5Δ1−100; or VP2D1 + VP2D2 + VP5Δ1−100 + VP7, all neutralised the homologous BTV-4(SPA2003/01) at higher titres (1.806–2.408) but (as expected) failed to neutralise BTV-8 ( Table SB431542 research buy 1). Neutralising antibody titres generated by Balb/c mice immunised with VP2D1 + VP2D2 + VP5Δ1–100 or VP2D1 + VP2D2 + VP5Δ1–100 + VP7 were not significantly different, but were significantly higher (P < 0.05) than those immunised with VP2D1 + VP2D2 ( Table 1). Neutralising antibody (NAb) titres of 1.806–2.017 were detected in mice immunised with VP2D1 + VP2D2; with 2.017–2.408 in those immunised with VP2D1 + VP2D2 + VP5Δ1–100 or VP2D1 + VP2D2 + VP5Δ1–100 + VP7 (Table 1), supporting previous studies indicating that VP5 may play a significant role in generation of NAbs [38], selleck [39] and [40]. There was no statistical difference between immunisation with VP2D1 + VP2D2 + VP5Δ1–100, or VP2D1 + VP2D2 + VP5Δ1–100 + VP7,

but a significant difference compared to immunisation with VP2D1 + VP2D2 only (P < 0.05) ( Table 1). Sera from IFNAR−/− mice immunised with recombinant VP2D1 + VP2D2, VP5Δ1–100 and VP7, ether singly or in different combinations, all reacted with

BTV-4 by ELISA (Table 1). The specificity of the antibodies was also confirmed by immunofluorescence (supplementary figure). Sera from non-immunised mice did not neutralise BTV-4 nor show old cross reactivity with BTV-4 ELISA. Mouse survival times p.i. provide a relative measure of protection afforded by vaccination. Blood samples collected on days 2, 3, 4, 5, 7, 10 and 12 p.i., and tested. Mice immunised with VP2D1 + VP2D2, VP2D1 + VP2D2 + VP5Δ1–100 or VP2D1 + VP2D2 + VP5Δ1–100 + VP7, then challenged with BTV-4, all survived until the end of the experiment on day 52 (12 days p.i.) (Fig. 2A). Two mice immunised with VP2D1 + VP2D2 were positive (Ct value of 34) on day 4 p.i. with BTV-4. Because no virus could be isolated from blood on KC cells or by plaque assay using BSR cells (possibly reflecting the presence of neutralising antibodies), we calculated PFU-equivalents using the formula linking Ct values to PFU numbers. A low PFU-equivalents/ml was calculated (∼0.3–9). Two mice in each group immunised with VP2D1 + VP2D2 + VP5Δ1–100, or VP2D1 + VP2D2 + VP5Δ1–100 + VP7, were also potentially viraemic on day 5 p.i. (Ct values ∼39), although no virus could be isolated on KC cells or by plaque assay on BSR cells (Fig. 2B).

The commercially available tablets were purchased from the local

The commercially available tablets were purchased from the local market. Stock solution of 1000 μg/mL was prepared by accurately weighing 5.00 mg of MMF, transferred into a 5.0 mL clean and dry volumetric flask, and dissolved in methanol. The primary standard solution of concentration of 10 μg/mL was prepared by taking 10 μL stock solutions and diluted to 1.0 mL with methanol. Further a series of working standard solutions of different concentrations were sequentially diluted to the required PF-01367338 datasheet volume. The LC/MS/MS analysis was carried out on Applied Biosystems API 3200 triple quadrupole mass spectrometer attached to LC 20 Series Shimadzu Corporation (Kyoto, Japan), equipped with pump (Shimadzu

LC-10AT VP), auto sampler (Shimadzu SIL-HTC), degasser (Shimadzu FCV-10AL VP) and system controller (Shimadzu SIL-HTC ver 6.03) in NISHKA Scientific and Research Laboratories, Hyderabad. The chromatographic SB431542 molecular weight analysis was performed under isocratic conditions using 75% acetonitrile containing 2 mM ammonium acetate at pH 5.0 at a flow rate of 600 μL/min and Chromosil ODS-3, C18, 4.6 × 50 mm, 2.5 μm column. The ionization was carried out

by ESI. The source heater temperature was maintained at 300 °C. The analysis was carried out in multiple reaction monitoring (MRM) mode for the transition m/z 434 → 114 at collision energy 30 V. The mass spectral analysis was carried out by direct infusion of 10 μg/mL solution of MMF in to the ESI source at a flow rate of 10 μL/min along with the mobile phase flow rate of 600 μL/min. The obtained mass spectrum showed m/z 434 as a major ion which can be attributed to the MH+ ion of the analyte. This ion was subjected to collision induced dissociation (CID) using nitrogen as a collision gas. The collision energy was tuned in such a way that the intensity of MH+ ion was reduced to a minimum of 20%. The obtained mass spectrum after CID showed m/z 114 as a major fragment. Hence the transition m/z 434 → 114 was used to monitor the analyte peak in LC/MS/MS analysis. The ESI mass

spectra of MMF obtained before and after fragmentation were presented in Fig. 2 and Fig. 3 and respectively. Intra/inter day precision was calculated at three different concentrations of working standard solution of reference MMF by taking measurements of six replicates at each concentration on different occasions. Mean, standard deviation (SD) and percent of relative standard deviation (%RSD) were calculated at each concentration and found to be within the acceptable limits. The results of intra day and inter day precision were presented in Table 1. In proposed method, accuracy was determined at three different concentrations of working standard sample solution of MMF (Tablet) by taking measurements of three replicates at each concentration. The proposed method was found to be highly accurate. The calculated %RSD of peak area, weight found and percent of weight found were found to be 2.382, 0.133 and 0.153; 1.