 Abu = Residu anorganik yang tersisa, baik

setelah pengapian maupun oksidasi secara

lengkap material organik dalam bahan
makanan.
◦ dry ashing (proximate analysis)
 whole grain (gandum), cereals and dried vegetables
◦ wet ashing (oxidation, preparation for elemental
analysis)
 meat and meat products
◦ microwave (low temperature ashing)
 volatile elements
 Ash content of fresh food is rarely >5%

2
 Sample (5-10 g) weighed into
crucible and pre-prepared
 Ignited in muffle furnace 12-18 hr, 550oC
◦ water & volatiles vapourised
◦ organic substances oxidised to water vapour,
carbon dioxide, and oxides of nitrogen
◦ minerals converted to oxides, sulphates,
phosphates, chlorides and silicates
◦ some elements may be partially lost through
volatilisation eg. Fe, Se, Pb, Hg
 Cool furnace and open door carefully as ash
may be fluffy (halus)
 Cool in desiccator and calculate ash weight
as percentage of original sample
3
 Wet oxidising of organic substances
 Place 1g dried sample of food in H2SO4
& HNO3
 Heated to 200C on hot plate in fume-
hood’ brown-yellow fume will evolve
◦ sample should become colourless
 Cool and transfer oxidised food
solution to 50 mL volumetric flask
 Make to volume with ultra pure water

4
 Totally automated
 May carry out dry or wet ashing
 Wet ashing is performed in open or closed
vessels (Teflon, quartz or Pyrex) which
withstand pressures of >1500psi
◦ acids may be heated past their boiling point
 ensures complete digestion in 30 min.
 permits use of nitric acid when normally we would
require sulfuric acid

5
 Time, temperature, pressure & microwave
power parameters are adjustable
◦ may ramp the temperature according to
preprogramming
 Dry ashing may reach up to 1200°C
◦ uses the same protocol and crucibles as muffle
furnace ashing
◦ generally 20 min. in a microwave oven is equal to
4hr in a muffle furnace

6
Destruksi sampel dalam wadah yang tertutup
pada oven microwave memiliki beberapa
keunggulan dibandingkan metode wadah
peleburan terbuka.
◦ Wadah terbuat dari polimer yang tahan terhadap
temperatur tinggi, dan kecil kemungkinan
mengandung kontaminan logam seperti pada
krus keramik atau gelas piala.
◦ wadah yang tertutup menghilangkan kesempatan
kontaminasi dari debu di udara.

Sampleprep_MJH 7
◦ Wadah yang tertutup, tekanan udara wadah mengurangi
penguapan, jadi dibutuhkan larutan yang tidak terlalu asam
untuk bahan dekstruksinya, mengurangi blanko.
◦ Wadah yang tertutup juga mengurangi hilangnya lebih
banyak spasies logam yang menguap, yang merupakan
masalah pada destruksi menggunakan wadah terbuka,
utamanya destruksi (pengabuan) kering.
◦ Kontrol elektronik pada microwave pendestruksi modern
memungkinkan kondisi destruksi yang reproducible. Sistem
otomatis mengurangi pekerjaan oleh operator. Terakhir,
kontrol pembuangan uap yang mengandung uap asam
dapat dinetralisir.

Sampleprep_MJH 8
Sampleprep_MJH 9
Sampleprep_MJH 10
Sampleprep_MJH 11
Destruksi dengan
microwave

Sampleprep_MJH 12
 Lima milliliters asam nitrat pekat ultra murni
ditambahkan ke masing-masing dan wadah
tertutup. Semua diproses dengan program (ramp)
100-600 W lebih dari 5 menit, pada 600 W selama
5 menit, dan pada 1000 W selama 10 menit.
Setelah 15 menit pendinginan, sampel dibuka dan
diencerkan dengan 50 atau 100 mL dengan
deionized warter, sebelum analisis dengan ICP-
AES.

Sampleprep_MJH 13
 Flame photometry and atomic absorbtion
spectroscopy
 EDTA complexation titration
 Redox reactions
 Precipitation titration
 Colorimetric methods
 Ion selective electrodes

14
 Milling and grinding with steel grinders
 Old glassware can contaminate samples
for micro-elemental analysis
◦ glass is acid washed & triple rinsed with ultra
pure water
 Solvents including water may contain high
amounts of minerals
◦ need pure reagents high in cost
◦ run reagent blank

15
• Formation of stable complexes of
metal ions with
ethylenediaminetetraacetic acid (EDTA)
– except alkali metals (Na)
• Via the presence of donor oxygen and
nitrogen atoms EDTA is able to form
six, five member chelate rings

