On- and off-line analysis by ICP-MS to measure the bioaccessible concentration of elements in PM10 using dynamic versions of the simplified bioaccessibility extraction test.

Two dynamic versions of the simplified bioaccessibility extraction test (SBET) were developed-an off-line procedure and an on-line procedure coupled directly to ICP-MS. Batch, on-line, and off-line procedures were applied to simulated PM10 samples prepared by loading NIST SRM 2711A Montana II Soil and BGS RM 102 Ironstone Soil onto 45-mm TX40 filters widely used in air quality monitoring. Three real PM10 samples were also extracted. A polycarbonate filter holder was used as an extraction unit for the dynamic procedures. Arsenic, Cd, Cr, Cu, Fe, Mn, Ni, Pb, and Zn were determined in the extracts using an Agilent 7700 × ICP-MS instrument. The residual simulated PM10 samples following application of the SBET were subjected to microwave-assisted aqua regia digestion and a mass balance calculation performed with respect to digestion of a separate test portion of the SRM. Leachates were collected as subfractions for the off-line analysis or continuously introduced to the nebuliser of the ICP-MS for the on-line analysis. The mass balance was generally acceptable for all versions of the SBET. Recoveries obtained with the dynamic methods were closer to pseudototal values than those obtained in batch mode. Off-line analysis performed better than on-line analysis, except for Pb. Recoveries of bioaccessible Pb relative to the certified value in NIST SRM 2711A Montana II Soil (1110 ± 49 mg kg-1) were 99, 106, and 105% for the batch, off-line, and on-line methods, respectively. The study demonstrates that dynamic SBET can be used to measure bioaccessibility of potentially toxic elements in PM10 samples.

Axillary dissection versus no axillary dissection in patients with breast cancer and sentinel-node micrometastases (IBCSG 23-01): 10-year follow-up of a randomised, controlled phase 3 trial.

BACKGROUND: We previously reported the 5-year results of the phase 3 IBCSG 23-01 trial comparing disease-free survival in patients with breast cancer with one or more micrometastatic (≤2 mm) sentinel nodes randomly assigned to either axillary dissection or no axillary dissection. The results showed no difference in disease-free survival between the groups and showed non-inferiority of no axillary dissection relative to axillary dissection. The current analysis presents the results of the study after a median follow-up of 9·7 years (IQR 7·8-12·7). METHODS: In this multicentre, randomised, controlled, open-label, non-inferiority, phase 3 trial, participants were recruited from 27 hospitals and cancer centres in nine countries. Eligible women could be of any age with clinical, mammographic, ultrasonographic, or pathological diagnosis of breast cancer with largest lesion diameter of 5 cm or smaller, and one or more metastatic sentinel nodes, all of which were 2 mm or smaller and with no extracapsular extension. Patients were randomly assigned (1:1) before surgery (mastectomy or breast-conserving surgery) to no axillary dissection or axillary dissection using permuted blocks generated by a web-based congruence algorithm, with stratification by centre and menopausal status. The protocol-specified primary endpoint was disease-free survival, analysed in the intention-to-treat population (as randomly assigned). Safety was assessed in all randomly assigned patients who received their allocated treatment (as treated). We did a one-sided test for non-inferiority of no axillary dissection by comparing the observed hazard ratios (HRs) for disease-free survival with a margin of 1·25. This 10-year follow-up analysis was not prespecified in the trial's protocol and thus was not adjusted for multiple, sequential testing. This trial is registered with ClinicalTrials.gov, number NCT00072293. FINDINGS: Between April 1, 2001, and Feb 8, 2010, 6681 patients were screened and 934 randomly assigned to no axillary dissection (n=469) or axillary dissection (n=465). Three patients were ineligible and were excluded from the trial after randomisation. Disease-free survival at 10 years was 76·8% (95% CI 72·5-81·0) in the no axillary dissection group, compared with 74·9% (70·5-79·3) in the axillary dissection group (HR 0·85, 95% CI 0·65-1·11; log-rank p=0·24; p=0·0024 for non-inferiority). Long-term surgical complications included lymphoedema of any grade in 16 (4%) of 453 patients in the no axillary dissection group and 60 (13%) of 447 in the axillary dissection group, sensory neuropathy of any grade in 57 (13%) in the no axillary dissection group versus 85 (19%) in the axillary dissection group, and motor neuropathy of any grade (14 [3%] in the no axillary dissection group vs 40 [9%] in the axillary dissection group). One serious adverse event (postoperative infection and inflamed axilla requiring hospital admission) was attributed to axillary dissection; the event resolved without sequelae. INTERPRETATION: The findings of the IBCSG 23-01 trial after a median follow-up of 9·7 years (IQR 7·8-12·7) corroborate those obtained at 5 years and are consistent with those of the 10-year follow-up analysis of the Z0011 trial. Together, these findings support the current practice of not doing an axillary dissection when the tumour burden in the sentinel nodes is minimal or moderate in patients with early breast cancer. FUNDING: International Breast Cancer Study Group.

