Alok S. Tripathi

Development and Validation of RP-HPLC Method for Simultaneous Estimation of Glimepiride and Sildenafil Citrate in Rat Plasma-Application to Pharmacokinetic Studies

A simple and sensitive method was developed for simultaneous estimation of Glimepiride (GLIM) and Sildenafil citrate (SIL) in rat Plasma by reverse phase high performance liquid chromatography (RP-HPLC). The drug samples were extracted by liquid-liquid extraction with 300 µl of acetonitrile and 5 ml of diethyl ether. Chromatographic separation was achieved on C18 column using methanol: water (85:15 v/v) as mobile phase at a flow rate of 1 ml/min and UV detection at 230 nm. The retention time of GLIM and SIL was found to be 2.5 and 4.0 min respectively with total run time of 7 min. The developed method was validated for accuracy, precision, linearity and recovery. The method was linear and found to be acceptable over the range of 100-12 000 ng/ml. The method was successfully applied for the analysis of rat plasma sample for application to pharmacokinetic.

Sildenafil, a phosphodiesterase type 5 inhibitor, attenuates diabetic nephropathy in STZ-induced diabetic rats.

Abstract Background: The present study evaluates the possible mechanism of sildenafil citrate (SIL) for the attenuation of renal failure in diabetic nephropathic (DN) animals. Methods: Diabetic nephropathy was induced by a single dose of streptozotocin (STZ) (60 mg/kg, i.p.) and confirmed by assessing the blood and urine biochemical parameters on the 28th day of its induction. The selected DN animals were treated with glimepiride (0.5 mg/kg, p.o.) and SIL (2.5 mg/kg, p.o.) for a period of 6 weeks. Biochemical parameters in blood and urine were estimated after the 29th and 70th day of the protocol for the estimation of the effect of SIL. Result: There were significant alterations in the blood and urine biochemical parameters in STZ-treated groups which confirmed DN. There was a significant decrease in the triglyceride level in the SIL-only-treated group on the 70th day of the protocol. The histopathology study also suggested that SIL treatment results in the improvement in the podocyte count in DN animals. Conclusions: The present study concludes that SIL improves the renal function by decreasing the triglyceride level and improving the podocyte count in DN animals.

Influence of Musa sapientum L. on pharmacokinetic of metformin in diabetic gastroparesis.

ObjectiveTo investigate the effect of Musa sapientum L. (MS) bark juice in diabetic gastroparesis and its effect on pharmacokinetic of metformin (MET).MethodsDiabetes was induced in rats by administering alloxan (120 mg/kg) saline solution and maintained for 8 week. All the 18 Sprague-Dawley rats were divided into three groups (n =6 in each group): normal control, diabetic control and MS bark juice. Assessment of diabetes was done by glucose oxidase-peroxidase method on the 3rd day of alloxan administration. The effects of MS bark juice (100 mL/kg) on gastric emptying time, intestinal transit time, contractility of fundus and pylorus as well as gastric acid secretion in chronic diabetic rats were observed after 8 weeks of alloxan administration. The effect of MS bark juice on the pharmacokinetic of orally administered single dose of MET (350 mg/kg) was evaluated on the 57th day of protocol. Any drugs that may reduce the blood glucose level or influence the fibrinolytic system were not used in this study.ResultsThe MS bark juice significantly reduced the blood glucose level in the diabetic rats (P<0.01). There was significant decrease in the pylorus motility and increase in the gastric emptying time, intestinal transit time, contractility of fundus, gastric acid secretion in the MS bark juice treated group (P<0.01). There was significant decrease in the time at which drug at a maximum concentration, half life of drug and increase in the maximum concentration of drug in the plasma of MET in MS bark juice treated group as compared to diabetic control group (P<0.01).ConclusionMS bark juice effectively manages diabetic gastroparesis and thereby improves the bioavailabilty of MET when administered with MS bark juice.

