PENGUJIAN MUTU BAHAN BAKU

DAN TITRASI KOMPLEKSOMETRI

Aliya Nur Hasanah
PHARMACEUTICAL ANALYSIS-PHYSICOCHEMISTRY LABORATORY
2019
PENGUJIAN MUTU BAHAN OBAT
 Setiap obat dan produk farmasi yang akan digunakan dan
diedarkan untuk dijual harus memenuhi syarat : khasiat,
keamanan dan kualitasnya (UU 36/2009, pasal 98
ayat 1)
 uji kualitas obat yang dilakukan di suatu
laboratorium pengujian kualitas/mutu( industri,
pemerintahan atau universitas) mengacu pada Farmakope
 Farmakope adalah sebuah buku yang berisi kumpulan
standar dalam bidang farmasi terutama bahan baku obat
serta sediaan jadinya, produk biologi, alat kesehatan,
metode analisis, prosedur dan instrumennya, bahan baku
pembanding, sediaan umum dan penerapan standar yang
berkaitan dengan standarisasi di bidang farmasi.
 PENGUJIAN MUTU BAHAN BAKU
OBAT

 Tujuan : menetapkan kesesuaian dengan persyaratan

bahan baku obat meliputi: identitas, atribut mutu,
kemurnian dan kadar.
 Cara : menggunakan metode, prosedur dan instrumen
yang tercantum dalam Farmakope.
 Kalau tidak tercantum dalam FI, maka dapat digunakan
persyaratan dari Farmakope lainnya seperti: USP, BP, JP,
EP, dll)
 Isi Monografi Farmakope

 Nama resmi farmakope (INN, generik) dalam bahasa Indonesia dan Latin.
 Rumus molekul, struktur dan nama kimia resmi dengan CAS number dan BM.
 Pernyataan kadar atau potensi bahan aktif dalam bahan baku atau sediaannya.
 Pemerian atau uraian dari segi organoleptik bahan.
 Kelarutan dalam berbagai pelarut.
 Identifikasi
 Syarat atribut mutu/tetapan fisika bahan
 Kemurnian
 Penetapan Kadar/Potensi
 Wadah dan Penyimpanan
1. SYARAT IDENTITAS

 Syarat identitas atau identitas baku adalah

pernyataan kualitatif yang harus dipenuhi untuk
membuktikan kebenaran, kesesuaian identitas dan
keotentikan senyawa aktif seperti yang tertera pada
etiketnya sehingga dapat dibedakan dengan senyawa/bahan
yang lain.
 Identifikasi adalah suatu cara untuk mengungkap
identitas dan membuktikan bahwa bahan yang diperiksa
mempunyai identitas yang sesuai dengan senyawa yang
tertera pada etiketnya.
 Cara melakukan identifikasi

Syarat identitas dapat diungkap dengan melakukan berbagai

uji identifikasi yang berdasar pada:
 Cara metode kimiawi : penggunaan pereaksi kimia yang
dapat bereaksi secara khas dengan senyawa yang diuji.
 Cara fisikokimia : menggunakan instrumen
 Cara kromatografi : KLT atau Kromatografi jenis lain
 Cara fisika : menggunakan tetapan Fisika seperti rotasi
jenis, indeks bias, jarak lebur, bobot jenis
 II. SYARAT ATRIBUT MUTU

1. Tujuan pengujian untuk menetapkan tetapan fisika yang dapat

digunakan sebagai atribut mutu (Atribut = parameter uji)
2. Tetapan fisika yang sering diuji adalah:
- titik/jarak lebur
- titik/jarak didih
- rotasi jenis
- indeks bias
3. Fungsi lain
a. dapat digunakan sebagai cara identifikasi (kalau murni)
b. dapat digunakan sebagai cara pengujian kemurnian (kalau tidak
murni)
III. SYARAT KEMURNIAN
Tujuan untuk membuktikan bahwa bahan bebas dari senyawa asing dan
cemaran atau mengandung senyawa asing dan cemaran pada batas tertentu.
Pengujian terhadap adanya senyawa asing dan cemaran dimaksudkan untuk
membatasi senyawa demikian sampai pada jumlah yang tidak mempengaruhi
artikel pada kondisi penggunaan biasa.