A novel two-step sequential bioaccessibility test for potentially toxic elements in inhaled particulate matter transported into the gastrointestinal tract by mucociliary clearance.

A novel two-step sequential extraction has been developed to assess the bioaccessibility of As, Cd, Cr, Cu, Fe, Mn, Ni, Pb and Zn in airborne particulate matter following inhalation and transport into the human gastrointestinal tract by mucociliary clearance. A new artificial mucus fluid (AMF) was used to determine the bioaccessible potentially toxic element (PTE) fraction in the upper airways, in sequence with the simplified bioaccessibility extraction test (SBET) or the stomach phase of the unified bioaccessibility method (gastric fluid only) (UBMG). Filter dynamic measurement system TX40 filters smeared with soil reference material (BGS RM 102) were used as test samples. Analysis was performed by ICP-MS. Comparison between results obtained for soil alone and when the soil was supported on TX40 filters indicated that the presence of the substrate did not affect the extraction efficiency, although a large Zn blank was detected. The sequential AMF→SBET extraction liberated similar amounts of Fe, Mn, Ni and Zn to the SBET alone; but significantly less Cd; and significantly more As, Cr, Cu and Pb. The sequential AMF→UBMG extraction liberated similar amounts of Cd, Cr, Mn and Zn to the UBMG alone, but significantly less As, Fe and Ni; and significantly more Cu and Pb. Enhanced extractability was due to the greater quantities of exchangeable ions and complexing agents present. Adoption of a two-step sequential extraction (AMF followed by either the SBET or the UBMG) is recommended because it is more representative of biological conditions and avoids overestimation or underestimation of bioaccessible PTE concentrations. Graphical Abstract Simulated PM10 sample: BGS RM 102 ironstone soil on TX40 filter.

Axillary dissection versus no axillary dissection in patients with sentinel-node micrometastases (IBCSG 23-01): a phase 3 randomised controlled trial.

