Acute toxicity tests and dyeing potentials of three azo dyes from Pyocyanin, produced by Pseudomonas aeruginosa from farmlands

The admiration for colours by man has made dyes to have very useful applications on a great variety of materials. Pyocyanin (PCN) or 5-methyl-1-hydroxyphenazine, a phenazine derivative biosynthesized by Pseudomonas aeruginosa isolated from the stressed soil of a farmland at Awka was used to prepare three azo dyes namely, pyocyanin azophenol, pyocyanin azo-1-naphthol and pyocyanin azo-2-naphthol. The PCN was treated with SOCl2 to convert it to the chloro analogue. Treatment of chloro analogue with NaCN in ethanol and subsequent acid hydrolysis gave the carboxylic acid from which the amide was obtained after treatment with (NH4)2CO3 and heat. The amide was converted to the amine by Hoffman’s amide degradation. The amine was diazotized and coupled with phenol, 1-naphthol and 2-naphthol to give the three azo dyes respectively. UV-VIS and infrared spectra of the compounds agreed with the assigned structures. Acute Toxicity (LD50) tests showed the compounds to be reasonably non-toxic. Dyeing potentials of the Pyocyanin (PCN), PCN azophenol, PCN azo-1-naphthol and PCN azo-2-naphthol by mordant dyeing on cotton, silk and nylon textile materials showed them to have blue, dark green, purplish-brown and red colours respectively. The dyes were reasonably fast to light, washing, rubbing, alkali and acid. Pyocyanin, Pyocyanin azophenol, Pyocyanin azo-1-naphthol and pyocyanin azo-2-naphthol produced from Pseudomonas aeruginosa isolated from farmland soil can be used effectively to dye cotton, silk and nylon textile materials.

Study of the Crude oil Properties of Oily seepages found in South Eastern Nigeria.

ASTM D-270 method was applied in obtaining well-head crude oil samples from Awoba 8T (Rivers State), Odidi 23L, and Odidi 46S (Delta State), Eremor 1L (Bayelsa State) and Oily Seepage samples from Ugwueme (Enugu State) and Anam (Anambra State). The0API gravity value of all the samples were 16.190 and 21.470 for Ugwueme seepage and Eremorcrude respectively, 22.470 for Odidi 46s crudeand 44.700, 43.840 and 34.760 for Anam seepage, Awoba 8t and Odidi 23L crudesrespectively, indicating heavy, medium and light crude oils respectively. All the well-head crudes and oily seepage samples studied in this work are onshore samples except Odidi 46s which is an offshore crude sample. The samples were subjected to liquid-liquid extraction using a mixture of n-hexane and dichloromethane (3:1). The extracted fractions were analyzed using GC-FID. The result of the analyses gave polycyclic aromatic hydrocarbons (PAHs) concentrations (mg/kg) of 7106.43 for Awoba 8T crude, 6767.41 for Odidi 23L crude, 5771.20 for Odidi 46S crude, 5191.21 for Eremor crude, 4908.04 for Ugwueme Seepage and 6983.16 for Anam seepage. The predominance of low molecular weight PAHs (LMWPs) over the high molecular weight PAHs (HMWPs) indicate that both the crude oil samples and the oily seepage samples were of Petrogenic Source. However, various ratios were obtained to identify the possible source of PAHs in the crude oil and seepage samples. Mixed sources of pyrogenic and petrogenic PAHs were also discovered with evidence of „pyrolytic‟ input which may suggest some geothermal-mediated processes.

