FORMULATION AND STABILITY EVALUATION OF KETOPROFEN LOADED VIRGIN COCONUT OIL BASED CREAMY EMULSION

Objective: To formulate and optimize a topical formulation; a virgin coconut oil (VCO) based Ketoprofen loaded creamy emulsion containing Tween 80® as the surfactant and to evaluate the stability of samples. Methods: In preformulatory studies optimization of the formulae was done using ternary phase diagrams with water titration method and emulsions were formulated using two methods; spontaneous emulsification and homogenization. Their stability was analyzed under visual observation to optimize the best formulae for Ketoprofen incorporated creamy emulsion. 2.5% w/w Ketoprofen topical formulations are available in the market. Results: Centrifugation provided more comparable data than visual observation. Phase separation was the main instability condition observed in unstable emulsions. Composition 23.60% VCO: 29.53% Tween 80®: 45.87% water was identified as the best optimized formulae in both with and without Ketoprofen formulations and all the samples with different Ketoprofen concentrations were stable for 14 days under centrifugation and visual observation stability studies. Conclusion: Homogenization was more effective in stable emulsion formation than spontaneous emulsification in VCO, Tween 80®,water emulsion. The best optimized formula was 23.60% VCO: 29.53% Tween 80®: 45.87% water

AWARENESS OF USAGE OF SUNSCREENS AMONG SCHOOL CHILDREN IN KANDY, SRI LANKA

Objective: The objective of this research was to evaluate the awareness of usage of sunscreens among school children in Kandy, Sri Lanka.Methods: School children (138) who do outdoor sports were recruited from six schools. Knowledge and attitudes towards the usage and application of sunscreens were assessed using a researcher administered questionnaire. Statistical analyses were done using Statistical Package for the Social Science (SPSS version-16).Results: Among the school children who do outdoor sports, 71 (51.4%) of participants used sunscreens and 67 (48.6%) of participants did not use sunscreens.Conclusion: According to the results most of the school children have a lack of knowledge, attitudes and practice towards the usage of sunscreens

Differential kinetic profiles and metabolism of primaquine enantiomers by human hepatocytes

BACKGROUND: The clinical utility of primaquine (PQ), used as a racemic mixture of two enantiomers, is limited due to metabolism-linked hemolytic toxicity in individuals with genetic deficiency in glucose-6-phosphate dehydrogenase. The current study investigated differential metabolism of PQ enantiomers in light of the suggestions that toxicity and efficacy might be largely enantioselective. METHODS: Stable isotope (13)C-labelled primaquine and its two enantiomers (+)-PQ, (−)-PQ were separately incubated with cryopreserved human hepatocytes. Time-tracked substrate depletion and metabolite production were monitored via UHPLC–MS/MS. RESULTS: The initial half-life of 217 and 65 min; elimination rate constants (λ) of 0.19 and 0.64 h(−1); intrinsic clearance (Cl(int)) of 2.55 and 8.49 (µL/min)/million cells, which when up-scaled yielded Cl(int) of 6.49 and 21.6 (mL/min)/kg body mass was obtained respectively for (+)- and (−)-PQ. The extrapolation of in vitro intrinsic clearance to in vivo human hepatic blood clearance, performed using the well-stirred liver model, showed that the rate of hepatic clearance of (+)-PQ was only 45 % that of (−)-PQ. Two major primary routes of metabolism were observed—oxidative deamination of the terminal amine and hydroxylations on the quinoline moiety of PQ. The major deaminated metabolite, carboxyprimaquine (CPQ) was preferentially generated from the (−)-PQ. Other deaminated metabolites including PQ terminal alcohol (m/z 261), a cyclized side chain derivative from the aldehyde (m/z 241), cyclized carboxylic acid derivative (m/z 257), a quinone-imine product of hydroxylated CPQ (m/z 289), CPQ glucuronide (m/z 451) and the glucuronide of PQ alcohol (m/z 437) were all preferentially generated from the (−)-PQ. The major quinoline oxidation product (m/z 274) was preferentially generated from (+)-PQ. In addition to the products of the two metabolic pathways, two other major metabolites were observed: a prominent glycosylated conjugate of PQ on the terminal amine (m/z 422), peaking by 30 min and preferentially generated by (+)-PQ; and the carbamoyl glucuronide of PQ (m/z 480) exclusively generated from (+)-PQ. CONCLUSION: Metabolism of PQ showed enantioselectivity. These findings may provide important information in establishing clinical differences in PQ enantiomers

