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Nutritional Analysis

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Submitted By darrielewis
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Nutritional Analysis of Horse Feed
Darrie Lewis
Partnered with Danielle Henry

Introduction Animal nutrition is essential for meeting livestock nutritional requirements in maintaining acceptable performance, growth, and development. Animal nutrition focuses on dietary needs of domestic animals in agriculture or food production. In order to properly give the right balance of nutritional needs, one must know what the animal need in order to properly grow and develop. The amount of feed one gives to an animal should include an overall balance of essential nutrients to ensure successful production. An optimal nutritious diet should include intakes of amino acids, fats, carbohydrates, vitamins, and minerals.
The importance and our overall objective of this lab was to determine the different nutritional aspects in animal feed. In our case, my group and I analyzed horse feed. We were to determine the amount and percentages of ether extract, protein analysis, NDF, acid detergent fiber, ADF, and ADL in an amount of feed.

Materials & Methods
Ether Extraction
Materials
Thimbles
Thimble stand and handler
Grinded sample
Condenser

Purpose
To purify samples of organic compounds that are solids at room temperature
Method
Attach the thimbles to the adapters. Weigh the sample (2g) into the thimble. Move the thimbles to the thimble stand. Put a defatted cotton plug on the top of the sample and place the thimbles into thimble supports attached to the holder. Insert the thimbles into the condensers. Raise the holder into the condensers. Move each knob to the boiling position. Insert the six extraction cups, tarred with boiling chips and loaded with 25-50mL of the solvent. Move the extraction mode knobs to the boiling position. Boil with the thimbles immersed for 15 minutes. Move the extraction mode knobs to the rinsing position for 30 minutes. After rinsing, close the condenser valves by turning a quarter turn. Wait 5 minutes. When almost all the remaining solvent is collected in the condenser, press the air button on the Service Unit. Wait 5 minutes. Close the evaporation valve and release the extraction cups.

Protein Analysis
Materials
Vario Max Cube CNS
Furnace
Grinded sample
Purpose
In feed and food analysis, it serves as a tool of assessing and expressing the broad nutritional value of a feed sample.

Methods
We used the Vario Max Cube CNS to analyze protein content of our samples. The furnaces were set to 1140°C, 850°C, and 800°C. We used a weight of 250mg of grinded sample.

Neutral Detergent Fiber (NDF)
Materials
Reagents: NDF, alpha-amylase, sodium sulfide, and acetone
Grinded sample
Digestion apparatus
Filtration device - ANKOM Tech F57
Impulse bag sealer
Desiccator
Purpose
The NDF is considered a close estimate of the total fiber constituents of feedstuffs, since it measures cellulose, hemicellulose, lignin, silica, tannins, and cutins

Method
Weigh filter bag (W1). Weigh 0.5g of sample (W2). Weigh one blank bag and include in digestion to determine blank bag correction (C1). Seal the bags. Spread sample uniformly. (We processed less than 20 bags) Add 100mL/bag of detergent solution. Add 20g of sodium sulfide to the solution in the vessel and add 4.0mL of heat stable alpha-amylase. After 75 minutes, open the drain valve and exhaust hot solution before opening lid. After the solution has been exhausted, close valve and open lid. Add approx. 2000mL of hot water and 4.0mL of alpha-amylase to the first and second rinse. Each rinse should last 3-5 minutes. Exhaust water and repeat rinse two more times. Remove filter bags from bag suspender and gently press out excess water. Place in beaker and soak in acetone. Allow bags to soak for 3 minutes. Press out excess acetone. Complete drying in oven at 105°C for at least 2 hours. Remove bags from oven, weigh pouch, and flatten to remove air. Cool and weigh bags (W3).

Acid Detergent Fiber
Materials
Grinded sample
Acid Detergent Solution (AD)
Acetone
Digestion apparatus
Filtration device – ANKOM Tech F57
Impulse bag sealer
Desiccator
Purpose
ADF is often used to calculate digestibility, total digestible nutrients (TDN) and/or net energy for lactation (NEl).
Methods
Weigh filter bag (W1). Weigh 0.5g of sample (W2). Weigh one blank bag and include in digestion to determine blank bag correction (C1). Seal the bags. Spread sample uniformly. (We processed less than 20 bags) Add 100mL/bag of detergent solution. Place bag suspender with samples into the solution in vessel. Set timer for 60 minutes. Close and seal lid vessel. After 60 minutes, open the drain valve and exhaust hot solution before opening the lid. Add approx. 2000mL of hot water (90-100°C). Lower lid but do not tighten. Rinse for 3-5 minutes. Exhaust water and repeat rinses for a total of three times. After final rinse, remove filter bags from the suspender and gently press out excess water. Place in beaker and cover with acetone. Allow bags to soak for 3 minutes. Spread bags out and allow acetone to evaporate. Complete drying in oven at 105°C for at least 2 hours. Cool and weigh bags (W3).

