Biomedical subjects

Raj Suryanarayanan

Publications and source records attributed to Raj Suryanarayanan.

At least 19 recordsLinked to original sources

PubMed2007

Influence of processing conditions on the physical state of mannitol--implications in freeze-drying.

PURPOSE: To study the effect of processing conditions on the physical state of mannitol during various stages of the lyophilization cycle of a protein formulation. MATERIALS AND METHODS: Mannitol and trehalose were used as the bulking agent and lyoprotectant, respectively. The physical state of mannitol during various stages of freeze-drying cycle, in the absence and presence of a model protein, was characterized using low temperature X-ray powder diffractometry (XRD) and differential scanning calorimetry (DSC). RESULTS: Mannitol did not crystallize even when the solution for lyophilization was cooled to -45 degrees C at a slow cooling rate of 1 degree C/min. Annealing facilitated mannitol crystallization, and in the absence of the protein, a mixture of delta-mannitol and mannitol hemihydrate was obtained at both low (-18 degrees C) and high (-8 degrees C) annealing temperatures. However, in the presence of protein, the high annealing temperature promoted delta-mannitol crystallization and inhibited formation of mannitol hemihydrate, while the low annealing temperature facilitated the formation of mannitol hemihydrate. Interestingly, the hemihydrate in the frozen solution was retained in the final lyophile, even when the primary and secondary drying temperatures were as high as -5 and 65 degrees C, respectively. CONCLUSIONS: The presence of protein as well as the processing conditions (annealing temperature and time, primary and secondary drying temperatures) influenced the physical form of mannitol in the final lyophile. The protein promoted formation of delta-mannitol while inhibiting the formation of mannitol hemihydrate. Since the physical form of mannitol was greatly influenced by the presence of protein, it will be prudent to conduct the preliminary lyophilization cycle development studies in the presence of the protein. If mannitol hemihydrate is formed during annealing, its dehydration may require high secondary drying temperature.

Calorimetry, Differential Scanning↗

PubMed2006

Investigation of the multi-step dehydration reaction of theophylline monohydrate using 2-dimensional powder X-ray diffractometry.

PURPOSE: (i) To study the dehydration kinetics of theophylline monohydrate using 2-dimensional (2D) powder X-ray diffractometry (XRD), and (ii) to investigate the effect of polyvinylpyrrolidone (PVP) on the dehydration pathway and kinetics. METHODS: Theophylline monohydrate (C(7)H(8)N(4)O(2).H(2)O; M) was recrystallized from aqueous PVP solutions of different concentrations. Dehydration kinetics was studied isothermally, at several temperatures, from 35 to 130 degrees C. The experimental set-up comprised of a high intensity X-ray source (synchrotron radiation or 8 kW rotating anode), a heating chamber, and a 2D area detector. Diffraction patterns were collected continuously, with a time resolution ranging from 40 ms to 30 s, over the angular range of 3 to 27 degrees 2theta. RESULTS: Dehydration of M resulted in either the stable (C(7)H(8)N(4)O(2); A), or the metastable anhydrate (A*), with the latter having a tendency to transform to A. The XRD technique allowed simultaneous quantification of M, A* and A during the dehydration reaction. The rate constants for individual reaction steps (M-->A*; M-->A and A*-->A) were determined by fitting the data to solid-state reaction models. In presence of PVP, there was a decrease in the magnitude of the rate constant associated with the M-->A transition, resulting in an increased build-up of A* in the product. The inhibitory effect of PVP on M-->A transition was more pronounced at lower dehydration temperatures, and was proportional to the concentration of PVP. CONCLUSIONS: Two dimensional powder X-ray diffractometry, using a high intensity source, is a powerful technique to study kinetics of rapid solid-state reactions. The inhibitory effect of excipients can have profound effect on phases formed during pharmaceutical processing.

Algorithms↗

PubMed2006

Ionization states in the microenvironment of solid dosage forms: effect of formulation variables and processing.

