Analytica Chimica Acta 383 (1999) 283±290

Determination of paracetamol in intact tablets by use

of near infrared transmittance spectroscopy
A. Eustaquioa, M. Blancoa,*, R.D. Jeeb, A.C. Moffatb
a
Departamento de QuõÂmica, Unitad de QuõÂmica AnalõÂtica, Universidad AutoÂnoma de Barcelona, E-08193 Bellaterra, Barcelona, Spain
b
Centre of Pharmaceutical Analysis, The School of Pharmacy, University of London, 29/39 Brunswick Square, London WC1N 1AX, UK

Received 10 July 1998; received in revised form 16 November 1998; accepted 16 November 1998

Abstract

Near infrared transmittance spectroscopy was used to determine the analgesic paracetamol in a pharmaceutical preparation
commercially available as tablets. Spectra were recorded on a dedicated instrument that measures the transmission of intact
tablets over the wavelength range 600±1900 nm. Spectral data were processed by using two multivariate calibration methods,
viz. stepwise multiple linear regression (SMLR) and partial least-squares regression (PLSR). The analgesic contents provided
by the two calibration methods were comparable and differed by less than 1% from the reference (UV spectrophotometric)
value. The calibration graphs constructed to determine the mean paracetamol content were used to analyse content uniformity
in the tablets. # 1999 Published by Elsevier Science B.V. All rights reserved.

Keywords: Near infrared transmittance spectroscopy; Intact tablet analysis; Stepwise multiple linear regression; Partial least-squares
regression; Content uniformity

1. Introduction The NIRS technique has proved a highly useful tool

The high industrial interest aroused by near infrared
spectroscopy (NIRS) in recent years is a direct result
of dramatic advances in this technique including the
increasing availability of fast-scan instruments that
enable measurements with little or no sample manip-
ulation and of software for implementing powerful
chemometric techniques. Also the increasing aware-
ness and acceptance of NIRS has facilitated the expe-
ditious analysis of many complex samples with
no need for reagents or solvents in a variety of ®elds
[1±3].
*Corresponding author. Fax: +34-93-581-24-77;
e-mail: iqan8@blues.uab.es
for controlling powdered pharmaceuticals (intact
powdered raw materials, intermediates and dosage
forms), which it can readily identify and quantify
[4]. The ability to analyse intact dosage forms (parti-
cularly tablets, which are the most widely used form)
has dramatically facilitated monitoring of every single
step in the production process as well as of the ®nal
form, all with minimal sample manipulation. On the
other hand, direct tablet analysis by NIRS has devel-
oped to a much lesser extent owing to technical
constraints [5]. The earliest applications of NIRS to
intact tablets were reported in 1988 [6]; the whole
surface of a double-re¯ecting aluminium sample con-
tainer was illuminated for the qualitative analysis
of tablets. Later applications have been primarily

