• Introduction of IC 940

Professional

Ion chromatography 1
CHROMATOGRAPHY

• KROMATOGRAFI ADALAH SUATU TEKNIK PEMISAHAN MOLEKUL BERDASARKAN

PERBEDAAN POLA PERGERAKAN ANTARA FASE GERAK DAN FASE DIAM UNTUK
MEMISAHKAN KOMPONEN (BERUPA MOLEKUL) YANG BERADA PADA LARUTAN.
 The principle of chromatography

take a piece of paper

make a little drop with «black» ink

just add water to it’s center

see the colors that make up

the «black» ink

Greek: Chroma (Colour), Grafein (to write)

Invented for separation of plant pigment (chlorophyll) 3

«chromatography» = wide range of physical chemical separation processes

 •

Ion Chromatography
is a chromatography process that separates
ions and polar molecules based on their
affinity
Ion
Chromatograph
y

4
 Ion chromatography

HPLC (UHPLC) IC
INTRODUCTION Sample 5 – 20 µL Variable volume until 1000 µL
System Stainless steel, 0 – 50 (100) MPa, typical 15 – PEEK, 0 – 35 MPa,
25 (90) MPa typical 10 – 20 MPa
Eluent Organic solvents, Ultrapure water with salt,
typical gradient profile typical isocratic method
Separatio Adsorptive effects Chemical reactions
n  fast with very sharp peaks  typical 15 min for 7 peaks
Detection UV Conductivity
• HPLC VS IC Specialty - Suppression
 •

Setup
of IC

Ion chromatography
General setup of an IC
Detector

Mobile phase
(Eluent)

Injection
valve
High pressure
pump

Separation
Separation
column
column
7
 •

Eluent in
IC Pemisahan terjadi pada fase diam di dalam kolom kromatogram
kolom

Standar air untuk eluen adalah water

type I (resistance > 18.2 MΩ*cm)

Ion chromatography
 Cations
 tartaric acid/dipicolinic acid

BEBERAPA  nitric acid

CONTOH ELUEN 

tartaric acid
citric acid
 sodium dihydrogene phosphate
 oxalic acid/ethylene diamine/acetone

Anions non suppressed IC Anions suppressed IC

 phthalic acid
 carbonate/bicarbonate
 salicylic acid

 p-hydroxybenzoic acid  hydroxide

 benzoic acid  Borate
 borate

 borate/Gluconate

 potassium hydroxide ION CHROMATOGRAPHY

9
 
Column • Anions: acetate, lactate, chloride, sometimes phosphate,
sulfate
10

useful for Metrosep Organic Acids – 100/7.8

Metrosep Organic Acids – 250/7.8

pharma Metrosep A Supp 5 – 250/4.0

Metrosep A Supp 5 – 100/4.0

• Cations: electrolytes such as sodium, potassium, calcium,

magnesium
Metrosep C 4 -100/4.0
(high capacity; detection of low K next to excess of Na
possible)

• UV/Vis applications
reversed-phase columns according to pharmacopoeia

Ion chromatography
Metrosep C4 – 100/4.0

Eluent 2.1 mmol/L HNO3 + 1.2

mmol/L DPA
Flow
0.9 mL/min

Temp
25°C

Injection
5 µL

Detection
Conductivity

Ion chromatography 11
 Ion
chromatography
Preparation

IC 940
1. Labu Ukur 250 ml, 1000ml, 2000 ml
2. Pipet Volume 10ml, 15ml, 20ml, 25ml, 30ml
3. HNO3 2.1 mmol/L, Dipicolinic 1.2 mmol/L, Acetone 5%
4. NaCl Pa, CaCl2 Pa, KCL Pa, Na Laktat Pa,
5. Sample RL
 Ion
chromatography
Preparation

IC 940 HNO3 65% 2.15 Mmol/L Dipicolinic Acid 1.2 mmol/L

BM HNO3 : 63 BM : 167,122
V : 2000 ml 2L
ρ HNO3 : 1,39
M : 1,2 mmol/L 0,0012 M
% HNO3 : 65

massa(gr): ?
M1.V1=M2.V2
massa : 401,0928 kg 0,401093 gr
M1 65%= 14,34127 Molar
V1= 15 ml
v2= 250 ml
M2 ? 2
M2= 0,860476 M
860,4762 Mmol/L
L
I
Perhitungan M1.V1=M2.V2
T
Eluen
V1= 2000 ml
Aceton 5%
M1= 2,15 Mmol/L
V1.M1=V2.M2
E
M2= 860,4762 Mmol/L
V1= 2000 ml R
V2= ?
M1= 5%
V2= 4,997233 ml
M2= 99,99 %
V2= ?
V2= 100,01 ml
 Ion
chromatography
Preparation

