Near-infrared spectroscopy and imaging for the monitoring of powder blend homogeneity.

In-process testing requirements for adequacy of mixing are established in 21 CFR 211.110(a)(3). Considering also, the U.S. Food and Drug Administration's draft guidance published in 1999 (Guidance for Industry, ANDAs: Blend Uniformity Analysis; http://www.fda.gov/cder/guidance/index.htm), the importance of when and how to perform blend uniformity analysis is obvious. Near-infrared (NIR) spectroscopy was used noninvasively, in this study, to monitor powder blend homogeneity. Powder mixtures consisting of salicylic acid and Fast-Flo lactose were blended in an 8-qt. V-Blender. Optical ports installed at six positions on the blender allowed spectral collection using fiber optics. A traditional thief probe was used to collect powder samples for ultraviolet (UV) reference analysis. The blender was stopped at preselected time points for collection of NIR and UV data. Several algorithms and sampling protocols were studied to identify an optimum methodology for blend homogeneity determination. The blending process was also monitored with an InSb imaging camera for comparison with the traditional NIR spectroscopy and UV reference data. Data analysis indicates that multiple sampling points were essential for accurate and precise estimation of mixing end points. Moreover, multiple runs of identical blends often display homogeneity at unique end points, thus demonstrating the potential advantage of monitoring every blend.

Multispectral imaging of tablets in blister packaging.

This experiment tested the hypothesis that using near-infrared (IR) imaging spectrometry on tablets through blister packs permits the identification and composition of multiple individual tablets to be determined simultaneously. Aspirin was selected for this study because its breakdown mechanism is well understood. Near-IR cameras were used to collect thousands of spectra simultaneously from a field of packaged aspirin tablets. Tablets were selected by a principal component analysis selection algorithm. Graphs of the columns of the transformation matrix showed that salicylic acid and acetylsalicylic acid in the samples were modeled by the principal components. The bootstrap error-adjusted single-sample technique chemometric-imaging algorithm was used to draw probability-density contour plots that revealed tablet composition. Choice of color was used to represent constituent identity, whereas intensity represented concentration. The percentage of usable pixels in the indium antimonide (InSb) array was 99.9%. The SEP was 0.06% of the tablet mass for both water uptake and salicylic acid production. The number of tablets that a typical near-IR camera can currently analyze simultaneously was also estimated to be approximately 1300.

Detection of formaldehyde-induced crosslinking in soft elastic gelatin capsules using near-infrared spectrophotometry.

The purpose of this research was to monitor the migration of formaldehyde from a polyethylene glycol (PEG) fill into the gelatin shell of a soft elastic gelatin capsule (SEGC) using near-infrared (NIR) spectrophotometry. SEGCs were filled with five solutions of aqueous formaldehyde in PEG (0, 0.05, 0.10, 0.20, and 0.40 v/v%), stored at ambient conditions for 48 hr, emptied, and scanned in NIR spectrophotometer. Principal component regression (PCR) was employed to analyze the spectra of the empty capsules. Good correlation was established (r2 = 0.988) when actual concentrations of formaldehyde in the PEG fill of the capsules were regressed against the principal component (PC) values from NIR spectra of the emptied and washed capsules. The loadings of the first PC describe a baseline shift in the spectra that arises from a change in water concentration. Lower PC loadings reveal the presence of signals at 1780 and 2200 nm that are not due to water absorbance, confirming the hypothesis that chemical bonds are formed during the formaldehyde-induced crosslinking of the gelatin in SEGCs. Gelatin crosslinking, initiated by formaldehyde migration from the PEG fill into the shell of an SEGC, was detected by NIR spectrophotometry. When NIR was coupled to principal component analysis, a linear relationship was found between the NIR spectra of empty SEGCs and the amount of crosslinking induced by concentrations of formaldehyde in the original fill material.

Mechanisms contributing to angiotensin II regulation of body weight.

Previous studies in our laboratory have implicated adipose tissue as a potential site for local angiotensin II (ANG II) synthesis. However, functions of ANG II in adipose tissue and the impact of ANG II on body weight regulation are not well defined. To study the effect of ANG II on body weight, a chronic ANG II infusion model was used. In study 1, a low dose of ANG II (175 ng.kg-1.min-1) was infused into rats for 14 days. Plasma ANG II levels were not elevated after 14 days of infusion. ANG II-infused rats did not gain weight over the 14-day protocol and exhibited a lower body weight than controls on day 8. Food intake was not altered, but water intake was increased in ANG II-infused rats. Blood pressure gradually increased to significantly elevated levels by day 14. Thermal infrared imaging demonstrated an increase in abdominal surface temperature. Measurement of organ mass demonstrated site-specific reductions in white adipose tissue mass after ANG II infusion. In study 2, the dose-response relationship for ANG II infusion (200, 350, and 500 ng.kg-1.min-1) was determined. Body weight (decrease), blood pressure (increase), white adipose mass (decrease), plasma ANG II levels (increase), and plasma leptin levels (decrease) were altered in a dose-related manner after ANG II infusion. In study 3, the effect of ANG II infusion (350 ng.kg-1.min-1) was examined in rats treated with the vasodilator hydralazine. Hydralazine treatment normalized blood pressure in ANG II-infused rats. The effect of ANG II to decrease body weight was augmented in hydralazine-treated rats. These results demonstrate that low levels of ANG II infusion regulate body weight through mechanisms related to increased peripheral metabolism and independent of elevations in blood pressure.

