Indian Journal of Chemical Technology
Vol. 12, March. 2005, pp. 229-231
Notes
Determination of heavy metals in selected
drug substances by inductively coupled
plasma mass spectrometry
A S R Krishna Murty*, U C Kulshresta, T Nageswara Rao
& M V N Kumar Talluri
Analytical Chemistry Division, Indian Institute of Chemical
Technology, Hyderabad 500 007, India
Received 20 May 2004; revised received 6 December 2004;
accepted 3 January 2005
Heavy metal contents in dicyclomineHCl, ethambutol, pyrazi-
namide and furazolidone drugs were determined by inductively
coupled plasma (ICP) - mass spectrometry (MS). The drugs were
analyzed for Ti, Cr, Mn, Fe, Co, Ni, Cu, Zn, Cd, Hg, Pb metals by
selecting a suitable isotope. Cr, Fe, Ti and Cu were observed to be
highest in dicyclomineHCl, ethambutol, pyrazinamide and fura-
zolidone respectively. Ni and Hg were absent in all the four drugs,
while traces of Cd were present in ethambutol and pyrazinamide.
Analytical results showed that ICP-MS method is useful for moni-
toring inorganic impurities present in such drugs.
Keywords: Inductivity coupled plasma, heavy metal determina-
tion, dicyclomineHCl, ethambutol, pyrazinamide,
furazolidone.
IPC Code: G01J 3/00; C22B 4/04
Impurity profiling of pharmaceutical products is a
major area of concern. Stringent regulations are laid
down for their effective use. The authors have re-
ported results of pharmaceutical analysis of some
compounds1-5. The impurities in a drug could be or-
ganic or inorganic in nature. Much is known about
organic impurities6-8, while the inorganic impurities9,10
are gaining importance recently. The inorganic impu-
rities i.e. metal contamination enter the bulk drug sub-
stances and intermediates through raw materials, cata-
lysts, reagents, solvents, various equipments used for
synthesis, etc. The metal ions entered have the ability
to decompose the materials of interest, which may
some times lead to potential toxic effects, in addition
to self-toxicity. It is therefore obvious that the metal
ion contents need to be monitored. Official Pharma-
copoeias11-13 describe heavy metal test in these drugs.
The method consists of precipitation of heavy metals
as sulphides and visual comparison of the colour with
______________________
*For correspondence (E-mail: akella@iict.res.in)
IICT Communication No: 040411
that obtained from similarly prepared solution of
standard lead solution. The elements respond to the
test by yielding different colours viz., white, yellow,
orange, black and dark brown14. Based on only colour
as parameter, it is difficult to give the identity for the
ion responsible for the colour. The procedure lacks
specificity, sensitivity and is time consuming with no
information about the recoveries. Several attempts
have been made to improve the procedure but they are
not of much advantage15,16, the main disadvantages
being their suitability for few elements and unequal
sensitivity.
Though the toxicity of trace metal ions like Hg, As,
Pb is known but their limits in pharmaceutical prod-
ucts have not been defined clearly11-13. Some of the
heavy metals are also required as micronutrients, but
above the necessary range, they cause toxicity. It is,
therefore, necessary to look for a suitable method for
the monitoring of these metals at very low concentra-
tion levels (ppm to ppb) either for single metal ion or
a mixture of metal ions as the case may be. AAS,
ICP-AES and ICP-MS are suitable techniques for this
propose17-19. Among these techniques, ICP-MS is the
most suited for multi-elemental analysis20. ICP-MS
finds extensive application due to very low detection
limits (ppb, ppt) for most of the elements of the peri-
odic table. The mass spectra of elements are simple
and, therefore, can provide quick access for qualita-
tive, semi-quantitative and quantitative analysis.
Equipment cost and non-availability of standard ref-
erence materials (SRMs) for pharmaceutical products
for ascertaining the accuracy21 are the main disadvan-
tages of ICP-MS analysis. Considering the importance
of trace elemental analysis in pharmaceutical prod-
ucts, four nitrogen containing compounds have been
selected and screened by ICP-MS. These compounds
were chosen on the basis of their ability to bind metal
ions present in contamination.
Experimental Procedure
Instrumentation
A Varian UltraMass 700 inductively coupled
plasma mass spectrometer (ICP-MS) was used
throughout the study. The instrumental conditions are
given below.
Plasma parameters: Coolant argon flow 15.5
230 INDIAN J. CHEM. TECHNOL., MARCH 2005

L/min; Auxiliary argon flow 1.40 L/min; Nebulizer Table 1 Heavy metal contents* in drug substances (ng/g)
argon flow 0.95 L/min; Power pump rate 1.2 KV, Element Iso- Di- Etham Pyrzi- Fura-
25rpm. tope cyclomin butol namide zolidone
Scanning parameters: Spacing 0.1 AMU; Dwell Ti 49 e HCl
7.9 19.2 38.2 6.4
time 200 s; Replicate per sample 5; Scans per repli- Cr 52 9.4 4.8 11.5 2.6
cate 25. Mn 55 3.5 6.8 3.7 4.2
Reagents and Materials Fea 57 6.6 26.6 1.9 3.0
Concentrated nitric acid (65%, trace select Ultra) Co 59 5.1 + + +
and water (for inorganic trace analysis) were pur- Cu 65 4.2 9.5 8.4 21.6
chased from Fluka, Switzerland. National Institute of Zn 66 4.1 1.5 0.6 0.7
Standards and Technology traceable plasma standards Cd 114 + 0.2 0.1 +
were purchased from J.T. Baker, USA. The pure
Pb 208 0.4 6.5 0.8 8.1
drugs were gifted by local manufacturers.
