The determination of the palladium content in

valacyclovir hydrochloride using an Agilent 720

ICP-OES according to USP 35

Application note
Pharmaceutical

Authors

Dharmendra Vummiti1 and Philip

Lowenstern2
1
Agilent Technologies, Gurgaon,
Haryana, India
2
Agilent Technologies, Mulgrave,
Victoria, Australia

Introduction
The control of impurities from catalysts or raw materials (e.g. plants,
animal proteins, rDNA) and excipients (stabilizers, fillers, binders, release
agents, flavors, colors and coatings) has always been a critical issue to the
pharmaceutical industry. Traces of inorganic impurities can not only be toxic,
but can also have less immediate effects such as reducing the stability and
shelf life of pharmaceutical products. The U.S. Food and Drug Administration
(FDA) and the British Pharmacopeia (BP) strongly advise that contamination
problems be fully investigated in a timely fashion.
For testing of pharmaceutical samples, laboratories Table 1. Method conditions used in the analysis of Pd using the Agilent 720
axial ICP-OES
require instruments that have the flexibility to handle
a wide range of sample matrices, while also meeting Parameter Setting
ever decreasing detection limit requirements. The RF power 1.35 kW
instrumentation must also be capable of delivering Plasma flow 15.0 L/min
accurate and precise results while meeting the strict
requirements of the regulations specified by the U.S. Auxiliary flow 2.25 L/min

Pharmacopeia (USP). Nebulizer flow 0.60 L/min

Replicate read time 3 sec

In this work, the active pharmaceutical ingredient (API)
Replicates 3
valacyclovir hydrochloride was analyzed. Valacyclovir
hydrochloride is an anti-viral drug used in the treatment Sample uptake delay 20 sec
of the herpes virus. Samples were analyzed for the Stabilization delay 15 sec
presence of the impurity palladium (Pd), which is a
Rinse time 20 sec
required analysis in the official monographs in USP 35.
Wavelength Pd 340.458 nm

Experimental Oxygen flow (via AGM 1) 7 units (200 - 300 mL/min)

Torch 1.4 mm ID injector

Instrumentation
Nebulizer Conikal
An Agilent 720 ICP-OES with axially viewed plasma
was used for the analysis. The Agilent 720 ICP-OES
features a custom designed Charge-Coupled Detector Sample preparation
(CCD) which provides true simultaneous measurement, A 98:2 mixture of dimethyl sulfoxide (DMSO) and
full wavelength coverage from 167 to 785 nm and hydrochloric acid (HCl) was used as the blank and
fast read-out enabling short sample analysis times. diluent (as defined in the official monograph relating to
The optical system is housed within a thermally the analysis of valacyclovir hydrochloride). Standards
stabilized environment at 35 °C and contains no were prepared from single element Pd standards and
moving parts, ensuring excellent long-term stability. diluted with the diluent.
The polychromator can be purged with either argon or
nitrogen gas for improved performance when measuring Three samples of the API were taken at random
at low UV wavelengths. The robust RF system of the 720 and 0.10 g of the sample was weighed into a 10 mL
ICP-OES is capable of handling a wide range of samples, standard flask, followed by the addition of the diluent
including challenging organic samples, such as dimethyl up to 10 mL. These samples were labeled Sample A,
sulfoxide (DMSO). Sample B and Sample C. Pd was added to Sample C at
concentrations of 0.05 ppm, 0.10 ppm and 0.20 ppm to
An Auxiliary Gas Module (AGM 1) was used to inject test spike recoveries.
oxygen into the plasma to prevent the buildup of carbon
on the torch injector from the incomplete combustion of
the carbon present in the organic sample matrix.

Method conditions used in the analysis are shown in

Table 1.