Near-IR and IR imaging in lipid metabolism and obesity.

Approximately one-third of Americans are classified as obese. There has long been an interest in drug therapies for obesity. Interest in obesity research and in drug interventions in obesity has greatly increased since the discovery of a protein named leptin, one of apparently many competing biological signals in energy metabolism. The complexity of the obesity problem demands new non-invasive and non-destructive methods for monitoring lipid metabolism and energy expenditure to study the competing biological signals and their effects. A new computer algorithm for spectrometric imaging of living subjects is used to remove artifacts arising from subject motion from spectra and images. The algorithm is sufficiently simple to be implemented easily in hardware for real-time video processing. Because the algorithm can be applied to images, thermogenesis and lipid metabolism in interscapular adipose tissue can be observed directly in unrestrained and unanesthetized subjects using an InSb focal plane array video camera. The accuracy and precision of temperature and spectral measurements are established using laboratory references and prototype drugs in test subjects.

Determination of extent of formaldehyde-induced crosslinking in hard gelatin capsules by near-infrared spectrophotometry.

PURPOSE: To predict the degree of crosslinking from formaldehyde-stressed hard gelatin capsules (HGCs) using near-infrared spectrophotometry (NIR). METHODS: HGCs were exposed to a 150 ppb atmosphere of formaldehyde for 2.25, 4.60, 9.42, 16.0 and 24.0 hours. The capsules were filled with fresh amoxicillin, placed in a 90 degrees conical reflector cone, and scanned in a NIR spectrophotometer. Principal component regression (PCR) was employed to analyze the spectra of the intact capsules. Dissolution profiles were then obtained for each experimental group. RESULTS: The dissolution of amoxicillin from the capsules at pH 1.2 was found to decrease with increasing time of exposure to the formaldehyde atmosphere. A set of principal components (PCs) was formed by a linear combination of the absorbance values at each wavelength scanned. A good correlation was established (r2 = 0.963) when PC values from the NIR spectra of the HGCs were regressed against percentage of amoxicillin dissolved at 45 minutes, at pH 1.2. Water content of the capsules was found to be the largest determinant in the variation between HGC spectra at each exposure time. CONCLUSIONS: NIR spectrophotometry, combined with PCR, was successful at not only predicting dissolution of HGCs exposed to formaldehyde, but also at determining which wavelengths contributed most to spectral variation of these stressed HGCs.

Near-infrared spectrophotometric monitoring of stroke-related changes in the protein and lipid composition of whole gerbil brains.

Strokes are a critical problem in the U.S. that affect more than 500,000 people annually. Research into the causes of stroke and testing of drug therapies to reduce ischemic and postischemic damage to the brain is frustrated by an inability to continuously follow the physical and chemical events that occur during ischemia and reperfusion in vivo. Near-IR spectrometry is used in this paper to observe stroke-induced changes in the lipids and proteins of whole brain samples and in intact subjects. The examination of whole brains is made possible by a combination of hardware and software techniques designed to make the sample presentation to the spectrometer more reproducible. Near-IR spectrophotometry of brain tissue discriminates between adult (3-4 months of age) and aged (18-20 months of age) brains as well as between brains exposed to 5- and 10-min ischemia. The near-IR analytical method has many applications in aging and stroke research, including the noninvasive determination of age from brain spectra obtained transcranially, simultaneous multicomponent analysis of lipids and proteins, and quantification of edema.

Near-IR imaging of atheromas in living arterial tissue.

A near-IR imaging system and parallel vector supercomputer are used with a fiber-optic probe to produce chemical maps of the intimal surface of living arteries. Spectrometric information collected at hundreds of near-IR wavelengths is assembled into color pictures of the lipoprotein and apolipoprotein composition of atheromas using a vectorized 3-D cellular automaton-based algorithm that operates in parallel. The nonparametric mathematics developed to identify and quantify the constituents of each voxel in the artery wall avoid the matrix factorizations that generate excess error in other pattern recognition methods and permit analysis in a wavelength space of over 1000 dimensions using fewer than 100 calibration samples. A surface feature resolution of 5.5 microns and depth resolution of 6.5 microns are achieved with the system. Data from the fiber optics confirm the injury hypothesis of lesion formation and the differing roles of HDL and LDL in cholesterol transport. In clinical studies, approximately 1/2 of human arterial lesions appear fibrous and contain little or no lipid. As such, these lesions would not be expected to regress in response to cholesterol-lowering agents such as lovastatin. Identification of lesion types in vivo will enhance the efficacy of treatment programs.

