PT.

Domas AgroInti Prima

Quality Control Department
TEST METHODS

GLYCEROL ASSAY OF GLYCERIN

TM-QC-309
BY TITRATION Effective date
March 01, 2010
Revision : 1
Prepared by: Reviewed by: Approved by:

Gusti Vitra Robby Sirait Syukri Kasim

QA/QC. Supervisor QA/QC. Superintendent QA/QC. Manager
Name/Signature/ Date Name/Signature/ Date Name/Signature/ Date

1. Principle
The determination is based on periodic acid
oxidizing poly alcohols containing three or more
adjacent hydroxyl groups, splitting the molecule
between the carbon atoms attached to these hydroxyl
groups. The end hydroxyl groups are oxidized to
aldehydes and the inner (HCOH) groups to formic
acid. The increase in acidity due to formic acid
production is titrated and equivalent amount of
glycerin is calculated. For glycerin, the reaction is as
follows :
1.Prinsip
Penentuan berdasarkan pada oksidasi asam
periodat terhadap poly alcohol yang mengandung tiga
atau lebih gugus hidroksil berdekatan, , pemecahan
molekul antara atom atom karbon yang terdapat
dalam group hydroksil. Gugus hidroksil ujung
dioksidasi menjadi aldehid dan gugus bagian dalam/
tengah (-HCOH-) menjadi asam formiat. Peningkatan
keasaman disebabkan hasil asam formiat yang dititrasi
dan setara dengan jumlah glycerin yang terhitung.
Untuk glycerin, reaksinya sebagai berikut:

H2C-OH H2C-OH
HC-OH + 2H5IO6  2HCHO + HCO2H + 2HIO3 + 5H2O HC-OH + 2H5IO6  2HCHO + HCO2H + 2HIO3 + 5H2O
H2C-OH H2C-OH
glycerin periodic formal formic iodic water glycerin asam formal asam asam air
acid dehyde acid acid periodat dehyde formiat iodat

Ethylene Glycol, trimethylene glycol, and other
impurities that can be associated with glycerin either
do not react or do not form formic acid as does
glycerin. The equation for oxidation of ethylene glycol
is :
Etilen glikol, trimetilen glikol, dan impurities yang lain
dapat berasosiasi dengan glycerin juga tidak bereaksi
atau tidak membentuk asam formiat seperti glycerin.
Persamaan untuk oksidasi dari etilen glikol adalah:

HOCH2CH2OH + H5IO6  2HCHO + HIO3 + 3 H2O HOCH2CH2OH + H5IO6  2HCHO + HIO3 + 3 H2O
Ethylene periodic formal iodic water Ethylene asam formal asam air
Glycol acid dehyde acid periodat dehid iodat

