Eugenia punicifolia (Kunth) DC, known as “pedra ume-caá” or “pitanguinha do cerrado”, is popularly used to treat infectious diseases like inflammation, and diabetes. Therefore, this study aimed to develop and validate an analytical method by high-performance liquid chromatography with diode array detector (HPLC-DAD) for the seasonal quantification of gallic acid (GA) and ellagic acid (EA), and to co-validate the method for quantification from the ethanol extract and fractions of the E. punicifolia leaves. Chromatographic separations were performed with a mobile phase gradient of acetonitrile/methanol/water acidified with 0.2% formic acid, a flow of 0.8 mL min 1, detection at 254 nm, and a C18 column (250 × 4.6 mm, 5 µm). Method validation parameters were determined following Brazilian legislation. The HPLC-DAD analytical method developed for the quantification of GA and EA was validated and demonstrated to be selective, linear, precise, accurate, robust, and without matrix effect. Accordingly, it is helpful for the analysis of these tannins in crude ethanol extracts and fractions from E. punicifoila leaves. Thus, our results may be of great help for quality control assessment and standardization of raw materials containing Eugenia punicifolia.

ABSTRACT In this work, we developed and validated a HPLC-PDA method for the quantification of hibalactone in Hydrocotyle umbellata L., Araliaceae, subterraneous parts extracts and optimized its ultrasound-assisted extraction. Chromatographic separations were carried out with an isocratic mobile phase of acetonitrile/methanol/water (10:65:25), a flow of 0.8 ml min−1, detection at 290 nm and C18 column (250 × 4.6 mm, 5 µm). The method validation parameters were determined according to Brazilian legislation. The optimization of the hibalactone ultrasound-assisted extraction was performed using Box–Behnken design and response surface methodology. The HPLC method for hibalactone quantification proved to be selective, linear, precise, accurate and robust, being useful for the analysis of hibalactone in H. umbellata subterraneous parts extracts. The optimal ultrasound-assisted extraction conditions were obtained with solid-to-liquid ratio of 1:5 g ml−1, ethanolic strength of 70% (v/v) and temperature of 65 °C. The results can provide support of the quality control and standardization of raw materials from H. umbellata.

Abstract Trembleya parviflora (D. Don) Cogn., Melastomataceae, also known as "quaresmeira-branca", is a subshrub that is commonly used to treat verminosis, scabies, dermatoses, rheumatism, vaginal infections, ulcerations and wounds. The aim of this work was to perform a morphological study of T. parviflora, evaluate the composition and chemical variability of the volatile oils from the leaves, perform phytochemical screening of the powder from the leaves and to define parameters for quality control of the plant material. Macroscopic characterization of T. parviflora was carried out by naked eye in Serra dos Pireneus, Pirenópolis, Goiás for 12 months. Volatile oils were subjected to hydrodistillation with Clevenger apparatus and analyzed by gas chromatography-mass spectrometry. Phytochemical screening and ash and volatile compound content determination were performed by conventional techniques. T. parviflora has simple, oppositely crossed and petiolate leaves. The inflorescence of this plant is a cyme. The presence of coumarins, steroids, triterpenes, flavonoids and tannins was observed. The total ash content was 4.05 ± 0.02%; the insoluble ash content was 0.10 ± 0.03%; and the volatile compound content was 9.53 ± 0.02%. The major compounds present in the volatile oils were α-terpineol (2.7–16.5%), α-pinene (0.6–25.4%), β-pinene (2.7–23.1%), sabinene (1.2–14.1%), acetoxyeudesman-4-α-ol (0.6–6.3%) and 2,4a-8,8-tetramethyldecahydrocyclopropanaphtalene (2.4–24.4). Two clusters were identified: Cluster I represented the period with low levels of rainfall, and Cluster II represented the period with high levels of rainfall. This study provides data that can be applied for the quality control of powdered leaves and is the first description of the chemical composition and variability of the volatile oils from the leaves of T. parviflora.