PURPOSE: Evaluation of the effect of formulation composition and processing variables on the microenvironment in solid dosage forms, based on ionization of indicator probes. MATERIALS AND METHODS: Sulfonephthalein indicators were intimately mixed with individual excipients, binary excipient mixtures or multi-component blends by the solvent deposition method. Diffuse reflectance visible spectroscopy of these solids provided a measure of indicator ionization extent. Indicator solution studies yielded equations relating solution pH to the ratio of the absorbance signals of the ionized to that of the unionized form, for each indicator. These equations and the spectral data of the indicator-treated solids were used to calculate an acidity function, 'pH(eq)' for the solids. The ionization of incorporated probes was also monitored during various stages of simulated pharmaceutical processing viz. wet and dry mixing. RESULTS: The pH(eq) provided a measure of the physicochemical environment experienced by the probe in the solid. The surface nature of formulation components and their surface area available for interaction influenced the overall properties of the final blend. The extent of probe ionization varied at different stages of a simulated wet mixing-drying process. The pH of the excipient suspension was not a good predictor of the probe ionization in the final dried solid. Indicator ionization is expected to be influenced by the microenvironmental acidity, polarity and ionic strength. Individual excipient properties contributed to the overall microenvironment in powder mixtures even when dry mixed at low water contents. CONCLUSIONS: The environment experienced by a drug in the final solid dosage form will be influenced by the nature of the excipients, the extent of their surfaces available for interaction, surface modification during processing and the amount and nature of solvent used.

Algorithms↗

PubMed2006

Solute crystallization in frozen systems-use of synchrotron radiation to improve sensitivity.

PURPOSE: To demonstrate the sensitivity of low temperature synchrotron X-ray diffractometry (SXRD) for detecting solute crystallization in frozen sodium phosphate buffer solutions. To determine the effect of annealing on solute crystallization in frozen solutions. MATERIALS AND METHODS: Sodium phosphate buffer solutions, at initial buffer concentrations ranging from 1 to 100 mM (pH 7.4) were cooled to -50 degrees C. The crystallization of disodium hydrogen phosphate dodecahydrate (Na(2)HPO(4) *12H(2)O) was monitored using a laboratory as well as a synchrotron source. At selected concentrations, the effect of annealing (at -20 degrees C) was investigated. RESULTS: With the laboratory source, solute crystallization, based on the appearance of one diagnostic peak with a d-spacing of 5.4 A, was evident only when the initial buffer concentration was at least 50 mM. In contrast, using SXRD, crystallization was detected at initial buffer concentrations down to 1 mM. In addition, the use of a high-resolution 2D detector enabled the visualization of numerous diffraction rings of the crystalline solute. At both 10 and 100 mM buffer concentration, there was no increase in solute crystallization due to annealing. CONCLUSION: By using synchrotron radiation, solute crystallization was detected with substantially increased sensitivity, making the technique useful for freeze-drying cycles of practical and commercial importance. Since numerous peaks of the crystalline solute appeared, the technique has potential utility in complex, multi-component systems.

Buffers↗

PubMed2006

Structure in dehydrated trehalose dihydrate--evaluation of the concept of partial crystallinity.

PURPOSE: (i) To use trehalose as a model compound to evaluate the concept of crystallinity in pharmaceuticals. (ii) To understand the structural nature of dehydrated trehalose dihydrate. MATERIALS AND METHODS: Trehalose dihydrate was dehydrated isothermally at several temperatures below 100 degrees C and the anhydrous product was characterized by XRD, DSC and water vapor sorption. RESULTS: XRD and DSC suggested that the dehydration product was a partially crystalline alpha-polymorphic form of anhydrous trehalose (T(alpha)). An increase in the temperature of dehydration resulted in a decrease in lattice order. In agreement with earlier findings, the ordered regions in the dehydrated lattice (T(alpha)) converted to the dihydrate at much lower RH values than amorphous trehalose. However, the lattice order in the dehydrated product dictated the RH at which this conversion was initiated--the higher the lattice order the lower this RH. The structural nature of these samples can be explained based on the one-state model of crystallinity. In dehydrated trehalose, there is a continuum in lattice order ranging from highly crystalline (T(alpha)) to a completely disordered (i.e., amorphous) state. CONCLUSION: The extent of lattice order in anhydrous trehalose T(alpha) was dictated by the kinetics of water removal from trehalose dihydrate. The partially crystalline nature of anhydrous trehalose produced by dehydration could be described on a continuous scale of lattice order based on the one-state model of crystallinity.

