Preformulation studies for the development of a microemulsion formulation from Ambrosia peruviana All, with anti-inflammatory effect

Abstract Natural products are considered an important source of the therapeutic arsenal currently available. Among these alternatives are the seeds of Ambrosia peruviana (altamisa), whose extract has shown an anti-inflammatory effect. The main objective of this work was to perform a preformulation study of Ambrosia peruviana seeds ethanolic extract, where the main factors that affect the physical, chemical, and pharmacological stability of the extract were evaluated, as well as a compatibility study by differential scanning calorimetry (DSC) analysis against different excipients. A dry extract was obtained by rotary evaporation of the seeds macerated with 96% ethanol. The anti-inflammatory activity was determined by measuring its effect on NO production in RAW 264.7 macrophages, stimulated with LPS. The results showed that the dry extract maintained its stability over time when stored at a temperature of 4 and 25ºC, demonstrating its biological activity, the content of phenolic compounds, and its physicochemical parameters remain practically invariable. However, when exposed to high temperatures (60 ºC) it was affected. The thermal analysis revelated that the behavior of most of the selected excipients and the dry extract was maintained, which indicates that it did not present incompatibilities, therefore they can be candidates for formulating a microemulsion.

The potential use of papaya and banana peels as substrate to enhance the pigment production of Gram-positive bacterial strain isolated from Holothuria (Lessonothuria) pardalis

Aims@#The microbial pigment can be the best promising alternative to replace synthetic colorant. However, due to the high cost of synthetic medium for microbial pigment production, there is a need to develop a new low-cost medium of bacterial pigment production. This study aims to investigate the potential of banana and papaya peels as alternative lowcost substrates for a carotenoid-producing bacterium, B12 strain (bacteria strain isolated from Holothuria (Lessonothuria) pardalis).@*Methodology and results@#B12 strain identified as an aerobic bacterium with non-motile, diplobacilli shaped and Grampositive bacteria. The fermentation was optimized with different parameters included the effect of temperature, time, concentrations, pHs, carbon and nitrogen sources to find the optimum relative pigment concentration produced by B12. The results showed that the B12 strain produced the highest relative pigment concentration measured at 450 nm when the strain was cultivated at 37 °C and pH 7 in the culture medium incorporated with the combination of dried papaya peels and banana peels (100% v/v with ratio 1:1) at 72 h of incubation. Lactose, peptone and yeast were observed as the best carbon and nitrogen sources to increase the pigment concentration of B12 strain. Stability of the pigment was studied at different physiochemical stress, and it showed the pigment obtained from dried papaya and banana substrates can tolerate and stable under stress condition.@*Conclusion, significance and impact of study@#This can be concluded that the combination of dried papaya and banana peels worked well as substrate and can be utilized as a fermentation medium to replace the synthetic medium which is more expensive and uneconomical for industry application. Besides, it also helps in managing waste and solving the pollution problem due to the increasing of biochemical oxygen demand (BOD) and chemical oxygen demand (COD).

Accelerated stability test of sustained-release thienorphine loaded microspheres for injection / 药学学报

In order to meet the clinical needs of long-acting sustained-release thienorphine, injectable thienorphine loaded microspheres were developed, and the accelerated stability study was carried out to explore the suitable storage and transportation conditions of the microspheres. Using poly(lactic-co-glycolic acid) (PLGA) as carrier material, 3 batches of microspheres were prepared in pilot scale with emulsion solvent evaporation method. By investigating the in vitro release of thienorphine loaded microspheres at 37, 45, 52, and 60 ℃, and applying the Arrhenius equation, the linear relationship between the release rate constant (lgk) and the temperature (1/T) was established to obtain the equation: lgk = -8.073/T + 24.35 (R2 = 0.985 3), which showed that the release of microspheres at high temperature can be used to predict the release in vitro at 37 ℃, and 52.0 ± 0.5 ℃ was selected as the accelerated release condition in vitro. The quality research methods were established to investigate the changes of critical quality attributes such as microsphere morphology, drug loading, particle size and distribution, polymer molecular weight, and the related substances under accelerated conditions. The difference factor f1 and similarity factor f2 were used to assess the similarity of release behavior under accelerated conditions. The results showed that under the accelerated experimental conditions of 25 ± 2 ℃ and relative humidity (RH) 60% ± 5%, the critical quality attributes of injectable thienorphine loaded microspheres had no significant change in 6 months, suggesting that the long-term storage condition could be 5 ± 3 ℃.

