Variability of internal standard method calibration factors estimated with a multifactorial Taguchi experiment in the analysis of alcoholic products by gas chromatography with flame ionization detection.

This study compares the variability of relative response factors (RRFs) using Taguchi's multifactorial analysis for two internal standard (IS) methods in gas chromatography (GC) for quality control of alcoholic products. Methods where either ethanol or pentan-1-ol was used as an IS were compared. For ten volatile substances prescribed by legislation, the RRFs of both methods were compared under 27 different experimental conditions. The influence of parameters (control factors) such as ethanol content in the matrix, analyte concentration, injected volume, injector temperature, split ratio, and flame ionization detector temperature was evaluated. The selected control factors had values at one of the three levels to cover the commonly used ranges of their settings in the measuring system and to characterize the majority of alcoholic products commonly analyzed in practice. The obtained results showed that the biggest differences in the variability of the results between the two methods were found for the most strictly controlled substances in alcoholic products, acetaldehyde, and methanol, where the application of ethanol as an IS provides clearly better results. For both methods, the way control factors affect the repeatability of GC measurements expressed in the form of relative deviation was also evaluated.

The extreme wet and large precipitation size increase carbon uptake in Eurasian meadow steppes: Evidence from natural and manipulated precipitation experiments.

The distribution of seasonal precipitation would profoundly affect the dynamics of carbon fluxes in terrestrial ecosystems. However, little is known about the impacts of extreme precipitation and size events on ecosystem carbon cycle when compared to the effects of average precipitation amount. The study involved an analysis of carbon fluxes and water exchange using the eddy covariance and chamber based techniques during the growing seasons of 2015-2017 in Bayan, Mongolia and 2019-2021 in Hulunbuir, Inner Mongolia, respectively. The components of carbon fluxes and water exchange at each site were normalized to evaluate of relative response among carbon fluxes and water exchange. The investigation delved into the relationship between carbon fluxes and extreme precipitation over five gradients (control, dry spring, dry summer, wet spring and wet summer) in Hulunbuir meadow steppe and distinct four precipitation sizes (0.1-2, 2-5, 5-10, and 10-25 mm d-1) in Bayan meadow steppe. The wet spring and summer showed the greatest ecosystem respiration (ER) relative response values, 76.2% and 73.5%, respectively, while the dry spring (-16.7%) and dry summer (14.2%) showed the lowest values. Gross primary production (GPP) relative response improved with wet precipitation gradients, and declined with dry precipitation gradients in Hulunbuir meadow steppe. The least value in net ecosystem CO2 exchange (NEE) was found at 10-25 mm d-1 precipitation size in Bayan meadow steppe. Similarly, the ER and GPP increased with size of precipitation events. The structural equation models (SEM) satisfactorily fitted the data (χ2 = 43.03, d.f. = 11, p = 0.215), with interactive linkages among soil microclimate, water exchange and carbon fluxes components regulating NEE. Overall, this study highlighted the importance of extreme precipitation and event size in influencing ecosystem carbon exchange, which is decisive to further understand the carbon cycle in meadow steppes.

Quantitative Analytical and Computational Workflow for Large-Scale Targeted Plasma Metabolomics.

Quantifying metabolites from various biological samples is necessary for the clinical and biomedical translation of metabolomics research. One of the ongoing challenges in biomedical metabolomics studies is the large-scale quantification of targeted metabolites, mainly due to the complexity of biological sample matrices. Furthermore, in LC-MS analysis, the response of compounds is influenced by their physicochemical properties, chromatographic conditions, eluent composition, sample preparation, type of MS ionization source, and analyzer used. To facilitate large-scale metabolite quantification, we evaluated the relative response factor (RRF) approach combined with an integrated analytical and computational workflow. This approach considers a compound's individual response in LC-MS analysis relative to that of a non-endogenous reference compound to correct matrix effects. We created a quantitative LC-MS library using the Skyline/Panorama web platform for data processing and public sharing of data. In this study, we developed and validated a metabolomics method for over 280 standard metabolites and quantified over 90 metabolites. The RRF quantification was validated and compared with conventional external calibration approaches as well as literature reports. The Skyline software environment was adapted for processing such metabolomics data, and the results are shared as a "quantitative chromatogram library" with the Panorama web application. This new workflow was found to be suitable for large-scale quantification of metabolites in human plasma samples. In conclusion, we report a novel quantitative chromatogram library with a targeted data analysis workflow for biomedical metabolomic applications.

Hyphenated liquid chromatography - diode array detection - charged aerosol detection - high resolution - multistage mass spectrometry with online hydrogen/deuterium exchange: One stop solution for pharmaceutical impurity profiling.

