In-line UV spectroscopy for the quantification of low-dose active ingredients during the manufacturing of pharmaceutical semi-solid and liquid formulations.

UltraViolet (UV) spectroscopy was evaluated as an innovative Process Analytical Technology (PAT) - tool for the in-line and real-time quantitative determination of low-dosed active pharmaceutical ingredients (APIs) in a semi-solid (gel) and a liquid (suspension) pharmaceutical formulation during their batch production process. The performance of this new PAT-tool (i.e., UV spectroscopy) was compared with an already more established PAT-method based on Raman spectroscopy. In-line UV measurements were carried out with an immersion probe while for the Raman measurements a non-contact PhAT probe was used. For both studied formulations, an in-line API quantification model was developed and validated per spectroscopic technique. The known API concentrations (Y) were correlated with the corresponding in-line collected preprocessed spectra (X) through a Partial Least Squares (PLS) regression. Each developed quantification method was validated by calculating the accuracy profile on the basis of the validation experiments. Furthermore, the measurement uncertainty was determined based on the data generated for the determination of the accuracy profiles. From the accuracy profile of the UV- and Raman-based quantification method for the gel, it was concluded that at the target API concentration of 2% (w/w), 95 out of 100 future routine measurements given by the Raman method will not deviate more than 10% (relative error) from the true API concentration, whereas for the UV method the acceptance limits of 10% were exceeded. For the liquid formulation, the Raman method was not able to quantify the API in the low-dosed suspension (0.09% (w/w) API). In contrast, the in-line UV method was able to adequately quantify the API in the suspension. This study demonstrated that UV spectroscopy can be adopted as a novel in-line PAT-technique for low-dose quantification purposes in pharmaceutical processes. Important is that none of the two spectroscopic techniques was superior to the other for both formulations: the Raman method was more accurate in quantifying the API in the gel (2% (w/w) API), while the UV method performed better for API quantification in the suspension (0.09% (w/w) API).

Batch statistical process control of a fluid bed granulation process using in-line spatial filter velocimetry and product temperature measurements.

Fluid bed granulation is a batch process, which is characterized by the processing of raw materials for a predefined period of time, consisting of a fixed spraying phase and a subsequent drying period. The present study shows the multivariate statistical modeling and control of a fluid bed granulation process based on in-line particle size distribution (PSD) measurements (using spatial filter velocimetry) combined with continuous product temperature registration using a partial least squares (PLS) approach. Via the continuous in-line monitoring of the PSD and product temperature during granulation of various reference batches, a statistical batch model was developed allowing the real-time evaluation and acceptance or rejection of future batches. Continuously monitored PSD and product temperature process data of 10 reference batches (X-data) were used to develop a reference batch PLS model, regressing the X-data versus the batch process time (Y-data). Two PLS components captured 98.8% of the variation in the X-data block. Score control charts in which the average batch trajectory and upper and lower control limits are displayed were developed. Next, these control charts were used to monitor 4 new test batches in real-time and to immediately detect any deviations from the expected batch trajectory. By real-time evaluation of new batches using the developed control charts and by computation of contribution plots of deviating process behavior at a certain time point, batch losses or reprocessing can be prevented. Immediately after batch completion, all PSD and product temperature information (i.e., a batch progress fingerprint) was used to estimate some granule properties (density and flowability) at an early stage, which can improve batch release time. Individual PLS models relating the computed scores (X) of the reference PLS model (based on the 10 reference batches) and the density, respectively, flowabililty as Y-matrix, were developed. The scores of the 4 test batches were used to examine the predictive ability of the model.

Evaluation of in-line spatial filter velocimetry as PAT monitoring tool for particle growth during fluid bed granulation.