TIME COURSE STUDY OF COMBINED N-ACETYL-P-AMINOPHENOL AND DICLOPHENAC INDUCED-HEPATOTOXICITY AND OXIDATIVE STRESS IN WISTAR RATS

The abuse of combined acetaminophen or N-acetyl-p-aminophenol (APAP) and diclofenac (DIC) due to their analgesics, anti-inflammatory and antipyretic properties is a predominant cause of hepatotoxicity and oxidative stress. This study investigated the time-course effects of APAP, DIC and their combination on biomarkers of hepatic function and oxidative stress in rats. Forty male Wistar rats were randomly divided into four groups of 10 animals each as follows; control (distilled water), APAP only, DIC only and APAP + DIC for 4 weeks. Indices of liver damage (serum ALT, alanine aminotransferase; AST, aspartate aminotransferase; ALP, alkaline phosphatase; GGT, gamma-glutamyl transferase and bilirubin) were measured. Oxidative stress biomarkers (MDA, malondialdehyde; NO, nitric oxide; CAT, Catalase activity; SOD, superoxide dismutase activity; GSH content, reduced glutathione), GR, glutathione reductase, and GST, glutathione-S-transferase) were also determined using spectrophotometric methods. Statistical analysis was done using one-way ANOVA with p < 0.05 considered significant. Acetaminophen and diclofenac caused marked liver damage as noted by time-dependent significant (p < 0.05) increased activities of serum ALT, AST, ALP, GGT, and bilirubin levels as well as significant (p < 0.05) increase in hepatic MDA and NO levels as compared to the control group. Hepatic GSH content, SOD, CAT, GPx, GST and GR activities were decreased significantly (p < 0.05) in all acetaminophen and diclofenac-treated groups compared to normal control in a time-dependent manner. These findings suggest that prolonged administration of diclofenac, acetaminophen or their combination may induce hepatotoxicity, oxidative stress and alteration of hepatic antioxidant status in a time-dependent manner.

CHEMICAL COMPOSITION OF SEEDS AND SEED OIL OF Afraegle paniculata (Rutaceae)

Afraegle paniculata (Schumach. & Thonn.) Engl. commonly known as Nigerian powder-flask is a plant of rutaceae family found in West Africa from Senegal to Nigeria. The chemical composition of seeds and seed oil of Afraegle paniculata were evaluated in this study. The seed oil was obtained by soxhlet extraction using n-hexane. Chemical composition analyses involved proximate, mineral element, physicochemical and phytochemical. The percentage mean value of the proximate analysis revealed that the seeds contained 28.81±0.02 % crude fat, 25.03±0.12 % crude protein, 10.90±0.03 % moisture, 3.11±0.01 % ash, 25.19±0.02 % crude fibre and 6.96±0.14 % carbohydrate. The mineral element analysis result showed that potassium (114.87 mg/l) was the predominant mineral element followed by magnesium (47.20±0.037 mg/l) and sodium (45.37±0.53 mg/l). Other minerals present were calcium (2.12±0.014 mg/l), iron (1.12±0.028 mg/l), copper (0.41±0.002 mg/l), zinc (0.39±0.094 mg/l) and manganese (0.29±0.005 mg/l). The oil was liquid at room temperature and golden yellow in appearance. The recorded pH, acid, peroxide, iodine, saponification, ester and oil yield values were 3.94, 1.29 mgKOH/g, 17.57 meq/kg, 42.24 mgI2/100g, 203.76 mgKOH/g, 202.47 mgKOH/g, 40.77 % w/w respectively . The presence of alkaloids, saponins, terpenoids, steroids and anthraquinone were evident in the phytochemical screening of the seed oil. Nutritional profile of A. paniculata seed could offer a scientific basis for use of the seeds and oils both in human diet and some commercial products. Keywords: Afraegle paniculata, proximate, mineral elements, physicochemical and phytochemical.