2.Scope. 2.Ruang lingkup

This method is used to determine the weight percent
of glycerin in a sample, and is especially useful in
analyses of samples containing oxidizable organic
impurities and certain hydroxylated compounds which
interfere with the dichromate and acetin procedures.
Trimethylene glycol and other polyalcohols in which
the hydroxyl groups are not adjacent do not react at
room temperature. Polyglycerols reduce periodic acid
but do not yield formic or other organic acid.
3.Validation Summary
This test method follows the current EP and USP
Monograph and therefore, requires no additional
validation.
3.1 USP/ EP Equivalency
Both monograph methods use similar chemistry
for the assay titration with slight differences in the
Page 1 of 6
Metoda ini digunakan untuk menentukan persen
berat glycerin dalam sampel, dan khususnya berguna
dalam menganalisa sampel yang mengandung
pengotor organik yang dapat dioksidasi dan senyawa
terhidroksil tertentu yang mengganggu dengan cara
kerja dikromat dan asetin. Trimetiln glikol dan
polialkohol lain yang mana gugus hidroksil yang tidak
berdekatan tidak bereaksi pada suhu kamar.
Poligliserol mengurangi asam periodat tetapi tidak
menghasilkan formiat atau asam organik lainnya
3.Ringkasan Pengesahan.
Test method ini mengikuti EP and USP Monograph
terkini dan karena itu, tidak diperlukan validasi
tambahan.
3.1 Kesamaan USP/ EP
Kedua monograf menggunakan metoda kimia
yang sama bagi titrasi assay dengan sedikit
PT. Domas AgroInti Prima
Quality Control Department
TEST METHODS
GLYCEROL ASSAY OF GLYCERIN
TM-QC-309
BY TITRATION
actual specific steps (e.g., incubation times, end point
detection). A limited equivalency exercise was
performed to demonstrate that both tests provide the
equivalent data. This is found below. Either test may
be performed to generate USP or EP assay results.
Sample Description USP Assay Result
N Mean
High-Purity Refined Glycerin 3 99.73
Diluted Refined Glycerin 3 96.38
Suitability of sodium meta periodate / sulphuric
acid solution for EP testing can be performed using
the USP test. The EP monograph specifies that
standardization of this solution is performed with the
addition of perchloric acid. We would hesitate to
implement the use of this chemical in a plant lab
unless no alternative was available. Perchloric acid is
a hazardous material causing burns when contact with
skin and also in respiratory passages upon inhalation.
Parallel testing of the EP sodium metaperiodate
solution using the USP method indicates the existing
method is sufficient to standardize the EP solution.
Ratio obtained was 0.752. ( The ratio of the volume of
0.1 N sodium thiosuphate required for the glycerin-
periodate mixture to that of the blank should be
between 0.750 and 0.765)
4.Reference
4.1 Current USP Monograph
4.2 Current EP Monograph
4.3 P&G Analytical Methods GCAS: 60065183, Issue7
5.Apparatus
5.1 Electronic Balance, accurate to 0.0001 g
5.2 pH meter, accurate to 0.01 pH units.
5.3 Dispenser bottle, 10 mL, 100mL.
5.4 Pipette, 50 mL, Class A Volumetric.
5.5 Sintered-glass filter, 600mL, Medium porosity.
5.6 Dosimat Titrator , 50 mL.
6.Reagents and Solutions
6.1 NaOH, 0.1N, Standardized Solution.
6.2 Sulfuric Acid 0.2N Solution.
6.3 Sodium meta-Periodate (NaIO4), GR
6.4 Ethylene Glycol.
6.5 Bromothymol Blue Indicator, 0.04%.
Page 2 of 6
Effective date
March 01, 2010
Revision : 1
perbedaan dalam tahapan tertentu (seperti masa
simpan, pengamatan titik akhir). Pengujian kesamaan
dilakukan untuk memperlihatkan bahwa kedua
pengujian memberikan data yang sama. Lihat data
berikut. Juga pengujian mungkin dilakukan untuk
memperoleh hasil USP atau EP.
EP Assay Result
Stdev N Mean Stdev
0.231 5 99.66 0.287
0.087 5 96.43 0.222
Kesuaian larutan sodium meta periodat / asam
sulfat bagi uji EP bisa dilakukan menggunakan uji
USP. Monograf EP menetapkan bahwa standardisasi
larutan ini dilakukan dengan penambahan asam
perklorat. Kita kurang suka menggunakan zat kimia ini
di lab pabrik keciali tidak ada lagi pilihan tersedia.
Asam perklorat bahan kimia berbahaya menyebabkan
luka bakar jika kontak dengan kulit dan juga saluran
pernapasan bila menghirupnya.
Pengujian yang sama dari larutan sodiummeta
periodat menggunakan metoda USP menunjukkan
metoda yang tersedia cukup untuk menstandarisasi
larutan EP. Perbandingan diperoleh 0.752. (
Perbandingan volume 0.1 N sodium tiosulfat
diperlukan bagi campuran glycerin-periodat terhadap
blanko harus antara 0.750-0.765)
4.Referensi
