Robert Mattes* is an applications scientist at FOSS

NIRSystems, Inc., 7703 Montpelier Rd., Laurel, MD 20723

tel. 301.680.7251, fax 301.236.0134,rmattes@foss-
nirsystems.com, and Denise Root is a marketing manager at
FOSS NIRSystems. Om Anand, Maria Gerald Rajan,
Namrata R. Trivedi, and Wen Qu are graduate students,
and Yingxu Peng, PhD, and Yichun Sun, PhD, are
postdoctorate fellows, Department of Pharmaceutical Sciences,
University of Tennessee, Memphis, TN.
*To whom all correspondence should be addressed.
Submitted: Nov. 11, 2006. Accepted: Jan. 17, 2007.
Key words: content uniformity, near-infrared, process analytical technology
Near-Infrared Assay and
Content Uniformity of Tablets
Robert Mattes, Denise Root, Om Anand, Maria Gerald Rajan, Namrata R. Trivedi, Wen Qu, Yingxu Peng, Yichun Sun
Near-infrared (NIR) assay and content uniformity
of tablets provide fast, accurate means of
monitoring tablet production that are in step with
FDAs process analytical technology initiative. The
authors discuss the process for testing a newly
released NIR tablet analyzer to determine
instrument precision and accuracy using
chlorpheniramine maleate tablets. The data show
promising results that could relieve laboratory
workload of high-performance liquid
chromatography analysis and bring analysis closer
to real time for process monitoring.
ear-infrared spectroscopy (NIRS) is an analytical
technique based on absorption measured in the
near-infrared region of the electromagnetic spec-
trum that is between the visible and the
mid-infrared (IR). The fundamental absorption bands of
functional groups occur in the mid-IR and are very strong.
Usually, potassium bromide pellets, mulls, or dilutions are
required to bring the absorbances within the linear range of
the mid-IR detector. The overtone absorptions of these
fundamental bands occur in the NIR spectral region and
allow direct measurement without sample preparation
because of the relative weakness of absorption. The OH,
CH, NH, and SH bonds have the strongest overtone
absorbances in the NIR region (1).
There is considerable interest in the ability to test solid-
dosage form samples more frequently than the 10 per batch
specified by the US Pharmacopeia monograph on content
uniformity. Interest has increased in using NIR for tablet
assay and content-uniformity testing because of concerns of
the European Union for better statistically based sampling
and the US Food and Drug Administrations initiative on
process analytical technology (PAT) for better under-
standing and monitoring of production (2). NIR can be
used as a rapid at-line analysis method to obtain processing
feedback in near real time during a tableting campaign.
Transmission NIRS through the tablet has been preferred to
reflectance NIRS because of heterogeneity within tablets (3,
4). The reflection NIRS technique may be used for coating
analysis, but for bulk tablet analysis, the transmission NIRS
technique may yield more consistent results.
Laboratory methods for tablet assay and content unifor-
mity are usually time-consuming because they routinely are
done by high-performance liquid chromatography (HPLC),
which requires lengthy calibration runs, the mixing of