In this study, the feasibility of spatial filter velocimetry (SFV) as process analytical technology tool for the in-line monitoring of the particle size distribution during top spray fluidized bed granulation was examined. The influence of several process (inlet air temperature during spraying and drying) and formulation variables (HPMC and Tween 20 concentration) upon the particle size distribution during processing, and the end product particle size distribution, tapped density and Hausner ratio was examined using a design of experiments (DOE) (2-level full factorial design, 19 experiments). The trend in end granule particle size distributions of all DOE batches measured with in-line SFV was similar to the off-line laser diffraction (LD) data. Analysis of the DOE results showed that mainly the HPMC concentration and slightly the inlet air temperature during drying had a positive effect on the average end granule size. The in-line SFV particle size data, obtained every 10s during processing, further allowed to explain and better understand the (in)significance of the studied DOE variables, which was not possible based on the LD data as this technique only supplied end granule size information. The variation in tapped density and Hausner ratio among the end granules of the different DOE batches could be explained by their difference in average end granule size. Univariate, multivariate PLS and multiway N-PLS models were built to relate these end granule properties to the in-line-measured particle size distribution. The multivariate PLS tapped density model and the multiway N-PLS Hausner ratio model showed the highest R(2) values in combination with the lowest RMSEE values (R(2) of 82% with an RMSEE of 0.0279 for tapped density and an R(2) of 52% with an RMSEE of 0.0268 for Hausner ratio, respectively).

[Culling of animals in large herds, experiences from the Netherlands]. / Keulung von Tieren in Grossbeständen, Erfahrungen in den Niederlanden.

The outbreaks of classical swine fever in 1997-1998 and foot- and mouth-disease in 2001 provided a lot of experiences in the culling of animals. These experiences, as well as the aspects of animal welfare and public acceptance are described. In the future these experiences will help to carry out culling in a more efficient way including improved aspects of animal welfare and public acceptance.

A self-expanding endoluminal graft for treatment of aneurysms: results through the development phase.

BACKGROUND: The results of two and a half years' experience of endoluminal treatment of aneurysmal disease (from March 1993 to December 1995) are reported. METHODS: The endoluminal grafts were individually made at Royal Perth Hospital. They are based on Dacron-covered stainless steel self-expanding 'Z' stents with Gianturco barbed stents (Cook Pty, Australia) for proximal anchorage for grafts within the aorta. RESULTS: Fourteen straight tube grafts (nine for aortic aneurysm, four for iliac aneurysm and one for subclavian aneurysm) and 24 bifurcate grafts were deployed; all were in patients considered high-risk for conventional repair. Seventy-two per cent of the straight tube grafts successfully excluded the aneurysm. The bifurcate grafts, in use since July 1994, successfully excluded the aneurysm in 88%. There were two delayed deaths from rupture after the grafts failed to exclude the aneurysms; two patients required conversion to open repair and survived; three patients have persistent endoleaks; and three of the bifurcate grafts subsequently occluded a graft limb but did not require further intervention. Ninety per cent of these complications occurred in the first half of the series (prior to January 1995). CONCLUSIONS: A learning and development curve was clearly apparent. The results thereafter compare favourably to those for open repair in similar high-risk groups, suggesting that these techniques hold promise for all patients with aneurysms.

The quality of blood used for transfusion.

The current risk of disease transmission through homologous blood transfusion has lead to a revival in the use of autologous blood. The development of a coagulopathy increases the usage of blood and blood products and therefore the risk of disease transmission. Blood salvaged at operation is subjected to physical and humoral activating factors. The potential systems to be activated are located either in the plasma or non red cell cellular elements. Homologous blood and blood salvaged at operation before and after washing was examined for activation of the plasma and cellular systems by measuring the presence of fibrin degradation products (xDP's), activated complement (C3a) and plasminogen activator and inhibitor activity from the plasma systems; and elastase, serotonin, lysophospholipids, leukotriene B4, lysoplatelet factor and phospholipase A2 from the cellular systems. The plasma systems were activated in the salvaged blood which had elevated levels of xDP's and C3a compared to the patient's blood (p less than 0.05). The products of cellular activation were also elevated in operatively salvaged blood (p less than 0.01 for all the above products). The levels xDP's were normal but the levels of C3a increased in stored blood. The levels of the cellular systems, elastase, serotonin and lysoplatelet activating factor, increased with the duration of storage of bank blood. Unwashed and aged stored blood contains potentially harmful products. Consideration should be given to removal of non red cell elements in blood stored for surgery.