0003-2670/99/$ ± see front matter # 1999 Published by Elsevier Science B.V. All rights reserved.
PII: S0003-2670(98)00815-0
284 A. Eustaquio et al. / Analytica Chimica Acta 383 (1999) 283±290
concerned with qualitative analyses [7]. Most reported
direct NIRS analyses of tablets are subject to two
hindrances, namely: (a) the lack of appropriate devices
for making reproducible sample measurements and (b)
the fact that measurements are made by re¯ectance, so
the information acquired corresponds to the sample
surface, which may lead to spurious results with
coated tablets.
These shortcomings are circumvented by recently
developed NIR instruments of improved design. Their
measuring modules provide more reproducible and
reliable measurements of individual tablets. Also, the
new instruments use transmission measurements (i.e.
the NIR beam passes through the whole sample thick-
ness). Using these instruments provides a number of
advantages, namely,
1. the ability to draw information for single tablets
from their individual spectra;
2. the ability to analyse coated tablets, the coating
thickness posing no special problem since the
spectrum is due to the whole tablet;
3. increased reproducibility in the measurements ±
those of individual tablets included ± by virtue of
the response being produced by a sizeable amount
of sample ± its entire thickness;
4. simple calibration models relative to those typi-
cally used with reflectance measurements ± the
recorded signal (absorbance) can be assumed to
obey the Lambert±Beer law since the improved
instrument design results in minimal scatter.
On the other hand, the most severe constraint on
transmission measurements is posed by the thickness
of typical tablets. Because the light beam must travel
through a large amount of product, only a small
fraction of incident light reaches the detector. The
high absorbances involved in this type of measure-
ment ± most often in excess of 4 absorbance units ±
have compelled instrument manufacturers to improve
existing commercial equipment by minimising losses
of incident light on its way to the detector (the
instrument design should also minimise stray light
reaching the detector, e.g. by passing around the sides
of the tablet sample) and using highly sensitive detec-
tors affording readings of much higher absorbances
than those usually recorded in other spectroscopic
techniques.
In this work, near infrared transmittance spectro-
scopy was tested for the analysis of tablets about 4 mm
thick. This type of sample is representative of many
commercially available tablets and consequently, if
thick tablets can be satisfactorily analysed, the NIR
technique should be applicable to virtually any other
type of tablet.
2. Experimental
2.1. Apparatus and software
Near infrared spectra were recorded on an NIRSys-
tem 6500 spectrophotometer (FOSS, NIRSystems)
equipped with an Intact Tablet Analyser that was in
turn furnished with an indium±gallium±arsenide
(InGaAs) detector which provides highly linear,
low-noise measurements of sample spectra and allows
processing of bands up to 3 absorbance units above the
baseline [8]. The spectrophotometric system was opti-
mised to measure tablet absorbances over the visible
(Vis) and near infrared (NIR) spectral ranges.
The equipment was controlled via the NSAS v. 3.52
software package (FOSS, NIRSystems), which
enables automated acquisition and processing of
NIR spectra. The same software was used to perform
SMLR computations.
PLS calibration was done by using the multivariate
calibration program Unscrambler v. 6.1, from CAMO
AS (Trondheim, Norway), which affords direct import
of NIR spectra.
The reference spectroscopic method was implemen-
ted on a Perkin-Elmer Lambda 15 UV/Vis spectrometer.
2.2. Samples
The specimens studied were circular cores about
4 mm thick consisting of paracetamol as active prin-
ciple (nominal content 84% m/m) and six different
excipients.
Twenty-®ve production batches containing 99±
102% of the nominal paracetamol value, and 10
batches of tablets made at a pilot plant were studied.
The latter allowed the content range to be extended to
90±110% of the nominal value.
The UV spectrophotometric procedure used for
reference was that of®cially recommended by the
British Pharmacopoeia [9].
 A. Eustaquio et al. / Analytica Chimica Acta 383 (1999) 283±290 285

2.3. Recording of NIR spectra
The instrument used permits the recording of
transmission spectra of intact tablets. The measuring
system was optimised by constructing a holder to
closely ®t a tablet in order to ensure maximal
reproducibility in the sample position and minimal
stray light. The holder, which held a single tablet
of the product being tested, was made as per the
instructions given in the instrument's user guide
[10].
Tablets were always placed in the holder the ¯at
side up (this was facilitated by the tablets having a
different mark on each side) and the holder inserted
into the instrument. The spectra (average of 32 scans)
for each of 20 tablets from each batch were recorded
from 600 to 1900 nm, using air as reference. First and
second derivative absorbance spectra were calculated
using the NSAS software with zero gap size and 10
and 20 segment sizes, respectively.
Fig. 1 shows the NIR spectra obtained in different
modes for the samples containing the extreme para-