Stability of hydroalcoholic extracts of two species of guaco; Mikania glomerata SPRENG. and Mikania laevigata SCHULTZ. (Asteraceae), by UHPLC-MS

It is important to study the stability of plant extracts used as active ingredients in phytotherapic medicine, as degradation of the active principles directly affects the efficacy and safety of these products. Therefore, a stability study of the hydroalcoholic extract of the species: Mikania glomerata and Mikania laevigata was conducted in order to determine the speed of degradation and shelf life of these extracts, which are incorporated in cough syrup in Brazil. Leaves of both species were dried in an oven or by lyophilization (freeze-dried). Hydroalcoholic extracts underwent both accelerated stability study of six months and long-term stability study for 12 months. Samples were stored at different temperatures and every three months were analysed by ultra-high performance liquid chromatography-mass spectrometry (UHPLC-MS) to monitor their chemical profile, quantifying coumarin and chlorogenic acid. For all conditions of the study, a reduction of the content of the chemical marker of this species, coumarin, greater than 5% was observed, so a shelf life of two years cannot be assigned to the hydroalcoholic extracts of these species as observed in commercial extracts.

Differences in percussion-type measurements of implant stability according to height of healing abutments and measurement angle / 대한치과보철학회지

PURPOSE: The purpose of this study was to evaluate the effect of healing abutment height and measurement angle on implant stability when using Periotest and AnyCheck. MATERIALS AND METHODS: 60 implants were placed into artificial bone blocks. After implant insertion, 2, 3, 4 and 5 mm healing abutments were installed on 15 specimens, respectively. Insertion torque value, implant stability test, Periotest value were measured. Insertion torque value was controlled between 45 – 55 Ncm. AnyCheck was used for measuring implant stability test and Periotest M was used for measuring Periotest value. Implant stability test and Periotest value were measured at the angles of 0 and 30 degrees to the horizontal plane. Measured values were analyzed statistically. RESULTS: Insertion torque value had no significant difference among groups. When healing abutment height was higher, implant stability test and Periotest value showed lower stability. Also when measurement angle was decreased, implant stability test and Periotest value showed lower stability. CONCLUSION: When measuring stability of implants with percussion type devices, measured values should be evaluated considering height of healing abutments and measurement angle.

Study on Quality Standard of Hirudo nipponica Freeze-dried Powder / 中国药房

OBJECTIVE:To establish the quality standard of Hirudo nipponica freeze-dried powder(called"freeze-dried powder"for short),and to provide reference for controlling its quality. METHODS:A total of 3 batches of freeze-dried powder were collected,identified and tested according to the requirements of H. nipponica stated in 2015 edition of Chinese Pharmacopoeia(part Ⅰ)(shorted for pharmacopoeia);the antithrombin activity was also analyzed. The maximum tolerated dose (MTD)was used to investigate the toxicity. The stability was determined by designing temperature,humidity and strong light exposure tests. RESULTS:In the TLC of test sample,the same red spots were found in the corresponding location of the control drug chromatogram,and the same orange-red fluorescence spots were shown under the UV light(365 nm). Average content of moisture in 3 batches of samples was 2.61%,and the levels of total ash,acid-insoluble ash,pH aflatoxin and antithrombin activity were 2.83%,0.38%,6.92,0.28 μg/kg and 257.0 U/g,respectively. The content of Pb,Cd,As and Cu were in line with the requirements of pharmacopoeia except that the content of Hg was slightly higher than lower limit of H. nipponica in pharmacopoeia. Results of MTD showed that no death and ADR was found in mice after giving 26.4 g/kg freeze-dried powder by the amount of crude drug,which was 58 times as large as the maximum dosage that the pharmacopoeia described. Under the condition of 20, 40 ℃ and strong light exposure [(4 500±500)Lx],the anticoagulase activity of freeze-dried powder decreased significantly over time,while the anticoagulase activity of freeze-dried powder stored at 40 ℃ for 6 months was in line with the requirements of pharmacopoeia. Under the condition of high humidity(relative humidity were 90%,75%),freeze-dried powder showed a strong hygroscopicity. CONCLUSIONS:Established quality evaluation standard for freeze-dried powder according to pharmacopoeia standard could be used to control its quality.