Fennema, 1996 p.625 16

 Adjust water sample pH to 10 by adding
buffer solution (NH4OH + Na2EDTA +
MgCl2) and Calmagite indicator solution
 Titrate with 0.01 EDTA to a blue endpoint
 This method is suitable to assess Ca in
ashed fruits and vegetables

17
 Add magnesium salt and enough EDTA to
bind all magnesium.
 In buffer solution the Ca replaces the Mg
bound to the EDTA.
 The free magnesium binds to Calmagite,
◦ pink magnesium Calmagite complex persists until
all Ca in the sample has been titrated with the EDTA.
 Excess EDTA removes Mg from Clamagite and
produces a blue endpoint

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 Revision!
◦ oxidation
 addition of oxygen OR removal of hydrogen
 removal of electrons = increase in positive charge
◦ reduction
 removal of oxygen OR addition of hydrogen
 addition of electrons = reduction in positive charge

19
 Food sample dried and ashed
 Dissolve ash in HCl and filter with rinsing
 Add aliquot of filtrate containing iron to
solution of hydroxylamine hydrochloride
 Add acetate buffer and colour developing
reagent such as 0.1% orthophenanthroline
 Dilute and read absorbance of colour at
510 nm
 To calculate iron content compare
absorbance of sample to standard curve
generated by known concentrations of
iron chloride
◦ prepared by dissolving analytical grade iron
wire in HCl
20
 ‘Mohr titration’ Based on formation of an
orange coloured solid, silver chromate
after silver from silver nitrate has
complexed with all available chloride
AgNo3 + NaCl  AgCl (Cl- is complexed)+ NaNo3
2AgCl + K2CrO4  Ag2CrO4 (orange all Cl- is complexed) +
2KCl
◦ butter is melted in boiling water in a conical flask
◦ potassium chromate solution is added
◦ titrated with silver nitrate until an orange brown
colour persists for 30 sec
◦ exact normality of silver nitrate solution
standardised against known amount of potassium
chloride
◦ % salt in food calculated from concentration &
titration volume of standardised silver nitrate 21
 Sample of food is ashed
 Add HCl and nitric acid
 Heat to dissolved ash, cool and make up to
volume in water
 Add 20 ml of molybdovanadate reagent to
aliquot of ash solution
 After colour development (10 min.) due to
formation of phosphomolybdovanadate
◦ read absorbance at 400 nm
 To calculate phosphorus content compare
absorbance of sample to standard curve
generated by known concentrations of
potassium dihydrogen phosphate
22
 Similar concept to pH electrodes that
measure H+ ions can be applied to other
ions and even dissolved gases
 Chemical composition of glass in electrode
is one means of making electrodes sensitive
to specific ions
◦ 71% SiO2, 11% Na2O and 18% Al2O3 for K
 A typical glass membrane sodium indicating
electrode operates in the range of 1 - 10-6
M
 Usually develop a calibration curve of
electrode potential (millivolts) developed in
standard solutions
◦ plot on semilog paper electrode potential vs
logarithms of the standard concentrations
23
 When sulphur dioxide or sulphur salts are
dissolved in water an equilibrium between
sulphur dioxide and a range of ions is
established as follows:
SO2 + H20 ↔ [H2SO3] ↔ H+ + HSO3- ↔ H+ + SO32-
sulphur sulphurous acid bisulphite sulphite
dioxide

24
 Pale coloured liquid foods or foods that can
be dispersed in water
◦ digest food with cold alkali (pH 13),
◦ acidify to produce un-dissociated sulphurous
acid
◦ sulphur dioxide reacts with standard iodine
solution
SO2 + I2 + 2H2O  2I- + 2H+ + H2SO4 + I 2
◦ excess iodine reacts with starch to give a dark blue
endpoint
 Foods not easily dispersed in water or
intensely coloured
◦ distilling of sulphur dioxide from the acidified
food
◦ titrating the colourless sulphur dioxide directly
with standard iodine solution as it distills over 25
 Levels of nitrites (NO2) & nitrates (NO3) in
foods are controlled by international
regulation
◦ have the potential to form toxic
nitrosamines and to interfere with infant
metabolism
 Aqueous extraction of the food
◦ to reduce nitrate loss, pH of extract is >5
 De-proteinisation of the extract at the
isoelectric point of the contaminating
proteins followed by filtration

26
 Extracted nitrates reduced to nitrites by
nicotinamide-adenine dinucleotide
phosphate (NADPH) in the presence of the
enzyme nitrate reductase
Nitrate + NADPH  nitrite + NADP+ + H2O
 The nitrites originally present in the sample
plus those converted from nitrates
converted to a diazo dye
Nitrite + sulphanilamide + NED  diazo dye
 Level of dye can then be determined
colorimetrically with reference to a standard
curve
 Nitrite and nitrates in food calculated as
nitrite
27