Adsorption↗

PubMed2006

Calculation of the penetration depth of X-rays in intact pharmaceutical film-coated tablets by microdiffractometry.

PURPOSE: (i) Develop a method to calculate the penetration depth of X-rays in intact film-coated tablets, and validate it using model bilayer tablets. (ii) Characterize the physical form of drug in intact pharmaceutical film-coated tablets by XRD. MATERIALS AND METHODS: An equation for the calculation of the penetration depth of X-rays, as a function of the incident angle, was derived. Model bilayer tablets were prepared to validate the calculation method. The upper layer of the tablets consisted only of microcrystalline cellulose, while the lower layer was a mixture of cerium oxide (10% w/w), blue dye (5% w/w) and microcrystalline cellulose. The total tablet thickness was 3,500 microm, with the upper layer thickness ranging from 200 to 700 microm. The diffracted intensity of a cerium oxide peak in the lower layer was determined using a microdiffractometer system (CuKalpha radiation) with a two-dimensional area detector. The calculated penetration depth of X-rays was compared with that determined by XRD. After validation of the XRD method, commercial ibuprofen tablets were characterized. RESULTS: The penetration depth calculated by the method developed in this study was, in general, in good agreement with that determined experimentally by XRD. In commercial ibuprofen tablets, the coating material exhibited peaks due to TiO2 (25.4 degrees 2 theta) and Fe2O3 (33.3 degrees). However, these did not interfere with the characteristic peak of ibuprofen (22.2 degrees). CONCLUSION: We developed a method for calculation of penetration depth of X-rays in film-coated tablets and validated it using the model bilayer tablets. This method enables the characterization of the active pharmaceutical ingredient in different regions (at different depths) of the film-coated tablet. Since the technique is nondestructive, the same tablet can be repeatedly analyzed during stability studies.

Absorption↗

PubMed2006

Impact of freeze-drying on ionization of sulfonephthalein probe molecules in trehalose-citrate systems.

"pH memory," i.e., correlation between pH of solution before freeze-drying and chemical reactivity in the freeze-dried state, has been reported in many systems. In this study, the "pH memory" is explored by comparing the extent of protonation of sulfonephthalein probe molecules, bromophenol blue, bromocresol green, and chlorophenol red, in aqueous solution in the pH range of 3.4-6.0 and in the resulting freeze-dried amorphous matrix (lyophile) containing trehalose and sodium citrate buffer. The protonation of the probe molecules was measured in the lyophiles by diffuse reflectance visible spectroscopy, and compared with that in the solution before drying. The protonation of the indicators in the amorphous matrix correlated with solution pH, that is, an increase in solution pH resulted in a progressive decrease in the indicator protonation in the corresponding lyophile. However, the protonation was consistently higher in the lyophile than in the corresponding solution. The Hammett acidity function of lyophiles was calculated based on the extent of protonation of the probe molecules. Protonation of the probe molecules and the Hammett acidity function depended not only on prelyophilization solution pH, but also on the residual water content and the presence of amorphous sugar in the lyophile.

Bromcresol Green↗

PubMed2005

Influence of the active pharmaceutical ingredient concentration on the physical state of mannitol--implications in freeze-drying.

