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Extending Calibrations for Near-Infrared Assay of Tablets Using

Synthetic Modeling and Variance from Placebos

Robert Mattes, Denise Root
FOSS NIRSystems, Inc.

The range of a near-infrared (NIR) calibration model for tablet assay is extended
using production “seed” spectra and synthetic spectra generated from placebos and
“pure” API spectra. This method resolves the problem of not being able to obtain a
range of production samples with +/- 15% of the label claim of API concentration on-
line. A synthetic model is developed using five real production samples and 70
samples synthesized from fractions of “pure” API spectra added to placebo spectra. A
synthetic calibration model with an R2 = 0.9987 and a standard error of calibration of
0.0037 mg was developed with this method for an API with 1.0 mg label claim in a
100 mg tablet.

Figure 1. Tablets in NIR instrument in Figure 4. The fanning out of the analytical
transmission mode. Figure 2. Raw calibration spectra with pure CPM Figure 3. Second derivative math region of the spectra where the CPM has a
The FDA initiative for process analytical technology (PAT) has created interest in
spectrum. pretreatment of calibration spectra. strong absorption band at 1138 nm
measuring pharmaceutical products upstream during processing for better quality and
process control (1). Near infrared has been a major analytical tool designated for PAT Discussion
analysis because it is fast, accurate, nondestructive and requires no sample
preparation. At-line solid dosage form analysis has become important for tablet assay Figure 2 shows the 75 raw NIR spectra from the calibration set and the
and content uniformity because pharmaceutical manufacturers would like to measure spectrum of the simulated pure CPM. By taking the second derivative of the
more frequently than the ten tablets per batch specified by the US Pharmacopoeia spectra, as shown in Figure 3, the baseline was normalized and the spectral
general chapter <905> on content uniformity (CU). features were enhanced so that the fanning out of the analytical region for
CPM was observed at 1138 nm. The second derivative causes the
Laboratory methods for tablet assay and content uniformity are routinely done by absorption bands to be inverted and increase in the downward direction.
HPLC and are usually time consuming requiring lengthy calibration runs, the mixing of The second derivative pretreatment was applied using a smoothing with a
buffers and the procurement and disposal of hazardous solvents. The analysis of ten segment of 10 and a gap of 0. The standard normal variate (SNV)
tablets for content uniformity could take hours and the results may not be available to pretreatment was applied to reduce scattering effects. An integral thickness
the tablet press operators or for batch release for many days or even weeks after the correction was applied as a math pre-treatment to correct for tablet
Figure 6. Predicted Residual Error
tablets are compressed. thickness and density variance from 1280 nm to 1350 nm.
Figure 5. Factor Loadings. Sum of Squares (PRESS)
Near infrared spectroscopic analysis is an empirically based technique that requires Figure 4 shows the expanded second derivative of the aromatic CH stretch
the prediction method developer to analyze a calibration set of samples that represent analytical band demonstrating the fanning out from 0.85 mg to 1.15 mg CPM
the natural variation in the solid dosage form (2). It is often difficult to obtain the proper (adjusted to 0.875 mg to 1.1838 mg from HPLC analysis). See Table I for
range of samples that would include slightly more than +/- 15% of the label claim. The real tablet values and the fractional adjustment that was made using the
process produces tablets normally distributed around the label claim and the model mean tablet value. The adjusted values are the constituent values that were
developer cannot obtain tablets that are more than a few percent away from the used for modeling.
nominal value. Therefore, to empirically develop a prediction model that can predict
samples that may be +/- 15% of the label claim without extrapolation, synthetic Partial least squares (PLS) regression was used to develop the prediction
methods are investigated. model. PLS uses principal component analysis and is a variation of principle
component regression (PCR). The correct number of principal components
This study discusses a method of obtaining a prediction method that uses only or factors determination is aided by the Vision® software supplied with the
placebo tablets and samples containing API from the product line. The natural instrument by calculating where the predicted residual error sum of squares
variation range from the product line is extended with synthetic samples created by (PRESS) reaches a minimum (3). Although the PRESS does not reach a
adding varying amounts of the pure active spectrum to the placebo spectra. Purely minimum until 13 factors, six were chosen as optimal allowing more than ten