buffers, and the procurement and disposal of volatile
solvents. Analyzing 10 tablets for content uniformity may
take hours, and the results may not be available to tablet-
press operators or for batch release for many days or even
weeks after the tablets are compressed. Statistical process
control (SPC) techniques can be applied while measuring
the tablets with NIR in real time during tableting so that
assay and content-uniformity problems can be detected
before they go beyond acceptable limits.
N
Figure 1: Examples of tablets being scanned in transmission
using the novel near-infrared spectrometer. In the insert on the
left is a tablet tray, and to the right is the standards tray
containing the wavelength standard (traceable to NIST SRM-
2035) and photometric standards.
ANALYTICAL METHODS
256361:3-column 6/15/07 2:50 PM Page 170
ANALYTICAL METHODS
Experimental
Five batches of tablets (0.25-in. diameter and 100-mg
weight) with 0 mg (placebo tablets), 0.1 mg, 0.5 mg, 1.0 mg,
and 2.0 mg of chlorpheniramine maleate (CPM) per tablet
were formulated and compressed on a tablet press (HT-AP
18 SS-U/I rotary tablet press, Elizabeth Hata International,
Inc., North Huntingdon, PA).
The NIR instrument used in the study was XDS
MasterLab (FOSS NIRSystems, Laurel, MD), which was
capable of automatically measuring multiple tablets after
they were positioned in a special tray (see Figure 1). In the
insert, a tablet tray is to the left, and to the right is the stan-
dards tray containing the wavelength standard (traceable to
NIST SRM-2035) for photometric standards. The universal
tablet tray used for this study had 20 positions for four
different tablet sizes and five positions for the 0.25-in.
diameter tablets under test. The tray was loaded twice to
scan all 10 tablets. The 10 tablets were scanned in less than
5 min, taking a reference spectrum before scanning each set
of five tablets. Spectra were collected in the transmission
mode from 800 nm to 1650 nm with 0.5-nm data intervals,
and 32 scans were coadded to produce a single spectrum.
HPLC analysis was run on each individual calibration and
validation tablet after spectra were collected with the NIR
instrument. The HPLC reference values and the NIR
spectra were used to develop the regression model.
Discussion
Figure 2 shows the raw NIR spectra from the calibration set
and a spectrum of pure CPM in green. The pure CPM
5.4000
5.1100
4.8200
4.5300
4.2400
3.9500
3.6600
3.3700
3.0800
2.7900
2.5000
800 846 893 939 986 1032 1079 1125 1171 1218 1264 1311 1357 1404
A
b
s
o
r
b
a
n
c
e
Wavelength (nm)
Figure 2: Raw spectra of calibration samples. The green spectrum
is of pure chlorpheniramine maleate in reflectance to show where
the absorption occurs at 1138 nm.
0.1725
0.1407
0.1088
0.0770
0.0452
0.0133
-0.0185
-0.0503
-0.0822
-0.1140
-0.1459
800 850 900 950 1000 1050 1100 1150 1200 1250 1300 1400 1450 1500
I
n
t
e
n
s
i
t
y
Wavelength (nm)
1550 1600 1650
Figure 3: Second derivative mathematical treatment of calibration
spectra. The absorption band at 1138 nm fans out from the
concentration of chlorpheniramine maleate.
8.8377
8.4304
8.0232
7.6159
7.2086
6.8014
6.3941
5.9869
5.5796
5.1724
4.7651
1123 1128 1133 1138 1143 1148 1153
A
b
s
o
r
b
a
n
c
e

