# Effect of Poultry and Goat Manures on the Nutrient Content of _Sesammum altatum_ Leaf Vegetables
## 2 Materials and Methods
### Proximate Analysis  
Shoots were examined for the protein, fibre and fat content from each treatment. _Crude proteins_: Two grams of dried sample powder were weighed and introduced into a 650 mL digestion flask along with 10 mL of distilled water. A digestion tablet along with 20 mL of concentrated H\({}_{2}\)O\({}_{4}\) was added to the flask to act as the solution catalyst. Boiling chips were added to the mixture and the digestion was given time until a colourless solution was obtained. The digestion solution was allowed to cool before it was diluted with distilled ammonia-free water up to 100 mL. The Kjeldahl flask was pre-washed using distilled water for use. A total of 10 mL of the digest's 100 mL solution was pipetted into a distillation flask with 90 mL of distilled water. Twenty millimetres of 40% NaOH were pipetted into the distillation flask. A 250 mL conical flask containing 10 mL boric acid solution along with some drops of the indicator was used. Ammonia was then purified from the solution. About 100 mL of ammonia was extracted from boric acid. After extraction, the 100 mL solution was titrated against 0.1 N HCl until a pink colour appeared. The titrated volume was then subtracted from the original sample's titrated volume. Values obtained from the above process were used to calculate the N content percentage of the sample using the formula:  
\[\% Nitrogen=\frac{millimetre\ acid\times normality\ standard\ acid}{weight\ of\ the\ sample\ in\ grams}\times 0.014\times 100\]  
The crude protein percentage was calculated by multiplying the nitrogen percentage by a conversion factor of 6.25. _Crude Fibre (Acid Detergent Fibre and Neutral Detergent Fibre)_: Crude fibre was estimated by 1.25% H\({}_{2}\)SO\({}_{4}\) and 1.25% NaOH solutions. Ether extract residue was introduced into a digestion flask along with a 200 mL boiling H\({}_{2}\)O\({}_{4}\) solution. Similarly, an anti-foaming agent was added and connected to the flask immediately with a condenser and heated for 30 min. After that time, the digestion flask was removed and filtered immediately for its content using linen cloth and then washed using boiling water till the acid was removed from the washings. After washing, the residue was again introduced into a flask containing 200 mL of boiling NaOH solution. The flask was then connected to a reflux condenser and heated for 30 min. After 30 min, the flask was removed, the precipitate content was filtered with a good crucible and then it was washed using boiling water. Filtered content was washed with 15 mL of 95% alcohol (ethanol) and it was dried along with the crucible at 110 \({}^{\circ}\)till a constant weight was achieved. The content and crucible were cooled in a desiccator and weighed. The crucible content was incinerated for 30 min in a furnace at 550 \({}^{\circ}\)C until the carbonaceous matter was consumed. It was cooled again in a desiccator and the final weight measurements were taken. The following formula was used to calculate crude fibre as the loss in weight:  
\[Crude\ fibre\ \%=\frac{wt.\ of\ dry\ crucible\ and\ sample-wt.\ of\ incinerated\ and\ ash}{sample\ weight}\times 100\]  
\(Crude\ fats\)The amount of fat contained was determined using petroleum ether as a reagent. Firstly, the weight of the bottle and lid to be used were ensured to be stable and sterilized. This was carried out by placing the bottle and lid in an incubator at 105 \({}^{\circ}\)C for 12 h. About 100 g of the sample was weighed and wrapped using a paper filter. The wrapped sample was extracted using a thimble and then transferred into a Soxhlet filled with about 250 mL of petroleum ether and then heated. The Soxhlet apparatus was connected and placed in water to cool down and then heated again for 14 h at a heating rate of 150 drops/min. The solvent was completely evaporated using a vacuum condenser and incubated at 80-90 \({}^{\circ}\)C until the bottle became dry. The lid of the bottle was transferred to the desiccator for cooling and the dried content of the bottle was reweighed.