Validation and statistical analysis of two high performance liquid chromatography methods for the determination of indinavir sulfate raw material and capsules

Two high performance liquid chromatography (HPLC) methods for the quantitative determination of indinavir sulfate were tested, validated and statistically compared. Assays were carried out using as mobile phases mixtures of dibutylammonium phosphate buffer pH 6.5 and acetonitrile (55:45) at 1 mL/min or citrate buffer pH 5 and acetonitrile (60:40) at 1 mL/min, an octylsilane column (RP-8) and a UV spectrophotometric detector at 260 nm. Both methods showed good sensitivity, linearity, precision and accuracy. The statistical analysis using the t-student test for the determination of indinavir sulfate raw material and capsules indicated no statistically significant difference between the two methods.

Stability-indicating methods for quantitative determination of zidovudine and stavudine in capsules

Zidovudine (AZT) and stavudine (D4T) are nucleoside reverse transcriptase inhibitors extensively used in human immunodeficiency virus (HIV) infected patients. In order to evaluate the quality of these drugs, two stability indicating HPLC methods were developed. The validated methods were applied in quantitative determination of AZT, D4T and their induced degradation products in capsule preparations. The stability studies were conducted at controlled temperature and relative humidity conditions based on the International Conference on Harmonization stability studies protocol for Zone IV areas. Easy sample preparation and low-cost make these methods especially useful for quality control and stability studies of AZT and D4T in drug products.

Supercritical CO2 recovery of caffeine from green coffee oil: new experimental solubility data and modeling

The caffeine solubility in supercritical CO2 was studied by assessing the effects of pressure and temperature on the extraction of green coffee oil (GCO). The Peng-Robinson¹ equation of state was used to correlate the solubility of caffeine with a thermodynamic model and two mixing rules were evaluated: the classical mixing rule of van der Waals with two adjustable parameters (PR-VDW) and a density dependent one, proposed by Mohamed and Holder² with two (PR-MH, two parameters adjusted to the attractive term) and three (PR-MH3 two parameters adjusted to the attractive and one to the repulsive term) adjustable parameters. The best results were obtained with the mixing rule of Mohamed and Holder² with three parameters.

Determination of cetirizine in tablets and compounded capsules: comparative study between CE and HPLC

A capillary electrophoresis (CE) method was developed and validated for determination of cetirizine dihydrochloride in tablets and compounded capsules. The electrophoretic separation was performed in an uncoated fused-silica capillary (40 cm x 50 μm i.d.) using 20 mmol L-1 sodium tetraborate buffer (pH 9.3) as background electrolyte, a hydrodinamic sample injection at 50 mBar for 5 s, 20 KV applied voltage at 25 °C, and detection at 232 nm. The proposed method was compared with the high performance liquid chromatographic (HPLC) method previously validated for this drug, and statistical analysis showed no significant difference between the techniques.

Losartan potassium dissolution test for drug release evaluation in pharmaceutical capsules using HPLC and UV spectrophotometry

This work describes the development and validation of a dissolution test for 50 mg losartan potassium capsules using HPLC and UV spectrophotometry. A 2(4) full factorial design was carried out to optimize dissolution conditions and potassium phosphate buffer, pH 6.8 as dissolution medium, basket as apparatus at the stirring speed of 50 rpm and time of 30 min were considered adequate. Both dissolution procedure and analytical methods were validated and a statistical analysis showed that there are no significant differences between HPLC and spectrophotometry. Since there is no official monograph, this dissolution test could be applied for quality control routine.

Development and validation of a dissolution test for diltiazem hydrochloride in immediate release capsules

This work describes the development and validation of a dissolution test for 60 mg of diltiazem hydrochloride in immediate release capsules. The best dissolution in vitro profile was achieved using potassium phosphate buffer at pH 6.8 as the dissolution medium and paddle as the apparatus at 50 rpm. The drug concentrations in the dissolution media were determined by UV spectrophotometry and HPLC and a statistical analysis revealed that there were significant differences between HPLC and spectrophotometry. This study illustrates the importance of an official method for the dissolution test, since there is no official monograph for diltiazem hydrochloride in capsules.

