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Trace level determination of Heterocyclic compounds in Active Pharmaceutical ingredients
by Ion exchange chromatography
Sankar Babu V.R., Thyagarajan. S
Standard preparation
Trace level determination of heterocyclic compounds piperadine, piperazine, n- Study III
methyl piperazine, morpholine in active pharmaceutical ingredients are quantified Piperidine
Selectivity : Mixed cations
Selectivity : Mixed cations
as cations using isocratic separation by ion-exchange chromatography with eluent3.0 mmol/L nitric acid + organic modifier with Metrosep C4 250/4.0 by non- Piperazine, n-methyl piperazine, Morpholine and Piperadine standards are prepared using Sigma-aldrich chemicals.
• 10 ppm mixed standards is prepared using its respective 1000 ppm standard.
A cyclic organic compound containing all carbon atoms in ring formation is referred to as a carbocyclic compound. If at least one atom other than carbon, forms a part Sample preparation
of the ring system then it is designated as a heterocyclic compound. Nitrogen, oxygen and sulfur are the most common hetero atoms.
About 100 mg of sample active pharmaceutical ingredients is taken in 10 mL Heterocyclic compounds are used as structural components and as base in Calibration curve :
pharmaceutical drugs manufacturing due to their high biological activities. Nitro- The sample solution is filtered through 0.2 μ membrane filter and injected into the Sample : Pioglitazone hydrochloride
Component : Piperidine
Sample : Mycophenolate mofetil
Calibration curve
carbon heterocyclic structures are found in various therapeutic agents, including numerous antihistamines, as well as anti-septic, anti-arrythmic, anti-rheumatic, andanti-biotic. Trace level determination of these compounds has become essential toovercome the toxicity effect and to control the impurity profile in active Chromatography condition
Pharmacological significance of heterocyclic compounds has led to analytical Study I : Analysis of Piperidine in Pioglitazone HCl
method development to quantify them in trace level. Methods by Capillary GC andcapillary electrophoresis are available for quantifying them from 10 (mg/L) •Column : Metrosep C4 250/4.0
onwards. The need is realized to develop the method for trace level (ug/L) •Flow rate : 0.9 mL/min ; Sample loop : 100 µL
determination of heterocyclic compounds.
Detection : Non-suppressed Conductivity Detection
In the present study, trace level determination of various heterocyclic compounds Reagents
Piperazine and n-Methyl piperazine
Selectivity : Mixed cations
in pharmaceutical drug matrices are carried out. Piperazine as impurity in n-methyl Mobile Phase : 4.5 mmol/L HNO + 3 % Acetone
Study III
Diluent : Mobile phase
Pioglitazone HCl API (insulin sensitizer) and Morpholine in Mycophenolate mofetil with ion exchange chromatography. Suitable mobile phase composition is selected Study II : Analysis of Piperazine and n-methyl piperazine
with nitric acid and different composition of organic modifiers.
Column : Metrosep C4 250/4.0
Flow rate : 0.9 mL/min ; Sample loop : 250 µL
Detection : Non-suppressed Conductivity Detection
Mobile Phase : 3 mmol/L HNO + 15 % Acetone,
Diluent : Mobile phase
Piperazine in n-methyl piperazine
n-methyl piperazine in piperazine
Study III : Analysis of Morpholine in Mycophenolate mofetil.
Simple cost effective ion chromatographic set up with non- •Column : Metrosep C4 150/4.0
Flow rate : 0.9 mL/min; Sample loop : 250 µL
Detection : Non-suppressed Conductivity Detection
Mobile Phase : 3 mmol/L HNO +1.0 mmol/L Crown ether ,

Diluent : Mobile phase
 Trace level analysis possibility.
Metrohm India Limited, Chennai, India.
Analysis of regional concentrations of piperidine in the brain by Mass fragmentography; Brain research, 188(1980) 291-294; Takeshi Miyata.
 Methods for the determination of mutagenic heterocyclic amines and their applications in environmental analysis; J. of chromatographic A, 774 (1997)121-142; Hiroyuki Kataoka.
e-mail :;
 Spectrophotometric determination of some pharmaceutical piperazine derivatives through charge-transfer and ion-pair complexation reactions; J. of Pharma. and Biomedical Analysis; 15;1997, 1679-1685; Abdel-Gawad.


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