X

1
0
-
5
Wavelength (nm)
Placebo
0.1 mg
0.5 mg
1.0 mg
2.0 mg
Figure 4: Shows the fanning out of the analytical region of the
spectrum where the chlorpheniramine maleate has a strong
absorption band at 1138 nm. Blue is placebo, red is 0.1 mg, brown
is 0.5 mg, dark blue is 1.0 mg, and pink is 2.0 mg.
2.9501
2.2068
1.4636
0.7204
-0.0228
-0.7660
-1.5093
-2.2525
-2.9957
-3.7369
-4.4822
1103 1124 1145 1166 1187 1206 1229 1250 1271 1292 1313 1334 1355 1376
Wavelength (nm)
A
b
s
o
r
b
a
n
c
e

X

1
0
-
5
Figure 5: PLS factor loadings around the 1138 nm absorption for
chlorpheniramine maleate and thickness-correction regions.
Thickness correction was applied between 1250 nm and 1350 nm.
256361:3-column 6/15/07 2:50 PM Page 171
spectrum was scanned in reflectance and multiplied by a
scaling factor to superimpose it over the transmission cali-
bration spectra. By taking the second derivative of the
spectra, as shown in Figure 3, the baseline was normalized
and the spectral features were enhanced so that the fanning
out of the analytical region for CPM was observed at 1138
nm. Figure 4 shows the expanded analytical band demon-
strating the linear response from 0.1 mg to 2.0 mg CPM.
Smoothing was done on the derivative with a segment of 10
and a gap of zero. A thickness correction was applied as a
math pretreatment to correct for tablet thickness and
density variance over the region of 12501350 nm. The raw
spectral variance of the calibration set is large over this
region, and the pure CPM has an absorption minimum as
seen in Figure 2. Thickness correction is a normalization
function offered in Vision software (Foss NIRSystems, Inc.)
as a spectral mathematical pretreatment used to correct for
path length variance. The integral correction factor was
calculated over the range of 12501350 nm with a unity-
scaling factor.
Integral correction factor =
0.1510
0.1214
0.0918
0.0622
0.0326
0.0030
-0.0266
-0.0562
-0.0858
-1154
-0.1450
1111 1132 1153 1174 1195 1216 1237 1258 1279 1300 1321 1342 1363 1384
Wavelength (nm)
I
n
t
e
n
s
i
t
y
Figure 6: PLS factor weights in the analytical region.
10.000
1.000
0.100
0.010
0.001
1 2 3 4 5 6 7 8 9 10
l
o
g

(
P
R
E
S
S
)
Wavelength (nm)
PRESS, using 6 Factors
Table II: Repeatability results for 0.1 mg and 0.5 mg
tablets of chlorpheniramine maleate (CPM).
Tablet in
number
one tray
position
NIR
Predicted
mg CPM
HPLC
mg CPM
Residual
Average precision
and average bias
for all 5 tablets
0.1 mg 0.103
Individual tablet
data not shown
Tablet 1a 0.099 0.004
Tablet 1a 0.102 0.001
Tablet 1a 0.107 0.004
Tablet 1a 0.104 0.001
Tablet 1a 0.109 0.006
Tablet 1a 0.107 0.004
Tablet 1a 0.114 0.011
Tablet 1a 0.107 0.004
Tablet 1a 0.101 0.002
Tablet 1a 0.114 0.011
Precision 0.0051 0.0039
Bias 0.0034 0.0018
0.5 mg
tablet
0.512
Tablet 1a 0.513 0.001
Tablet 1a 0.526 0.014
Tablet 1a 0.527 0.015
Tablet 1a 0.534 0.022
Tablet 1a 0.521 0.009
Tablet 1a 0.511 0.001
Tablet 1a 0.529 0.017
Tablet 1a 0.528 0.016
Tablet 1a 0.52 0.512 0.008
Tablet 1a 0.517 0.512 0.005
Precision 0.007442 0.0055
Bias 0.0106 0.0057
*NIR is near-infrared; HPLC is high-performance liquid chromatography.
Table I: Prediction equation statistics.
Number of
factors
R
2
SEC PRESS F-statistic Xval
6 0.9998 0.0119 0.0095 24093 0.0148
*R
2
is the coefficient of determination. SEC is standard error of
calibration; PRESS is predicted residual error sum of squares; and Xval
is the standard error of cross validation using the leave-one-out method.
Figure 7: Predicted residual error sum of squares (PRESS) plot for