Enantioselective metabolism of primaquine by human CYP2D6

BACKGROUND: Primaquine, currently the only approved drug for the treatment and radical cure of Plasmodium vivax malaria, is still used as a racemic mixture. Clinical use of primaquine has been limited due to haemolytic toxicity in individuals with genetic deficiency in glucose-6-phosphate dehydrogenase. Earlier studies have linked its therapeutic effects to CYP2D6-generated metabolites. The aim of the current study was to investigate the differential generation of the CYP2D6 metabolites by racemic primaquine and its individual enantiomers. METHODS: Stable isotope (13)C-labelled primaquine and its two enantiomers were incubated with recombinant cytochrome-P450 supersomes containing CYP2D6 under optimized conditions. Metabolite identification and time-point quantitative analysis were performed using LC-MS/MS. UHPLC retention time, twin peaks with a mass difference of 6, MS-MS fragmentation pattern, and relative peak area with respect to parent compound were used for phenotyping and quantitative analysis of metabolites. RESULTS: The rate of metabolism of (+)-(S)-primaquine was significantly higher (50% depletion of 20 μM in 120 min) compared to (−)-(R)-primaquine (30% depletion) when incubated with CYP2D6. The estimated V(max) (μmol/min/mg) were 0.75, 0.98 and 0.42, with K(m) (μM) of 24.2, 33.1 and 21.6 for (±)-primaquine, (+)-primaquine and (−)-primaquine, respectively. Three stable mono-hydroxylated metabolites, namely, 2-, 3- and 4-hydroxyprimaquine (2-OH-PQ, 3-OH-PQ, and 4-OH-PQ), were identified and quantified. 2-OH-PQ was preferentially formed from (+)-primaquine in a ratio of 4:1 compared to (−)-primaquine. The racemic (±)-primaquine showed a pattern similar to the (−)-primaquine; 2-OH-PQ accounted for about 15–17% of total CYP2D6-mediated conversion of (+)-primaquine. In contrast, 4-OH-PQ was preferentially formed with (−)-primaquine (5:1), accounting for 22% of the total (−)-primaquine conversion. 3-OH-PQ was generated from both enantiomers and racemate. 5-hydroxyprimaquine was unstable. Its orthoquinone degradation product (twice as abundant in (+)-primaquine compared to (−)-primaquine) was identified and accounted for 18–20% of the CYP2D6-mediated conversion of (+)-primaquine. Other minor metabolites included dihydroxyprimaquine species, two quinone-imine products of dihydroxylated primaquine, and a primaquine terminal alcohol with variable generation from the individual enantiomers. CONCLUSION: The metabolism of primaquine by human CYP2D6 and the generation of its metabolites display enantio-selectivity regarding formation of hydroxylated product profiles. This may partly explain differential pharmacologic and toxicologic properties of primaquine enantiomers