Cara/metode pengujian : uji batas, kemurnian kromatografi, susut pengeringan,

kadar air, sisa pemijaran, kelarutan, zat mudah menguap, zat mudah terarangkan

Sumber : cemaran dan senyawa asing (bahan baku, hasil antara, hasil urai,
wadah, lingkungan dll.
IV. SYARAT KADAR
Tujuan untuk menetapkan kadar senyawa aktif dalam bahan yang diuji. Adanya
batas-batas dan toleransi, tidak merupakan suatu dasar untuk menyatakan bahwa
bahan yang hampir mendekati kemurnian 100 %, melampaui kualitas farmakope.

Spesifikasi dari ukuran tertentu peralatan wadah dan istrumen untuk penetapan
kadar adalah rekomendasi.Yang penting tingkat ketelitiannya paling sedikit sama
dengan alat tersebut.

Dalam melaksanakan penetapan kadar jumlah satuan takaran yang digunakan tidak
boleh lebih kecil dari yang telah ditetapkan.
TITRASI KOMPLEKSOMETRI
 DEFINITION

TITRIMETRIC METHODS THAT ARE BASED

UPON COMPLEX FORMATION
REACTIONS
Application

1) Calcium determination in food using EDTA titration

AOAC Method 968.31
2) Water hardness (Ca, Mg)
3) Ca or Al content of drugs such as calcium pantothenate or
alumina.
4) Metal content in every kind of matrix

Typical kit for testing for water hardness

in household water.
 Terms in Complex-Formation Reaction

 Ligand : an ion or molecule that forms a covalent bond

with a cation by donating a pair of electron, which are
then shared by the ligand and the cation
 Coordination number : number of covalent bonds it tends
to form with electron donor species
 Complex : compound formed when metal ion combines
with a molecule which can donate electron
 Chelate : cyclic complex formed when a cation is bonded
by two or more donor groups contained in single ligand
KIND OF COMPLEXING AGENT
 CHEMISTRY AND PROPERTIES OF EDTA

 Ethylenediaminetetraacetic acid, or EDTA is an

aminocarboxylic acid
 EDTA is a lewis base
 Has six binding sites (the four carboxylate groups and the
two amino groups)
 The resulting metal-ligand complex, in which EDTA forms
a cage-like structure around the metal ion is very stable
 All metal-EDTA complexes have a 1:1 stoichiometrically
Structure of the EDTA (as Y4-)
Six coordinate metal-EDTA complex
Reaction of EDTA with metal ion :

Mn+ + Y4- MY(n-4)+

Example :
Formation of the silver and aluminium complexes
The constant value refers to the equilibrium
involving the spesi Y4- with the metal ion :
 EQUILIBRIUM CALCULATION INVOLVING
EDTA

 A titration curve for the reaction of a cation with

EDTA consists of a plot a pM as a function of
reagent volume
 Values for pM computed by assuming that the
equilibrium concentration of M is equal to its
analytical concentration
 For calculating [Y4-] we need 4 (Y4- fraction)
 CT is the molar concentration of uncomplexed EDTA
 To get [Y4-] in equilibrium and beyond, we used
equation involving constant KMY and 4 :

K’MY is the conditional formation constant, describes

equilibrium relationship only at the pH for which 4 is aplicable
 Conditional formation constant (K’MY) depends on pH, it becomes
smaller at lower pH level and the complex becomes less stable
 to maintain a constant pH, we must add a buffering agent
 If one of the buffer’s components forms a metal-ligand complex,
then EDTA must compete with the ligand for the metal
Titrations

EDTA solution

Sample + indicator+buffer
Detection of the End
Point
 Using indicator

 Example indicator : EBT (eriochrome black T)

contains three ionizable protons.

can be used for the titration of Mg 2+ with EDTA.

a small amount of indicator is added to the sample solution and it forms a red
complex with part of the Mg 2+; the color of the uncomplexed indicator is blue.