4.1 Current USP Monograph
4.2 Current EP Monograph
4.3 P&G Analytical Methods GCAS: 60065183, Issue 7
5.Peralatan
5.1 Timbangan elektonik, berketelitian 0.0001 g
5.2 pH meter, berakurasi 0.01 pH unit.
5.3 Botol Dispenser, 10 mL, 100mL
5.4 .Pipet 50 mL, kelas A Volumetrik.
5.5 Saringan Sintered-glass, 600mL, porositas
medium
5.6 Dosimat Titrator, 50 mL
6 Larutan dan Reagensia.
6.1 Larutan Standar NaOH 0.1N.
6.2 Larutan Asam Sulfat 0.2N
6.3 Sodium meta-periodate, (NaIO4), GR
6.4 Etilen Glikol
6.5 Indikator Bromthymol Blue 0.04%.
PT. Domas AgroInti Prima
Quality Control Department
TEST METHODS
GLYCEROL ASSAY OF GLYCERIN
TM-QC-309
BY TITRATION
6.6 Potassium Iodide 16.5% w/v in water.
6.7 Starch Indicator 0.5% Solution.
6.8 Hydrochloric Acid concentrated
6.9 Sodium Thiosulfate 0.1N Solution.
6.10 pH Buffer solution, pH 4.0, 7.0 and 10.0
6.11 Phenol phthalein indicator.
7.Facilities – N/A
8.Preparation of Solutions
8.1 Preparation of Special Reagent
8.1.1 Ethylene glycol,
Using a graduated cylinder, add 100 mL of ethylene
glycol to 100 mL of distilled water in a 250 mL beaker
and thoroughly mix.
8.1.2 Sodium meta periodate USP Preparation
 Using a 1000 mL volumetric flask, dissolve 60 g of
sodium meta-Periodate (NaIO4) in about 600 mL
water containing 60 mL of 0.2 N sulfuric acid.
 Dilute to volume with distilled water.
 Do Not Heat to dissolve sodium meta-periodate. If
the solution is not clear, vacuum filter through a
sintered glass filter. Store in a light-resistant or amber
glass container. Solution is good form 1 month.
8.1.3 Preparation Sodium metaperiodate Solution EP
 Using a 1000 ml volumetric flask, dissolve 21.4 g
sodium meta periodate in about 600 ml distilled water.
 Dilute to volume with distilled water. Mix throughly
 Do Not Heat to dissolve sodium meta-periodate. If
the solution is not clear, vacuum filter through a
sintered glass filter. Store in a light-resistant or amber
glass container. Solution is good for 1 month.
8.1.4 Solution Suitability Test
 Pipette 10 mL of the periodate solution into a 250
mL volumetric flask, dilute to volume with distilled
water and thoroughly mix with a stir bar and stir plate.
 Weight 0.5 g  0.05 g of glycerin into a 400 mL
beaker.
 Using a graduated cylinder, add 50 mL of distilled
water to the glycerin.
 Using a class A volumetric pipette, add 50 mL of
the diluted periodate solution to the glycerin and water
mixture.
 Prepare a blank in the same manner, but without glycerin. Letakkan larutan ke-dalam ruang gelap
the glycerin. Place the solution in a dark cabinet for 30 selama 30 menit.
minutes.
 After 30 minutes, use a graduated cylinder to add 5  Setelah 30 menit, gunakan gelas ukur untuk
mL of hydrochloric acid to both beakers and swirl to menambahkan 5 mL HCl pada kedua beaker dan
mix.
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Effective date
March 01, 2010
Revision : 1
6.6 KI 16.5% w/v dalam air.
6.7 Larutan indicator Starch 0.5%.
6.8 HCl konsentrat.
6.9 Larutan Sodium thiosulfat 0.1N.
6.10 Larutan Buffer pH 4.0, 7.0 dan 10.0.
6.11 Indikator fenol ftalein
7.Fasilitas – T/A
8.Persiapan Larutan
8.1 Penyiapan Reagent Khusus
8.1.1 Etilen glikol
Gunakan gelas ukur untuk menambahkan 100 mL
etilen glikol ke-dalam 100 mL air demin. pada beaker
250 mL dan aduk.
8.1.2 Persiapan Sodium meta periodat USP
 Gunakan labu ukur 1000 mL, larutkan 60 g sodium
meta periodate (NaIO4) dengan air sekitar 600 mL
yang mengandung 60mL asam sulfat 0.2N.
 Encerkan dengan air demin sampai batas volume.
 Jangan Panaskan untuk melarutkan sodium meta-
periodate. Jika larutan tidak jernih, saring lewat
saringan sintered-glass vakum. Simpan pada wadah
gelas yang berwarna gelap. Larutan bagus sebulan.
8.1.3 Persiapan larutan metaperiodat EP
 Larutkan 21.4 g sodium metaperiodat dengan 600
ml air suling dalam labu taker 1000 ml.
 Encerkan sampai tanda dengan air suling. Campur
dengan baik.
 Jangan Panaskan untuk melarutkan sodium meta-
periodate. Jika larutan tidak jernih, saring lewat
saringan sintered-glass vakum. Simpan pada wadah
gelas yang berwarna gelap. Larutan bagus sebulan.
8.1.4 Uji Kesesuaian Larutan
 Pipet 10 mL larutan periodat masukkan ke-dalam
labu ukur 250 mL, encerkan dengan sampai batas
dan aduk dengan batang pengaduk atau piringan
pengaduk hingga homogen
 Timbang 0.5 g  0.05 g glycerin kedalam beaker
400 mL.
 Gunakan gelas ukur, tambahkan 50 mL air ke
dalam glycerin.
 Gunakan pipet volumemetri kelas A , tambahkan
50 mL larutan periodat kedalam campuran glycerin-air.
