ASH ANALYSIS

Introduction
◼ Ash:
◼ Refer to inorganic (mineral) residue remaining after
the combustion or complete acid-facilitated oxidation
of organic matter in food
◼ Estimate the total mineral content in foods
◼ Minerals are not destroyed by heating, and they
have a low volatility
◼ Ash content of most fresh foods rarely > 5%, but
processed food (e.g.: dried beef = 12%)
◼ Pure oils & fats contain no / little ash
 Importance of Ash
◼ Nutritional labeling –concentration and type of
mineral normally stipulated on the label
◼ Quality – depends on the concentration and type of
mineral esp. appearance, texture, taste or stability
◼ Microbiological stability- high mineral content may
retard the growth of certain microorganisms
◼ Nutrition – essential to healthy diets (Ca, K, P, Na)
but some are toxic mineral (lead, aluminum,
mercury)
◼ Processing- will effect the physicochemical
properties of food
◼ 3 major types of ashing method:
◼ Dry ashing
– muffle furnace, 500 – 600oC (or microwave)
- water and volatiles are vaporized
- organic substances burned in the presence of oxygen
to from CO2 and oxides of N2
- the remaining minerals are converted to oxides,
sulfates, phosphates, chlorides ans silicates
- elements such as Fe, Se, Pb and Hg may partially
volatilize (thus, must use other ashing method)
◼ Wet ashing
– a procedure used to chemically oxidized and
remove organic substances using strong acids,
oxidizing agent, or combination thereof.
- HCl, H2SO4,HNO3, HClO4 (perchloric acid)
- use lower temperature- thus less mineral was
loss due to volatility (preferable for mineral
analysis)
 Sample Preparation For
Ashing
◼ 2-10 g sample is sufficient
◼ Particle size reduction (milling, grinding), representative
◼ Avoid micro-elemental contamination (water, glass, metal
from grinder)
- use acid wash, deionized water, sample blank
◼ Plant materials : over-dried, grinding, ashing
◼ Meat, sugar, syrups should be evaporated to dryness on a
steam bath or infrared lamp, prior to ashing
 Dry Ashing

◼ Principle: incineration of sample at high temp. (above 525oC), in a

muffle furnace
◼ Selection of crucible highly depends on sample and furnace
temperature
◼ Quartz : resistant to acids & halogen, but not alkali at high temperature

◼ Porcelain: crack with rapid temperature changes, inexpensive

- usually the crucible of choice
◼ Steel: resistant to acids & alkalis, inexpensive, but may be a source of
contaminants (Ni & Cr)
◼ Platinum: very inert, too expensive for routine use for large number of
sample
◼ All crucibles should be labeled for identification (2B pencil for porcelain
crucible)
Type of crucible for dry
ashing
Muffle Furnace for Dry
Ashing
 Advantages of Dry Ashing
◼ Safe and inexpensive method, less sample required, no
added reagents or blank subtraction
◼ sample recovery is high
◼ Can handle a large number of crucibles at one time
◼ Little attention is needed once ignition begins
◼ Resultant ash can be used for:
◼ Analysis of individual elements
◼ Acid insoluble ash
◼ Water-soluble & insoluble ash
 Disadvantages of Dry Ashing

◼ Time consuming (12 – 18 hr or overnight)

◼ Expensive equipment –electrical cost
◼ Loss of volatile elements: As, B, Cd, Cr, Cu,
Fe, Pb, Hg, Ni, P, V and Zn
◼ May have interaction between crucibles and
elements
 Procedure

5 – 10 g sample in a crucible (pre-dry if

very moist)
Place in muffle furnace

Ignite 12 – 18 hr (or overnight), 550oC

Open door carefully Turn off, wait until temp drop to <250oC
to avoid loss of fluffy
ash
Quickly transfer crucibles to a desiccator
 Ash content calculated as,

◼ % ash (dry basis)

= wt after ashing (g) - wt of empty crucible (g) x 100

= wt after ashing (g) - wt of empty crucible (g) x 100

original sample weight x dry matter coefficient (%)

If moisture content =80%, dry matter coefficient = 0.8

 Special Applications
◼ Completely oxidized sample: white in colour
◼ If C still present, add several drops of H2O or HNO3, then re-
ash
◼ If C persists such as in high sugar sample;
◼ 1) suspend the ash in water
◼ 2) filter through ashless filter paper
◼ 3) dry the filtrate
◼ 4) place paper & dried filtrate in muffle
◼ High fat sample: extract fat or burn off the fat- before ashing
◼ Prevent spattering (jam/jellies): mixed sample with cotton
wool
◼ Glycerin, alcohol & hydrogen accelerate ashing
◼ Salt rich foods – may require separate ashing for water-
insoluble components and salt-rich water extract (use crucible
cover to avoid spattering)
◼ Cereals: an alcoholic solution of magnesium acetate can
accelerate ashing. Must use a blank
 Question:

◼A grain was found to contain 11.5%

moisture. A 5.2146 g sample was placed into
a crucible (28.5053 g after tare). The ashed
crucible weighed 28.5939 g. Calculate the
percentage ash on:
a) an as-received basis (wet basis)
b) a dry basis
 Wet basis (sample with moisture)
Dry basis (sample without moisture=dry matter)