synthetic spectra lack the scattering variance of real tablets and would not create a calibration samples for each factor and considering possible degrees of
robust prediction model. The placebos, which are often available from clinical studies, freedom. Figure 8. Validation of synthetic
Figure 7. Factors=6, R2=0.9987,
provide the real scattering and spectral variance seen in a calibration set. Calibration equation CPMsyncal2.
SEC=0.0037, xVal=0.004.
Figure 5 is a plot of the principal component loadings around the 1138 nm
Experimental absorption band for CPM. The loadings appear spectra-like and are not
noisy, indicating good modeling attributes for the factors chosen. The
The NIR instrument used in the study was XDS MasterLab (FOSS NIRSystems, loadings indicate regions of high correlated variance throughout the
Laurel, MD) which is capable of automatically measuring multiple tablets after they spectrum for each factor. Figure 6 is a plot of the PRESS. The resulting
Table I. HPLC and fraction adjusted by mean tablet value.
are positioned in a special tray (Figure 1). Spectra were collected in the transmission model had a multiple correlation coefficient (R2) value of 0.9987 and a
mode from 800 nm to 1650 nm with 0.5 nm data intervals and 32 scans were co- Table II. Predicted values for the validation set.
standard error of calibration (SEC) of 0.0037 mg. The one-left-out cross
Tablet ID HPLC mg || Fraction Adjusted
added to produce a single spectrum. validation demonstrates good predictability with a standard error of cross
Model: CPMsyncal2 SEP=0.0104, Bias=0.0139
validation of 0.004mg (very close to the SEC).
Tablet #1 1.017 || 0.85 0.875 Tablet ID NIR mg HPLC mg Residual
Ten chlorpheniramine maleate (CPM) tablets of nominal 1.0 mg and ten placebo
Tablet #2 1.032 || 0.90 0.926 Tablet #6a 1.024 1.035 0.011
tablets of the same excipient composition were scanned on the MasterLab Figure 7 shows the NIR predicted CPM concentrations versus the HPLC
Tablet #3 1.034 || 0.95 0.978 Tablet #7a 0.994 1.009 0.015
transmission instrument. The tablets from a 1.0 mg CPM batch were sent for HPLC results for each tablet in the calibration set using the synthetic calibration
Tablet #4 1.040 || 1.00 1.029 Tablet #8a 1.017 1.039 0.022
analysis after NIR scanning. The spectra of the first five of the 1.0 mg tablets were equation CPMsyncal2. The other five tablets scanned (tablet 6a through
Tablet #5 1.024 || 1.05 1.081 Tablet #9a 1.026 1.037 0.011
averaged and the placebo spectra were averaged. The other five tablets measured tablet 10a) on the MasterLab instrument, were predicted for validation.
mean value 1.029 || 1.10 1.132 Tablet #10a 0.990 1.000 0.010
were saved for a validation set. The average placebo spectrum was subtracted from Figure 8 shows the NIR predictions of the validation set versus the Lab
1.15 1.184
spectrum of the average of the five1.0 mg tablets in the FOSS Vision® software. This (HPLC) CPM value seen in Table II. The standard error of prediction (SEP)
resulted in a simulated transmission spectrum of pure CPM. This spectrum could not of the validation set compared to the HPLC data was 0.0104 mg and the References
be obtained in transmission because the pure CPM would have had to be pressed as bias was 0.0139 mg. The SEP is significantly larger than the SEC because
a tablet. the synthetic calibration samples have reduced error except for the seed 1) FDA, Guidance for Industry: PAT-a framework for innovative pharmaceutical development, manufacturing,
samples. The SEP is low at only about 1% of batch label claim. and quality assurance, September, 2004.
The pure CPM spectrum was multiplied by factors to obtain a range of pure spectra 2) R.A.Mattes, et al., Pharmaceutical Technology, 4, 2007.
from 85% to 115% in 5% increments of label claim. The average HPLC value for the Conclusion 3) Chemometric Techniques for Quantitative Analysis, R. Kramer, Marcel Dekker, Inc. 1998.
five real tablet spectra was calculated. The average value was multiplied by each 4) Chemometrics: A Practical guide, K. R. Beebe, R. J. Pell, M. B. Seasholtz, John Wiley & Sons, 1998.
fraction from .85 to 1.15 to correct for the HPLC versus nominal tablet CPM values. A method for generating a prediction model from placebo and seed batch
The fractional pure spectra from 85% to 115% were then added to the ten individual samples from the production line with synthetic samples generated from Acknowledgements
placebo spectra resulting in 70 spectra. The five real tablet spectra were added to the average placebo and batch samples to increase the sample range was
synthetic calibration set as “seed” spectra resulting in 75 spectra in the total demonstrated. A prediction model with an R2 of 0.9987 and a standard error Thanks to Om Anand, Maria Gerald Rajan, Namrata R. Trivedi, Wen Qu, Yingxu Peng, Yichun Sun who
calibration set. of calibration (SEC) of 0.0037 mg was developed. The standard error of were at the University of Tennessee, Department of Pharmaceutical Sciences at the time the CPM tablets
prediction (SEP) on the validation set was 1% of label claim at 0.0104 mg. were made and analyzed them with HPLC.