BACKGROUND: For patients with breast cancer and metastases in the sentinel nodes, axillary dissection has been standard treatment. However, for patients with limited sentinel-node involvement, axillary dissection might be overtreatment. We designed IBCSG trial 23-01 to determine whether no axillary dissection was non-inferior to axillary dissection in patients with one or more micrometastatic (≤2 mm) sentinel nodes and tumour of maximum 5 cm. METHODS: In this multicentre, randomised, non-inferiority, phase 3 trial, patients were eligible if they had clinically non-palpable axillary lymph node(s) and a primary tumour of 5 cm or less and who, after sentinel-node biopsy, had one or more micrometastatic (≤2 mm) sentinel lymph nodes with no extracapsular extension. Patients were randomly assigned (in a 1:1 ratio) to either undergo axillary dissection or not to undergo axillary dissection. Randomisation was stratified by centre and menopausal status. Treatment assignment was not masked. The primary endpoint was disease-free survival. Non-inferiority was defined as a hazard ratio (HR) of less than 1·25 for no axillary dissection versus axillary dissection. The analysis was by intention to treat. Per protocol, disease and survival information continues to be collected yearly. This trial is registered with ClinicalTrials.gov, NCT00072293. FINDINGS: Between April 1, 2001, and Feb 28, 2010, 465 patients were randomly assigned to axillary dissection and 469 to no axillary dissection. After the exclusion of three patients, 464 patients were in the axillary dissection group and 467 patients were in the no axillary dissection group. After a median follow-up of 5·0 (IQR 3·6-7·3) years, we recorded 69 disease-free survival events in the axillary dissection group and 55 events in the no axillary dissection group. Breast-cancer-related events were recorded in 48 patients in the axillary dissection group and 47 in the no axillary dissection group (ten local recurrences in the axillary dissection group and eight in the no axillary dissection group; three and nine contralateral breast cancers; one and five [corrected] regional recurrences; and 34 and 25 distant relapses). Other non-breast cancer events were recorded in 21 patients in the axillary dissection group and eight in the no axillary dissection group (20 and six second non-breast malignancies; and one and two deaths not due to a cancer event). 5-year disease-free survival was 87·8% (95% CI 84·4-91·2) in the group without axillary dissection and 84·4% (80·7-88·1) in the group with axillary dissection (log-rank p=0·16; HR for no axillary dissection vs axillary dissection was 0·78, 95% CI 0·55-1·11, non-inferiority p=0·0042). Patients with reported long-term surgical events (grade 3-4) included one sensory neuropathy (grade 3), three lymphoedema (two grade 3 and one grade 4), and three motor neuropathy (grade 3), all in the group that underwent axillary dissection, and one grade 3 motor neuropathy in the group without axillary dissection. One serious adverse event was reported, a postoperative infection in the axilla in the group with axillary dissection. INTERPRETATION: Axillary dissection could be avoided in patients with early breast cancer and limited sentinel-node involvement, thus eliminating complications of axillary surgery with no adverse effect on survival. FUNDING: None.

Towards better understanding of an industrial cell factory: investigating the feasibility of real-time metabolic flux analysis in Pichia pastoris.

BACKGROUND: Novel analytical tools, which shorten the long and costly development cycles of biopharmaceuticals are essential. Metabolic flux analysis (MFA) shows great promise in improving our understanding of the metabolism of cell factories in bioreactors, but currently only provides information post-process using conventional off-line methods. MFA combined with real time multianalyte process monitoring techniques provides a valuable platform technology allowing real time insights into metabolic responses of cell factories in bioreactors. This could have a major impact in the bioprocessing industry, ultimately improving product consistency, productivity and shortening development cycles. RESULTS: This is the first investigation using Near Infrared Spectroscopy (NIRS) in situ combined with metabolic flux modelling which is both a significant challenge and considerable extension of these techniques. We investigated the feasibility of our approach using the industrial workhorse Pichia pastoris in a simplified model system. A parental P. pastoris strain (i.e. which does not synthesize recombinant protein) was used to allow definition of distinct metabolic states focusing solely upon the prediction of intracellular fluxes in central carbon metabolism. Extracellular fluxes were determined using off-line conventional reference methods and on-line NIR predictions (calculated by multivariate analysis using the partial least squares algorithm, PLS). The results showed that the PLS-NIRS models for biomass and glycerol were accurate: correlation coefficients, R2, above 0.90 and the root mean square error of prediction, RMSEP, of 1.17 and 2.90 g/L, respectively. The analytical quality of the NIR models was demonstrated by direct comparison with the standard error of the laboratory (SEL), which showed that performance of the NIR models was suitable for quantifying biomass and glycerol for calculating extracellular metabolite rates and used as independent inputs for the MFA (RMSEP lower than 1.5 × SEL). Furthermore, the results for the MFA from both datasets passed consistency tests performed for each steady state, showing that the precision of on-line NIRS is equivalent to that obtained by the off-line measurements. CONCLUSIONS: The findings of this study show for the first time the potential of NIRS as an input generating for MFA models, contributing to the optimization of cell factory metabolism in real-time.