PURPOSE: The aim of this study was to investigate the effect of the concentration of the active pharmaceutical ingredient on the physical state of mannitol in frozen aqueous systems. METHODS: A human monoclonal antibody was used as the model protein. Mannitol and sucrose were used as the bulking agent and the lyoprotectant, respectively. The thermal behavior of frozen mannitol-sucrose solutions during and after annealing, in the absence and presence of the protein, were characterized by low-temperature powder X-ray diffractometry and differential scanning calorimetry. The influence of the protein on the crystallization behavior of mannitol was also evaluated. RESULTS: The excipient concentration had a pronounced effect on the glass transition temperature of maximally freeze-concentrated amorphous phase (T(g)'). At fixed excipient compositions, the protein had no effect on the T(g)' if the protein concentration was < or =20 mg/ml. However, at higher protein concentrations, there was a marked increase in T(g)' as a function of protein concentration. The inhibitory effect of the protein on mannitol crystallization was concentration dependent and was directly evident from X-ray diffractometry experiments. Annealing facilitated both mannitol nucleation and crystal growth even in the presence of the protein. CONCLUSIONS: The ratio of mannitol to sucrose and the protein concentration have an impact on the T(g)' and may therefore influence the primary drying temperature. The protein inhibits both the nucleation and growth of mannitol crystals and this effect seems to be concentration dependent. The presence of the protein and the protein concentration dictate the processing conditions, i.e., annealing time, annealing temperature, and primary drying temperature.

Calorimetry, Differential Scanning↗

PubMed2005

Quantification of crystallinity in substantially amorphous materials by synchrotron X-ray powder diffractometry.

PURPOSE: The aim of this study was to develop a highly sensitive powder X-ray diffraction (XRD) technique for quantification of crystallinity in substantially amorphous pharmaceuticals, utilizing synchrotron radiation and a 2-D area detector. METHODS: Diffraction data were acquired at the European Synchrotron Radiation Facility (France) using a 2-D charge-coupled device detector. The crystallization of amorphous sucrose was monitored in situ, isothermally at several temperatures in the range of 90 to 160 degrees C. An algorithm was developed for separation of the crystalline and amorphous intensities from the total diffraction pattern. RESULTS: The synchrotron XRD technique allowed powder diffraction patterns to be recorded with a time resolution of 40 ms. The gradual crystallization of sucrose is analogous to a series of physical mixtures with increasing content of the crystalline component. The in situ crystallization approach circumvented the problem of inhomogeneity in mixing--a potentially serious issue at extreme mixture compositions. The estimated limit of detection of crystalline sucrose in an amorphous matrix was 0.2% w/w, a considerable improvement over the reported value of approximately 1% w/w with a conventional XRD. CONCLUSION: High-intensity XRD can discern subtle changes in the lattice order of materials. The first evidence of crystallization can serve as an indicator of the potential physical instability of the product.

Crystallization↗

PubMed2005

Partially crystalline systems in lyophilization: I. Use of ternary state diagrams to determine extent of crystallization of bulking agent.

Two model ternary systems: water-glycine-raffinose and water-glycine-trehalose were investigated to determine the extent of glycine crystallization in frozen solutions. The use of such partially crystalline systems allows primary drying to be carried out substantially above the collapse temperature. Differential scanning calorimetry (DSC) and variable temperature X-ray diffractometry (XRD) were used to monitor phase transitions in frozen systems as well as to determine the T'g. Aqueous solutions containing different glycine to carbohydrate weight ratios were first cooled to -60 degrees C and then warmed to room temperature. In both raffinose and trehalose systems, when the initial glycine to sugar (raffinose pentahydrate or trehalose dihydrate) ratio was <1, glycine crystallization was not detected. When the ratio was >or=1, partial glycine crystallization was observed during warming. The presence of amorphous glycine caused the T'g to be substantially lower than that of the solution containing only the carbohydrate. To determine the extent of glycine crystallization, the solutions were annealed for 5 h just above the temperature of glycine crystallization. The T'g observed in the second warming curve was very close to that of the carbohydrate solution alone, indicating almost complete glycine crystallization. These studies enabled the construction of the water-rich sections of the raffinose-glycine-water and trehalose-glycine-water state diagrams. These diagrams consist of a kinetically stable freeze-concentrated solution and a doubly unstable glassy region, which readily crystallizes during cooling or subsequent warming. In addition, there is an intermediate region, where during the experimental timescale, there appears to be hindered glycine nucleation but unhindered crystal growth. To obtain substantially crystalline glycine in the frozen solutions, the glycine to carbohydrate ratios should be >or=1.