Development and validation of a method for the analysis of Ochratoxin A in roasted coffee by liquid chromatography/electrospray-mass spectrometry in Tandem (LC/ESI-MS/MS)

A method using LC/ESI-MS/MS for the quantitative analysis of Ochratoxin A in roasted coffee was described. Linearity was demonstrated (r = 0.9175). The limits of detection and quantification were 1.0 and 3.0 ng g-1, respectively. Trueness, repeatability and intermediate precision values were 89.0-108.8%; 2.4-13.7%; 12.5-17.8%, respectively. To the best of our knowledge, this is the first report in which Ochratoxin A in roasted coffee is analysed by LC/ESI-MS/MS, contributing to the field of mycotoxin analysis, and it will be used for future production of Certified Reference Material.

Evaluation of hard gelatin capsules and hydroxypropyl methylcellulose containing ampicillin

This study aims to develop and evaluate formulations containing ampicillin in capsules of gelatin and hydroxypropyl methylcellulose (HPMC). Two formulations (A and B) were developed. The final product quality was evaluated by testing for quality control and the results were in agreement with the Brazilian Pharmacopoeia. The formulations with HPMC capsules showed lower percentages of drug dissolved (99.67%, HPMC-A and 87.70%, HPMC-B) than the gelatin (100.18%, GEL-A and 101.16% GEL-B). Because of the delay of the ampicillin release observed in the dissolution profiles, it becomes necessary to evaluate the drugs that can be conditioned in the HPMC capsules.

Performance characteristics of high performance liquid chromatography, first order derivative UV spectrophotometry and bioassay for fluconazole determination in capsules

The bioassay, first order derivative UV spectrophotometry and chromatographic methods for assaying fluconazole capsules were compared. They have shown great advantages over the earlier published methods. Using the first order derivative, the UV spectrophotometry method does not suffer interference of excipients. Validation parameters such as linearity, precision, accuracy, limit of detection and limit of quantitation were determined. All methods were linear and reliable within acceptable limits for antibiotic pharmaceutical preparations being accurate, precise and reproducible. The application of each method as a routine analysis should be investigated considering cost, simplicity, equipment, solvents, speed, and application to large or small workloads.

Espectroscopia no infravermelho médio e análise sensorial aplicada à detecção de adulteração de café torrado por adição de cascas de café

Mid-infrared spectroscopy and chemometrics were used to identify adulteration in roasted and ground coffee by addition of coffee husks. Consumers' sensory perception of the adulteration was evaluated by a triangular test of the coffee beverages. Samples containing above 0.5% of coffee husks from pure coffees were discriminated by principal component analysis of the infrared spectra. A partial least-squares regression estimated the husk content in samples and presented a root-mean-square error for prediction of 2.0%. The triangular test indicated that were than 10% of coffee husks are required to cause alterations in consumer perception about adulterated beverages.

Description of epidemics of coffee rust with neural networks

The objective of this paper was to evaluate the potential of neural networks (NN) as an alternative method to the basic epidemiological approach to describe epidemics of coffee rust. The NN was developed from the intensities of coffee (Coffea arabica) rust along with the climatic variables collected in Lavras-MG between 13 February 1998 and 20 April 2001. The NN was built with climatic variables that were either selected in a stepwise regression analysis or by the Braincel® system, software for NN building. Fifty-nine networks and 26 regression models were tested. The best models were selected based on small values of the mean square deviation (MSD) and of the mean prediction error (MPE). For the regression models, the highest coefficients of determination (R²) were used. The best model developed with neural networks had an MSD of 4.36 and an MPE of 2.43%. This model used the variables of minimum temperature, production, relative humidity of the air, and irradiance 30 days before the evaluation of disease. The best regression model was developed from 29 selected climatic variables in the network. The summary statistics for this model were: MPE=6.58%, MSE=4.36, and R²=0.80. The elaborated neural networks from a time series also were evaluated to describe the epidemic. The incidence of coffee rust at four previous fortnights resulted in a model with MPE=4.72% and an MSD=3.95.

