FST306

FOOD ANALYSIS

LABORATORY REPORT

EXPERIMENT 6:
DETERMINATION OF ASH BY TOTAL ASH METHOD
AND ACID-INSOLUBLE ASH METHOD

DATE OF EXPERIMENT: 4th NOVEMBER 2024

DATE OF SUBMISSION: 14th JANUARY 2025

GROUP: AS1163A3

PREPARED FOR: DR. SAFIYYAH BINTI SHAHIMI

PREPARED BY:

Name Student ID IC
1. Nur Iman Athirah binti Ahmad Asri 2023614652 051217-14-0718
2. Nurul Izzah binti Mohd ‘Asri 2023614808 051220-06-0364
3. Siti Aisyah Faqihah binti Norazman 2023644712 050715-14-1006
4. Najwa Hani binti Zamri 2023623056 050621-10-0932
INTRODUCTION
Ash contents are more important in the food industry than people may think. They can alter
various key features of foodstuffs including nutritional value, shelf-life, appearance, texture,
and taste. As a result, these characteristics affect several aspects of your food quality. Not
only does it impact the enjoyment of your meal, but also the safety, freshness, and nutrition of
the food. Ash content refers to the minerals and inorganic substances left after heating to
extremely high temperatures to remove moisture, volatiles, and all organic materials.
Following this, the leftover product is known as ash; otherwise known as inorganic materials.
These inorganic materials include the salts of calcium, magnesium, sodium, potassium,
manganese, zinc and iron.

Measuring the ash content in food is important for a few reasons. Ash content can affect
different characteristics of food including physiochemical and nutritional properties. Ash
determination of food samples is part of the proximate analysis necessary for nutritional
evaluation. This ensures the safety of foods, making sure there are no toxic minerals present.
The ash content in food can also impact the taste, texture and stability of foods so it is vital to
know the mineral content for quality control purposes. Ash content provides vital information
towards the nutritional value of food and can be used as an indicator on how processed
different food items are. More processed foods tend to have a higher ash content when
compared to natural foods, which contain a lower ash content. For example, pure oil and fats
can have an ash content as low as 0%, whereas processed dried meats can contain up to 12%
ash content. Processed foods are gaining popularity due to their affordability and scalability.

In this experiment, ash will be determined by using two methods, total ash method and
acid-insoluble ash method. Both samples will be placed in a previously ignited, cooled, and
tared porcelain crucible and then incinerated in a muffle furnace at 550°C until carbon-free.
Crucibles are transferred to a desiccator and cooled, and the weight loss is used to determine
the percentage of ash. Meanwhile, the other method will be determining acid-insoluble ash by
dissolving ash in dilute hydrochloric acid (10% m/m), the liquid filtered through an ashless
filter paper and thoroughly washed with hot water. The filter paper is then ignited in the
original dish, cooled and weighed. The difference between two methods is that total ash
quantifies the total amount of ash remaining after incineration of the sample without adding
sulfuric acid. Determination of ash content is required to ensure the quality and purity of a
given material and to check it for regulatory compliance.

OBJECTIVES
1. To determine the total ash and acid insoluble of ash of food sample using dry ashing
method.
2. To illustrate the use of a muffle furnace.

APPARATUS
Porcelain dish, Bunsen burner, tongs, spatula, filter paper and watch glass.

EQUIPMENT
Analytical balance, Muffle furnace and oven.

PROCEDURE OF TOTAL ASH METHOD

1. The porcelain dish was dried in oven at 105°C for 3 hours. The porcelain dish was cooled
it in a desiccator and weighed it after it had reached room temperature.
2. 5.0g of homogenized food sample was weighed into a porcelain dish. If the sample
contained high moisture, dried the sample in the oven at 130°C for one day.
3. Charred the sample gently over a Bunsen burner until it had ceased smoking.
4. The muffle furnace was placed and heated at 550°C for 3 hours.
5. The ashed was sample until a whitish or greyish ash was obtained.
6. The dish was removed, cooled in desiccator and weighed it after it had reached room
temperature.
7. Replaced it in the muffle furnace and continued heating until a constant weight was
obtained.
8. The total ash content of food sample was calculated.