Validated Stability-indicating assay method for determination of Ilaprazole in bulk drug and tablets by high performance liquid chromatography

H metal ion pollution is a serious environment problem that has attracted more and more attentions in recent years. Mercury ion (Hg2+) and lead ion (Pb2+) are two of the most toxic metallic pollutants even at an extremely low concentration. Copper ion (Cu2+), an essential micronutrient element for human life, can cause adverse health effects when present in high concentrations. Considerable efforts have been devoted to the detection of heavy metal ions due to their high toxicity towards the ecosystem and human health. Unlike the traditional detection technologies, we have developed some simple and sensitive biosensors without the requirement of sophisticated instrumentation and skilled personnel. With the combination of DNAzyme, DNA machine and lateral flow biosensor, the visual instrument-free method offers a point-of-use solution for heavy metal ion analysis and provides a basis for the future work aiming at the development of household devices for sensitive detection of various analytes.Purpose: The aim of our present work was to develop and validate a reverse phase high-performance liquid chromatography (RP-HPLC) method for the determination of Decitabine (DCB). The developed method was further applied to observe the degradation of DCB under various stress conditions. Methods: Chromatographic separation was achieved on C18, 250 × 4.6 mm, particle size 5 μm, Agilent column, using ammonium acetate (0.01M) as mobile phase with flow rate of 1mL/min and injection volume was 20 μL. Quantification was carried out with UV detector at 230 nm with a linear calibration curve in the concentration range of 10–100 μg/mL based on peak area. Thus, developed method was validated for linearity, accuracy, precision, and robustness. Results: Linearity was found to be in the range between 10–100 μg/mL with a significantly higher value of correlation coefficient r = 0.9994. The limits of detection (LOD) and the limits of quantification (LOQ) were found to be 1.92μg/mL and 5.82 μg/mL respectively. Moreover, validated method was applied to study the degradation profile of DCB under various stress degradation conditions. Examination of different stress conditions on degradation of DCB showed that its degradation was highly susceptible to oxidative condition as 31.24% of drug was degraded. In acidic and alkaline conditions, the drug was degraded by 21.03% and 12.16% respectively, while thermal and photolytic condition causes least degradation, i.e. 0.21% and 0.3% respectively. Conclusion: The proposed method was found to be sensitive, specific and was successfully applied for the estimation of DCB in bulk drug, and lipid based nanoparticles.Abstract: A validated stability-indicating HPLC method has been reported for the determination of Ilaprazole in bulk drug and tablet. The drug was subjected to the various stress conditions as per the ICH guidelines. The degradation behavior of Ilaprazole was studied under hydrolytic, oxidative, photolytic and thermal conditions and was found to be unstable in almost all conditions except under alkaline and photolytic conditions. The separation of drug and its degraded products was carried out on Kinetex C-18 100A (5µ, 250×4.6 mm) column. The initial mobile phase composition used was Acetonitrile and water in the ratio 50:70v/v for 1 min then changed to 70:30v/v in next 6 min and finally equilibrated back to initial composition in 14min. The method was applied for the determination of Ilaprazole in marketed tablet formulation. The detection was carried at 305 nm using PDA detector with a flow rate of 1.0ml/min and injection volume 20µl. The validation of developed method was performed for linearity, accuracy, precision, selectivity and specificity and robustness.In the present work orodispersible tablets of Ondansetron hydrochloride were formulated by effervescent and sublimation method using factorial design. In the effervescent method, crospovidone (1.31%-4.68%) was used as superdisintegrant whereas citric acid (7.95%-18.04%) and sodium bicarbonate (5.22%-28.77%) were used as effervescent agent. In the sublimation method, crospovidone (1.58%-4.41%) was used as superdisintegrant whereas camphor (6.34%-17.35%) was used as subliming agent. The prepared batches of tablets were evaluated for hardness, friability, drug content uniformity, wetting time, disintegration time and In Vitro drug release pattern. Total 15 batches were formulated by effervescent method and 9 batches were formulated by sublimation method. Disintegration time of the formulations prepared by effervescent method was found between 13-36 secs whereas formulations prepared by sublimation method showed disintegration time between 7-39 secs. Among all the formulations, the formulation (S6) prepared by sublimation method using crospovidone 4% and camphor 8% was found to have the minimum disintegration time (7 seconds). Finally, it was concluded that Orodispersible tablet of Ondansetron Hydrochloride can be successfully formulated using effervescent and sublimation method and can be used as a novel drug dosage form for pediatric and geriatric with improved patient compliance and enhanced bioavailability.I this presentation, the interaction between cilostazol and two different cyclodextrins (β-CD and DM-β-CD) is studied by using LC. The capacity factors (k) of cilostazol were monitored in the presence of increasing concentrations of β-CD or DM-β-CD from the reduction of the retention time (tR). It was observed that cilostazol forms a 1:1 inclusion complex with β-cyclodextrin (β-CD) and dimethyl-β-cyclodextrin (DM-β-CD) at 250C, 370C and 450C. The interaction of cilostazol with DM-β-CD was more efficient and the highest the formation constant (K) was found for DM-β-CD (23.82M-1) at 250C. Moreover, the values of K decreased as the system temperature increased. To obtain the information on the mechanism of cilostazol affinity for β-CD and DM-β-CD, the thermodynamic parameters of the complexation (ΔG, ΔH, and ΔS) were studied. Finally, a comparison of the K values obtained for the two different cyclodextrins revealed that the K values of the complexation are dependent upon the structure of the host molecule. The change in the thermodynamic parameters suggested that the complexation could proceed spontaneously (ΔG<0) along with the releasing of heat (ΔH<0) and the decrease of entropy (ΔS<0).T light scattering of single nanoparticle, such as gold and silver nanoparticles, is stable and efficient, hence the applications of single nanoparticle-based dark-field microscopic imaging have attracted extensive attention. Both the changes of single nanoparticle scattering imaging color and scattering intensity can be used as effective detection signal in analytical chemistry. In the author’s group, to study the localized surface plasmon resonance (LSPR) light scattering of single nanoparticle, first the shape effect of single silver nanoparticle on the scattering light color and refractive index sensitivities was investigated. It was found that particles with a large radial ratio or a tip structure always had a higher sensitivity which could be due to the LSPR maxima at long wavelengths and the strong electric field intensity distribution. Then single nanoparticle-based RGB analytical method was established by coding the colors of the scattering light of individual nanoparticles with the RGB system, and the imaging date was manipulated with the IPP software. In addition, the scattering light intensity of single gold nanoparticle was digitized and expressed as digital information through the aforementioned software. Based on the RGB system and digitized method, it was established new scattering analytical method which had been certified in our other energy transfer experiment. Furthermore, the single nanoparticle-based dark-field microscopic imaging was also used to monitoring some chemical reactions. Real-time monitoring of the etching process of gold nanoparticles by iodine, in situ growth of single Ag@Hg nanoalloys and photochemical reaction were successfully achieved by dark-field microscopic imaging. In addition to the chemical reactions, the single nanoparticle-based dark-field microscopic imaging has a potential to real-time monitoring the intracellular biochemical reactions and infection process of bacteria and virus which may provide a new method for the diagnosis of the disease.