 Buat blanko dengan cara yang sama, tetapi tanpa
kocok sampai homogen.
PT. Domas AgroInti Prima
Quality Control Department
TEST METHODS
GLYCEROL ASSAY OF GLYCERIN
TM-QC-309
BY TITRATION
 Use a graduated cylinder to add 10 mL of the
potassium iodide solution to both beakers and swirl to
mix.
 Place the solution in a dark cabinet for an
additional 5 minutes.
 Removes the beakers from the cabinet. Using a
graduated cylinder, add 100 mL of distilled water to
each beaker.
 Titrate both solutions with 0.1N sodium thiosulfate.
 As the end-point is approached, add 3 mL of starch
solution and continue to titrate to the starch end-point.
 Record the volumes of titrant used. The ratio of the
volume of 0.1N sodium thiosulfate required for the
glycerin-periodate mixture to that of the blank should
be between 0.750 and 0.760.
 Make fresh Sodium meta-Periodate if the current
solution does not pass the test.
8.2 Procedure
8.3 Preparation of Sample – N/A
8.4 Preparation of Equipment – N/A
8.5 Instrument Calibration – N/A
8.6 Operation
8.6.1 USP Parameter
 Accurately weight 0.40g  0.05g of sample to the
nearest 0.0001 g into a 400 mL beaker.
 Using a graduated cylinder, add 50 mL of distilled
water to the beaker.
 Prepare a blank by adding 50 mL of distilled water
to a 400 mL beaker.
 Add 5-7 drops of bromothymol blue indicator to
both the sample and blank.
 Acidify both the sample and the blank by adding
0.2N H2SO4 drop-wise to a definite green or greenish-
yellow color. Neutralize both the sample and the blank
by adding 0.05N NaOH drop-wise to a definite blue
color, free from any green color. This step is critical to
assure reproducible results.
 Using a volumetric pipette, add 50 mL of sodium
meta-periodate solution to each beaker. This volume
is critical. Swirl to thoroughly mix.
 Place a watch glass on the beakers and place in a
dark cabinet for 30 minutes, at room temperature.
Light has been found to affect the oxidation reaction.
Failure to protect from strong light may cause high
results.
 At the end of the 30 minutes, use a graduated
cylinder to add 10 mL of ethylene glycol solution to
both beakers and swirl to thoroughly mix. Return the
beakers to cabinet for an additional 20 minutes.
Thorough mixing of sample after addition of ethylene
glycol is necessary for reproducible results.
Page 4 of 6
Effective date
March 01, 2010
Revision : 1
 Gunakan gelas ukur untuk menambahkan 10 mL
larutan KI pada ke-dua beaker dan kocok hingga
homogen.
 Letakkan larutan pada ruang gelap dan biarkan 5
menit.
 Keluarkan beaker dari ruang gelap. Dengan
menggunakan gelas ukur 100 mL tambahkan air
demin. ke-dalamnya.
 Titrasi kedua larutan dengan sodium thiosulfat 0.1N
 Bila titik akhir tercapai tambahkan 3 mL larutan
starch dan teruskan titrasi sampai titik akhir starch-nya.
 Catat volume penitrasinya. Rasio volume dari
sodium thiosulfat 0.1N yang diperlukan untuk
campuran glycerin periodat dengan blanko harus
diantara 0.750 dan 0.760.
 Buat larutan sodium meta periodat yang baru jika
larutan tidak layak pengujian.
8.2 Prosedur
8.3 Persiapan Sampel – T/A
8.4 Persiapan Alat – T/A
8.5 Kalibrasi Instrument – T/A
8.6 Operasional
8.6.1 Parameter USP
 Timbang teliti 0.40 g  0.05 g sampel hingga desimal
0.0001 g ke-dalam beaker 400 mL.
 Dengan gelas ukur tambahkan 50 mL air demin. ke-
dalam beaker.
 Buat blanko dengan menambahkan 50 mL air
demin. ke-dalam 400 mL beaker.
 Tambah 5 – 7 tetes indicator bromthymol blue pada
ke-dua sampel dan blanko.
 Asamkan keduanya dengan menambahkan asam
sulfat 0.2N tetes demi tetes sampai warnanya hijau
atau hijau ke-kuningan. Netralkan keduanya dengan
menambahkan NaOH 0.05N tetes demi tetes sampai
terjadi warna biru, tidak ada warna hijau. Langkah ini
kritikal untuk memastikan hasilnya.
 Dengan pipet volumetric tambahkan 50 mL larutan
sodium meta periodat pada masing-masing beaker.
Volume ini juga sangat kritikal. Aduk hingga merata.
 Letakkan kaca arloji di-atas beaker dan simpan
dalam ruang gelap selama 30 menit pada suhu
ruangan. Cahaya dapat mempengaruhi reaksi
oksidasi. Kegagalan untuk melindungi dari cahaya
terang menyebabkan hasil yang besar.
 Pada 30 menit terakhir tambahkan 10 mL etilen
glikol dengan gelas ukur pada ke-dua beaker dan
aduk hingga merata. Kembalikan beaker tersebut ke-
dalam ruang gelap selama 20 menit. Pencampuran
yang merata pada sampel setelah penambahan etilen
glikol sangat diperlukan untuk hasilnya.
PT. Domas AgroInti Prima
Quality Control Department
TEST METHODS