2
 Results Sample analysis
Each API sample was measured in triplicate. The
Detection limits concentration results for Pd are corrected for the
The Instrument Detection Limit (IDL) is expressed dilution factor applied during sample preparation
as three times the standard deviation of nine (Table 3). The results show the excellent precision and
replicate measurements of the blank. The Instrument stability that can be obtained on the 720 ICP-OES when
Quantitation Limit (IQL) is expressed as ten times the analyzing this challenging matrix.
standard deviation of nine replicate measurements
of the blank. The IDL and IQL for Pd in the DMSO:HCl Table 3. Results for the triplicate analysis of three samples of the API.
Results are shown as the concentration in the sample.
matrix blank are shown in Table 2.
Label Number of determinations Pd 340.458 nm
Table 2. Instrument Detection Limit and Instrument Quantification Limits for Concentration µg/g
Pd in the DMSO:HCl matrix, reported in ppb (ppm)

Element IDL (ppb) IQL (ppb) Sample A Measurement-1 (0.10039 g/10 mL) 3.59

Pd 1.0 3.5 Measurement-2 (0.10014 g/10 mL) 3.56

Measurement-3 (0.10009 g/10 mL) 3.65

Calibration linearity
Average, µg/g (ppm) 3.60
The USP official monograph requires that the correlation
coefficient to be not less than (NLT) 0.999. To test Stdev 0.04
this requirement, a calibration was performed using RSD % 1.25
a diluent blank and six Pd standards, spanning the
Sample B Measurement-1 (0.10065 g/10 mL) 3.33
concentration range from 0.01 ppm to 0.50 ppm. The
resulting calibration curve is shown below in Figure 1 Measurement-2 (0.10063 g/10 mL) 3.25
and shows excellent linearity over the calibration range. Measurement-3 (0.10001 g/10 mL) 3.30
The correlation coefficient obtained in the experiment Average, µg/g (ppm) 3.29
was 0.999743, easily meeting the requirement of NLT
0.999. Stdev 0.04

RSD % 1.27
Pd 340.458 Calibration Curve
5000 Sample C Measurement-1 (0.09800 g/10 mL) 0.43
4500
4000 Measurement-2 (0.10025 g/10 mL) 0.44
y = 8591.8x + 123.39
3500
Intensity (c/s)

3000 Measurement-3 (0.1005 g/10 mL) 0.43

2500 Average, µg/g (ppm) 0.43
2000
1500 Stdev 0.01
1000
500 RSD % 1.53
0
0.00 0.10 0.20 0.30 0.40 0.50
Concentration (ppm)
Figure 1. Calibration curve for Pd obtained on the 720 ICP-OES

3
Spike recoveries Conclusion
Recoveries of the Pd spikes in Sample C are shown in
Table 4. Recoveries for the three spikes of increasing This work has demonstrated the ability of the
Pd concentration were within 100% to 108%, which is Agilent 720 ICP-OES with axially viewed plasma to
well within the 80% to 120% limits required in the USP accurately measure Pd in a tough organic matrix to
monograph. the requirements of the USP 35 monograph relating to
the analysis of the API valacyclovir hydrochloride. This
Table 4. Recoveries for Pd spikes in Sample C. Results are shown as the
concentration in the solution includes:
Sample ID Pd 340.458 nm
Result µg/mL (ppm) • Excellent sensitivity with an IDL of 1.0 ppb and IQL
3.5 ppb
Spike-1: 0.05 ppm Sample C 0.0043

Sample C-Spike-1: 0.05 ppm 0.0547 • A linear calibration curve for Pd with a correlation
Spike amount 0.050 coefficient of 0.999743, meeting the analytical
requirement of NLT 0.999
Spike sample 0.050

Recovery (%) 100.9 • Spike recoveries ranging from 100% to 108%, well
Spike-2: 0.1 ppm Sample C-Spike-2: 0.10 ppm 0.1070 within the limits of 80% to 120%
Spike amount 0.100

Spike sample 0.103

Recovery (%) 102.7

Spike-3: 0.2 ppm Sample C-Spike-3: 0.20 ppm 0.2207

Spike amount 0.200

Spike sample 0.216

Recovery (%) 108.2

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© Agilent Technologies, Inc. 2013

Published April 22, 2013
Publication number: 5991-2151EN