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APPENDIX 1

LABORATORY PRODUCTION OF LIVER ETRACT AND LIVER DIGEST

FROM FRESH BOVINEPANCREASE OBTAINED FROM AN ABATTOIR

IN ZARIA.

Initial preparation of liver and pancrease for extraction using aseptic techniques. The

hands were washed with soap and disinfected with a hand disinfectant mixture (chlorhexidine and

isopropyl) before proceeding. The following were then disinfected with boiled water either by pouring

boiled water on the surface twice or by filling containers halfway with boiled water with the lids on and

kept for 10minutes:

1. Mortar and pestle

2. Cutting boards and knives

3. All plastic and metal containers

4. Working bench surfaces

The following were further disinfected with 10% hypochlorite solution (HOCL) which is 35,000ppm

solution of chlorine. Ten percent (10%) HOCL solution was made by adding 100ml of concentrated jik

bleach to 900ml of tap water:

1. Bench surfaces

2. Mortar and pestle

3. Cotton board and knives

4. All plastic and metal containers

Ten percent (10%) HOCL was poured on bench surfaces and cleaned with cotton wool soaked in

the solution. The containers were filled with 10% HOCL until they were half-full and closed with theirs

lids. The fluid was swirled around so that it touched all the surfaces of the containers and they were

then left on the bench for 10 minutes. The chlorine solution was discarded and the containers were

rinsed thrice with boiled tap water. The liver and pancreas were washed with detergent using and

cooled tap water and rinsed thoroughly to remove all traces of soap. The liver and pancreas were

immersed in hypochlorite solution in separate containers previously disinfected and half-filled with

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10%HOCL solution. This completed the surface disinfected. The surface disinfected liver and pancreas

were placed on the previously disinfected cutting board and minced,(cut in very tiny bits),using a

previously disinfected knife.

Extraction of Liver and Pancreas Pastes.

The minced liver and pancreas were each placed separately into a disinfected and rinsed mortar

and pounded into smooth pastes. A disinfected small metal cup was used to fetch the minced liver and

pancreas into a previously disinfected large plastic container in each case. The number of cups of

minced liver and pancreas was noted.

Boiled and cooled tap water was added to the liver and pancreatic pastes in covered containersin the following ratio:

Liver: 1 part of liver pastes to four parts of boiled and cooled tap water.

Pancreas: 1 part of pancreatic paste to 3 parts of boiled and cooled tap water.

The suspensions were kept in covered containers and held in the laboratory for 3hours at 300+/_

10C to

effect extraction. After this period, decantation and filtration of extracts was done. The pH of the liver

extract was adjusted to neutrality and then boiled in 200ml, 300ml and 400ml portions which were

pasteurized, cooled and stored in the refrigerator.

Preparation of Liver Digest

Pancreatic extract was added to the extracted liver paste to effect digestion of the liver paste. One

volume of pancreatic extract was added to one volume of liver paste.

The suspension was incubated at 30-350C for 4 hours with occasional agitation to effect thorough mixing

of the suspension. As digestion progresses, the suspension became acidic due to formation of acidic

fermentation products. The pH of the suspension was monitored and the tendency of high acidity was

counteracted by the addition of 40% KOH to the suspension, using Bromothymol blue color standard to

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guide the pH adjustment. After 4hour of digestion with pancreatic enzymes, the process was terminated

by boiling the suspension. The boiled suspension was filtered and final adjustment to neutrality was

carried out. The liver digest was bottled in 200ml, 300ml and 500ml portions in plastic containers,

pasteurized cooled and stored in the refrigerator.

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