GLYCEROL ASSAY OF GLYCERIN

TM-QC-309
BY TITRATION
 After 20 minutes remove the beakers from cabinet
and dilute to 300 mL mark with distilled water.
 While stirring on a stir plate, titrate the blank to pH
6.5  0.1 and the samples to pH 8.1  0.1 with 0.1N
NaOH. Record volume of NaOH used to nearest 0.01
mL and the normality of the titrant.
 Calculate % glycerin and report to nearest 0.1%.
8.6.2 EP Parameters
 Accurately weigh 0.075 ± 0.005 g of sample the
nearest 0.0001 g into a 600 ml beaker. Record sample
weight.
 Using graduated cylinder, add 45 mL of distilled
water to the beaker.
 Prepare blank by adding 45 mL of distilled water to
a 400 ml beaker.
 Add 25 mL of sodium metaperiodate EP solution to
both of sample and blank.
 Allow to stand 15 minutes and protect for light.
 Add 5 mL of a 50% w/v solution of ethylene glycol.
 Allow to stand for 20 minutes and protect from light.
 Titrate with standardized 0.1 N sodium hydroxide
solution using 0.5 mL of 0.1% phenol phthalein as
indicator.
 Record volume of NaOH used to the nearest 0.01
mL.
Effective date
March 01, 2010
Revision : 1
 Setelah 20 menit keluarkan beaker dari kabinet dan
encerkan sampai batas 300 mL dengan air demin
 Pada saat pengadukan diatas piring pengaduk,
titrasi blanko sampai pH 6.5  0.1 dan sampel pada pH
8.1  0.1 dengan NaOH 0.1N. Catat volumenya hingga
0.01 mL dan normalitas dari peniter.
 Hitung % glycerin dan laporkan hingga 0.1 %.
8.6.2 Parameter EP
 Dengan teliti timbang 0.075 ± 0.005 g sampel
medekati 0.0001 g kedalam beaker 600 mL. Catat
berat sampel
 Dengan menggunakan gelas ukur, tambahkan 45
mL air suling kedalam beaker
 Siapkan blanko dengan menambahkan 45 mL air
suling kedalam beker 400 mL.
 Tambahkan 25 mL larutan sodium metaperiodat EP
kedalam sampel dan blanko.
 Simpan dalam ruang gelap selama 15 minit.
 Tambahkan 5 mL larutan ethylene glycol 50% (w/v).
 Simpan lagi selama 20 minit dalam ruang gelap.
 Tambahkan 0.5 mL indicator fenol ftalein dan titrasi
dengan larutan standard NaOH 0.1 N sampai titik
akhir titrasi.
 Catat volume NaOH terpakai pada ketelitian 0.01
mL.