Let say 100 g sample, use 100 g to calculate wet basis

But for dry basis, use weight of dry matter only (88.5 g in this
case)

a) Weight of ash = (28.5939 - 28.5053) g = 0.0886 g ash

% of ash (wet basis) = (0.0886 g/5.2146 g) x 100%
=1.7%

b) Water content =11.5%

Dry matter = 100% - 11.5% = 88.5%
Dry matter (%) = Weight of dried sample x100 %
Weight of initial sample
88.5 = (a/5.2146) x 100
a = 88.5 x 5.2146
100
= 4.6149 g

Ash (%) in dry basis = Weight of ash x 100

Weight of dried sample
= 0.0886 x 100
4.6149
= 1.92%
 Wet Ashing
◼ Sometimes called wet oxidation or wet digestion
◼ Used mainly for more volatile minerals such as Fe,
Cu, Zn, P
◼ Principle: oxidizing organic substances by using
strong and oxidizing agents acids (nitric, perchloric,
sulfuric acid) or their combinations, with heating.
◼ Advantages:
◼ Mineral stay in aqueous solution. Thus, little or no loss
from volatilization because of the lower temp. (350°C)
◼ Time used is shorter than dry ashing (10 min to few
hours)
 Wet Ashing
◼ Disadvantages:
◼ Need constant operator attention
◼ Use corrosive reagent (HNO3, HClO4 & H2SO4)
◼ A special perchloric acid hood is required
◼ Only small numbers of samples at a time
◼ Single acid does not give complete & rapid
oxidation, need to combine with H2SO4 or HClO4
– diff. combination for diff .samples
◼ Very hazardous
◼ Do not determine the total ash content
 Procedure
1 g sample

Prepare blank of 3 ml of H2SO4 followed by 5 ml of HNO3

+ 3 ml of H2SO4 followed by 5 ml of HNO3 to the sample

Heat the sample on a hot plate (200oC). Brown yellow fumes - white fumes
and finally the sample turn dark.

Remove the flask from the hot plate, slowly add 3-5 ml HNO3

Continue heating to allow HNO3 to boil off. Once the colour is clear to straw
yellow, proceed with boiling to reduce the volume. Cool down

Transfer to an appropriate volumetric flask and dilute with ultrapure water

 Microwave Ashing
◼ Can be used for both dry & wet ashing
Wet ashing
◼ Working with open or closed vessels for ashing a variety
of samples
◼ Closed vessels can work at high pressure and
temperature
◼ Advantages : reduce time of ashing greatly to 40 min = 4
h time ashing with a conventional muffle furnace
◼ Wet ashing in a closed system is rapid & safe
◼ Disadvantages : Not possible to analyze as many
sample as in a muffle furnace
◼ Dry ashing
- Can reach a temperature of 1200oC
 Other Ash Measurements

◼ Soluble & Insoluble Ash in Water

◼ As an index of the fruit content of preserves & jellies
◼ A lower ash in the water-soluble fraction is an indicator
of higher fruit content in the product
◼ Dry ashing → Weigh the total ash → add 10 ml
deionized water → cover & heat to boiling → filter &
rinse with hot deionized water → dry & re-ash filter paper
at least 30 min → weigh & calculate as % water
insoluble ash.
◼ amount of water-soluble ash is determined by drying the
filtrate
 Other Ash Measurement..

◼ Ash Insoluble in Acid

◼ Refers to ash that is insoluble in acids.
◼ Determination of the surface contamination
(silicates) of fruits & vegetables, wheat & rice
coatings, primarily from soil, and generally not
tested in food
◼ Silicates remain insoluble in acid
◼ Weigh total ash → add 25 ml 10% HCl → cover
& boil → filter with hot distilled water → dry & re-
ash → weigh & calculate as %
 Other Ash Measurements..

◼ Alkalinity of ash
◼ Alkaline ash: Ca, Mg, K, Na (fruits & veg.)
◼ Acid ash: P, S, Cl (meat, some cereals)
◼ The alkalinity of ash has been used as a quality
index of fruit & fruit juices
◼ Total ash → add 0.1 N HCl → add boiling water
if necessary & warm on a water bath → cool →
titrate the excess HCl with 0.1N NaOH (methyl
orange as indicator)→ express as ml of 1N acid
/ 100 g sample.
Factors to consider when analysing ash:

1. High fat sample can be solvent extracted

first to remove fat
2. High moisture can be pre dried first
3. Alcohol, magnesium acetate, and glycerol
may speed ashing
4. Spattering can be controlled with lids on
the crucibles
5. Wet ashing is often used as prep for
mineral analysis
 Question

What is the amount of total ash content (in dry basis) in

1.2026 gram of biscuit (containing 14% water) when the
weight of total ash obtained was 0.0183 gram? Show your
calculation.
Dry matter =100 – Moisture
= 100 – 14
= 86%

Dry matter (%) = Weight of dried sample x100 %

Weight of initial sample
86 = a x 100
1.2026
a = 86 x 1.2026
100
= 1.0342 g
Ash (%) in dry basis = Weight of ash x 100
Weight of dried sample
= 0.0183 x 100
1.034
= 1.77%

Ash (%) in wet basis = Weight of ash x 100

Weight of wet sample
= 0.0183 x 100
1.2026
= 1.52%