Effect of extracts from citric biomass, rusted coffee leaves and coffee berry husks on Phoma costarricencis of coffee plants

Phoma leaf spot, caused by Phoma costarricensis poses a serious threat to coffee (Coffea arabica) production, especially in the highlands of the state of Minas Gerais, Brazil. Extracts of citric biomass, coffee berry husks and coffee leaves severely affected by rust caused by Hemileia vastatrix, were evaluated against P. costarricensis. In an in vitro assay, aqueous extracts of rusted leaves and berry husks plus the commercial extracts based on citric biomass named Ecolife® and Agromil® were tested at various dilutions on the mycelial growth inhibition of P. costarricensis. In vivo, coffee seedlings maintained in glasshouse, were sprayed with these extracts seven days before inoculation of P. costarricensis. Only extracts from citric biomass had inhibitory effects on the fungus. In vivo, Ecolife® (5 ml/l), Agromil® (5 g/l) and the aqueous extract of rusted coffee leaves (dilution 1:6) reduced Phoma leaf spot. Both, Ecolife® and the extract of rusted coffee leaves were significantly more effective in reducing the area under the lesion progress curve when applied at lower doses, indicating a possible effect on the induction of resistance.

Vírus da mancha anular do cafeeiro (Coffee ringspot virus - CoRSV): influência na qualidade da bebida e na produção de grãos de café

A mancha anular do cafeeiro, causada pelo Coffee ringspot virus (CoRSV) que é transmitido pelo ácaro Brevipalpus phoenicis (Geijskes) (Acari: Tenuipalpidae), tem sido observada em altas incidências em várias regiões cafeeiras do Estado de Minas Gerais. O CoRSV causa manchas cloróticas arrendondadas ou irregulares nas folhas, caules e frutos. Foi feita uma avaliação do efeito da infecção de frutos do cafeeiro pelo CoRSV na qualidade da bebida por meio de teste bioquímico e de degustação, e também na eventual perda de peso nos grãos. Testes revelaram que grãos provenientes de frutos de café infectados pelo CoRSV apresentavam menor teor de açúcares redutores e maior condutividade elétrica. Houve também depreciação na qualidade de bebida gerada pelos frutos infectados por meio do teste de degustação (teste de xícara). O peso médio dos grãos provenientes de frutos manchados foi cerca de 5% menor do que dos grãos de frutos sem sintomas.

Incubation period of Hemileia vastatrix in coffee plants in Brazil simulated under climate change

Risk analysis of climate change on plant diseases has great importance for agriculture since it allows the evaluation of management strategies to minimize future damages. This work aimed to simulate future scenarios of coffee rust (Hemileia vastatrix) epidemics by elaborating geographic distribution maps using a model that estimates the pathogen incubation period and the output from three General Circulation Models (CSIRO-Mk3.0, INM-CM3.0, and MIROC3.2.medres). The climatological normal from 1961-1990 was compared with that of the decades 2020s, 2050s and 2080s using scenarios A2 and B1 from the IPCC. Maps were prepared with a spatial resolution of 0.5 × 0.5 degrees of latitude and longitude for ten producing states in Brazil. The climate variables used were maximum and minimum monthly temperatures. The maps obtained in scenario A2 showed a tendency towards a reduction in the incubation period when future scenarios are compared with the climatological normal from 1961-1990. A reduction in the period was also observed in scenario B1, although smaller than that in scenario A2.

GFP-fluorescent protein on the study of blister spot in coffee plants

In Brazil, Colletotrichum gloeosporioides is associated with a complex of symptoms in coffee culture. Although this pathogen had its pathogenesis observed and identified, its importance has still been questioned due to its several endophytic forms, raising doubts as to the real importance of the pathosystem. The aim of this study was to demonstrate, by using an isolate transformed with the gene gfp, the infection and colonization capability of C. gloeosporioides in coffee seedlings. After the fourth day of inoculation, manifestation of symptoms as punctual necrosis could be observed, which progressed during the evaluation period, culminating in the death of seedlings. Epifluorescence microscopy confirmed the presence of the pathogen in the seedlings, as well as the visualization of internal colonization of tissues, acervulus formation and conidium production, confirming that it was responsible for the observed symptoms.