PROCEDURE ACID-INSOLUBLE ASH METHOD

1. The ash obtained above was boiled with 25ml diluted HCl (HCl: H2O=1:2.5) for 5
minutes.
2. If necessary, the dish wash covered with watch glass to prevent spattering.
3. The insoluble matter was collected by filtering through an ashless filter paper and washed
with hot water until it free from acid.
4. The filter paper was ignited with the residue in the original dish.
5. The acid-insoluble ash obtained was calculated.

RESULTS AND DATA ANALYSIS

Table 1: Determination of total ash

Weight of
Weight of Weight of
Sample Trial crucible + % Total ash
sample (g) crucible (g)
ash (g)
1 (ain) 5.0144 51.4667 51.4688 0.0419
2 (ainul) 5.0103 43.5030 43.5226 0.3912
Oat 3 (nina) 5.0207 50.0460 50.0787 0.6513
Mean ± 0.3615±0.47
SD 34
1 (wajihah) 5.0344 56.8598 56.9281 1.3567
2 (amani) 5.0848 40.1368 40.1412 0.0865
Jacob biscuit 3 (iman) 5.0180 38.6627 38.7557 1.8533
Mean ± 1.0988±1.42
SD 75

Table 2: Determination of Acid-Insoluble ash

Weight of
crucible + % Acid-
Weight of Weight of
Sample Trial acid- insoluble
ash (g) crucible (g)
insoluble ash
ash (g)
1 0.0021 50.0674 50.0400 -1304.76
2 0.0190 43.5030 43.4912 -62.11
Oat 3 0.0327 50.0460 50.0457 -0.9174
Mean ± -455.93 ±
SD 865.56
1 0.0683 56.8598 55.4766 -2025.18
2 0.0044 40.1368 38.8409 -29452.27
Jacob biscuit 3 0.0930 38.6627 38.6674 5.054
Mean ± -10490.80 ±
SD 19512.57
CALCULATION
Determination of total ash (% Total ash).
Sample: Oat
Trial 1
(51.4688 − 51.4667)
× 100 = 0.0419%
5.0144
Trial 2
(43.5226 − 43.5030)
× 100 = 0.3912%
5.0103
Trial 3
(50.0787 − 50.0460)
× 100 = 0.6513%
5.0207
Mean
(0.0419 + 0.3912 + 0.6513)
= 0.3615%
3
SD

√((0.04192 + 0.39122 + 0.65132 ) − 0.36152)

= 0.4734
3−1
Sample: Jacob biscuits
Trial 1
(56.9281 − 56.8598)
× 100 = 1.3567%
5.0344
Trial 2
(40.1412 − 40.1368)
× 100 = 0.0865%
5.0848
Trial 3
(38.7557 − 38.6627)
× 100 = 1.8533%
5.0180
Mean
(1.3567 + 0.0865 + 1.8533)
= 1.0988%
3
SD
 √((1.35672 + 0.08652 + 1.85332 ) − 1.09882)
= 1.4275
3−1

Determination of Acid-insoluble Ash (% Acid-insoluble ash)

Sample: Oat
Trial 1
(50.0400 − 50.0674)
× 100 = −1304.76%
0.0021
Trial 2
(43.4912 − 43.5030)
× 100 = −62.11%
0.0190
Trial 3
(50.0457 − 50.0460)
× 100 = −0.9174%
0.0327
Mean
(−1304.76) + (−62.11) + (−0.9174)
= −455.93%
3
SD

√(((−1304.76)2 + (−62.11)2 + (−0.9174)2 ) − (−455.93)2 )

= 865.56
2
Sample: Jacob biscuits
Trial 1
(55.4766 − 56.8598)
× 100 = −2025.18%
0.0683
Trial 2
(38.8409 − 40.1368)
× 100 = −29452.27%
0.0044
Trial 3
 (38.6674 − 38.6627)
× 100 = 5.054%
0.0930
Mean
(−2025.18) + (−29452.27) + 5.054
= −10490.80%
3
SD

√(((−2025.18)2 + (−29452.27)2 + (5.054)2) − (−10490.80)2 )