for factors used in the model. Where PRESS reaches a minimum is
usually considered the maximum number of factors. The PRESS
using six factors was 0.0095.
256361:3-column 6/15/07 2:50 PM Page 172
The correction factor equals the 0.5-
nm increment at which the data were
collected times the sum of the y-axis
values (S
i
) and the y-axis value plus 1
(S
i+1
), divided by 2. Then, the original
spectrum was divided by this correc-
tion factor throughout the analytical
region of 11201380 nm.
Partial least squares (PLS) regres-
sion was used to develop the
prediction model. PLS uses principal
component analysis and is a variation
of principal component regression
(PCR). The correct number of prin-
cipal components or factors was
determined by the Vision software
supplied with the instrument by deter-
mining where the predicted residual
error sum of squares (PRESS) reaches
a minimum (5).
Figures 5 and 6 are plots of the PLS
factor loadings and weights around the
1138-nm absorption band for CPM.
The loadings and weights appear
spectra-like and are not noisy, indi-
cating good modeling attributes for
the factors chosen. Figure 7 is a plot of
the PRESS leading to a model with
eight factors. The model chosen used
only six of these factors, trading
decreased error for robustness (6). The
PRESS for six factors was 0.0095. The
resulting model had a multiple corre-
lation coefficient (R
2
) value of 0.9998
and a standard error of calibration
(SEC) of 0.0119. The one-left-out
cross-validation demonstrates good
predictability with a standard error of
cross-validation of 0.0148.
Table I contains the model statistics
for the CPM prediction equation. Table
II is the repeatability results for five
tablets measured 10 times each of
nominal 0.1 mg CPM and 10 tablets of
0.5 mg CPM. The same tablet placed in
the same tray position was scanned 10
times. Data for tablet tray position
(number 1) are shown, and only the
combined statistical results are shown
for the other four tablets from each
dosage level. The average precision for
the nominal 0.1-mg level was 0.0039.
The precision for the nominal 0.5-mg
level was 0.0055. The bias was 0.0018
for the lower-level CPM and 0.0057 for
the higher level. Table III contains the
results from scanning the ten 0.1 mg
CPM tablets for content uniformity.
Table IV contains the results from scan-
ning the ten 0.5-mg CPM tablets for
content uniformity. The Vision soft-
ware has a convenient routine analysis
method for calculating content unifor-
mity automatically. Figures 8 and 9 are
X control charts for the 0.1-mg and 0.5-
mg CPM content uniformity tests.
These charts are for SPC, plotting target
label claim and 15% control limits.
The HPLC results showed that some of
the nominal 0.5-mg CPM tablets were
as high as 0.53 mg CPM.
Figure 10 shows the NIR predicted
CPM concentrations versus the HPLC
results for each tablet in the calibration
set. One tablet was left out of the cali-
bration set at each level for
prediction-model validation. Figure 11
shows the NIR predictions of the vali-
dation set versus the HPLC results for
CPM on each tablet.
Better precision and accuracy can be
achieved with a training set designed
with smaller increments around the
target label claim. Tablets from on-line
press processing can be scanned and
sent to the laboratory for HPLC analysis
and selected for calibration samples to
cover the range using a few extra pilot-
batch samples needed to extend the
range to 15% of label claim.
0.115
0.100
0.085
X Control chart
LCL
UCL