9.Quality Control (System Suitability) Requirement 9.Persyaratan Pengendalian Mutu (Kesesuaian

Sistim)
Calibrate the pH meter prior to each set of analyses Kalibrasi pH meter sebelum menganalisa sampel
as described in the calibration procedure. A blank seperti diterangkan pada prosedur kalibrasi. Blank
must be run with every batch of samples. harus dijalankan untuk setiap kelompok sampel.

10.Calculations. 10.Perhitungan

10.1 Actual Percent Glycerin 10.1 Aktual Persen Glycerin

[ Vs – Vb ] x N x 9.209 [ Vs – Vb ] x N x 9.209
% Glycerin = ------------------------------- % Glycerin = -------------------------------
SW (g) SW (g)

10.2 Percent Glycerin on Anhydrous Basis 10.2 Persen Glycerin tanpa air

[ Vs – Vb ] x N x 9.209 [ Vs – Vb ] x N x 9.209
% Glycerin = ------------------------------------- % Glycerin = ---- -------------------------------
SW (g) – [(%H2O/100/SW)] SW (g) – [(%H2O/100/SW)]

Where Dimana
Vs = Volume of NaOH consumed by sample Vs = Volume NaOH diperlukan oleh sampel
Vb = Volume of NaOH consumed by blank Vb = Volume NaOH diperlukan oleh blanko
N = Normality of NaOH N = Normalitas NaOH
SW = Sample weight (g) SW = Berat sampel (g)
9.209 = Milliequivalent of glycerin x 100 to adjust for 9.209 = Milliequivalent glycerin x 100 menghitung
percentage persentase

10.3 Reporting Requirements 10.3 Persyaratan Pelaporan

Page 5 of 6
PT. Domas AgroInti Prima
Quality Control Department
TEST METHODS

GLYCEROL ASSAY OF GLYCERIN

TM-QC-309
BY TITRATION Effective date
March 01, 2010
Revision : 1
Both monographs express the limits for glycerin Kedua monograf menyatakan batas glycerin tanpa air.
anhydrous basis.
 USP/EP : If % glycerin is between 99.0 and 101.0%  USP/EP : Jika % glycerin diantara 99.0 dan
inclusive, the sample passes 101.0%, sampel lulus.
 EP only : If %glycerin is between 98.0 and 101.0%  Hanya EP : Jika %glycerin diantara 98.0 dan
inclusive, the sample passes. 101.0%, sampel lulus.

10.Diagram/ Graphs/ Traces – N/A 10.Diagram/ Grafik/ Lampiran – T/A

11.Reason for Issue or Revision 11.Alasan Penerbitan atau Perubahan

11.1 May 01, 2005 – New issue 11.1 May 01, 2005 – Penerbitan baru
11.2 Mar. 01, 2010 – Change organization 11.2 Mar. 01, 2010 – Perubahan organisasi

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