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ANALISIS PROKSIMAT

Table 7.1 Methods of analysis for water

Procedure Applicability Limitations Capital Selected references
costs
Physical removal of water
Air oven at Most foods, Caramelization of Low AOAC International,
100 105°C except those rich sugars, degradation of 2002; Anklam, Burke
in sugars and fats unsaturated fats, loss of and Isengard, 2001;
other volatiles Nielsen,  1998
Vacuum oven Most foods Loss of volatiles Low As above
at 60°C
Freeze- Most foods Slow, residual water in Medium As above
drying samples
Microwave Medium or high Charring Low As above
oven moisture
Dean & Stark Foods high in Safety of solvents used Low As above
distillation volatiles
Chemical reactivity
Karl Fischer Low moisture,   Low As above
hygroscopic
foods
Physical methods
NMR Most foods Need for calibration High Bradley, 1998; Hester
with specific food and Quine,  1976
NIR Established for Need for extensive High Williams, 1975
cereals and some calibration with specific
other foods food. Particle size
dependence
Chromatography
GLC Meat and meat   High Reineccius and Addis, 
products 1973
GSC Some meat   High Khayat,  1974
products
Notes: 
References selected provide detailed procedures, evaluations or reviews.
NMR = nuclear magnetic resonance; NIR near infrared reflectance; GLC = gas—liquid
chromatograpphy; GSC = gas—solid chromatography.
Low, Medium, High capital costs are described in the text.
Penetapan kadar air
Ada banyak metoda yang dipakai untuk penetapan kadar air, selain yang klasik (penguapan
pada 100-105oC), yaitu physical reactivity, chemical reactivity, dan kromatografi. Masing-
masing untuk bahan yang berbeda. Misal untuk bahan tepung, metoda klasik bisa dipakai
tetapi untuk produk daging, keju.
Kelompok 1: kajilah metoda klasik, cari AOAC International, 2002; Anklam, Burke and
Isengard, 2001; Nielsen,  1998, bagaimana prosedur sebenarnya penetapan kadar air dengan
cara ini, apa kelebihan dan kekuranganannya
Kelompok 2: kajilah metoda Dean and Stark Distillation, dan cocoknya untuk produk apa,
kelebihan dan kekurangannya, perhitungannya

Penetapan Kadar Protein

Table 7.4 Methods of analysis for amino acids

Capital
Procedure Applicability Limitations Selected references
costs

Ion-exchange All foods Hydrolytic losses of High  AOAC International, 2002;

chromatography after more labile  amino De Geeter and
acid hydrolysis acids and slow Huyghebaert,1992.
release of branched
chain amino acids

High-performance  All foods  As above  High  As above

liquid chromatography
after acid hydrolysis

Gas chromatography  Most foods  Choice of derivatives  Medium  As above

after acid hydrolysis is critical to high
and
derivatization

(Sulphur amino acids)  Most foods  Hydrolytic losses  High  As above
Acid hydrolysis
after oxidation of
sulphur amino acids.

(Tryptophan) Alkaline  Most foods  Hydrolytic losses of  High Moore and Stein, 1948;
hydrolysis and  ion- other amino acids  Landry and Delhave, 1993
exchange
chromatography

(Tryptophan, S amino Most foods Low  Blackburn, 1968;

 
acids) Colorimetry   Christie & Wiggins, 1978

(Available lysine) Most foods Low Carpenter,1960;

 
Colorimetry   Booth,1971
Metoda Kjeldahl hanya menetapkan total protein, untuk beberapa makanan, sekarang
trendnya adalah mengetahui susunan asam amino.
Kelompok 3: kajilah cara penetapan asam amino lisin menggunakan 2,4
fluorodinitrobenzene menggunakan metoda Carpenter. Yang dikaji: prinsip reaksi,
alat/metoda/pereaksi, cara perhitungan. Sedikit informasi:

Lysine can become nutritionally unavailable under certain conditions that lead to the e-amino
group reacting with carbohydrate. This reaction reduces the biological value of the protein.
Using the Carpenter method (1960) available lysine can be measured by its reaction with 2,4-
fluorodinitrobenzene. This method has been the subject of many modifications (Williams,
1982). HPLC separation of e-DNP lysine is described by Peterson and Warthesen (1979).