IC 940

1. Pipet 10 ml ke dalam 250 ml ad sampai tanda batas (std 50% )

2. Pipet 15 ml ke dalam 250 ml ad sampai tanda batas (std 75% )
Preparasi 3. Pipet 20 ml ke dalam 250 ml ad sampai tanda batas (std 100% )
Larutan 4. Pipet 25 ml ke dalam 250 ml ad sampai tanda batas (std 125% )
Standart 5. Pipet 30 ml ke dalam 250 ml ad sampai tanda batas (std 150% )
 Ion
chromatography
Preparation

IC 940
1. Pipet 20 ml sample RL ke dalam 250 ml ad sampai tanda batas

Preparasi
Larutan
Sample
IC 940

1. BASELINE/STAR
HW
2. INJECT
SAMPLE/STD
3. PEMBUATAN KURVA
STD

4. PEMBACAAN
SAMPLE
IC 940

1. BASELINE/STAR
HW
IC 940
(BASE
LINE)

1. BASELINE/
- Pilih Equilbrium,
- Methode: Rl-3DU
- Start HW
-Tunggu Sampai stabil
30 menit
IC 940
(BASE
LINE)

1. BASELINE/
- Pilih Equilbrium,
- Methode: Rl-3DU
- Start HW
-Tunggu Sampai stabil
30 menit
IC 940
(INJECT
SAMPLE)

1. Pilih Single
Determination

2. Isi kolom sesuai

kebutuhan
3. Klik Start
4. Masukan Selang
kedalam sample
5. Masukan injector,
Tarik Hingga 1ml
6. Klik Continue
7. Lepas Injector
IC 940
(Pembuatan Kurva)

- Setting Komponen

1. Inject Standart 50%,

75%, 100%, 125%, 150%
IC 940
(Pembuatan Kurva)

-Setting Komponen

1. Inject Standart 50%,

75%, 100%, 125%, 150%
IC 940
(Pembuatan Kurva)

- Setting Komponen

1. Pilih DataBase
2. Klik Kanan Pada
STD yg sudah di
proses
3. Klik Reproses
IC 940
(Pembuatan Kurva)

- Setting Komponen

1. Pilih Components,
klik components
table,klik Na
2. Arahkan kursor di
Peak paling kiri
3. Pilih Update
retention time
4. Pilih Update
IC 940
(Pembuatan Kurva)

- Setting Komponen

1. Pilih Components,
klik components
table,klik K
2. Arahkan kursor di
Peak Tengah
3. Pilih Update
retention time
4. Pilih Update
IC 940
(Pembuatan Kurva)

- Setting Komponen

1. Pilih Components,
klik components
table,klik Ca
2. Arahkan kursor di
Peak Paling Kanan
3. Pilih Update
retention time
4. Pilih Update
IC 940
(Pembuatan Kurva)

- Setting Komponen

1. Pilih reprocessing
2. Pilih From Std of
Reprocessing Table
IC 940
(Pembuatan Kurva)

-Setting Event

1. Pilih Integration
2. Pilih Events
3. Klik No.1 arahkan
kursor ke Peak paling
kiri, batas warna
merah, cek RT dan
masukan kedalam
Tabel (start)
IC 940
(Pembuatan Kurva)

-Setting Event

1. Pilih Integration
2. Pilih Events
3. Klik No.2 arahkan
kursor ke Peak ke
batas warna merah,
cek RT dan masukan
kedalam Tabel (Ends)
IC 940
(Pembuatan Kurva)

-Setting Event

1. Lakukan Hal serupa

sampai urutan ke 6,
sesuai Peak yang
terbaca
IC 940
(Pembuatan Kurva)

-Setting Event

1. Pilih Reprocessing, klik

From standarts Of
Reprocessing table
2. Klik Ok
IC 940
(Pembuatan Kurva)

-Setting Event

1. Cek Kurva,
Jika RSD<1% =>kurva std
oke