ABSTRACT Objective: To present our technique of ureteroileal bypass to treat uretero-enteric stric- tures in urinary diversion. Materials and Methods: One hundred and forty-one medical records were reviewed from patients submitted to radical cystectomy to treat muscle-invasive bladder cancer between 2013 and 2015. Twelve (8.5%) patients developed uretero-enteric anastomotic stricture during follow-up. Five patients were treated with endoscopic dilatation and double J placement. Four were treated surgically with standard terminal-lateral im- plantation. Three patients with uretero-enteric anastomotic stricture were treated at our institution by “ureteroileal bypass”, one of them was treated with robotic surgery. Results: All patients had the diagnosis of uretero-enteric anastomotic stricture via computerized tomography and DTPA renal scan. Time between cystectomy and diag- nosis of uretero-enteric anastomotic stricture varied from five months to three years. Mean operative time was 120±17.9 minutes (98 to 142 min) and hospital stay was 3.3±0.62 days (3 to 4 days). Mean follow-up was 24±39.5 months (6 to 72 months). During follow-up, all patients were asymptomatic and presented improvement in ure-terohydronephrosis. Serum creatinine of all patients had been stable. Conclusions: Latero-lateral ureter re-implantation is feasible by open or even robotic surgery with positive results, reasonable operation time, and without complications.

ABSTRACT Campomanesia adamantium (Cambess.) O. Berg., Myrtaceae, is a plant popularly used for its anti-inflammatory, anti-diarrhoeal and urinary antiseptic activities. The aims of this study were to obtain the crude ethanolic extract and the hexane, dichloromethane, ethyl acetate, aqueous and concentrated aqueous tannin fractions from C. adamantium leaves, perform biomonitored fractionation to isolate and identify chemical compounds, study the chemical composition of the volatile oils of the leaves and flowers and test the antimicrobial activity of the ethanolic extract, fractions, isolated substances and volatile oils. Phytochemical screening and chromatographic and spectrometric techniques were used. Volatile oils were isolated by hydrodistillation in a Clevenger apparatus and analyzed by gas chromatography/mass spectrometry. The antimicrobial activity was tested by a broth microdilution test. The component stictane-3,22-diol was isolated and identified from the hexane fraction, while valoneic and gallic acid were isolated and identified from the concentrated aqueous tannin fraction. The major constituents of the volatile oils of the leaves were verbenene (13.91%), β-funebrene (12.05%) and limonene (10.32%), while those of the volatile oils of the flowers were sabinene (20.45%), limonene (19.33%), α-thujene (8.86%) and methyl salicylate (8.66%). Antibacterial activity was verified for the hexane fraction, while antifungal activity was observed for the aqueous fraction and concentrated aqueous tannin fraction and for vanoleic acid. These results may justify the popular use of C. adamantium.

ABSTRACT Costus spiralis is a Brazilian native plant used in popular medicine, but the safety of this therapeutic use needs investigation. So, the aim of this study was to evaluate the cytogenotoxic and antigenotoxic effects of C. spiralis leaves or stems aqueous extracts on Allium cepa root cells. Moreover, a phytochemical screening and an antioxidant and antifungal activities evaluation were performed. C. spiralis aqueous extracts presented cytotoxicity, but no mutagenicity was observed. When the antigenotoxicity was evaluated, C. spiralis leaves aqueous extract presented preventive and modulatory effects on A. cepa root cells, reducing the sodium azide cytogenotoxic effects. In contrast, C. spiralis stems aqueous extract enhanced the sodium azide cytogenotoxicity in some conditions. The phytochemical screening revealed the presence of phenolic compounds in C. spiralis. When total phenolic content was determined, the leaves presented 73% more phenolic content than stems. Corroborating this data, C. spiralis leaves antioxidant potential was 30% higher than C. spiralis stems. However, these extracts did not present antifungal activity against Candida spp. In conclusion, empirical utilization of C. spiralis aqueous extracts should be avoided. Moreover, the cytotoxic effect of C. spiralis leaves and stems can play an important role in anticancer therapy and must be deeply studied.

ABSTRACT Spiranthera odoratissima A. St.-Hil., Rutaceae, known as “manacá” is a shrub native of the Brazilian Cerrado. Their leaves and roots are popularly used to treat rheumatism, infection and abdominal pain. This study analyzed the chemical composition of volatile oils from leaves of S. odoratissima and verified the seasonal variability of its chemical composition. The volatile oils were obtained by hydrodistillation using a Clevenger type apparatus and analyzed by gas chromatography coupled to mass spectrometry. The main chemical components found in samples of volatile oils were β-caryophyllene, bicyclogermacrene, δ-cadinene, amorphous-4,7(11)-diene, α-epi-muurolol, α-cadinol, α-muurolol and γ-cadinene. The hierarchical clustering identified three groups: the first was characterized by α-epi-muurolol, the second by amorphous-4,7(11)-diene and the third group was characterized by α-muurolol. The discriminant canonical analysis was used to differentiate between clusters on the basis of oil composition. The results suggest that the rainfall presented a relationship with the chemical composition of the volatile oil. This is the first study conducted on the seasonal behavior of the chemical constituents in volatile oil from leaves of S. odoratissima.

