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Organisationseinheit der BAM
The current practise of focusing the periodic retesting of composite cylinders primarily on the hydraulic pressure test has to be evaluated as critical - with regard to the damage of the specimen as well as in Terms of their significance. This is justified by micro damages caused to the specimen by the test itself and by a lack of informative values. Thus BAM Federal Institute of Materials Research and Testing (Germany) uses a new approach of validation of composite for the determination of re-test periods. It enables the description of the state of a population of composite cylinders based on destructive tests parallel to operation.
An essential aspect of this approach is the prediction of residual safe service life. In cases where it cannot be estimated by means of hydraulic load cycle tests, as a replacement the creep or burst test remains. As a combination of these two test procedures BAM suggests the 'slow burst test SBT'. On this a variety of about 150 burst test results on three design types of cylinders with plastic liners are presented. For this purpose both, the parameters of the test protocol as well as the nature and intensity of the pre-damage artificially aged test samples are analysed statistically. This leads first to an evaluation of the different types of artificial ageing but also to the clear recommendation that conventional burst tests be substituted
totally if indented for assessment of composite pressure receptacles.
The current practice of periodic retesting of composite pressure receptacles primarily based on the hydraulic pressure test has to be evaluated as critical.
This is justified by micro damages caused to the specimen by the test
itself and by a lack of informative values. Thus, BAM Federal Institute of Materials Research and Testing uses a new validation approach in the case of retest periods of more than 5 years. It enables the description of the condition of a specific design type series based on destructive tests parallel to operation. With respect to this approach it is explained how the statistical evaluation of sample tests and their extrapolation to end of life or at least to the time of next retesting can be achieved. In addition, it is explained how the reliability limits may be linked to the size and pressure level of the respective composite pressure receptacle. The frame of this safety Approach includes the influence of the maximum working pressure on the safety assessment.
One topic of this approach is the introduction of the working diagram
essential for safety assessments that allows graphical evaluation of load cycle strength by mean value, scatter and survival rate of samples as well as instruction how to use it. For this, it is explained how the number of filling cycles and the reduction of the respective hydraulic residual cycle strength interacts with the newly introduced value of specific filling sensitivity.
Slow burst testing of samples as a method for quantification of composite cylinder degradation
(2014)
The current practise to focus periodic retesting of composite cylinders primarily on the hydraulic pressure test has to be evaluated as critical. The test itself always causes a certain amount of micro damage to the cylinders but does not necessarily deliver sufficient and evaluable information. Thus BAM Federal Institute of Materials Research and Testing (Germany) moves the focal point to a new approach for validation of composite cylinders, based on destructive sample tests parallel to operation. Statistical assessment of results of these destructive tests is employed for the estimation of remaining safe service life, based on reliability demands. The estimated service life is also used for the determination of re-test periods of the examined population of composite cylinders.
An essential aspect of this approach is the validation of current residual strength and its prediction at any point of service life. In cases of gas cylinders with very high cycle strength, residual strength cannot be quantified statistically by means of hydraulic load cycles. As a replacement, creep tests or burst tests may be employed. BAM suggests the 'slow burst test SBT' as a combination of these two test procedures. This is a compromise between the practicability of the (conventional) burst test and the practical relevance of sustained loads during service, to be tested in creep rupture tests. In this paper, a variety of 99 burst results of a cylinder design type used for breathing apparatus (CFRP with PE-liner) is evaluated. The influence of test procedure parameters and nature and intensity of artificial ageing on the test sample strength are analysed statistically. This leads to an evaluation of different procedures of artificial ageing and the recommendation to substitute conventional burst tests by slow burst tests for the assessment of composite pressure receptacles.
Developers interested in high pressure storage of hydrogen for mobile use increasingly rely on composite cylinders for onboard storage or transport of dangerous goods. Thus, composite materials and systems deserve special consideration. History gives interesting background information important to the understanding of the current situation as to regulations, codes and standards.
Based on this review, origins of different regulations for the storage of hydrogen as dangerous good and as propellant for vehicles will be examined. Both categories started out using steel and sometimes aluminium as cylinder material. With composite materials becoming more common, a new problem emerged: vital input for regulations on composite pressure systems was initially derived from decades of experience with steel cylinders. As a result, both regulatory fields suffer somewhat from this common basis. Only recent developments regarding requirements for composite cylinders have begun to go more and more separate ways. Thus these differences lead to some shortcomings in regulation with respect to composite storage systems.
In principle, in spite of separate development, these deficits are in both applications very much the same: there are uncertainties in the prediction of safe service life, in retesting procedures of composite cylinders and in their intervals. Hence, different aspects of uncertainties and relevant approaches to solutions will be explained.