Effect of particle properties of powders on the generation and transmission of Raman scattering.

Transmission Raman measurements of a 1 mm thick sulfur-containing disk were made at different positions as it was moved through 4 mm of aspirin (150-212 µm) or microcrystalline cellulose (Avicel) of different size ranges (<38, 53-106, and 150-212 µm). The transmission Raman intensity of the sulfur interlayer at 218 cm(-1) was lower when the disk was placed at the top or bottom of the powder bed, compared to positions within the bed and the difference between the sulfur intensity at the outer and inner positions increased with Avicel particle size. Also, the positional intensity difference was smaller for needle-shaped aspirin than for granular Avicel of the same size. The attenuation coefficients for the propagation of the exciting laser and transmitted Raman photons through the individual powders were the same but decreased as the particle size of Avicel increased; also, the attenuation coefficients for propagation through 150-212 µm aspirin were almost half of those through similar sized Avicel particles. The study has demonstrated that particulate size and type affect transmitted Raman intensities and, consequently, such factors need to be considered in the analysis of powders, especially if particle properties vary between the samples.

Comparison of the determination of a low-concentration active ingredient in pharmaceutical tablets by backscatter and transmission Raman spectrometry.

A total of 383 tablets of a pharmaceutical product were analyzed by backscatter and transmission Raman spectrometry to determine the concentration of an active pharmaceutical ingredient (API), chlorpheniramine maleate, at the 2% m/m (4 mg) level. As the exact composition of the tablets was unknown, external calibration samples were prepared from chlorpheniramine maleate and microcrystalline cellulose (Avicel) of different particle size. The API peak at 1594 cm(-1) in the second derivative Raman spectra was used to generate linear calibration models. The API concentration predicted using backscatter Raman measurements was relatively insensitive to the particle size of Avicel. With transmission, however, particle size effects were greater and accurate prediction of the API content was only possible when the photon propagation properties of the calibration and sample tablets were matched. Good agreement was obtained with HPLC analysis when matched calibration tablets were used for both modes. When the calibration and sample tablets are not chemically matched, spectral normalization based on calculation of relative intensities cannot be used to reduce the effects of differences in physical properties. The main conclusion is that although better for whole tablet analysis, transmission Raman is more sensitive to differences in the photon propagation properties of the calibration and sample tablets.

Quantitative analysis of powder mixtures by Raman spectrometry: the influence of particle size and its correction.

Particle size distribution and compactness have significant confounding effects on Raman signals of powder mixtures, which cannot be effectively modeled or corrected by traditional multivariate linear calibration methods such as partial least-squares (PLS), and therefore greatly deteriorate the predictive abilities of Raman calibration models for powder mixtures. The ability to obtain directly quantitative information from Raman signals of powder mixtures with varying particle size distribution and compactness is, therefore, of considerable interest. In this study, an advanced quantitative Raman calibration model was developed to explicitly account for the confounding effects of particle size distribution and compactness on Raman signals of powder mixtures. Under the theoretical guidance of the proposed Raman calibration model, an advanced dual calibration strategy was adopted to separate the Raman contributions caused by the changes in mass fractions of the constituents in powder mixtures from those induced by the variations in the physical properties of samples, and hence achieve accurate quantitative determination for powder mixture samples. The proposed Raman calibration model was applied to the quantitative analysis of backscatter Raman measurements of a proof-of-concept model system of powder mixtures consisting of barium nitrate and potassium chromate. The average relative prediction error of prediction obtained by the proposed Raman calibration model was less than one-third of the corresponding value of the best performing PLS model for mass fractions of barium nitrate in powder mixtures with variations in particle size distribution, as well as compactness.