Abstract The Pterodon genus comprises two native species in Brazil, known as “sucupira-branca” or “faveira”. Their fruits have long been used in Brazilian natural medicine, mainly for the treatment of infections and inflammations. The pharmacological properties of these fruits have often been linked with vouacapane diterpenoids. This review evaluated the scientific research in the period from 1973 to February 2017, aiming to answer how difficult it still is to develop a scientifically supported product based on Pterodon vouacapanes. Therefore, this paper reviews purification, identification, and quantification methods applied to vouacapane diterpenoids from Pterodon, as well as the performance of these phytochemicals in pharmacological tests described in the literature. Data analysis results support conventional notions that suggest vouacapane diterpenoids from Pterodon have anti-inflammatory properties. However, the studies carried out so far still represent partial assessment of the vouacapane activities and further studies need to be completed. Pterodon diterpenoids have also been associated with larvicidal, leishmanicidal, cardiovascular, and antitumor activities, which reinforces the genus' potential as a source of phytomedicines. Some remaining gaps about the reviewed activities were mentioned, while trends and perspectives for future research were proposed.

ABSTRACT Obtaining artemisinin and its derivatives is very costly, which limits access to low-income people. Some hydroalcoholic extract of Artemisia annua L., Asteraceae, which has shown comparable antimalarial activity could be an alternative to the purified compound, especially if the cultivars have higher content of artemisinin. The objective of this study was to evaluate the effects of the extraction parameters (ethanol graduation, previous shaking time in an ultrasound bath and drug/solvent ratio) on the yield of artemisinin in the liquid extract obtained by percolation from A. annua and then optimize the extraction efficiency of this compound. The Box–Behnken (33) factorial design was used in association with response surface methodology. The derivatization reaction of artemisinin was used in an analytical method which was submitted to validation, after reaching the specification of the selectivity, linearity, precision, accuracy and robustness. Detection and quantification limits were 1.3 and 4.0 µg/ml, respectively. The largest amount of this compound of interest was obtained without any ultrasound bath, with an ethanol graduation of 95% and a drug/solvent ratio of 2%. Drug/solvent ratio was the factor which most influenced extraction efficiency. The maximum range of artemisinin yield was 1.21%. Information obtained in this study can be used for future approaches to determining and extracting artemisinin from A. annua.

ABSTRACT Trembleya phlogiformis DC., Melastomataceae, is a shrub whose leaves are used as a dye for dyeing wool and cotton. The present article aimed to carry out the morphological description of the species, to study the chemical composition of volatile oils from the leaves and flowers and the seasonal variability from the leaves during a year. Macroscopic characterization was carried out with the naked eye and with a stereoscopic microscope. Volatile oils were isolated by hydrodistillation in Clevenger apparatus and analyzed by gas chromatography/mass spectrometry. The major components of the volatile oil of T. phlogiformis flowers were: n-heneicosane (33.5%), phytol (12.3%), n-tricosane (8.4%) and linoleic acid (6.1%). It was verified the existence of a large chemical variability of the volatile oils from the leaves of T. phlogiformis over the months, with the majority compound (oleic alcohol, ranging from 5.7 to 26.8%) present in all samples. A combination of Cluster Analysis and Principal Component Analysis showed the existence of three main clusters, probably related to the seasons. The results suggested that the volatile oils of T. phlogiformis leaves possess high chemical variability, probably related to variation associated with rainfall and the variation in the behavior of specimens throughout the year. This research provides insights for future studies on the volatile oils obtained from the T. phlogiformis leaves and flowers, mainly related to biological markers of applications monitored in the leaves and flowers of this species.