Molecular Modeling a Tool for Postulating the Mechanism of Drug Interaction: Glimepiride Alters the Pharmacokinetics of Sildenafil Citrate in Diabetic Nephropathy Animals

T X decoction, a kind of traditional herb medicines, has been prescribed frequency in the top five for treating asthma, bronchitis, chill and allergic rhinitis for more than hundreds of year in Korea. Nevertheless, no such study has been published yet related to the interaction of X decoction and we need to study for this issue as much as we prescribed. Furthermore, we evaluated activity of natural active compounds by different manufacturer method in X decoction and understood the multicomponent interactions, such as herb-herb interaction. The profiling of individual herb and decoction of X decoction was carried out using ultra-high performance liquid chromatography coupled with a quadrupole time of flight mass spectrometry (UPLC-QTOF-MS). Active compounds from X decoction were identified by multivariate analysis and compared activities between two groups. Also, we evaluated anti-inflammatory effect of X decoction investigating the protein expression of iNOS and COX-2 on LPS-induced macrophage RAW 264.7 cells. Chromatogram of individual herbs and decoction of X decoction were detected in positive, negative polarity mode and UV-PDA mode. After multivariate statistical analysis, we found good clustering and many active compounds were identified by comparing fragmentation patterns and plant databases. In this study, activity of active compound related to anti-inflammatory effect was significantly higher in decoction of X decoction than individual herb of X decoction. The result was in accordance with western blot by inhibiting the protein expression of iNOS on LPS-induced macrophage RAW 264.7 cells, in vitro. These results suggest that decoction of X decoction may have anti-inflammatory effect than individual herbs of X decoction.OBJECTIVE To determine the association of ABCB1 polymorphism G2677T with anti-emetic efficacy in patients treated with ondansetron for preventing postoperative nausea and vomiting. STUDY DESIGN A clinical trial. PLACE AND DURATION OF STUDY Combined Military Hospital, Rawalpindi and Institute of Biomedical and Genetic Engineering, Islamabad, from 2012 to 2013. METHODOLOGY Four mg ondansetron was administered intravenously 30 minutes before the end of surgery. A total of 246 patients with the complaints of nausea and vomiting and 244 patients without nausea and vomiting were analyzed for G2677T polymorphism using PCR-RFLP method. Results were described as frequency percentages and chi-square test with significance at p < 0.05. RESULTS The patients with TT genotype had significantly lower incidence of postoperative nausea and vomiting during the first 2 hours (p < 0.001) and between 2 - 24 hours after surgery as compared to other genotypes (p < 0.001). The patients with GG genotypes had significantly higher incidence of this complaint (p=0.014). CONCLUSION Polymorphism of ABCB1 has an association with responsiveness for ondansetron. There is a role for genetics in the management of PONV.