Kelompok 4: kajilah metoda HPLC untuk mengetahui susunan asam amino. Prinsip HPLC,
berdasarkan apa asam amino terpisah, dan bagaimana bisa keluar hasilnya.(lihat di table
untuk referensi, tetapi boleh dari mana saja)

Kelompok 5: kajilah bagaimana menentukan kadar asam amino yang mengandung sulfur
(lihat di table, referensi yang bisa dipakai adalah: Blackburn, 1968;
  Christie & Wiggins, 1978) tidak harus ini tetapi bisa dilihat dimana saja bagaimana cara
penetapan kelompok asam amino dengan sulfur ini

Kelompok 6: kajilah penetapan triptofan, referensinya yang kolom paling kanan

(Tryptophan) Alkaline  Most foods  Hydrolytic losses of  High Moore and Stein, 1948;
hydrolysis and  ion- other amino acids  Landry and Delhave, 1993
exchange
chromatography

Kelompok 7: cari jurnal yang meneliti kandungan flavonoid (boleh flavonoid yang mana
saja: quercetin, kampferol, myricetin, luteolin, apigenin)pada bahan makanan, presentasikan
kembali , singkat saja, isinya: apa khasiat flavonoids tersebut, metoda penetapan flavonoid
tersebut, hasilnya kandungan flavonoidsnya berapa/kesimpulan. (sertakan URL jurnal
tersebut)

Kelompok 8: sama seperti kelompok 7 tetapi untuk phytoestrogen

Kelompok 9: sama seperti kelompok 7 tetapi untuk isoflavons dan coumestrol

Kelompok 10: sama seperti kelompok 7 tetapi untuk lignan

Flavonoids. A rapid method based on reversed-phase HPLC with UV detection was

developed by Hertog, Hollmann and Venema (1992) for the quantitative determination of
five major flavonoid aglycones (quercetin, kaempferol, myricetin, luteolin and apigenin) in
freeze-dried vegetables and fruits, after acid hydrolysis of the parent glycosides. More
recently Merken and Beecher (2000) published a gradient HPLC method with photodiode
array detection for 17 prominent monomeric flavonoid aglycones representing all of the five
common classes of flavonoids.

Phytoestrogens. The main plant compounds with known or suspected estrogenic activity are
lignans, isoflavones, coumestans and resorcyclic acid lactones (Price and Fenwick, 1985).
The modes of estrogenic action are discussed by Clarke et al. (1996). The major
isoflavonoids are genistein, daidzein, formononetin, biochanin A and glycitein. Genistein,
daidzein and glycitein occur in foods as their glycosides, all of which are biologically
inactive. The free aglycones are formed by metabolic action of the human gut microflora,
although this hydrolysis varies considerably from person to person (Xu et al., 1994). The
total bioactivity is represented by the analysis of aglycones; however, this potential activity is
represented by analysis of the conjugates and aglycones separately. The most active plant
estrogen known is coumestrol (a coumestan); zearalenone is a potent resorcyclic acid lactone
formed as a secondary metabolite of fungal species, mainly Fusarium (and is thus regarded
as a contaminant). The lignans matairesinol, secoisolariciresinol, pinoresinol and
isolariciresinol are potent phytoestrogens and are precursors of the mammalian lignans,
enterolactone and enterodiol.

Given the very large number of plant compounds with estrogenic activity and the question of
whether to analyse both the conjugates and the free forms or only the aglycones (after
hydrolysis), many methods of analysis are in existence and there is little agreement on which
method is best. No method is available to separate and quantify all bound and free
compounds of interest in this category. Probably the most comprehensive method for the
aglycones is the isotope dilution gas-chromatographic–mass spectrometric method of
Adlercreutz and coworkers (Mazur et al., 1996), which analyses daidzein, genistein,
biochanin A, formononetin, coumestrol, secoisolariciresinol and matairesinol, but not
glycitein, as silyl derivatives. The method is expensive and needs access to mass
spectrometry (MS). Another comprehensive method for foods that analyses daidzein,
genistein, biochanin A, formononetin, coumestrol, secoisolariciresinol and matairesinol, but
not glycitein, uses an HPLC-MS method originally developed for plasma and urine (Horn-
Ross et al., 2000; Coward et al., 1996; Horn-Ross et al,. 1997; Barnes et al., 1998).