Isolation and Characterization of Secondary Metabolite from the Stem Bark Extract of Allophylus africanus Beauv (Sapindaceae)

Isolation of secondary metabolites from the stem bark of Allophylus africanus Beauv (Sapindaceae) was carried out. The plant has a wide distribution in West Africa, with Nigeria (Mambila Plateau, Calabar) inclusive. It has been used in communities for both its medicinal and economic importance. The stem bark of A. africanus was extracted by percolation using chloroform as solvent and concentrated by evaporation at room temperature. Single spot with an Rf value of 0.87 was obtained at a solvent mixture of hexane: chloroform at ratio 2:8 from finger column chromatography . Characterization using spectra analysis which include FT-IR, 1H-NMR, 13C-NMR, H-H COSY, H-C COSY, 135 DEPT and mass spectroscopy. Spectroscopic studies of A. Africanus stem bark extract led to the elucidation of the structure of the compound. FT-IR spectrum indicates that the compound is an aromatic derivative. Aromatic protons where seen at the 1H-NMR chemical shift values of 7.25 ppm. Olefinic protons at 5.124, 5.146 and 6.162 ppm appeared as doublets of doublets of doublets. The compound 1-((E)- 7(((1E,5E)-6-((2,3-dimethyl-4-(((E)-3,4,5,6-tetramethyldeca-7,9-dien-2-yl)oxy)pentyl)oxy)-3,4- dimethylhexa-1,5-dien-1-yl)oxy)hept-4-en-1-yl)-4-propylbenzene was isolated.

GC-MS ANALYSIS OF PHYTOCHEMICALS FROM THE EXTRACT OF Hibiscus sabdariffa GROWN IN NORTHERN NIGERIA

Hibiscus sabdariffa, also known as Roselle belongs to Malvacae family. It is popularly known in Nigeria as Zobo. It has been used traditionally to cure certain ailments like cold, due to its rich pharmacological potential. Hence this study aimed at validating the pharmacological potential of Hibiscus sabdariffa by identification of the compounds present in its leaves using GC-MS technique. The extract was obtained by heating the leaves in ethanol under reflux. The ethanol extract was then subjected to GC-MS analysis. The constituents were identified by comparing their mass fragmentation (MS) pattern with those gathered in the library of NIST-MS and with those reported in literature. The graph obtained from the study revealed the presence of nine (9) organic compounds in Hibiscus sabdariffa leaf extract (HSLE) namely cyclohexane carboxylic acid ester, cyclopropane carboxylic acid methyl ester, hexanoic acid-4-octyl ester, hexadeca-2-11-dienoic acid, n-hexadecanoic acid, oleic acid, octadecanoic acid, E-13-docosenoic acid and E-11-hexadecanal. Oleic acid has the highest peak value with the retention time of 20.711 which makes it the most abundant fraction and the active ingredient in the HSLE. The presence of some of these bioactive compounds has proved the scientific evidences for the antihypertensive and anti-inflammatory properties of the plant.

CATALYTIC DETOXIFICATION OF AQUEOUS CYANIDE USING COPPER CROWN ELECTROSPUN CHITOSAN NANOFIBER

Owing to the common phenomenon of cyanide toxicity, this study focused on oxidative catalytic detoxification of aqueous cyanide in the presence of copper-crown chitosan nanofiber. To overcome the difficulty in electrospinning chitosan, chitosan-salicylaldehyde was initially synthesized via Schiff base condensation, electrospun, and post neutralised into chitosan nanofiber. The copper-crown nanofiber was prepared using the impregnation method. The nanofiber material was characterized using FT-IR, SEM, TGA and DSC. The observed pseudo first-order rate constant for the catalytic H2O2 oxidation of aqeous cyanide with copper-crown nanofiber as catalyst was found to be 16 times greater than the oxidation with only copper catalyst. In addition, the reaction was found to be dependent on the size of the copper-crown nanofiber, pH, temperature, and H2O2/CN-. The reusability of the copper-crown nanofiber catalyst makes the process economically and potentially viable for commercial application.

