Ollection from the plant was performed in January, 2009. The stem-bark was
Ollection from the plant was completed in January, 2009. The stem-bark was collected from Itak- Ikot Akap village in Ikono Nearby Government Location of Akwa Ibom State. The plant was collected by an Herbalist Mr. Okon Etefie attached to Pharmacognosy Division in the MMP-12 manufacturer University of Uyo, and identified by a Botanist named Dr (Mrs.) Margret Bassey of Botany Division in the University of Uyo. A voucher specimen (UUH 998), wasNwidu et al., Afr J Tradit Complement Altern Med. (2014) 11(two):257-dx.doi.org/10.4314/ajtcam.v11i2.five deposited at the University Herbarium. The stem-bark was air-dried and powdered. The pulverized plant material had been stored at space temperature until made use of. Preparation and extraction of plant materials The stem-bark collected was air-dried and pulverized making use of harmer mill. The powder plant supplies have been weighed applying weighing balance (BG 4000). 5 hundred grams of your stem-bark was weighed and immersed in 3 x 500 ml of ethanol (99.8 ) for 72hrs. The soaked extract was shaken twice daily. The supernatant had been filtered working with Whatman filter paper (pore sizes-20-25. The filtrate of ethanol solvent was lowered in volume practically to dryness within a rotatory evaporator (BUCCHI USA), at 40 oC. The residue from filtration procedure had been air-dried for 24hrs, and subjected towards the same procedure for three successive time. Immediately after which the extract was dried beneath a flow of nitrogen till continual weight was obtained. The yield was 43.4 . The extract was stored in an air tight container inside a refrigerator until used. Before pharmacological assay, a sample of extract was dissolved in distilled water and applied for the animal experiments.Finger Print Analysis The chromatographic fingerprint with the C. lutea stem-bark extract was established working with a Jasco (Tokyo, Japan), liquid chromatograph equipped with a PU-2089, quaternary solvent pump, a MD-2010 PAD, and an AS-2055, autsampler injector using a 20 L sample loop. The analytical column was a Phenomenex Synergi Hydro RP18, (250 four.six mm i.d.; 4 m), equipped having a Phenomenex safety guard column (4.0 2.0 mm i.d.). The mobile phase composition was: water (eluent A), and metanol (eluent B), each containing 0.05 of TFA. The gradient program was linear beginning with 0 B to 100 B in 60 min. The flow price was 1.0 mL/min. EZChrom Elite Data Program application (Chromatec, Idstein, Germany) was applied for both the operation of detector and for information processing. The stem-bark extract (2 mg), was dissolved in two mL methanol, filtered through a 0.45-m membrane polytetrafluoroethylene (PTFE), filter (Millex), resuspended in 3 mL of water and 20 L was surrendered to HPLC analysis.Phytochemical Analysis The ESE of C. lutea was subjected to qualitative chemical screening working with typical procedure to reveal glycosides, polyphenols and saponins (Trease and Evans, 2001).Elemental analysis from the plant stem-bark The elemental element of ESE stem-bark of C. lutea was elucidated working with the technique of Dahlquist Knoll, (1978) as reported for the C. lutea leaf fractions (Nwidu et al., 2012d).Determination of ionic content of plant stem-bark This determination was carried out by potentiometric titration as previously reported for leaf fractions (Nwidu et al., 2012d).Animals Swiss albino mice weighing involving 25-30 g, and adult albino rats (100-150 g), of both sexes had been obtained in the δ Opioid Receptor/DOR Biological Activity Faculty of Pharmacy Animal Home, University of Uyo, Uyo, Nigeria. Each of the animals have been housed in typical cages beneath laboratory situation in Depa.