Calorimetry, Differential Scanning↗

PubMed2005

Partially crystalline systems in lyophilization: II. Withstanding collapse at high primary drying temperatures and impact on protein activity recovery.

In an accompanying article we have described the construction of the water-rich sections of raffinose-glycine-water and trehalose-glycine-water state diagrams. In this study, we use the information obtained from the state diagrams to identify the minimum weight fraction of the crystalline component in glycine-carbohydrate systems necessary to withstand collapse at high primary drying temperatures. We also determine the impact of primary drying, substantially above T'g, on the recovery of lactate dehydrogenase (LDH) activity. Ambient and variable temperature X-ray powder diffractometry and differential scanning calorimetry were used to characterize the frozen and freeze-dried systems. Aqueous solutions with glycine to carbohydrate (raffinose pentahydrate or trehalose dihydrate) weight ratios ranging from 0.2 to 2.0 were freeze dried. The protein formulations contained 20 mM citrate buffer (pH 6.0) and LDH (20 microg/mL). A glycine to anhydrous raffinose weight ratio >or=1.18 and a glycine to anhydrous trehalose weight ratio >or=1.56 were necessary to withstand macroscopic collapse in the system, when the primary drying was carried out at a product temperature at least 10 degrees C above the T'g. The recovery of LDH activity was almost complete in the reconstituted lyophile whether the primary drying was carried out above T'g (-10 degrees C) or below T'g (-32 degrees C). Thus, by judiciously combining crystalline and amorphous components, it was possible to primary dry at temperatures substantially above the T'g.

Chemistry, Pharmaceutical↗

PubMed2005

Raffinose crystallization during freeze-drying and its impact on recovery of protein activity.

PURPOSE: To study i) phase transitions in raffinose solution in the frozen state and during freeze-drying and ii) evaluate the impact of raffinose crystallization on the recovery of protein activity in reconstituted lyophiles. METHODS: X-ray powder diffractometry (XRD) and differential scanning calorimetry (DSC) were used to study the frozen aqueous solutions of raffinose pentahydrate. Phase transitions during primary and secondary drying were monitored by simulating the entire freeze-drying process, in situ, in the sample chamber of the diffractometer. The activity of lactate dehydrogenase (LDH) in reconstituted lyophiles was determined spectrophotometrically. RESULTS: Raffinose formed a kinetically stable amorphous freeze-concentrated phase when aqueous solutions were frozen at different cooling rates. When these solutions were subjected to primary drying without annealing, raffinose remained amorphous. Raffinose crystallized as the pentahydrate when the solutions were annealed at a shelf temperature of -10 degrees C. Primary drying of these annealed systems resulted in the dehydration of raffinose pentahydrate to an amorphous phase. The phase separation of the protein from the amorphous raffinose in these two systems during freeze-drying resulted in a significant reduction in the recovery of LDH activity, even though the lyophile was amorphous. CONCLUSIONS: Annealing of frozen aqueous raffinose solutions can result in solute crystallization, possibly as the pentahydrate. The crystalline pentahydrate dehydrates during primary drying to yield an amorphous lyophile. Raffinose crystallization during freeze-drying is accompanied by a significant loss of protein activity.

Calorimetry, Differential Scanning↗

PubMed2005

Crystallization of cephalothin sodium during lyophilization from tert-butyl alcohol-water cosolvent system.