ANTIBACTERIAL PHOTODYNAMIC THERAPEUTIC STUDIES OF METALLATED PORPHYRIN AGAINST CHRONIC WOUND COLONISING BACTERIAL ISOLATES

Wound infections have become life threatening as a result of treatment failures caused by multi-drug resistant pathogens. The search for newer compounds potent against antibiotic resistant bacteria associated with wounds is crucial. Hence this study investigated the application of antibacterial photodynamic therapy using meso tetra-(4-phenyl) porphyrin (TPP), metallated with zinc, tin and silver (ZnTPP, SnTPP and AgTPP), meso tetra-(4-sulphonatephenyl) porphyrin (TPPS) and the corresponding metallo meso tetra-(4-sulphonatephenyl) porphyrin (MTPPS) as photosensitizers. The in-vitro toxicity and photo-toxicity properties on four chronic wound colonizing multi-drug resistant bacterial strains: Staphylococcus aureus, Klebsiella sp., Proteus sp., and Escherichia coli were assessed using agar well diffusion method. Photo-toxicity of the compounds was investigated using 100 Watt tungsten lamp. Inhibitory activity of porphyrins tested against these bacterial strains showed Staphylococcus aureus to have both lowest (11±0.0 mm) and highest (33±1.1 mm) susceptibility to SnTPPS and ZnTPPS respectively. The sequence of data also showed appreciable improvement in the antimicrobial activities of five metalloporphyrins (SnTPP, AgTPP, ZnTPPS, SnTPPS and AgTPPS) exposed to light rays than when tested against bacterial strains in dark condition. ZnTPPS exhibited the best activity with improved photo-toxic activities against all bacterial strains (Staphylococcus aureus 33±1.1 mm, Klebsiella sp. 32±0.7 mm, Proteus sp. 28±0.7 mm and Escherichia coli 30±1.4 mm) examined in this study.

Comparative Analysis on the Phytochemical, Proximate and Mineral Composition of the Seeds and Peels of Lime

Comparative Analysis on the Phytochemical, proximate and mineral composition of the seeds and peels of lime was carried out. The phytochemical, and proximate screenings were carried out using standard methods. The mineral analysis was carried out by using Atomic Absorption Spectrophotometer. The proximate analysis revealed that the seeds contain Moisture 8.20%, Ash 12.09%,Crude fat 8.50%,Crude Protein 1.93%,Crude fibre 20.71%, and Carbohydrate 48.58% while the peels contain Moisture10.60%, Ash 15.27%, Crude fat14.67%, Crude Protein 2.30%, Crude fibre23.36% and Carbohydrate 33.80%.The phytochemical analysis revealed the presence of alkaloid, saponins, flavonoids, terpenoids, phenols, and volatile oils in both the seeds and peels of the lime. In addition, the seeds contain steroids, while the peels contain tannins, balsams and anthraquinone. The mineral analysis revealed that the seeds contains Ni(0.17mg/100g),Pb(0.01mg/100g),Mn (0.20mg/100g),Zn(0.57mg/100g)Cr(0.17mg/100g),Mg(9.02mg/100g),Ca(27.02mg/100g),Cd(0.01mg/100g), Fe(2.55mg/100g) and Cu(4.81mg/100g) while the peels contain Ni(0.15mg/100g),Pd(0.21mg/100g),Mn(0,21mg/100g),Zn(0.23mg/100g),Cr(0.13mg/100g),Mg(7.61mg/100g),Ca(100.22mg/100g),Cd(0.01mg/100g),Fe(1.67mg/100g) and Cu(0.21mg/100g).The lime seeds and peels can serve as potential sources of drugs and nutrition with the seeds having higher concentration of minerals.

Determination of Physico-chemical properties of selected tropical timbers in Anambra State, South Eastern Nigeria

The analyses on chemical properties of some tropical timbers have been studied. The physical properties of these timbers varied in the pH(4.26 – 6.85) , moisture content(13%-37%), specific gravity(0.16-0.45), charring temperature (61 – 116o C)and porosity index(1.17-2.08%).Also the solubility behaviors of these timbers were mostly insoluble in hot and cold water, NaOH and ethanol and slightly soluble in concentrated HCl and H2SO4 respectively. The Qualitative results show heavy presence of flavonoids, saponins, tannis, glycosides, steroids and terpenoids in some of the timbers.Quantitative determinations were carried out and results show that the woods contain chemical constituents which are useful to both humans and industries.