Validity of particle size analysis techniques for measurement of the attrition that occurs during vacuum agitated powder drying of needle-shaped particles.

Analysis of needle-shaped particles of cellobiose octaacetate (COA) obtained from vacuum agitated drying experiments was performed using three particle size analysis techniques: laser diffraction (LD), focused beam reflectance measurements (FBRM) and dynamic image analysis. Comparative measurements were also made for various size fractions of granular particles of microcrystalline cellulose. The study demonstrated that the light scattering particle size methods (LD and FBRM) can be used qualitatively to study the attrition that occurs during drying of needle-shaped particles, however, for full quantitative analysis, image analysis is required. The algorithm used in analysis of LD data assumes the scattering particles are spherical regardless of the actual shape of the particles under evaluation. FBRM measures a chord length distribution (CLD) rather than the particle size distribution (PSD), which in the case of needles is weighted towards the needle width rather than their length. Dynamic image analysis allowed evaluation of the particles based on attributes of the needles such as length (e.g. the maximum Feret diameter) or width (e.g. the minimum Feret diameter) and as such, was the most informative of the techniques for the analysis of attrition that occurred during drying.

Calibration model transfer in mid-infrared process analysis with in situ attenuated total reflectance immersion probes.

Process applications of mid-infrared (MIR) spectrometry may involve replacement of the spectrometer and/or measurement probe, which generally requires a calibration transfer method to maintain the accuracy of analysis. In this study, direct standardisation (DS), piecewise direct standardisation (PDS) and spectral space transformation (SST) were compared for analysis of ternary mixtures of acetone, ethanol and ethyl acetate. Three calibration transfer examples were considered: changing the spectrometer, multiplexing two probes to a spectrometer, and changing the diameter of the attenuated total reflectance (ATR) probe (as might be required when scaling up from lab to process analysis). In each case, DS, PDS and SST improved the accuracy of prediction for the test samples, analysed on a secondary spectrometer-probe combination, using a calibration model developed on the primary system. When the probe diameter was changed, a scaling step was incorporated into SST to compensate for the change in absorbance caused by the difference in ATR crystal size. SST had some advantages over DS and PDS: DS was sensitive to the choice of standardisation samples, and PDS required optimisation of the window size parameter (which also required an extra standardisation sample). SST only required a single parameter to be chosen: the number of principal components, which can be set equal to the number of standardisation samples when a low number of standards (n < 7) are used, which is preferred to minimise the time required to transfer the calibration model.

Improved method for kinetic studies in microreactors using flow manipulation and noninvasive Raman spectrometry.

A novel method has been devised to derive kinetic information about reactions in microfluidic systems. Advantages have been demonstrated over conventional procedures for a Knoevenagel condensation reaction in terms of the time required to obtain the data (fivefold reduction) and the efficient use of reagents (tenfold reduction). The procedure is based on a step change from a low (e.g., 0.6 µL min(-1)) to a high (e.g., 14 µL min(-1)) flow rate and real-time noninvasive Raman measurements at the end of the flow line, which allows location-specific information to be obtained without the need to move the measurement probe along the microreactor channel. To validate the method, values of the effective reaction order n were obtained employing two different experimental methodologies. Using these values of n, rate constants k were calculated and compared. The values of k derived from the proposed method at 10 and 40 °C were 0.0356 ± 0.0008 mol(-0.3) dm(0.9) s(-1) (n = 1.3) and 0.24 ± 0.018 mol(-0.1) dm(0.3) s(-1) (n = 1.1), respectively, whereas the values obtained using a more laborious conventional methodology were 0.0335 ± 0.0032 mol(-0.4) dm(1.2) s(-1) (n = 1.4) at 10 °C and 0.244 ± 0.032 mol(-0.3) dm(0.9) s(-1) (n = 1.3) at 40 °C. The new approach is not limited to analysis by Raman spectrometry and can be used with different techniques that can be incorporated into the end of the flow path to provide rapid measurements.

