Vol.:(0123456789) 1 3
Journal of the Iranian Chemical Society
https://doi.org/10.1007/s13738-019-01831-y
ORIGINAL PAPER
First report on clorprenaline HCl degradation products: identifcation
and characterization by LC–MS/MS
Krunal J. Prajapati
1
· Charmy S. Kothari
1
Received: 2 May 2019 / Accepted: 16 December 2019
© Iranian Chemical Society 2019
Abstract
Clorprenaline hydrochloride (CLOR), β
2
adrenergic agonist, was used for the therapeutic treatment for bronchial asthma. It
is important to know the stability behavior of the drug in various degradation conditions because the drug is incompatible
with strong oxidizing agent and strong base. The main objective of the study was to identify and characterize degradation
products of clorprenaline HCl by LC–MS/MS. The mass-compatible method was developed, validated and further used
for LC–MS/MS. In reverse-phase high-performance liquid chromatography (RP-HPLC), the resolution was achieved using
Phenomenex C
18
(250 × 4.6 mm, 5 µ) column and ammonium acetate bufer (5 mM), 0.1% TEA, pH 6.5 with acetic acid/
methanol (70:30, v/v) as mobile phase. The fow rate was 1 mL/min. The wavelength was selected for UV detection at 215 nm.
Diferent forced degradation conditions such as hydrolytic (acidic and basic), oxidative, thermal and sunlight have been
applied according to ICH Q1A (R2) guidelines to derive stability of the drug. Further, the method was validated as per ICH
Q2 (R1) guidelines. The degradation product was characterized by LC–MS/MS using ESI in positive ion mode. Potential
degradation product DP (36.87% of total degradation) was found in basic condition, while no degradation was observed in
acidic, neutral, sunlight, oxidation and thermal conditions. The stability indicating analytical method was found to be novel,
simple, specifc, selective, sensitive, accurate, precise and robust for the determination of clorprenaline HCl.
Keywords Clorprenaline HCl · Identifcation · Characterization · Stability indicating analytical method · LC–MS/MS
Abbreviations
TEA Trimethylamine
UV Ultraviolet spectroscopy
ICH International Council for Harmoni-
sation of Technical Requirements
for Pharmaceuticals for Human
Use
DP Degradation product
HPLC High-performance liquid
chromatography
% RSD % Relative standard deviation
HCl Hydrochloric acid
NaOH Sodium hydroxide
H
2
O
2
Hydrogen peroxide
AR Analytical reagent
PVDF Polyvinylidene difuoride
PDA Photodiode array detector
RBF Round-bottom fask
NTP Number of theoretical plates
LOD Limit of detection
LOQ Limit of quantitation
RT Room temperature
ESI-ion trap MS/MS Electrospray ionization-ion trap
mass spectrometry
TIC Total ion chromatogram
List of symbols
mm Millimeter
μ Micron
mM Millimolar
mL/min Milliliter per minute
nm Nanometer
μL Microliter
µg/mL Microgram per milliliter
r Correlation coefcient
λ Wavelength
°C Celsius
t
R
Retention time
* Charmy S. Kothari
charmy.kothari@nirmauni.ac.in; charmyshah@gmail.com
1
Department of Pharmaceutical Analysis, Institute
of Pharmacy, Nirma University, Sarkhej-Gandhinagar
Highway, Post: Chandlodia, Via: Gota, Ahmedabad,
Gujarat 382 481, India