Quantitative models for drug shelf-life estimation / 药物评价研究

Shelf-life estimation is the core of stability tests and critical to the final label claim on the package of new drug.Firstly,this paper depicts the methods of ICH Q1E for shelf-life estimation,compares it to the counterpart in the newest version of Chinese Pharmacopeia (Ch.P 2015,IV-9001),and proves its compliance.Secondly,this paper also unfolds the latent assumptions and hinged limitations of the quantitative methods in the guidelines.Finally,the empirical distributions of the shelf-life estimators are obtained via Monte-Carlo simulation to facilitate better understanding of properties of the estimators.It is shown that the approach that takes the average as final shelf-life estimate is likely to further underestimate the true shelf-life comparing to the ICH Q1E approach,and hence puts the manufacturers in unnecessary disadvantage.

Validity problems between pharmaceutical preparations and pharmaceutical raw materials in the production process / 中国药学杂志

OBJECTIVE: To identify the relationship between API validity and chemical agent validity and discuss the problem of legal characterization about the behavior of the use of API beyond validity in the production process of chemical agents.

Shelf-life of a 2.5 percent sodium hypochlorite solution as determined by arrhenius equation

Accelerated stability tests are indicated to assess, within a short time, the degree of chemical degradation that may affect an active substance, either alone or in a formula, under normal storage conditions. This method is based on increased stress conditions to accelerate the rate of chemical degradation. Based on the equation of the straight line obtained as a function of the reaction order (at 50 and 70 ºC) and using Arrhenius equation, the speed of the reaction was calculated for the temperature of 20 ºC (normal storage conditions). This model of accelerated stability test makes it possible to predict the chemical stability of any active substance at any given moment, as long as the method to quantify the chemical substance is available. As an example of the applicability of Arrhenius equation in accelerated stability tests, a 2.5 percent sodium hypochlorite solution was analyzed due to its chemical instability. Iodometric titration was used to quantify free residual chlorine in the solutions. Based on data obtained keeping this solution at 50 and 70 ºC, using Arrhenius equation and considering 2.0 percent of free residual chlorine as the minimum acceptable threshold, the shelf-life was equal to 166 days at 20 ºC. This model, however, makes it possible to calculate shelf-life at any other given temperature.

Testes acelerados de estabilidade são indicados para avaliar, em um curto período de tempo, o grau de degradação química que poderá afetar uma substância química, isoladamente ou quando inserida em uma fórmula, sob condições normais de armazenamento. Este método está fundamentado na intensificação das condições de estresse para acelerar a velocidade de degradação química. Baseando-se na equação da reta obtida e na ordem de reação determinada (a 50 e 70 ºC) e usando a equação de Arrhenius, a velocidade de reação foi calculada para a condição de temperatura de 20ºC (condições normais de armazenamento). Este modelo de teste acelerado de estabilidade torna possível a predição da estabilidade química de qualquer substância, em qualquer tempo, desde que o método de quantificação da substância química esteja disponível. Como exemplo da aplicabilidade da equação de Arrhenius em teste acelerado de estabilidade, uma solução de hipoclorito de sódio a 2,5 por cento foi analisada por ser quimicamente instável. A quantificação do cloro residual livre foi determinada através de titulação iodométrica. A partir dos dados obtidos decorrentes das amostras submetidas às temperaturas de 50 e 70 ºC e com o emprego da equação de Arrhenius, o tempo de prateleira obtido foi de 166 dias em temperatura de 20 ºC, considerando como limite inferior a concentração de 20 mg/mL de cloro residual livre. Este modelo, entretanto, possibilita o cálculo de tempo de prateleira em qualquer outra temperatura de interesse.