Bacterial monitoring in vials using a spectrophotometric assimilation method.

Aseptic-filling processes are often used with fragile parenteral products that might be destroyed by terminal autoclaving. However, aseptic filling is not as effective as autoclaving in reducing contamination. As a result, time-consuming microbiological methods and turbidimetry are employed currently as product inspection techniques, but these processes can destroy the product and might not detect low levels of contamination. Thus, near-infrared (IR) light scattering was evaluated in this study as a new method for determining low levels of contamination noninvasively and nondestructively. A new parallel mathematical technique was used in conjunction with near-IR spectrophotometry to detect successfully contamination by several species of bacteria through intact glass vials. Using the near-IR method, products can be evaluated without introducing contamination, preserving the sample vial for dispensing or evaluation by another method.

Spectrophotometric prediction of the dissolution rate of carbamazepine tablets.

A near-infrared (IR) spectrophotometer, integrating optics, and parallel-vector supercomputer are employed to develop a mathematical model that predicts the dissolution rate of individual intact tablets from near-IR spectra (r2 = 0.985). Each tablet can be analyzed nondestructively by the spectrophotometer in less than 1 min. The model permits hundreds of near-IR wavelengths to be used in the determination of dissolution rate, leading to increased accuracy.

Near-infrared spectrometry of microorganisms in liquid pharmaceuticals.

Biotechnology and pharmaceutical research have created a number of new and potentially life-saving drugs. Many of these drugs are formulated as injectable products. Some drug products do not survive autoclaving or other means of terminal sterilization. An aseptic filling process is typically used to sterilize such products, but it is less reliable than autoclaving, making detection of unsterile units even more essential. Invasive microbiological methods and turbidimetry are currently employed as inspection techniques. These processes are time-consuming, destroy product, and may not detect low levels of contamination. Near-IR light scattering is proposed as a new method of determining low levels of contamination noninvasively and nondestructively. The method is used successfully in the current study to detect contamination by a species of yeast, mold, and bacteria in intact plastic infusion bags at levels as low as three colony-forming units per milliliter for yeast. By use of the near-IR method, each injectable unit can be evaluated with its integrity maintained, allowing the product to be dispensed or evaluated by another analytical method.

Nondestructive near-infrared analysis of intact tablets for determination of degradation products.

Near-infrared spectrometry was used in this study to examine intact aspirin tablets in order to demonstrate the usefulness of the technique as a nondestructive method of quality control. Unique sampling optics were used to simultaneously illuminate the entire surface of the tablets, including the top, bottom, and side. Changes in individual tablet spectra were correlated to (a) the time that the tablets spent in a hydrator, (b) the mass of water absorbed by the tablets, and (c) the mass of salicylic acid formed by base-catalyzed hydrolysis of acetylsalicylic acid. A prediction equation for each of these three parameters was constructed using near-infrared spectral reflectance values obtained from intact tablets. Prediction errors were low for (a) the time that tablets spent in the hydrator (+/- 19 h over a period of 168 h), (b) the mass of water absorbed (+/- 0.04% of tablet mass), and (c) the mass of salicylic acid formed (+/- 0.04% of tablet mass).

Near-infrared spectroscopic determination of residual moisture in lyophilized sucrose through intact glass vials.

A rapid, noninvasive, and nondestructive method for determining moisture in sealed freeze-drying vials is described. The method, based on near-infrared spectrometry, used a novel fiber-optic diffuse-reflectance probe to make remote reflectance measurements from 1100 to 2500 nm through the bottom of glass vials. The correlation of the method to results obtained by Karl Fischer analysis was good (r2 = 0.97). The moisture content of sucrose, a common cryoprotectant, was determined with an error of 0.27% using a single sample scan.

Assessment of the feasibility of determination of cholesterol and other blood constituents by near-infrared reflectance analysis.

Near-infrared reflectance spectrometry of blood serum can yield values for serum cholesterol that correlate reasonably well (r = 0.96) with those from common reference analytical methods. However, the variability of serum can cause ostensibly validated calibrations to fail on new samples. The determination of blood components such as cholesterol and triglycerides by near-infrared reflectance is complicated by their low concentrations, the variety of forms in which they appear, and by the natural variability of the blood matrix. These difficulties, when combined with the problems encountered in obtaining a representative sample from a given individual, can make it almost impossible to select, by a regression procedure, a wavelength combination that is characteristic of the complete blood matrix. The failure of the regression process to find characteristic wavelengths generates a false-sample problem in which even small changes at the analytical wavelengths produce a grossly unreliable cholesterol or triglyceride determination.