Fig. 1. Absorbance, first-derivative (absorbance) and second-derivative (absorbance) near infrared transmittance spectra for three different
samples: (A) and (C) sample with a paracetamol content equal to 90% and 110%, respectively, of the nominal value; (B) production sample.
286 A. Eustaquio et al. / Analytica Chimica Acta 383 (1999) 283±290
cetamol contents examined as well as that for a normal
production sample. There are signi®cant differences
on the spectra around 1400±1500 nm; these differ-
ences have been only observed in overdosed pilot
plant samples and are not correlated to the paraceta-
mol content in the samples. Its source is unknown.
2.4. Data processing
The mean paracetamol content was determined
from the mean spectrum for each batch (i.e. the mean
of the 20 individual spectra recorded). Paracetamol
was quanti®ed by using two different multivariate
calibration methods, viz. inverse stepwise linear mul-
tiple regression (SMLR) and partial least-squares
regression (PLSR).
With the SMLR method, each model was optimised
in terms of the number of wavelengths used in order to
avoid over®tting; an F statistical criterion was applied
to the sum of the squares of the residuals for the
calibration samples in the models [11]. The different
models thus constructed were compared by estimating
the goodness of ®t from the multiple correlation
coef®cient
s
Pn 
…C NIRi ÿ 
C LABi † 2
R ˆ Pniˆ1 ; (1)
…CLABi ÿ C  LABi †2
iˆ1
where CNIR is the paracetamol content calculated by
using the NIR method and C LAB the mean paracetamol
content in the calibration samples provided by the
reference method. For accurate comparison, this para-
meter must be corrected for the number of degrees of
freedom:
 
2 mÿ1
Raj ˆ 1 ÿ …1 ÿ R2 †; (2)
mÿn
where n is the number of coef®cients in the equation
and m the number of calibration samples used. While
R2 increases and approaches unity as the number of
terms is increased, R2aj tends to level off after the
optimum model has been reached.
PLS models were constructed by cross-validation,
using as many segments as samples were included in
the calibration matrix. The optimum number of PLS
components was chosen according to the criterion of
Haaland and Thomas [12]. The calibration procedure
was applied to centred, unscaled data.
The results provided by the different models tested
were compared via the relative standard error of
prediction (RSEP), given by [13]
sP
n
iˆ1 P …CLABi ÿ CNIRi †2
RSEP …%† ˆ n 2
 100; (3)
iˆ1 CLABi
where CLAB and CNIR are the paracetamol content deter-
mined by the reference and NIR methods, respectively.
3. Results and discussion
Available samples were split into two different sets
for calibration and validation, respectively. The cali-
bration set consisted of 20 batches with paracetamol
contents that evenly spanned the range from 90% to
110% of the nominal value; it included production and
prepared at the pilot plant samples. The remaining 15
batches were used as the validation set and spanned the
range from 95% to 105% of the nominal paracetamol
content; it also included both types of samples.
3.1. Determination by stepwise multiple linear
regression
The wavelengths used in constructing the SMLR
model were chosen in the ascending mode. The start-
ing wavelength range was 600±1350 nm ± the region
from 1350 to 1900 nm was excluded owing to the high
absorbances and noise associated with it.
Results were compared on the basis of two different
criteria for wavelength selection, namely:
A. Automatic selection. The wavelength resulting
in the highest possible correlation between
absorbance and concentration was chosen and
the equation was successively expanded with
new terms by incorporating those previously
unused wavelengths that met the same criterion.
The number of wavelengths used in each model
was optimised in terms of the above-mentioned
F criterion.
B. Imposing a first wavelength based on a
chemical criterion. The first wavelength used
was that where differences between the spectra
for the samples containing the extreme con-
centrations of paracetamol (Fig. 1) were max-
imal, the software was allowed to automatically
select successive wavelengths as in A.
 A. Eustaquio et al. / Analytica Chimica Acta 383 (1999) 283±290 287

Table 1
Relative standard error of prediction for the calibration set (RSEP(C)), validation set (RSEP(V)) and R2aj as obtained by using different SMLR
methods

Mode R2aj RSEP(C) (%) RSEP(V) (%) Wavelengths (nm)

Absorbance A 0.989 0.6 0.8 1206, 1186

B 0.989 0.6 0.8 1206, 1186

First derivative A 0.989 0.6 0.7 900

B 0.991 0.6 0.8 1190, 886

Second derivative A 0.989 0.6 1.0 1212, 850

B 0.990 0.6 0.9 1202, 820
A: automatic wavelength selection; B: first wavelength based on chemical criterion (see text for more details).