Lipolytic enzyme and phospholipid level changes in intraoperative salvaged blood.

Autotransfusion is becoming increasingly popular, mainly because it eliminates the risk of disease transmission. One of the techniques available is intra-operative blood salvage and retransfusion with or without washing of the collected blood. The blood collected during this process is subjected to a variety of chemical and physical insults which can alter the normal composition of the plasma by activating plasma and cellular homeostatic mechanisms. In this study, we measured the plasma levels of total phospholipids, lysolecithin and non-esterified fatty acids, and the lipolytic enzymes phospholipase A2 (PLA2) and lipase in the salvaged blood before and after washing. In the unwashed salvaged blood the mean levels of PLA2, non-esterified fatty acids and lysophospholipids increased by 144, 96 and 149%, respectively, while those of total phospholipids and lipase did not change to any extent. All these substances were reduced to well below the patients circulating plasma levels by washing the collected blood. The changes indicate that the lipid profile of salvaged blood is significantly altered and that potentially dangerous substances such as PLA2 and its metabolites, lysolecithin and non-esterified fatty acids, are present in increased amounts. Washing the blood is recommended prior to reinfusion.

Levels of complement factor C3 and its activated product, C3a, in operatively salvaged blood.

In intra-operative blood salvage the collected blood is exposed to traumatized tissue and a synthetic circuit. Because of this exposure it was expected that the complement system, which is a contact system present in plasma, would be activated in salvaged blood. To determine whether this occurs, the plasma levels of the complement factor C3 and its activated fragment, C3a, were measured in intra-operative salvaged blood before and after washing. Samples were obtained from patients undergoing aortic surgery where intra-operative salvage was used. In the unwashed salvaged blood, the level of C3 fell (mean C3: 0.33 g/L) and the level of C3a increased (mean C3a: 1994 ng/mL) compared with the patient circulating levels of C3 and C3a (mean C3: 0.70 g/L, mean C3a: 855 ng/mL) respectively. Washing of the collected blood reduced the C3a level (mean: 346 ng/mL) to the patient's level and reduced C3 to the lower detection limit of the test (less than 0.2 g/L). The raised level of C3a and the reduced level of C3 confirm that the complement system is activated and imply that other complement factors are also activated.

Elastase levels in salvaged blood and the effect of cell washing.

Blood was salvaged from the operating field of 16 orthopaedic and vascular operations and processed by the Shiley Dideco cell saver for retransfusion. Plasma elastase levels in the salvaged blood were used as a marker of white cell lysosomal granule release. The plasma level of this enzyme was measured by the technique described by Dreher et al. using the immuno-activation (IMAC) system (Merck-Darmstadt, Federal Republic of Germany). The levels of the enzyme rose dramatically in the salvaged blood and were reduced to preoperative levels after washing in the cell saver. Washing of the salvaged blood before retransfusion is effective in reducing the plasma levels of white cell lysosomal contents which have the potential for producing harmful systemic effects.

D-dimer levels in blood salvage for autotransfusion.

Autotransfusion of operatively salvaged blood is an increasingly attractive and used practice. The fear of precipitating a coagulopathy, however, has retarded the acceptance of autotransfusions and the efficiency and convenience of banked homologous blood has proved too competitive in recent years. The risk of transmitting diseases with bank homologous blood has seen a resurgence in the development of autotransfusion. The current alternatives to pre-operative autologous banking or dilution are the reinfusing of filtered salvaged blood, and of blood which has been further processed by washing which involves the extra cost of time, personnel and equipment. Washing removes possible harmful products, but also removes coagulation factors. D-Dimer levels were estimated by monoclonal antibody techniques in salvaged blood before and after washing in 10 patients undergoing aortic surgery and in 10 u of homologous banked blood. The D-Dimer levels in the unwashed blood were increased 85 times, but were normal in the washed blood. D-Dimers are an indication of activation of the coagulation and fibrinolytic systems and the presence of fibrin degradation products.