Estabilidade e estudo de penetração cutânea in vitro da rutina veiculada em uma emulsão cosmética através de um modelo de biomembrana alternativo / Stability and in vitro penetration study of rutin incorporated in a cosmetic emulsion through an alternative model biomembrane

A rutina é empregada como antioxidante e na prevenção da fragilidade capilar. Estudos de penetração in vitro através da pele humana seria a situação ideal, entretanto, há dificuldades de sua obtenção e manutenção de sua viabilidade. Entre os demais modelos de membrana, a muda de pele de cobra se apresenta como estrato córneo puro, fornecendo barreira similar ao humano e é obtida sem a morte do animal. Os objetivos desta pesquisa foram desenvolver e avaliar a estabilidade de uma emulsão cosmética contendo rutina e, como promotor de penetração cutânea, o propilenoglicol; e avaliar a penetração e a retenção cutânea in vitro da referida substância ativa da formulação, empregando um modelo de biomembrana alternativo. A emulsão foi desenvolvida com rutina e propilenoglicol, ambos a 5,0 por cento p/p. Quantificou-se a rutina das emulsões por espectrofotometria a 361,0 nm, método previamente validado. A penetração e retenção cutânea in vitro foram realizadas em células de difusão vertical com muda de pele de cobra de Crotalus durissus, como modelo de biomembrana alternativo, e água destilada e álcool etílico absoluto 99,5 por cento (1:1), como fluido receptor. O experimento foi conduzido em um período de seis horas, a 37,0 ± 0,5 ºC e agitação constante de 300 rpm. Empregou-se o método espectrofotométrico validado a 410,0 nm para a quantificação da rutina após penetração e retenção cutânea. A emulsão não promoveu a penetração cutânea da rutina através da muda de pele de C. durissus, retendo 0,931 ± 0,0391 mg de rutina/mg de muda de pele de cobra. Nas condições de armazenamento a 25,0 ± 2,0 ºC; 5,0 ± 0,5 ºC e 45,0 ± 0,5 ºC, a emulsão apresentou-se quimicamente estável durante 30 dias. De acordo com os resultados, a emulsão não favoreceu a penetração cutânea da rutina, mas apenas sua retenção no estrato córneo de C. durissus, condição considerada estável no período de 30 dias.

Rutin is employed as antioxidant and to prevent the capillary fragility and, when incorporated in cosmetic emulsions, it must target the action site. In vitro cutaneous penetration studies through human skin is the ideal situation, however, there are difficulties to obtain and to maintain this tissue viability. Among the membrane models, shed snake skin presents itself as pure stratum corneum, providing barrier function similar to human and it is obtained without the animal sacrifice. The objectives of this research were the development and stability evaluation of a cosmetic emulsion containing rutin and propylene glycol (penetration enhancer) and the evaluation of rutin in vitro cutaneous penetration and retention from the emulsion, employing an alternative model biomembrane. Emulsion was developed with rutin and propylene glycol, both at 5.0 percent w/w. Active substance presented on the formulation was quantified by a validated spectrophotometric method at 361.0 nm. Rutin cutaneous penetration and retention was performed in vertical diffusion cells with shed snake skin of Crotalus durissus, as alternative model biomembrane, and distilled water and ethanol 99.5 percent (1:1), as receptor fluid. The experiment was conducted for six hours, at 37.0 ± 0.5 ºC with constant stirring of 300 rpm. Spectrophotometry at 410.0 nm, previously validated, determined the active substance after cutaneous penetration/retention. Emulsion did not promote rutin cutaneous penetration through C. durissus skin, retaining 0.931 ± 0.0391 mg rutin/mg shed snake skin. The referred formulation was chemically stable for 30 days after stored at 25.0 ± 2.0 ºC, 5.0 ± 0.5 ºC and 45.0 ± 0.5 ºC. In conclusion, it has not been verified the active cutaneous penetration through the model biomembrane, but only its retention on the Crotalus durissus stratum corneum, condition considered stable for 30 days.