Calibration of multiplexed fiber-optic spectroscopy.

Large-scale commercial bioprocesses that manufacture biopharmaceutical products such as monoclonal antibodies generally involve multiple bioreactors operated in parallel. Spectra recorded during in situ monitoring of multiple bioreactors by multiplexed fiber-optic spectroscopies contain not only spectral information of the chemical constituents but also contributions resulting from differences in the optical properties of the probes. Spectra with variations induced by probe differences cannot be efficiently modeled by the commonly used multivariate linear calibration models or effectively removed by popular empirical preprocessing methods. In this study, for the first time, a calibration model is proposed for the analysis of complex spectral data sets arising from multiplexed probes. In the proposed calibration model, the spectral variations introduced by probe differences are explicitly modeled by introducing a multiplicative parameter for each optical probe, and then their detrimental effects are effectively mitigated through a "dual calibration" strategy. The performance of the proposed multiplex calibration model has been tested on two multiplexed spectral data sets (i.e., MIR data of ternary mixtures and NIR data of bioprocesses). Experimental results suggest that the proposed calibration model can effectively mitigate the detrimental effects of probe differences and hence provide much more accurate predictions than commonly used multivariate linear calibration models (such as PLS) with and without empirical data preprocessing methods such as orthogonal signal correction, standard normal variate, or multiplicative signal correction.

Studies of particle drying using non-invasive Raman spectrometry and particle size analysis.

The evaporation of methanol from needle-shaped particles of cellobiose octaacetate (COA) has been studied directly in a jacketed vacuum drier using in situ measurements by Raman spectrometry. A design of experiments (DoE) approach was used to investigate the effects of three parameters (method of agitation, % solvent loss on drying and jacket temperature), with the intention of minimising the drying time and extent of particle attrition. Drying curves based on Raman signals for methanol and COA in the spectra of the wet particles indicated the end of drying and revealed three stages in the drying process that could be used to monitor the progress of solvent removal in real time. Off-line particle size measurements based on laser diffraction were made to obtain information on the extent of attrition, to compare with the trends revealed by the Raman drying curves. The study demonstrated that non-invasive Raman spectrometry can be used to study the progress of drying during agitation of particles in a vacuum drier, allowing optimisation of operating conditions to minimise attrition and reduce drying times. Although a correlation between particle size and off-line Raman measurements of COA was demonstrated, it was not possible to derive equivalent information from the in situ Raman spectra owing to the greater effects of particle motion or bulk density variations of the particles in the drier.

Systematic prediction error correction: a novel strategy for maintaining the predictive abilities of multivariate calibration models.

The development of reliable multivariate calibration models for spectroscopic instruments in on-line/in-line monitoring of chemical and bio-chemical processes is generally difficult, time-consuming and costly. Therefore, it is preferable if calibration models can be used for an extended period, without the need to replace them. However, in many process applications, changes in the instrumental response (e.g. owing to a change of spectrometer) or variations in the measurement conditions (e.g. a change in temperature) can cause a multivariate calibration model to become invalid. In this contribution, a new method, systematic prediction error correction (SPEC), has been developed to maintain the predictive abilities of multivariate calibration models when e.g. the spectrometer or measurement conditions are altered. The performance of the method has been tested on two NIR data sets (one with changes in instrumental responses, the other with variations in experimental conditions) and the outcomes compared with those of some popular methods, i.e. global PLS, univariate slope and bias correction (SBC) and piecewise direct standardization (PDS). The results show that SPEC achieves satisfactory analyte predictions with significantly lower RMSEP values than global PLS and SBC for both data sets, even when only a few standardization samples are used. Furthermore, SPEC is simple to implement and requires less information than PDS, which offers advantages for applications with limited data.

Non-invasive analysis in micro-reactors using Raman spectrometry with a specially designed probe.