Calibration equations were derived from spectra than ttab (2.14, Pˆ0.05, dfˆ14), the proposed proce-
subjected to various treatments (absorbance, ®rst deri- dure exhibits no statistically signi®cant differences
vative, second derivative). from the reference method.
Table 1 gives the wavelengths used for the models
and their corresponding errors of prediction; in the 3.2. Determination by partial least-squares
absorbance spectral mode, criterion B was equivalent regression
to criterion A. It can be seen that R2aj was similar for all
the models examined, and therefore that with the Although PLSR is a whole-spectrum technique,
smallest RSEP(V) was chosen. This was one of the quantitations based on it can bene®t from careful
simplest models, using ®rst derivative (absorbance) selection of spectral wavelengths. We examined two
spectra and a single wavelength. different ranges, viz. 600±1350 and 600±1900 nm.
Table 2 gives the results obtained for the validation Although the high end of the spectrum exhibited high
samples using the optimum model. The results were absorbances and considerable noise, we assayed using
compared to the reference values using a paired PLSR to model the system in order to determine
samples Students' t test. As texp (0.354) was smaller whether the broader range would lead to improved
results.
Table 2
Table 3 shows the errors of prediction for the
Paracetamol contents (% m/m) in the validation samples provided models tested. As can be seen, the predictive capacity
by the optimum SMLR and PLSR models was not affected by the spectral treatment used; how-
ever, the models constructed from whole spectra were
Sample [Paracetamol]LAB [Paracetamol]SMLR [Paracetamol]PLSR
generally more complex (they used more PLS com-
3 80.3 79.2 79.3 ponents); this can be ascribed to small baseline shifts
6 83.9 84.6 85.0
among spectra, shifts that were minimised by deriva-
9 84.1 84.2 83.8
11 84.2 85.2 84.6 tion. Models constructed by applying Savitzky±Golay
13 84.2 84.5 84.7 smoothing (9 data points, third-order polynomial) to
15 84.3 84.5 84.6 the spectra were also tested; however, their results are
17 84.4 85.0 85.1 not reported because they did not improve on those of
19 84.4 84.8 84.7
the other models in any way.
21 84.7 83.4 83.5
23 84.9 84.7 84.8 As the prediction capability is comparable in all
25 85.1 84.5 84.4 models assayed, the best PLS model was selected on
27 85.1 84.6 84.5 the basis of its simplicity. In this case, the simplest
28 85.5 85.0 85.0 model needed only one PLS-component and was
30 85.7 85.5 84.7
calculated using the shorter spectral range and ®rst-
33 88.5 88.5 88.5
derivative spectra. The results for the samples in the
288 A. Eustaquio et al. / Analytica Chimica Acta 383 (1999) 283±290

Table 3
Relative standard error of prediction calculated for the calibration set (RSEP(C)) and validation set (RSEP(V)) as obtained by using different
PLSR models

Range (nm) No. PLS-comp. RSEP(C) (%) RSEP(V) (%)