Accelerated chemical stability data of O/W fluid emulsions containing the extract of Trichilia catigua Adr. Juss (and) Ptychopetalum olacoides Bentham

Development of topical dosage forms requires physical, physicochemical and chemical assays that provide, as soon as possible, the formulation with the best stability profiles. This study evaluated the stability of O/W fluid emulsions, by total flavonoids determination, expressed in rutin, containing the standardized extract of Trichilia catigua Adr. Juss (and) Ptychopetalum olacoides Bentham. Samples were evaluated for 90 days stored at 24.0 ± 2.0 ºC, 5.0 ± 0.5 ºC and 40.0 ± 0.5 ºC, following a protocol for the assessment of accelerated chemical stability assay, also known as Normal Stability Test. A sensitive UV-spectrophotometric method at 361.0 nm was previously validated for the determination of the active substance. By Normal Stability Test, the O/W fluid emulsions presented acceptable chemical stability, for at least 90 days, when the samples were stored at 24.0 ± 2.0 ºC and 5.0 ± 0.5 ºC. The storage condition at 40.0 ± 0.5 ºC has accelerated the degradation process of the total flavonoids, consequently, those O/W emulsions containing this kind of natural active substance or a similar preparation must not be stored at elevated temperatures.

O desenvolvimento de formas farmacêuticas tópicas necessita ensaios físicos, físico-químicos e químicos que selecionem rapidamente a formulação de melhor desempenho de estabilidade. Este estudo avaliou a estabilidade de emulsões O/A fluidas, por meio da determinação de flavonóides totais, expressos em rutina, contendo o extrato padronizado de Trichilia catigua Adr. Juss (e) Ptychopetalum olacoides Bentham. As amostras foram armazenadas a 24,0 ± 2,0 ºC; 5,0 ± 0,5 ºC e 40,0 ± 0,5 ºC durante 90 dias e foram avaliadas segundo o protocolo para a determinação da estabilidade acelerada, conhecida como Teste de Estabilidade Normal. A quantificação da substância ativa foi determinada por espectrofotometria na região do ultravioleta a 361,0 nm, previamente validado. Após os ensaios de estabilidade, as emulsões O/A fluidas apresentaram estabilidade adequada, pelo menos, no período de 90 dias, quando armazenadas a 24,0 ± 2,0 ºC e 5,0 ± 0,5 ºC. A condição de armazenamento a 40,0 ± 0,5 ºC acelerou a cinética de degradação dos flavonóides totais, expressos em rutina, portanto, preparações possuindo esta categoria de substância ativa natural ou formulações similares não devem ser armazenadas em temperaturas elevadas.

Apply the accelerated stability test to predict the self-life of national standard serum tetanus antitoxin

Accelerated stability test has been applied for predicting self-life of national standard serum tetanus anti-toxin (SAT). Samples of the tetanus antitoxin were kept at - 20°C, 4°C, 22°C, 37°C and 45°C for 6 months and their potency was estimated by using the toxin neutralization method on mice. There was relationship between the potency change and the self- life of serum tetanus antitoxin by using Arrhenius equation. The results demonstrated that the self-life of the national standard serum tetanus antitoxin was 9 years and 6 months when stored at -20°C.