An optical interface has been designed to maximise the sensitivity and spatial resolution required when Raman spectrometry is used to monitor a reaction in a micro-reactor, revealing advantages over a conventional commercial probe. A miniature aspheric lens was shown to be better than microscope objectives to focus the probing laser beam onto the sample. The diameters of the exciting and collection optical fibres were also shown to have a significant influence on sensitivity and the signal-to-background ratio, with 62.5 microm diameter 0.28 numerical aperture (NA) fibres found to be best for analysis of liquids in the 150 microm deep channel in the micro-reactor used. With a spectral measurement time of 2 s, it was shown that the probe could monitor the progress of an esterification reaction in real time and quickly optimise the reagent flow rates. The fast response time revealed features related to short-term pump instabilities and micro-reactor rheology effects that would not have been identified without rapid real-time measurements.

Non-invasive monitoring of the mixing of pharmaceutical powders by broadband acoustic emission.

Broadband acoustic transducers, including an intrinsically safe device, were assessed for non-invasive monitoring of aspirin, citric acid or Avicel mixing in a bench scale convective blender. The frequency information content of the acoustic emission (AE) spectra depends on the response characteristics of the transducers, which vary depending on the design. As acoustic waves generated from the impact of particles propagated through and around the glass mixing vessel, comparable spectra were obtained from different locations on the glass. The intensity of AE increased as the impeller speed, mass of powder or density of the particles was increased. AE also increased with particle size, with a relatively greater increase in intensity at lower frequencies. Mixing profiles were generated in real time from the change in the integrated intensity over selected frequency ranges on addition of aspirin to Avicel; the AE signal initially increased and then came to a plateau as the mixture became homogeneous. The average plateau signal was plotted against concentration for three different particle size ranges of aspirin in Avicel; for aspirin concentrations <21% m/m the increase in the AE was relatively small with no discernable effects of the aspirin particle size; however, for >21% m/m aspirin, there was a proportionally greater increase in AE, and particle size effects were more obvious. The study has shown that AE is relatively easy to measure non-invasively during powder mixing, but has poorer sensitivity than NIR spectrometry for detection of effects caused by addition of secondary compounds, especially at smaller particle sizes.

A rural perspective on minimally invasive parathyroidectomy: optimal preoperative imaging and patient outcomes.

BACKGROUND: Our retrospective review of prospectively collected data evaluated the efficacy of minimally invasive parathyroidectomy (MIP) and compared preoperative imaging modalities in a rural referral centre. METHODS: Patients with a diagnosis of primary hyperparathyroidism underwent surgeon-performed ultrasound (SUS) and technetium-99 m sestamibi (MIBI). Radiologist-performed ultrasound (RUS) was sought when the diagnosis remained in doubt. Four-dimensional computed tomography (4DCT) first replaced RUS in mid-2014, then MIBI as a frontline modality in 2015. MIP was conducted if possible and bilateral neck exploration (BNE) when localization remained doubtful. Treatment was evaluated by histopathology and serum parathyroid hormone and calcium levels at 6 weeks. RESULTS: A total of 122 of 165 (73.9%) glands were removed by MIP and 43 of 165 (26.1%) by BNE. Of 15 cases with non-localizing preoperative investigations, one patient had a negative BNE. A total of 160 of 165 (97.0%) patients underwent a successful operation, with five (3.0%) suffering persistent post-operative hypercalcaemia. SUS had a sensitivity of 79.4% (131/165) and a positive predictive value (PPV) of 97.0% (131/135). MIBI had a sensitivity of 60.0% (81/135) and a PPV of 95.3% (81/85). RUS produced a sensitivity of 65.5% (76/116) and PPV of 98.7% (76/77). When used as a second-line modality, 4DCT had a sensitivity of 76.9% (10/13) and PPV of 100%. The sensitivity and PPV were 85.7% (18/21) and 94.7% (18/19) after 4DCT's promotion to first-line use. CONCLUSION: MIP can be safely performed in rural centres of adequate volume. We recommend that operations be guided by SUS with routine use of an adjunctive modality. Our study has seen 4DCT replace MIBI in this regard.