A novel method that optimizes the screening for antibody-secreting hapten-specific hybridoma cells by using flow cytometry is described. Cell clones specific for five different haptens were analyzed. We selectively double stained and analyzed fixed hybridoma cells with fluorophore-labeled haptens to demonstrate the target-selectivity, and with a fluorophore-labeled anti-mouse IgG antibody to characterize the level of surface expression of membrane-bound IgGs. ELISA measurements with the supernatants of the individual hybridoma clones revealed that antibodies from those cells, which showed the highest fluorescence intensities in the flow cytometric analysis, also displayed the highest affinities for the target antigens. The fluorescence intensity of antibody-producing cells corresponded well with the produced antibodies' affinities toward their respective antigens. Immunohistochemical staining verified the successful double labeling of the cells. Our method makes it possible to perform a high-throughput screening for hybridoma cells, which have both an adequate IgG production rate and a high target affinity.
Gas cylinders made from composite materials receive growing popularity in applications where light weight is of advantage. At the same time manufacturers are interested in cutting cost and weight through material reduction for better acceptance of a product, e.g. H2-vehicles. This requires a better understanding of safety relevant properties.
The safety determination of current standards is mainly based on the minimum burst pressure of a few specimens. Various research projects were conducted aiming at reducing minimum burst pressure requirements without compromising safety. No satisfying results were found. While looking at reliability aspects it could be concluded, that the minimum burst pressure of unused specimens is not a satisfying criterion for safety during service life.
This paper introduces first ideas for a method to determine one aspect of reliability of composite gas cylinders, employing probabilistic analysis of burst pressures of a sample of specimens. This can create potential for saving material cost while granting a higher safety level than the current method. Additionally, degradation over service time can be assessed.
The conventional hybridoma screening and subcloning process is generally considered to be one of the most critical steps in hapten-specific antibody production. It is time-consuming, monoclonality is not guaranteed, and the number of clones that can be screened is limited. Our approach employs a novel hapten-specific labeling technique of hybridoma cells. This allows for fluorescence-activated cell sorting (FACS) and single-cell deposition and thereby eliminates the above-mentioned problems. A two-step staining approach is used to detect antigen specificity and antibody expression: in order to detect antigen specificity, hybridoma cells are incubated with a hapten−horseradish peroxidase conjugate (hapten−HRP), which is subsequently incubated with a fluorophore-labeled polyclonal anti-peroxidase antibody (anti-HRP−Alexa Fluor 488). To characterize the expression of membrane-bound immunoglobulin G (IgG), a fluorophore-labeled anti-mouse IgG antibody (anti-IgG−Alexa Fluor 647) is used. Hundreds of labeled hybridoma cells producing monoclonal antibodies (mAbs) specific for a hapten were rapidly isolated and deposited from a fusion mixture as single-cell clones via FACS. Enzyme-linked immunosorbent assay (ELISA) measurements of the supernatants of the sorted hybridoma clones revealed that all hapten-specific hybridoma clones secrete antibodies against the target. There are significant improvements using this high-throughput technique for the generation of mAbs including increased yield of antibody-producing hybridoma clones, ensured monoclonality of sorted cells, and reduced development times.
The non-steroidal anti-inflammatory drug (NSAID) diclofenac (DCF) is an important environmental contaminant occurring in surface waters all over the world, because, after excretion, it is not adequately removed from wastewater in sewage treatment plants. To be able to monitor this pollutant, highly efficient analytical methods are needed, including immunoassays. In a medical research project, monoclonal antibodies against diclofenac and its metabolites had been produced. Based on this monoclonal anti-DCF antibody, a new indirect competitive enzyme-linked immunosorbent assay (ELISA) was developed and applied for environmental samples. The introduction of a spacer between diclofenac and the carrier protein in the coating conjugate led to higher sensitivity. With a test midpoint of 3 mg L−1 and a measurement range of 1–30 mg L−1, the system is not sensitive enough for direct analysis of surface water. However, this assay is quite robust against matrix influences and can be used for wastewater. Without adjustment of the calibration, organic solvents up to 5%, natural organic matter (NOM) up to 10 mg L−1, humic acids up to 2.5 mg L−1, and salt concentrations up to 6 g L−1 NaCl and 75 mg L−1 CaCl2 are tolerated. The antibody is also stable in a pH range from 3 to 12. Cross-reactivity (CR) of 1% or less was determined for the metabolites 40-hydroxydiclofenac (40-OH-DCF), 5-hydroxydiclofenac (5-OH-DCF), DCF lactam, and other NSAIDs. Relevant cross-reactivity occurred only with an amide derivative of DCF, 6-aminohexanoic acid (DCF-Ahx), aceclofenac (ACF) and DCF methyl ester (DCF-Me) with 150%, 61% and 44%, respectively. These substances, however, have not been found in samples. Only DCF-acyl glucuronide with a cross-reactivity of 57% is of some relevance. For the first time, photodegradation products were tested for cross-reactivity. With the ELISA based on this antibody, water samples were analysed. In sewage treatment plant effluents, concentrations in the range of 1.9–5.2 mg L−1 were determined directly, with recoveries compared to HPLC-MS/MS averaging 136%.
Concentrations in lakes ranged from 3 to 4.4 ng L−1 and were, after pre-concentration, determined with an average recovery of 100%