Sample number
m
g
Figure 8: Statistical process control chart of 0.1-mg chlorpheniramine maleate content uniformity. The X control chart plots the target label
claim with 15% control limits. UCL is the upper control limit, and LCL is the lower control limit.
Figure 9: Statistical process control chart of 0.5-mg chlorpheniramine maleate content uniformity. The X control chart plots the target label
claim with 15% control limits. UCL is upper control limit, and LCL is lower control limit.
0.575
0.500
0.425
X Control chart
LCL
UCL

Sample number
m
g
256361:3-column 6/15/07 2:50 PM Page 173
Table III: Content uniformity test results for ten 0.1-mg
chlorpheniramine maleate tablets.
Test level 1
Sample Target Test result*
Percentage of
target
Pass or fail
1 0.100 0.091 91.491 Pass
2 0.100 0.099 98.655 Pass
3 0.100 0.103 103.421 Pass
4 0.100 0.101 101.487 Pass
5 0.100 0.100 99.782 Pass
6 0.100 0.099 98.910 Pass
7 0.100 0.101 101.139 Pass
8 0.100 0.100 99.987 Pass
9 0.100 0.100 99.649 Pass
10 0.100 0.102 101.616 Pass
* Relative standard deviation is 3.2%.
Table IV: Content uniformity test results for ten 0.5-mg
chlorpheniramine maleate (CPM) tablets.
Test level 1
Sample Target Test result*
Percentage of
target
Pass or fail
1 0.500 0.523 104.642 Pass
2 0.500 0.520 104.094 Pass
3 0.500 0.530 106.097 Pass
4 0.500 0.525 104.947 Pass
5 0.500 0.529 105.804 Pass
6 0.500 0.526 105.106 Pass
7 0.500 0.525 104.950 Pass
8 0.500 0.529 105.877 Pass
9 0.500 0.524 104.717 Pass
10 0.500 0.531 106.248 Pass
* Relative standard deviation is 0.7%.
Conclusion
Near-infrared assay and content uniformity of tablets
provides a fast, accurate means of monitoring tablets for
production that is in step with the US Food and Drug
Administrations process analytical technology initiative.
The data show promising results that could relieve labora-
tory workload of high-performance liquid chromatography
analysis and bring analysis closer to real time for process
monitoring. Ten tablets can be analyzed in 5 min. The
software provided with the instrument is used for data
collection and developing prediction models. The software
also provides dedicated routine analysis methods for
content-uniformity analysis yielding results in percent label
claim and percent relative standard deviation as well as
passfail indication.
The average repeatability result for five different tablets
measured 10 times of nominal 0.1-mg chlorpheniramine
maleate was 0.0039 with a bias of 0.0018. The repeatability
result for 5 tablets of 0.5 mg chlorpheniramine maleate was
0.0055 with a bias of 0.0057. Better precision and accuracy
can be achieved with a training set designed with smaller
increments around the target label claim.
References
1. Handbook of Near-Infrared Analysis, D.A. Burns and E.W.
Ciuczak, Eds.(Marcel Dekker, Inc., New York, NY, 2001).
2. US Food and Drug Administration, Guidance for Industry: PAT
A Framework for Innovative Pharmaceutical Development,
Manufacturing, and Quality Assurance, (FDA,Rockville, MD, Sept.
2004).
3. P.J. Larkin, E. Fruhling, and C. Longfellow, Comparison of
Fourier Transform (FT) and Grating Based NIR Spectrometers
for Content Uniformity of Pharmaceutical Solid Dosage Forms,
Am. Pharm. Review. 9 (6), 102109 (2006).
4. Q. Ji et al., Rapid Content Uniformity Determination of Low-
Dose TCH 346 Tablets by NIR, Am. Pharm. Review 9 (5), 2026
(2006).
5. R. Kramer, Chemometric Techniques for Quantitative Analysis,
(Marcel Dekker, Inc., New York, NY, 1998).
6. K.R. Beebe, R.J. Pell, and M.B. Seasholtz, Chemometrics: A Prac-
tical Guide, (John Wiley & Sons, Hoboken, NJ, 1998). PT
3.0
2.6
2.2
1.8
1.4
1.0
0.6
0.2
-0.2
-0.6
-1.0
-1.0 -0.6 -0.2 0.2 0.6 1.0 1.4 1.8 2.2 2.6 3.0
HPLC CPM mg
N
I
R

P
r
e
d
i
c
t
e
d

C
P
M

m
g
Calibration Set: NIR Predicted vs Lab Data
Figure 10 (Left): Calibration set. Predicted
versus high-performance liquid
chromatography (HPLC). CPM is
chlorpheniramine maleate.
2.0
1.8
1.6
1.4
1.2
1.0
0.8
0.6
0.4
0.2
-0.0
-0.2
-0.4
-0.6
-0.8
-1.0
-1.0 -0.7 -0.4 -0.1 0.2 0.5 0.8 1.1 1.4 1.7 2.0
HPLC CPM mg
N
I
R

P
r
e
d
i
c
t
e
d

C
P
M

m
g
Validation Set: NIR Predicted vs Lab Data
Figure 11 (Right): Validation set. One tablet
was left out of the calibration set at each
level for production-model validation.
CPM is chlorpheniramine maleate.
Reprinted from PHARMACEUTICAL TECHNOLOGY, April 2007 Printed in U.S.A.
256361:3-column 6/15/07 2:51 PM Page 174
7703 Montpelier Road
Laurel, MD 20723
U. S. A.
T: 301.680.9600
F: 301.236.0134
E: info@foss-nirsystems.com
www.foss.dk
256361:3-column 6/15/07 2:51 PM Page 175