Absorbance 600±1350 3 0.6 0.8

600±1900 5 0.6 0.8

First derivative 600±1350 1 0.7 0.8

600±1900 3 0.6 0.8

Second derivative 600±1350 2 0.6 0.8

600±1900 2 0.6 0.8

validation set obtained by this model are given in Table 4

Table 2. Again, a paired samples Students' t test Paracetamol contents (% m/m) in validation samples obtained by
using a calibration set consisting of pilot plant samples only
revealed that the proposed procedure is not signi®-
cantly different from the reference method (texpˆ Sample [Paracetamol]LAB [Paracetamol]PLSR
0.136, ttabˆ2.14, Pˆ0.05, dfˆ14). 5 83.5 85.0
The simplicity of the optimum model, which uses a 6 83.9 85.3
single PLS component, suggested that the calibration 7 84.0 85.6
set could be constructed from only samples made at 8 84.1 85.1
10 84.1 84.6
the pilot plant permitting a decreased number of 12 84.2 85.1
samples for calibration. To verify this hypothesis, 13 84.2 85.1
we constructed a new PLSR model using the previous 14 84.3 85.2
optimum parameters (viz. the 600±1350 range and 15 84.3 85.0
®rst-derivative spectral mode) and the 10 spectra for 16 84.3 85.0
17 84.4 85.4
samples made at the pilot plant. Table 4 shows the
18 84.4 84.8
paracetamol contents in the 25 production batches 19 84.4 85.0
obtained with this new calibration model; the RSEP 20 84.5 85.1
was 0.9%, indicating that the predictive capacity is 21 84.7 83.8
somewhat lower than the model which included pro- 22 84.8 85.1
23 84.9 85.1
duction samples in the calibration set. Although the 24 85.0 84.9
pilot plant samples are very similar to the production 25 85.1 84.7
samples, the predictive capacity of the model is 26 85.1 85.0
improved if variability in the production samples is 27 85.1 84.8
introduced in the calibration. This agrees with pre- 28 85.5 85.3
29 85.6 85.2
vious studies using calibrations including synthetic 30 85.7 85.0
[14] or overdosed samples [15,16]. 31 85.8 85.3
RSEP (%) 0.9

3.3. Content uniformity

A measuring system capable of recording spectra
for individual tablets provides an additional advan-
tage, viz. the ability to simultaneously determine,
using the same calibration, the content in individual
tablets. Fig. 2 shows the spectra of 20 individual
tablets from a single production batch.
The European Pharmacopoeia [17] test for unifor-
mity of content requires the speci®ed active principle
in 10 randomly chosen tablets to be measured.
Although the test is strictly only required for tablets
in which the active principle is less than 2 mg or less
than 2% (m/m), we have applied the ``spirit'' of the
test to the paracetamol tablets. Using the ®rst 10
 A. Eustaquio et al. / Analytica Chimica Acta 383 (1999) 283±290 289

Fig. 2. Near infrared transmittance spectra for 20 individual paracetamol tablets from a production batch.

Table 5 models provided the same coef®cient of variation

Study of content uniformity in a production batch
(CV). The relative paracetamol contents obtained in
SMLR PLSR all the determinations were all within the accepted
Tablet limits of the standard (viz. 85±115% of the nominal
[Parac.]SMLR Nominal [Parac.]PLSR Nominal
(%) value (%) (%) value (%) value).
1 84.6 100.5 84.6 100.5
2 84.3 100.1 84.3 100.1
3 84.6 100.5 84.8 100.7 4. Conclusions
4 84.5 100.4 84.7 100.6
5 85.0 101.0 85.0 101.0 The results obtained in this work reveal that the
6 84.5 100.4 84.7 100.6 direct analysis of intact tablets by near infrared trans-
7 84.4 100.3 84.8 100.7
mittance spectroscopy is an effective way of quantify-
8 84.3 100.1 84.3 100.1
9 84.5 100.4 84.7 100.6 ing this type of end product. The ensuing methodology
10 84.3 100.1 84.4 100.3 is accurate enough to ascertain whether the content in
CV (%) 0.3 0.3 the active principle assayed is within the speci®ed
range and can thus be used to control the process. In
addition, a single calibration allows the simultaneous
individual spectra previously recorded we evaluated determination of the mean content and content uni-
them by using the optimum SMLR and PLSR models. formity in a production batch.
Table 5 shows the per cent contents obtained relative The two calibration techniques tested, SMLR and
to the nominal value. As can be seen, both calibration PLSR, provide quite satisfactory results and exhibit a
290 A. Eustaquio et al. / Analytica Chimica Acta 383 (1999) 283±290
comparable predictive ability. The simplicity of the
SMLR and PLSR calibration models used suggests
that transmittance measurements are highly correlated
with the analyte content since the typical scatter in
diffuse re¯ectance measurements is negligible here.
Acknowledgements
The authors are grateful to Spain's DGICyT (Pro-
ject PB96-1180) for funding this work. A. Eustaquio
also acknowledges additional funding from Spain's
Ministry of Education and Science in the form of an
FPI grant. The authors also wish to thank Sano® for
kindly supplying the production and pilot plant sam-
ples, and A.D. Trafford for providing the reference
values for all the samples.
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