PURPOSE: Because cephalothin sodium (I) does not crystallize readily when freeze-dried from aqueous solutions, organic cosolvents were used to increase the crystallinity of lyophilized I. METHODS: Compound I was lyophilized from water-organic cosolvent (5% w/w) systems of each ethanol, ispropanol, and tert-butyl alcohol (TBA). RESULTS: When frozen solutions of I (10% w/w) in each of these cosolvent systems was characterized by DSC, the presence of cosolvent in the freeze-concentrate was evident. Moreover, the presence of the cosolvent accelerated the solute crystallization. This observation was based on the XRD of these systems during the various stages of freeze-drying. High initial solute concentration and annealing of frozen solutions facilitated the formation of a highly crystalline lyophile. The accelerated crystallization is attributed to supersaturation in cosolvent systems, facilitating nucleation during freezing with subsequent growth during annealing. Lyophiles obtained from water-isopropanol and water-ethanol systems collapsed, while the use of TBA as a cosolvent yielded a friable and pharmaceutically elegant cake, containing fine needle-shaped crystals of I. Gas chromatography revealed a residual TBA concentration of approximately 0.001% w/w in the crystalline lyophiles. In general, residual cosolvent levels were higher in lyophiles with lower crystallinity. CONCLUSIONS: TBA-water was found to be a suitable freeze-drying medium to promote crystallization of I and yielded a lyophile with desirable product characteristics.

Cephalothin↗

PubMed2004

Solid-vapor interactions: influence of environmental conditions on the dehydration of carbamazepine dihydrate.

The goal of this research was a phenomenological study of the effect of environmental factors on the dehydration behavior of carbamazepine dihydrate. Dehydration experiments were performed in an automated vapor sorption apparatus under a variety of conditions, and weight loss was monitored as a function of time. In addition to lattice water, carbamazepine dihydrate contained a significant amount of physically bound water. Based on the kinetics of water loss, it was possible to differentiate between the removal of physically bound water and the lattice water. The activation energy for the 2 processes was 44 and 88 kJ/mol, respectively. As expected, the dehydration rate of carbamazepine dihydrate decreased with an increase in water vapor pressure. While dehydration at 0% relative humidity (RH) resulted in an amorphous anhydrate, the crystallinity of the anhydrate increased as a function of the RH of dehydration. A method was developed for in situ crystallinity determination of the anhydrate formed. Dehydration in the presence of the ethanol vapor was a 2-step process, and the fraction dehydrated at each step was a function of the ethanol vapor pressure. We hypothesize the formation of an intermediate lower hydrate phase with unknown water stoichiometry. An increase in the ethanol vapor pressure first led to a decrease in the dehydration rate followed by an increase. In summary, the dehydration behavior of carbamazepine dihydrate was evaluated at different vapor pressures of water and ethanol. Using the water sorption apparatus, it was possible to (1) differentiate between the removal of physically bound and lattice water, and (2) develop a method for quantifying, in situ, the crystallinity of the product (anhydrate) phase.

Anhydrides↗

PubMed2004

Influence of processing-induced phase transformations on the dissolution of theophylline tablets.

The object of this investigation was to evaluate the influence of (1) processing-induced decrease in drug crystallinity and (2) phase transformations during dissolution, on the performance of theophylline tablet formulations. Anhydrous theophylline underwent multiple transformations (anhydrate --> hydrate --> anhydrate) during processing. Although the crystallinity of the anhydrate obtained finally was lower than that of the unprocessed drug, it dissolved at a slower rate. This decrease in dissolution rate was attributed to the accelerated anhydrate to hydrate transformation during the dissolution run. Water vapor sorption studies proved to be a good predictor of powder dissolution behavior. While a decrease in crystallinity was brought about either by milling or by granulation, the effect on tablet dissolution was pronounced only in the latter. Tablet formulations prepared from the granules exhibited higher hardness, longer disintegration time, and slower dissolution than those containing the milled drug. The granules underwent plastic deformation during compression resulting in harder tablets, with delayed disintegration. The high hardness coupled with rapid anhydrate --> hydrate transformation during dissolution resulted in the formation of a hydrate layer on the tablet surface, which further delayed tablet disintegration and, consequently, dissolution. Phase transformations during processing and, more importantly, during dissolution influenced the observed dissolution rates. Product performance was a complex function of the physical state of the active and the processing conditions.