Isoflavones and coumestrol. For the USDA–Iowa State University Isoflavones Database

(2002), the reference method adopted was the linear gradient method of Murphy et
al. (1997), which separates daidzein, genistein, glycitein and their conjugates in soy-based
infant formulas. Hutabarat, Greenfield and Mulholland (2000) have published a rigorously
validated isocratic HPLC method for genistein, daidzein, formononetin, biochanin A and
coumestrol (but not glycitein), while King and Bignell (2000) have published an HPLC
method for daidzin, genistin, glycitin and their aglycones. A collaborative trial published by
Klump et al. (2001) led to a recommendation to adopt as first action AOAC Method No.
2001.10 for the determination of isoflavones in soy and selected foods containing soy. This
method uses reversed-phase liquid chromatography to separate and measure genistein,
glycitein and daidzein and their glucosides, and also produces values for total isoflavones
expressed as aglycones.

Lignans. Meagher et al.  (1999) measured isolariciresinol, pinoresinol, secoisolariciresinol

and matairesinol using HPLC with photodiode array detection, and Liggins, Grimwood and
Bingham (2000) have published a GC-MS method for the determination of matairesinol,
secoisolariciresinol and shonanin in foods as trimethylsylyl derivatives.
Total antioxidant activity. There is growing interest in ways to represent the total
antioxidant activity of foods. A number of methods have been used but no standards exist and
at this stage the inclusion of values for total antioxidant activity in foods in databases is not
recommended. The topic is fully reviewed by Frankel and Meyer (2000).

Kelompok 11: cari jurnal penetapan kadar lemak/lipid dengan “mixed solvent extraction”.
Presentasikan kembali meliputi : prinsip/metoda. Diaplikasikan untuk makanan apa. Hasil
seperti apa

Kelompok 12: cari jurnal penetapan kadar lemak/lipid GLC (Gas Liquid Chromatography)
Presentasikan kembali meliputi : prinsip/metoda. Diaplikasikan untuk makanan apa. Hasil
seperti apa

Table tentang penetapan lemak/lipid/asam lemak/triasilgliserol

Table 7.5 Methods of analysis for lipids

Selected
Procedure Application Limitations Capital costs
references

Total fat

Continuous  Low moisture foods Incomplete extraction Low Sullivan and

extraction  (single solvent) from many foods. (dry Carpenter, 1993
analytical samples)
Time consuming. 
Extracts cannot be 
used for fatty acid
studies

Acid hydrolysis All foods except Some hydrolysis of Low AOAC

dairy and high sugar lipids. Extracts  cannot International,
products be used for fatty acid 2002;  Sullivan and
studies Carpenter, 1993

High Ngeh-Ngwainbi,
Acid hydrolysis and Cereal foods (NLEA
  Lin and Chandler,
capillary GLC compliant)
1977

Mixed solvent Rapid, efficient for Complete extraction Low Bligh and
extraction many foods.  Extract from most foods.  Dyer,1959;
can be used for fatty Extractsoften need Hubbard etal.,
acid measurements clean-up  1977

Alkaline hydrolysis Dairy foods Validated for dairy Low AOAC

 International,
foods only
2002

NIR Established for Requires extensive High Hunt et al., 1977a

cereals calibration against
other methods

Triacylglycerols

Range of All foods Free fatty acids can Medium Gurr, Harwood
chromatographic interfere.  TLC checks and  Frayn, 2002
methods useful

Fatty acids
GLC All foods after Validated for most High See above
transmethylation foods

HPLC Under development Not found to have High See above

advantages over GLC at
present

Transfatty acids
Availability of authentic
GLC with infrared  Medium to
All foods standards for  some See above
analyses High
isomers

Infrared absorption All foods Some interference High See above

GLC All foods Capillary techniques are High/medium See above

required

Notes: GLC = gas-liquid chromatography; NLEA = United States Nutrition Labeling and Education
Act NIR = near infrared reflectance; TLC = thin-layer chromatography; HPLC = high-performance
liquid chromarography.