Determination of MDA by HPLC in Blood of Levofloxacin Treated Rats

Background Because of its structure and complex manufacturing process, every biotherapeutic product (BTP; medicinal products made by or derived from living organisms and produced by biotechnology) adheres to stringent quality assurance and control requirements, in addition to extensive nonclinical and clinical study data. Similarly, copy products of original biotherapeutics (termed as “biosimilars”) are subjected to equally strict regulatory control. BTPs have been registered in Malaysia since the 1990s; however, registration of biosimilars started only in 2008.Introduction: Anaphylaxis is a serious, rapid and potentially life-threatening allergic response involving IgE or IgG. Clinacanthus nutans, a small native shrub found in tropical Asia possess analgesic, anti-inflammatory and anti-viral activities and traditionally used for skin rashes, insect and snake-bites. In Thailand, alcoholic C. nutans extracts has been used topically for skin rashes, a symptom of allergy. Aim: To justify that C. nutans can treat skin rashes; this study investigated the anti-allergenicity of C. nutans extracts. Methods: Cytotoxicity of C. nutans extracts was evaluated by MTT on RBL-2H3. The most active C. nutans extract was determined by IgE-mediated mast cell degranulation. Acute toxicity of C. nutans aqueous extract was evaluated using female Sprague Dawley rats at 5000 mg/kg. Anti-allergenicity of C. nutans aqueous extract was studied by IgE-mediated passive systemic anaphylaxis (PSA). The release of preformed mediator (β-hexosaminidase) as well as newly synthesized mediators (TNF-α, IL-4 and LTC4) was evaluated. Results: C. nutans extracts were not cytotoxic up to 1 mg/ml (ethanolic) and 6 mg/ml (aqueous). In vitro, C. nutans aqueous extract was able to inhibit the release of preformed mediators but not newly synthesized mediators at 5 mg/ml. The ethanolic extracts were not able to inhibit all mediators tested. At 5000 mg/kg, C. nutans aqueous extract was non-toxic to the rats; no significant difference observed haematologically and biochemically. In vivo, C. nutans aqueous extract did not inhibit mediators of IgE-mediated PSA at 500 mg/kg and 2000 mg/kg. Conclusion: C. nutans aqueous extract was most active but could not inhibit mediators of IgE-mediated PSA. As anaphylaxis could be mediated by IgE orIgG, we postulate that C. nutans aqueous extract may exhibit its anti-allergenicity in IgG-mediated pathway.Accelerating wound healing is now considered as a principle clinical treatment and increasing the quality and speed of healing which has always been emphasized by the scientists. Propolis and honey are natural bee products with wide range of biological and medicinal properties. This study was aimed to determine the synergistic effect of honey and propolis in wound healing of rat skin. A total of 75 Wistar rats weighing 200-250 gr were placed under general anesthesia and sterile conditions. Then a square shape wound with 1.5*1.5 mm dimension was made on the back of the neck. Animals were randomly divided into control, honey, propolis, combined honey propolis and phenytoin 1% groups, respectively. Rats were randomly divided into the following groups: 4th, 7th and, 14th days of treatment in each period of study. Wound area in the experimental group was covered once daily with a fixed amount of thyme honey, propolis, propolis and honey and phenytoin cream (1%), the control group did not receive any treatment. For histological studies, during the fourth, seventh and fourteenth days rats were sacrificed and samples were taken from the wound and adjacent skin. After histological staining fibroblast, neutrophils, macrophages and vascular sections were counted in the wound bed. The macroscopic and microscopic evaluations showed that the percentage of wound healing on different days in the experimental and control groups were significant (P<0.05). The macroscopic and microscopic evaluation showed that the percentage of wound healing on different days in combined propolis and honey experimental group was significantly different from the control group (Multivariate ANOVA test) (P<0.05). Combined application of propolis and honey on the open wound healing in rats has a synergistic effect.Children are more susceptible to medication errors than adults. Medication administration process is the last stage in the medication treatment process and most of the errors detected in this stage. Little research has been undertaken about medication errors in children in the Middle East countries. This study was aimed to evaluate how the paediatric nurses adhere to the medication administration policy and also to identify any medication preparation and administration errors or any risk factors. An observational, prospective study of medication administration process from when the nurses preparing patient medication until administration stage (May to August 2014) was conducted in Saudi Arabia. Twelve paediatric nurses serving 90 paediatric patients were observed. 456 drug administered doses were evaluated. Adherence rate was variable in 7 steps out of 16 steps. Patient allergy information, dose calculation, drug expiry date were the steps in medication administration with lowest adherence rates. 63 medication preparation and administration errors were identified with error rate 13.8% of medication administrations. No potentially life-threating errors were witnessed. Few logistic and administrative factors were reported. The results showed that the medication administration policy and procedure need an urgent revision to be more sensible for nurses in practice. Nurses’ knowledge and skills regarding to the medication administration process should be improved. Keywords—Double checking, Medication administration errors, Medication safety, Nurses.

Amylin dual action: a second gluco regulatory β-cell hormone, treatment and cause for the diabetes

Amylin also known as islets of amyloid polypeptides (IAPP), is a secretary product of pancreatic β -cells, co-synthesized and co-secreted with insulin. It is a glucose regulatory β -cell hormone with a central action on food intake; it inhibits glucagon secretion, delays gastric emptying and acts as a satiety agent. It has been proposed to have a role in the management of diabetes. Pramlintide a synthetic version of amylin is used in glycemic control of diabetes mellitus. Accumulation of amyloid on β- cell often reflects the severity of diabetes. This review is primarily focused on dual role of amylin: induction as well as treatment of Diabetes.

Development of health supplement probiotic in relation to physical strength and biological activity in experimental animal.

A probiotic nutritive health supplement having a healthy nutritive value was formulated to enhance body function. A 90 days study proved the formulated health supplement to improve the body weight with a maintained normal level of cholesterol and triglycerides in blood which is needful for the proper cardiac function. The supplement also maintained the blood parameters to avoid any illness in the body. In the present study, the health supplement was able to maintain the Haemoglobin level, RBC Count, WBC Count, Platelet Count in Sprague Dawley rats. It is also found that the supplement maintained the cardiac rhythm in the stressful condition and increased in the time of swimming in the force swimming test which indicates an increase in the physical strength.