ABSTRACT Hyptis carpinifolia Benth., Lamiaceae, is a species known popularly as "rosmaninho" and "mata-pasto", and leaves are employed in Brazilian folk medicine to treat colds, flu, and rheumatism. The aim of this study was to perform a morphological description of H. carpinifolia and to evaluate the seasonal chemical variability of the leaf essential oils during 12 months. Macroscopic characterization of H. carpinifolia was carried out with the naked eye and with a stereoscopic microscope. Essential oils were isolated from leaves by hydrodistillation in Clevenger apparatus and analyzed by gas chromatography/mass spectrometry. Major compounds were found to be 1,8-cineole (39.6-61.8%), trans-cadina-1(6),4-diene (2.8-17.5%), β-caryophyllene (4.4-10.0%), prenopsan-8-ol (4.2-9.6%) and β-pinene (2.9-5.3%). Results of essential oils compositions were processed by cluster analysis and principal component analysis. Data showed high variability in the concentration of the components. Besides, there was a seasonal variability of chemical composition, probably related mainly to the rainfall regime.

Abstract Hortia oreadica Groppo, Kallunki & Pirani, Rutaceae, known as “para-tudo”, “quina”, and “quina-do-campo”, is used in traditional medicine locally to treat stomach pain and fevers. The aims of this study were: analyze the chemical composition of essential oils from leaves, flowers and fruits of H. oreadica and verify the seasonal variation of the chemical components of essential oils from leaves. The essential oils were obtained by hydrodistillation using a Clevenger type apparatus and analyzed by GC/MS. The major components found in the samples of the essential oils were the amorpha-4,7(11)-diene (29.27% – flowers, 20.26% – fruits, 27.66–37.89% – leaves), bicyclogermacrene (23.28% – flowers, 20.64% – fruits, 14.71% to 31.37% – leaves). This work represents the first study of the chemical composition of essential oils from leaves, flowers and fruits and seasonal variation in the essential oils from leaves of H. oreadica.

This study aimed to impact of different extraction methods on the quality of Dipteryx alata Vogel, Fabaceae, extracts from fruits. The major compounds found were the lipids 38.9% (w/w) and proteins 26.20% (w/w). The residual moisture was 7.20% (w/w), total fiber 14.50% (w/w), minerals 4.10% (w/w) and carbohydrate 9.10 % (w/w). The species studied has great potential in producing oil, but the content and type of fatty acids obtained is dependent on the method of extraction. The Blingh & Dyer method was more selective for unsaturated fatty acids and Shoxlet method was more selective for saturated fatty acids. The tannin extraction by ultrasound (33.70 % w/w) was 13.90% more efficient than extraction by decoction (29 % w/w).

Syzygium jambos (L.) Alston, Myrtaceae, occurs in tropical regions and is a widespread medicinal plant used to treat several diseases, such as hemorrhage, dysentery, diabetes, inflammation, diabetes and gastrointestinal disorders. Leaf essential oils of ten specimens of S. jambos collected from two localities of Central Brazilian Cerrado were investigated by GC-MS. Soil and foliar nutrients were analyzed to determine the mineral compositions. The aims of this study was to evaluate the influence of environmental factors on chemical composition of leaf essential oils of S. jambos. Studies on the influence of environmental factors over composition of essential oils are important because they contribute data for its cultivation, harvest and establish parameters to essential oil components. The data were analyzed using stepwise Multiple Regression and Cluster Analysis, and the results suggest that the main factor capable to influence the chemical composition of leaf essential oils was the collection period and the collection site had a minor effect. The results also suggest that the leaf essential oils composition was influenced mainly by foliar nutrients (N, Mn, Co, Fe, S and Mg) and soil nutrients (Na, Al, S and H+Al). The compound with the best model obtained was the (E)-caryophyllene, with a coefficient of determination equal 0.8113.

Myrcia tomentosa (Aubl.) DC., Myrtaceae, found in Central Brazilian Cerrado and popularly known as "goiaba-brava", belongs to the Myrcia genus, which has several species with medicinal properties such as: hypoglycemic, diuretic, hypotensive, antidiarrheal, antimicrobial and antitumor. The present study aimed to analyzed the environmental influence on concentrations of phenolic metabolites in M. tomentosa leaves. Compounds assayed in the leaves were: total phenols, tannins by protein precipitation, hydrolysable tannins and total flavonoids and mineral nutrients, while soil fertility was also analyzed, all over during one year. The results were submitted to Pearson Correlation Analysis and stepwise Multiple Regression Analysis to investigate the relationship between phenolics and environment data. Analysis of variance and Cluster Analysis allowed indicated a high variability in samples from different sites. The results obtained suggests that content of phenolics from M. tomentosa leaves are influenced by environmental factors, particularly some foliar nutrients (N1, Ca1 and Mn1), soil nutrients (Ca s and Ks) and Rainfall.