Acid Detergent Lignin (ADL)
Materials
Grinded sample
Daisy incubator

Methods
Weigh filter bag (W1). Weigh 0.5g of sample (W2). Weigh one blank bag and include in digestion to determine blank bag correction (C1). Seal the bags. Spread sample uniformly. Perform ADF determinations using Fiber Analyzer. After performing ADF determinations, place samples into the incubator jar. Add 500mL of 72% H2SO4 to cover bags. Place jars into Daisy. Turn rotator on. After 3 hours, pour off H2SO4 and rise with tap water to remove all acid. Repeat rinses until pH is neutral. Rinse with approx. 250mL of acetone for 3 minutes to remove water. Complete drying in oven at 105°C for at least 2 to 4 hours. Remove bags from oven and place directly into MoistureStop. Weigh pouch and flatten to remove air. Cool and weigh bags (W3).

Results
Ether Extract Sample I.D. | Cup # | Empty Cup (g) | Sample Wt. (g) | Cup + Fat (g) | % Fat | | | | | | | 5 | 23 | 24.876 | 2.00 | 24.990 | 5.70% | 6 | 34 | 23.630 | 2.016 | 23.748 | 5.85% |

Dry Matter/Ash Cru # | Empty Cru. (g) | Sample wt. (g) | Dry S + Cru (g) | Furnace (g) | % Dry Matter | % Ash | | | | | | | | K8 | 19.728 | 2.027 | 21.530 | 19.865 | 88.9% | 6.76% | K18 | 18.490 | 2.024 | 20.289 | 18.630 | 88.9% | 6.92% |

Protein Analysis Sample I.D. (mg) | % Protein | | | G1 – 1: 253.80 | 12.028% | G1 – 2: 250.80 | 12.016% |

Blank Bag Correction Lab | NDF | ADF/ADL | | | | Oven-Dried (C1) | 0.5 | 0.5 |

NDF Sample I.D. | Filt. bag wt.(g) | Sample wt. (g) | Dry Sample wt. (g) | % NDF | | | | | | G1 #1 | 0.484 | 0.595 | 0.531 | 48.57% | G1 #2 | 0.505 | 0.540 | 0.519 | 49.35% |

ADF Sample I.D. | Filt. bag wt.(g) | Sample wt. (g) | Dry Sample wt. (g) | % ADF | | | | | | G1 #1 | 0.513 | 0.593 | 0.486 | 41.06% | G1 #2 | 0.511 | 0.543 | 0.505 | 46.87 |

ADL Sample I.D. | Filt. bag wt.(g) | Sample wt. (g) | Dry Sample wt. (g) | % ADL | | | | | | G1 #1 | 0.513 | 0.593 | 0.486 | 38.70% | G1 #2 | 0.511 | 0.543 | 0.505 | 45.95% |

Calculations
Ether Extract
#5 24.990-24.8762.00 x 100%=5.70%
#6 23.748-23.6302.016 x 100%=5.85%

Dry Matter
#K8 21.530-19.728 21.755-19.728 x 100%=88.9%
#K18 20.289-18.490 20.514-18.490 x 100%=88.9%

Ash
#K8 19.865-19.728 21.755-19.728 x 100%=6.76%
#K18 18.630-18.49020.514-18.490 x 100%=6.92%

NDF
#1 (0.531-0.484 x 0.5) 0.595 x 100%=48.57%
#2 (0.519-0.505 x 0.5) 0.540 x 100%=49.35%

ADF
#1 (0.50-0.513 x 0.5) 0.593 x 100%=41.06%
#2 (0.510-0.511 x 0.5) 0.543 x 100%=46.87%

ADL
#1 (0.486-0.513 x 0.5) 0.593 x 100%=38.70%
#2 (0.505-0.511 x 0.5) 0.543 x 100%=45.95%

Discussion
After completing all procedures, we were able gather our results. We were given equations for different labs and we had to use those results collected to help figure out the percents that were asked for. The amount of ether extract in horse feed averaged to be 5.78% fat. It was determined protein in horse averaged to be 12.02% protein. Neutral Detergent Fiber (NDF) in horse feed averaged to be 48.96%. Acid Detergent Fiber (ADF) in horse feed averaged to be 43.97%. Acid Detergent Lignin (ADL) in horse feed averaged to be 42.33%.

Conclusion
In conclusion, we were able to learn the different components of a feed or how to find them. What we learned through the lab and our data analysis, the majority of horse feed is dry matter. Dry matter is what remains after all of the water is evaporated out of a feed. This included grain and fresh, or dried forages. Dry matter is an indicator of the amount of nutrients that are available to the animal in a particular feed. We learned that the minority of horse feed contains fat. This is accurate because fats are needed in small proportions. Some labs were short and some were time consuming. The equipment we use was heavy machinery so wouldn’t be able to do these procedure at home. Overall, these labs were interesting.

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