2
= 19512.57
DISCUSSION
Ashing is the mineralization procedure used in analytical chemistry to preconcentrate trace
chemicals before chemical analysis. All non-aqueous residue that is left over after a sample is
burned and is primarily made up of metal oxides is referred to as ash. Ash, which is primarily
composed of salty, inorganic components, is one of the components used in the proximate
analysis of biological materials. It contains metal salts, which are crucial for procedures that
require for ions like sodium (Na+), potassium (K+), and calcium (Ca2+). Additionally, it
contains trace minerals that are necessary for special molecules like hemoglobin and
chlorophyll.

In this experiment, two food samples are used which is oats and Jacob biscuits were
analyzed for total ash and total acid-insoluble ash content. The percentage amount of ash and
total acid-insoluble ash content that present in the two samples after it go through incineration
at high temperature in oven was calculated. Ash is alluded to inorganic excess after the water
and organic matter was removed. A common procedure for determining the total ash content
in the majority of food samples is dry ashing. Dry ashing methods use high temperature
muffle furnace that able to maintain the temperature between 500°C and 600°C. Weighing the
crucible and the homogenized food sample to around 5g was the first step in the experiment.
The crucible was placed in the muffle furnace at 550°C until the sample particles converted
into whitish or greyish ash after the appropriate weights were completed. It was permitted to
cool at room temperature in a desiccator once the presence of ash was noticed. It was
weighed twice using an analytical balance, and the results were then collected.

As a results, oats had an average total ash concentration of 0.3615% ± 0.4734, but
Jacob biscuits had a substantially higher total ash content of 1.0988% ± 1.4275. This
discrepancy is consistent with the food samples' expected composition. Oats have a reduced
ash percentage because they are a natural grain that has undergone minimal processing and
have less inorganic additions or fortifications. Jacob biscuits, on the other hand, are a
processed food product, and the addition of substances such baking powders, salts, and other
additions that are high in minerals accounts for their increased ash level. These components
are frequently added in the production process to improve shelf life, flavor, or texture, which
raises the total amount of inorganic residue left over after combustion.
 The average percentage of acid-insoluble ash for oats and Jacob biscuits was -1304.76%
± 2599.34 and -10409.80% ± 19512.57, respectively. In addition to being substantially
negative, these numbers deviate greatly from any theoretical expectations. The siliceous
components, such sand or soil, that do not dissolve when treated with acid should be
represented as acid-insoluble ash; these values should be a minor positive fraction of the total
ash content. Significant errors during experiments are indicated by the observed negative
results. Inaccuracies in the weighing procedure, such as incorrectly recording the weight of
the crucible and ash residue, could be one likely cause of these incorrect numbers. Such
discrepancies may also result from insufficient burning of the organic waste, contamination
during acid treatment, or loss of ash during transfer. These errors may also be compounded
by the tiny sample sizes that were used, which could lead to disproportionately significant
errors in estimated percentages from even small measurement errors.

Nevertheless, the acid-insoluble ash values for both samples showed significant
deviations, with negative results being recorded. Theoretically, this is impossible because
total or acid-insoluble ash content should always result in positive readings.

According to research, the total ash content is the entire amount of mineral residue
that remains after all organic matter has burned completely at high temperatures. This
procedure guarantees that only inorganic materials remain after all volatile and organic
components have been burnt off. The siliceous residues that make up the acid-insoluble ash, a
subset of the total ash, are insoluble in hydrochloric acid. Theoretically, the total ash
percentage should always be higher than the acid-insoluble ash percentage, as the latter
represents only a specific fraction of the total inorganic residue. The experiment's
significantly negative acid-insoluble ash results indicate that either the weights were not
measured accurately, or the ash was lost during the process of determining acid-insoluble ash.
This could occur if the material was lost during the acid digestion process or if the ash was
not handled appropriately during transfer.

Their different compositions also help to explain why oats and Jacob biscuits have
different amounts of ash. Because they are a whole grain, oats have a lower total ash level
because they mostly contain naturally occurring minerals like potassium, magnesium, and
phosphorus. In contrast, Jacob biscuits are made with extra components which increase their
mineral content, like calcium from leavening agents and sodium from salt. Greater sample
variability or potential inconsistencies throughout the experimental process are indicated by
the significantly increased standard deviation in the Jacob biscuit results.