as soon as all the free Mg 2+ is titrated, the EDTA displaces the indicator from
magnesium, causing a change in the color from red to blue.
MgIn- + H2Y2- MgY 2- + HIn 2- + H+
red colorless colorless blue
The metal indicator complex must be less stable than that of
the metal-EDTA complex, or else the EDTA will not displace it
from the metal
The metal indicator complex must not be too weak, or the
EDTA will start replacing it at the beginning of the titration
The metal-indicator complex should be 10-100 times less -
stable than the metal-titrant complex
Metal Ion Indicators
Metal ion indicators are compounds whose color changes when they
bind to a metal ion. Useful indicators must bind metal less strongly than
EDTA does.
A typical titration is illustrated by the reaction of Mg2+ with EDTA, using
Eriochrome black T as the indicator.
MgIn + EDTA  MgEDTA + In
(red) (colorless) (colorless) (blue)

Structure and molecular model of Eriochrome Black T(left) and Calmagite (right).
 most indicator for complexation titration are organic
dyes, form stable complexes with metal ions
 Most of these dyes (metalochromic indicators) are
weak acids or weak bases
 Conditional formation constant for metal-indicator
depends on pH
EDTA Titration Techniques
 Direct titration: analyte is titrated with standard EDTA.
 Back titration: a known excess of EDTA is added to the
analyte.
 Displacement titration: For metal ions that do not
have a satisfactory indicator.
 Techniques of EDTA Titration
 Direct titration
the solution containing the metal ion to be determined is buffered
to the desired pH and titrated directly with the standard EDTA
solution. It may be necessary to prevent precipitation of the
hydroxide of the metal by addition of some auxiliary complexing
agent, such as tartrate or citrate or triethanolamine
 Back titration
many metals cannot be titrated directly; thus they may precipitate
from the solution in the pH range necessary for the titration or
they may form inert complexes, or a suitable metal indicator is not
available. In such cases an excess of standard EDTA solution is
added, the resulting solution is buffered to the desired pH and the
excess of the EDTA is back titrated with a standard metal ion
solution; a solution of zinc chloride or sulphate or of magnesium
chloride or sulphate is often used for this purpose. The end point is
detected with the aid of the metal indicator which responds to the
zinc or magnesium ions introduced in the back titration
 Replacement or substitution titration

May be used for metal that do not react (react unsatisfactory) with metal indicator, or
for metal ion which form EDTA complexes that are more stable than those of other
metals such as magnesium and calcium.

Example :
In titration of calcium. In the direct titration of calcium ions, solochrome black gives
poor end poin; if magnesium is present, it is displaces from its EDTA complex by
calcium and improved end point results
 There is excess EDTA, and
vitually all the metal ion is in the
form Myn-4

There is exactly as much EDTA as

metal in the solution. [Mn+ ] =
[EDTA]

In this region, there is excess Mn+

left in solution after the EDTA has
been consumed.

Three regions in an EDTA titration illustrated for reaction of 50.0mL of 0.050

0M Mn+ with 0.050 0M EDTA, assuming Kf’ = 1.15  1016.
1. Derive the titration curve for 40 mL of 0,015 M Fe2+
with 0,03 M EDTA in a solution buffered to pH 7,0.
Calculate pFe values after the addition of 0 ; 15 ; 20
and 25 mL of titrant. KMY value for Fe-EDTA 2,1 x 1012
and Y4- for pH 7 = 4,8 x 10-4
2. Calculate the concentration of Mg 2+ in a solution
prepared by mixing 100 mL of 0,03 M Mg 2+ with
100 mL of 0,05 M EDTA. The mixture is buffer to
a pH of 3.
4 at pH 3 = 2,5 x 10-4. KMg-Y4- = 4,9x 108
3. Calculate volume of 0,05 M EDTA needed to titrate
27,16 mL of 0,0741 M Mg(NO3)2
4. A solution contains 1,694 mg of CoSO4 (155,0 g/mol)
per mililiter. Calculate the volume of 0,08640 M EDTA
needed to titrate a 25mL aliquot of this solution
5. A solution contains 1,694 mg of CoSO4 (155,0 g/mol)
per mililiter. Calculate the volume of 0,009450 M Zn2+
needed to titrate the excess reagent after addition of
50 mL of 0,008640 M EDTA to a 25 mL aliquot of this
solution