Cellulose↗

PubMed2004

Preformulation studies to meet the challenges in the manufacture of betaine solid dosage form.

The objectives were (1). to perform solid-state characterization of anhydrous betaine (A) and betaine monohydrate (M), (2). to develop a pressure differential scanning calorimetric (DSC) technique for the quantification of M when present as a minor component in a mixture with A and, (3). to study the effect of annealing of A on the kinetics of A --> M transition. X-ray powder diffractometer (XRD), DSC, thermogravimetric analyzer (TGA), and an automated moisture sorption apparatus were used to characterize the phases. DSC at an elevated pressure of 200 psi enabled quantification of M in mixtures of A and M. Humidity-controlled TGA allowed study of the kinetics of A --> M transition. Automated moisture studies showed that A has a strong tendency to sorb water (at RH >or= 20%, 25 degrees C) and convert to M. When M was subjected to DSC at ambient pressure, the endotherms due to dehydration and vaporization of water overlapped. Pressure DSC enabled separation of these two thermal events. In mixtures of A and M, the enthalpy of dehydration (deltaH(d)) of M could be used for its quantification. A linear relationship was obtained when deltaH(d) was plotted as a function of the weight fraction of M in the mixture. The limits of detection and quantification of M in A were 0.15% and 1.5% w/w, respectively. The kinetics of water uptake by the annealed as well as the unannealed A, could be best described by the Avrami-Erofeev model (three-dimensional nucleation and growth). The calculated rate constant (k) of unannealed A (0.075 +/- 0.002 min(-1)) was significantly higher than that of annealed A (0.052 +/- 0.004 min(-1)). DSC at elevated pressure was a sensitive technique for quantification of M when present as a mixture with A. Annealing of A decelerated the A --> M phase transition reaction, possibly by increasing the degree of crystallinity of A.

Betaine↗

PubMed2004

Infrared spectroscopic studies of protein formulations containing glycine.

Glycine is extensively used as an excipient in protein formulations. However, it absorbs significant infrared (IR) radiation in the conformationally sensitive amide I region (1700-1600 cm(-1)) of proteins. Furthermore, glycine can form a number of polymorphs, as well as an amorphous phase. Each of these forms possibly exhibits a different IR absorption spectrum. Accurate subtraction of glycine signals, in order to obtain reliable amide I spectra, was found to be possible only if the protein-to-glycine ratio was >/=1:1. In those cases, the solid-state conformation of the protein could be determined. In addition, a new method for estimating the degree of crystallinity of freeze-dried glycine is described, using IR bands in the 1350-1300 cm(-1) region.

Chemistry, Pharmaceutical↗

PubMed2004

Quantification of glycine crystallinity by near-infrared (NIR) spectroscopy.

The object of this investigation was to use near-infrared (NIR) spectroscopy for quantification of glycine crystallinity. Glycine samples, with different degrees of crystallinity, were obtained by physically mixing different proportions of crystalline beta-glycine with amorphous glycine. NIR spectra were obtained, directly from samples in glass vials, over the wavelength range of 1100-2500 nm. A partial least squares (PLS) model was developed to correlate the NIR spectral changes with the degree of crystallinity. Using this model, a standard error of calibration (SEC) of 2.1% was obtained with an r(2) value of 0.996. Cross validation was used to test the precision of the quantitative model, resulting in a standard error of prediction (SEP) of 3.2%. These results indicate that NIR spectroscopy is well suited to the measurement of glycine crystallinity in lyophilized products. Employing the PLS model, the crystallinity of glycine in freeze-dried sucrose-glycine mixtures was evaluated. At a sucrose to glycine ratio >4, glycine crystallization during lyophilization was inhibited. Conversely, at ratios < or =0.67, glycine remained substantially crystalline. At intermediate compositions, the glycine was partially crystalline.

Crystallization↗