The results obtained in this experiment can be compared with the theoretical
expectations for ash determination. As acid-insoluble ash represents only a fraction of the
total ash content, total ash values should theoretically be higher than acid-insoluble ash
values. Since the ash content indicates the amount of inorganic residue in the sample, both
methods should yield positive results. The observed variances, especially the negative acid-
insoluble ash values, emphasize how crucial accuracy is in sample preparation, weighing, and
following procedures. Incomplete combustion of the samples during the total ash assessment
may also be the cause of these inaccuracies, as unburned organic residues may be present and
obstruct subsequent acid digestion.

Strict adherence to experimental precaution steps is essential to improve the accuracy

and reliability of results. To ensure that all organic material burns completely, the furnace
temperature should be kept at the recommended level, which is usually 550°C. The crucibles
must be allowed to cool in a desiccator before to weighing to avoid moisture absorption that
could skew the readings. Other than that, careful handling is required to prevent ash loss
during transfers during the acid-insoluble ash determination process, and the equipment used
needs to be carefully cleaned to avoid contamination. It is essential to weigh samples and
crucibles precisely both before and after ashing, and using a calibrated analytical balance can
reduce inaccuracies. To prevent the occurrence of unburned organic residues, it is imperative
that the samples in the furnace be completely burned. Therefore, to avoid material loss
throughout the acid-insoluble ash determination process, the ash must be handled carefully
and undergo a complete acid digestion. Larger sample sizes may reduce the effect that minor
measurement errors have on the final result. By implementing these measures, the ash content
determination's precision and dependability can be significantly improved, yielding
meaningful data for food sample analysis.
CONCLUSION
To sum up, the total ash and acid-insoluble ash methods work on the quality of foods,
determining the amount of total minerals and residue left which is not soluble in acids.
Though the experiment did not produce a positive result, these methods are important for
determining impurities and in ensuring conformance with regulations. Problems encountered
highlight the importance of accuracy and troubleshooting during laboratory procedures. It is
necessary to repeat the experiment with an improved methodology and calibrated apparatus.
These methods form the backbone of analytical chemistry and food safety.

QUESTIONS

1. How do you know when the ash is free from carbon?

The ash is free from carbon when the ash become whitish or greyish after being heated in
the muffle furnace.

2. Why is the sample needed to be heated over a Bunsen burner before heating in a muffle
furnace?
The sample needed to be heated over Bunsen burner before hetaing in a muffle furnace
because to accelerate the heating process and to eliminate water content, volatile
compounds and substances easily thermodegradable before the calcining in a muffle
furnace.
REFERENCES
1. Determination of Ash content | Food and Drink | PrepASH. (2024, April 9). Precisa.
Retrieved from: https://www.precisa.com/article/ash-content-in-food-analysis/

2. Rowanjenkins. (2024, September 10). Sulfated Ash and Total Ash Determination.
MaTestLab. Retrieved from:
https://matestlabs.com/blog/sulfated-ash-total-
ash/#:~:text=Total%20ash%20quantifies%20the%20total,check%20it%20for%20regulato
ry%20compliance.

3. Jadess Lorraine Fusio. (2015, August 9). Experiment 2 - Analysis of ash. Forever hungry.
Retrieved from:
https://foreverhungry101.weebly.com/blog/experiment-2-analysis-of-ash

4. MicroChem's Experiments. (2020, July 2). Determination of ash content (Total

Minerals)_A complete procedure (AOAC 942.05). YouTube. Retrieved from:
https://youtu.be/n2Qwb8Pw8YE?si=OoH8TjALOJrt_2hy

5. MicroChem's Experiments. (2021, January 4). Determination of acid insoluble Ash_A

complete procedure (AOAC 941.12 & pH. Int. (WHO), 2019. YouTube. Retrieved from:
https://youtu.be/JecqdeVUTpE?si=NvCk_-3fI9D_6bNY