Advanced calibration strategy for in situ quantitative monitoring of phase transition processes in suspensions using FT-Raman spectroscopy.

There is an increasing interest in using Raman spectroscopy to identify polymorphic forms and monitor phase changes in pharmaceutical products for quality control. Compared with other analytical techniques for the identification of polymorphs such as X-ray powder diffractometry and infrared spectroscopy, FT-Raman spectroscopy has the advantages of enabling fast, in situ, and nondestructive measurements of complex systems such as suspension samples. However, for suspension samples, Raman intensities depend on the analyte concentrations as well as the particle size, overall solid content, and homogeneity of the solid phase in the mixtures, which makes quantitative Raman analysis rather difficult. In this contribution, an advanced model has been derived to explicitly account for the confounding effects of a sample's physical properties on Raman intensities. On the basis of this model, a unique calibration strategy called multiplicative effects correction (MEC) was proposed to separate the Raman contributions due to changes in analyte concentration from those caused by the multiplicative confounding effects of the sample's physical properties. MEC has been applied to predict the anhydrate concentrations from in situ FT-Raman measurements made during the crystallization and phase transition processes of citric acid in water. The experimental results show that MEC can effectively correct for the confounding effects of the particle size and overall solid content of the solid phase on Raman intensities and, therefore, provide much more accurate in situ quantitative predictions of anhydrate concentration during crystallization and phase transition processes than traditional PLS calibration methods.

In situ monitoring of the seed stage of a fermentation process using non-invasive NIR spectrometry.

Non-invasive NIR spectrometry has been used to monitor in situ the seed stage of a streptomyces fermentation process. The main spectral change occurred at 7263 cm(-1) in the 1st derivative spectrum, and from comparison with off-line NIR spectra acquired of components present in the fermentation broth, can be attributed to biomass. The biomass signal was constant for the first 20 h of the seed stage, after which it decreased before increasing again. The time at which the minimum occurred in the NIR profile was either the same or slightly earlier than that at which a maximum in the carbon dioxide evolution rate (CER) occurred. The changes observed for the biomass signal in the NIR spectra can be attributed to growth and then fragmentation of mycelia, which indicates a change in metabolic activity. Hence, it may be possible to use NIR spectrometry in situ to determine the optimum transfer time for the seed stage of a fermentation process. Spectra were also acquired of the final stage of the same fermentation process. The variation in the biomass signal in the NIR spectra was more complicated in the final stage owing to changes in stir rate, and biomass concentration and morphology.

Effects of particle size and cohesive properties on mixing studied by non-contact NIR.

A scaled-down convective blender was used along with non-invasive NIR spectrometry to study the mixing of citric acid, aspirin, aspartame or povidone with microcrystalline cellulose. NIR mixing profiles were generated in real time using measurements at the 2nd overtone wavelength of the added compounds. Trends demonstrated previously for aspirin were confirmed for additions of citric acid: the magnitude of the 2nd overtone NIR measurements is less affected by changes in particle size than that of the 1st overtone; the peak-to-peak noise of the 2nd overtone NIR mixing profile increases with the particle size of the added compound. The study has demonstrated the usefulness of continuous NIR measurements for rapid evaluation of the mixing process when deciding the best particle size of microcrystalline cellulose to mix with compounds of different particle shape and cohesive properties. Smaller particle sizes of microcrystalline cellulose (53-106 microm) were better for aspirin (212-250 microm), whereas larger particles (212-250 microm) were better for aspartame (212-250 microm). The characteristics of the compounds also need to be considered when deciding the order of addition of secondary compounds when mixed with microcrystalline cellulose. The time required to achieve a uniform mixture was much less when povidone was added before aspirin, rather than vice versa.