Ni and Hg were not detected in any drug.
Preparation of standards + Not detected
Calibration standards were made by diluting * Average of five determinations. (RSD <5%)
plasma standards in nitric acid (5%v/v). Two- a Total Fe
multielement calibration standards of 200 and 500 Table 2 Heavy metal contents* in drug substances (ng/g) es-
ng/mL were prepared by directly diluting the plasma timated after dissolution in dil. HNO3 (5% v/v)
stock standard solution. Element Dicyclomine Ethambu- Pyrazi- Furazo-
HCl tol namide lidone
Preparation of samples Ti 0.2 + + +
Accurately weighed sample (~0.1 g) was taken in a Cr + 0.3 2.5 1.6
100 mL-beaker along with 5 mL of conc. nitric acid Mn 0.6 + + 0.1
and kept overnight. The solution was heated at Fe 2.4 23.9 0.8 0.6
60-80 C by adding H2O2 dropwise until all the or-
Cu + 2.4 5.0 11.8
ganic matter was destroyed. Then cooled to room
temperature and made up to 100 mL with distilled Zn 1.9 + + 0.6
water. A blank was also prepared in the same manner. Pb + + + 8.0
Co, Ni, Cd and Hg were not detected in any drug
ICP-MS analysis + Not detected.
Each element was analyzed by ICP-MS by select- * Average of five determinations (RSD<5%)
ing a suitable isotope, free from interference.
butol is perhaps due to its ability to chelate with the
metal ions23 resulting due to contamination. The high-
Results and Discussion
est amount of Cu (21.6 ppb) in furazolidone might be
In the present study, dicyclomineHCl, ethambutol,
due to the use of copper cathode in the preparation of
pyrazinamide and furazolidone drugs were analyzed
furazolidone as it involves electrolytic reduction of
for trace levels of heavy metal ion contents. All these
3-nitro-2-oxazolidone at copper cathode in 10% sul-
compounds contain nitrogen, which can bind the
phuric acid24. The relatively higher values for Cr
heavy metal ions through the lone pair of electrons on
(11.5 ppb) and Cu (8.4 ppb) in the case of pyrazina-
nitrogen. The results of estimation of heavy metal
mide could be understood because pyrazoic acid is
contents are given in Table 1. In a separate attempt,
used as raw material in pyrazinamide synthesis with
the samples of the drugs were directly dissolved in
copper chromite as catalyst25. However, the higher
dil. HNO3 (5%v/v) and analyzed. In these samples
value of Ti could not be explained at this moment.
very low concentrations of metals were obtained (Ta-
ble 2), indicating the interference of organic matter. Accuracy and Precision
Thus, the opted HNO3 / H2O2 method of digestion for True values for heavy metal contents of the sam-
destroying organic matter22, was found to be satisfac- ples studied are not reported for comparing accuracy
tory. Comparatively higher Fe value in case of etham- of the present method. In the absence of true values
the accuracy is usually assessed by running any stan-
 NOTES 231

Table 3 Concentration of various trace elements (ng/ml) in 3 Husain S, Krishna Murty A S R & Narsimha R, Indian
NIST 1643b (water reference sample) in comparison with cer- Drugs, 26 (1989) 557.
tified values 4 Husain S, Krishna Murty A S R, Ravi Prasad P & Seker R,
Indian Drugs, 32 (1995) 336.
Element ICP-MS Results Certified values 5 Husain S, Krishna Murty A S R & Seker R & Ravi Prasad P,
Cr 19.300.84 18.60.70 Indian Drugs, 32 (1995) 574.
Mn 28.101.10 28.02.10 6 Nageswara Rao R & Nagaraju V, J Pharm Biomed Anal, 33
(2003) 335.
Fe 101.385.10 99.03.30 7 Roy J, AAPS Pharm Sci Tech, 3 (2002) 6.
Co 26.120.64 26.00.90 8 Lehr G J, Barry T L, Petzinger G, Hanna G M & Zito S W, J
Ni 48.301.90 49.01.90 Pharm Biomed Anal, 19 (1999) 373.
9 Lewen N, Mathew S, Schenkenberger M & Raglione T, J
Cu 22.520.72 21.90.80 Pharm Biomed Anal, 35 (2004) 739.
Cd 20.520.91 20.01.0 10 Pranda M & Vytras K, J Pharm Biomed Anal, 14 (1996) 765.
Zn 65.651.20 64.792.47 11 Pharmacope Europenne, Deuxime Edition, Maisonneuve
S.A., (57-Sainte-Ruffine, France), 1980.
Pb 24.092.50 23.72.1
12 British Pharmacopoeia, vol. 2 (Her Majestys Stationery
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13 The United States Pharmacopoeia, The National Formulary,
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present studies, NIST 1643b (Trace elements in Water Twin Brook Parkway, Rockville, MD 20852), 1990.
Reference Sample from National Institute of Stan- 14 Moine I, Recherche des Mtaux lourds les mdicaments,
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19 Skoog D A & West D M, Fundamentals of Analytical
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