Enantioselective metabolism of primaquine by human CYP2D6

BACKGROUND: Primaquine, currently the only approved drug for the treatment and radical cure of Plasmodium vivax malaria, is still used as a racemic mixture. Clinical use of primaquine has been limited due to haemolytic toxicity in individuals with genetic deficiency in glucose-6-phosphate dehydrogenase. Earlier studies have linked its therapeutic effects to CYP2D6-generated metabolites. The aim of the current study was to investigate the differential generation of the CYP2D6 metabolites by racemic primaquine and its individual enantiomers. METHODS: Stable isotope (13)C-labelled primaquine and its two enantiomers were incubated with recombinant cytochrome-P450 supersomes containing CYP2D6 under optimized conditions. Metabolite identification and time-point quantitative analysis were performed using LC-MS/MS. UHPLC retention time, twin peaks with a mass difference of 6, MS-MS fragmentation pattern, and relative peak area with respect to parent compound were used for phenotyping and quantitative analysis of metabolites. RESULTS: The rate of metabolism of (+)-(S)-primaquine was significantly higher (50% depletion of 20 μM in 120 min) compared to (−)-(R)-primaquine (30% depletion) when incubated with CYP2D6. The estimated V(max) (μmol/min/mg) were 0.75, 0.98 and 0.42, with K(m) (μM) of 24.2, 33.1 and 21.6 for (±)-primaquine, (+)-primaquine and (−)-primaquine, respectively. Three stable mono-hydroxylated metabolites, namely, 2-, 3- and 4-hydroxyprimaquine (2-OH-PQ, 3-OH-PQ, and 4-OH-PQ), were identified and quantified. 2-OH-PQ was preferentially formed from (+)-primaquine in a ratio of 4:1 compared to (−)-primaquine. The racemic (±)-primaquine showed a pattern similar to the (−)-primaquine; 2-OH-PQ accounted for about 15–17% of total CYP2D6-mediated conversion of (+)-primaquine. In contrast, 4-OH-PQ was preferentially formed with (−)-primaquine (5:1), accounting for 22% of the total (−)-primaquine conversion. 3-OH-PQ was generated from both enantiomers and racemate. 5-hydroxyprimaquine was unstable. Its orthoquinone degradation product (twice as abundant in (+)-primaquine compared to (−)-primaquine) was identified and accounted for 18–20% of the CYP2D6-mediated conversion of (+)-primaquine. Other minor metabolites included dihydroxyprimaquine species, two quinone-imine products of dihydroxylated primaquine, and a primaquine terminal alcohol with variable generation from the individual enantiomers. CONCLUSION: The metabolism of primaquine by human CYP2D6 and the generation of its metabolites display enantio-selectivity regarding formation of hydroxylated product profiles. This may partly explain differential pharmacologic and toxicologic properties of primaquine enantiomers

Мониторинг воспроизводства и влияния фекального прогестерона на репродуктивную цикличность циклов самбарского оленя Шри-Ланки (Rusa unicolor unicolor)

Sambar deer hinds, estrus, progesterone, estrous cycle, Sri LankaThis study examines the length of the estrous cycle in 16 Sambar deer hinds in National zoological gardens in Dehiwala and Kegalle, Sri Lanka (NZGDK) assessed with the use of changes in progesterone concentrations, along with the changes in the profile of this hormone and by the visual estrus manifestations. The objectives of the present study were to characterize ovarian activity throughout the estrous cycle and the non-pregnant luteal phase of captive sambar deer in Sri Lanka. These objectives were achieved with the use of radioimmunoassay (RIA) to measure fecal concentrations of progesterone and visual estrus manifestation. Fecal samples were collected from non-pregnant sambar deer hinds (aged 2–4 years)over the period of six months on daily basis, both during breeding and non-breeding seasons. Estrous cycles were recorded in non-pregnant females, based on fecal progesterone concentrations. The average estrous cycle length was 26.1±2.08 days (mean ± SEM) and 2.10 ± 0.51 days in the inter-luteal phase.The average fecal progesterone concentrations attained the peak mid-luteal values of 2.74 ng mL–1. There appeared to be variation in fecal progesterone amplitude between animals and between dates, but the low frequency of sampling prohibited confirmation of trends. Behavioral estrus was detected only when the average progesterone concentrations were less than 0.07 ng mL–1. However, not all periods of depressed progesterone secretion were associated with the observed estrus. Behavioral estrus was detected in hinds when progesterone concentrations were less than 0.07 ng mL–1; a subsequent rise in progesterone indicated ovulation taking place at this time