Finite Element Analysis and Torsional Stability Testing of Reference (Metaphyseal and Diaphyseal Fixation) and Experimental (Metaphyseal Fixation Only) Cementless Femoral Stems / 대한정형외과학회잡지

PURPOSE: To report the details of the findings of finite element analysis and torsional stability testing of a reference stem (fixation in both metaphysis and diaphysis) and an experimental stem (fixation in the metaphysis only). MATERIALS AND METHODS: Finite element ABAQUS software (version 5.5) was used for all analyses. A Newton Raphson iterative solution scheme was used to calculate the nodal displacements and to solve the contact equations. Finite element models of the reconstructed proximal femur were developed. A model of bone geometry was developed from the digitized sections of a right adult human femoral specimen. Two femoral stems (reference and experimental stems) were designed in accordance with the bone geometry. Rotational micromotion of the implant relative to the proximal femoral cortical surface was measured using a single, linearly variable differential transducer (LVDT). Three piezoelectric transducers were used to detect the displacement of the stem in the femur. RESULTS: Under all interface conditions, the compressive stresses of the coating surface were below 1.5 MPa. Shear stresses in the two friction models were below 0.5 MPa, and below 1 MPa in the bonded model. The stress exerted over the cortex in the experimental model was 50% of that in the reference model. The relative displacement of the stem in the coated region was less than 0.05 mm, but it increased distally in a linear fashion and was 0.45 mm at the stem tip. A stress concentration in the proximal femoral cancellous bone was noticeably higher in the experimental model than in the reference model. However, the overall characteristics of stress transfer were not changed by stem shape modification. Experimental stem was found to have significantly less rotational micromotion and total permanent rotational displacement values (between 10 and 29 N.m) than the reference stem. CONCLUSION: The torsional stability of the experimental stem is enhanced by: increasing stem thickness in the anteroposterior plane; adding a lateral flare to the stem; and a congruent fit between the proximal medial portion of the stem and calcar femorale. A role of short and tapered distal stem of the experimental stem was negligible in providing with the stem stability and, therefore, the femoral distal stem can be removed. Distal stem removal can minimize stress shielding related bone resorption and can avoid thigh pain.

Study on the Stability Test of Diedahuoxue Film Former / 中国药房

AIM:To predetermine the period of efficacy of Diedahuoxue film former METHODS:The chemical stability of Diedahuoxue film former was studied by the constant temperature accelerated test according to the principle of chemical kinetics Using the emodin in Diedahuoxue film former as an index,its content was determined by spectrophotometry The period of efficacy of Diedahuoxue film former was determined RESULTS:The results showed that the decomposition of emodin in Diedahuoxue film former accorded with the primary reaction CONCLUSION:At room temperature(25℃),it is predetermined that the period of efficacy of Diedahuoxue film former is 1 10 years

Preparation and stability test of Lyophiled Royal Jelly Soft Capsule / 中成药

AIM:To prepare the Lyophiled Royal Jelly Soft Capsule and study its stability and Influential factors.METHODS:The suspending agent and processing method were optimized using sedimentation volume rate as the index.Soft capsules were prepared and product stability under high temperature and high humidity environment was studied according to the determination of the content of 10-HAD by HPLC.RESULTS:The finished product yield in pilot test was more than 90%,the soft capsule products stored in cold were stable,while those stored under room temperature or high temperature and high humidity were unstable with a noticeable decrease in quality.Water content in capsule shell affects the 10-HDA content of the finished product.CONCLUSION:The preparative process is feasible and the products should be storaged in cold enviroment.

Form Stability Test on Gastric Aspirate and Respiratory Distress Syndrome in Newborn Baby

Form stability test on gastric aspirate was performed on 102 cases of newborn within 30 minutes afterbirth, who were admitted to Special Care Unit of St. Benedict Hospital from Sep. 1977 to Aug. 1979, and following results were obtained. 1) There was no differnce between amniotic fluid and gastric aspirate in control group. 2) In all newborn infants with positive form stability test, respiratory distress syndrome was not developed. 3) Respiratory distress syndrome was observed in 75% by Clements method and 55% by Transwell's method in negative form stability test group. 4) The frequency of negative foam stability test and incidence of respiratory distress syndrome were higher in the group of younger gestational age, smaller body weight and lower Apgar score at birth respectively.