Hyphenation of different analytical techniques has always been advantageous in structural characterization as it saves time, money and resources. In the pharmaceutical sector, chromatography-based impurity profiling, including identification, characterization, and quantification in drug substances or finished products, is of utmost importance to comply with quality, patient safety and regulatory requirements. These impurities are monitored using LC-UV/DAD and identified and/or characterized using HRMS and MS/MS. LC analysis usually yields the area percent purity of the targeted peak, however, this is not sufficient for pharmaceutical purposes; where the regulatory requirement is to report impurities in percent weight by weight. Unfortunately, the non-availability of impurity standards and relative response factors at an early stage of drug development, risks the product quality due to the inability of the method to differentiate percent purity, and percent weight by weight. Hence, there is a need for a distinctive way of determining the relative response factor. In the current study, a unique hyphenation has been employed by integrating LC with DAD, CAD, and HRMSn with hydrogen-deuterium exchange. The LC flow, post-DAD detection has been diverted to CAD with an inverse gradient for relative response factor determination and MS Orbitrap for exact mass, and MSn fragmentation. A separate infusion pump has been incorporated to infuse D2O on a need basis, which can perform partial hydrogen deuterium exchange for determining the number of labile hydrogens in the impurity structure. This hyphenation has been validated with four model compounds and a total of nineteen chromatographic peaks. The technique provides ample information for their qualitative analysis along with percent weight-by-weight values, which fulfils the regulatory requirements and can be used as one-stop solution for impurity profiling.

Machine Learning-Assisted Identification and Quantification of Hydroxylated Metabolites of Polychlorinated Biphenyls in Animal Samples.

Laboratory studies of the disposition and toxicity of hydroxylated polychlorinated biphenyl (OH-PCB) metabolites are challenging because authentic analytical standards for most unknown OH-PCBs are not available. To assist with the characterization of these OH-PCBs (as methylated derivatives), we developed machine learning-based models with multiple linear regression (MLR) or random forest regression (RFR) to predict the relative retention times (RRT) and MS/MS responses of methoxylated (MeO-)PCBs on a gas chromatograph-tandem mass spectrometry system. The final MLR model estimated the retention times of MeO-PCBs with a mean absolute error of 0.55 min (n = 121). The similarity coefficients cos θ between the predicted (by RFR model) and experimental MS/MS data of MeO-PCBs were >0.95 for 92% of observations (n = 96). The levels of MeO-PCBs quantified with the predicted MS/MS response factors approximated the experimental values within a 2-fold difference for 85% of observations and 3-fold differences for all observations (n = 89). Subsequently, these model predictions were used to assist with the identification of OH-PCB 95 or OH-PCB 28 metabolites in mouse feces or liver by suggesting candidate ranking information for identifying the metabolite isomers. Thus, predicted retention and MS/MS response data can assist in identifying unknown OH-PCBs.

Accurate Determination of the Degree of Deacetylation of Chitosan Using UPLC-MS/MS.

The mole fraction of deacetylated monomeric units in chitosan (CS) molecules is referred to as CS's degree of deacetylation (DD). In this study, 35 characteristic ions of CS were detected using liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS/MS). The relative response intensity of 35 characteristic ion pairs using a single charge in nine CS samples with varying DDs was analyzed using 30 analytical methods. There was a good linear relationship between the relative response intensity of the characteristic ion pairs determined using ultrahigh performance (UP) LC-MS/MS and the DD of CS. The UPLC-MS/MS method for determining the DD of CS was unaffected by the sample concentration. The detection instrument has a wide range of application parameters with different voltages, high temperatures, and gas flow conditions. This study established a detection method for the DD of CS with high sensitivity, fast analysis, accuracy, stability, and durability.

Relative response factors and multiple regression models in liquid chromatography to quantify low-dosed components using alternative standards-proof of concept: total Δ9-THC content in cannabis flowers using CBD as reference.

A classical quantitative analysis in liquid chromatography is performed using either a one-point calibration or a calibration line, prepared using a reference standard of the compound(s) of interest. However, in some cases, adequate reference standards may be very expensive, rare to obtain, or have limited shelf-life properties. Also, in herbal matrices, multiple compounds could be necessary to be quantified, needing a whole series of different (related) reference standards. In these cases, the use of relative response (sometimes called relative correction factors) factors (RRFs) towards reference standards, different of the compound to be quantified, gained attraction. This study performed a comparison of the use of RRFs and linear relative response factor models (LRRFM) for the quantification of targeted low-dosed compounds using an alternative standard, since it is known that classical RRFs often fail in lower concentration ranges. For this purpose, the determination of the total Δ9-tetrahydrocannabinol (Δ9-THC + Δ9-THC-A) content in dried cannabis flowers, using UHPLC-DAD, was used as a case study. A chromatographic method was implemented and validated, and the use of classical calibration lines, classical RRF, and the LRRFM was applied and compared, with special focus on the concentration around 0.2% (w/w) total Δ9-THC, the legal limit (in most European countries) in these products. Results showed that the newly presented and validated LRRFM approach outperformed the classical RRFs, especially in the low-concentration ranges and that concentrations obtained with the LRRFM were in accordance with the interpolation results obtained with a calibration line.

Evaluation of the variation in relative response factors of GC-MS analysis with the internal standard methods: Application for the alcoholic products quality control.

This paper focuses on the evaluation of variation of relative response factors (RRFs) obtained by two internal standard (IS) methods that are used to control the quality of alcoholic products. A standard IS method using 1-pentanol was compared with an "Ethanol as IS" method. The variation of RRF values for both methods was determined using standard solutions based on 20, 40 and 96% ethanol-water matrices. For this purpose, solutions of the ten most abundant volatile compounds were analysed at four different concentrations (250, 500, 1000 and 5000 mg L-1 absolute alcohol, AA) within these matrices. Each solution was measured four times by gas chromatography-mass spectrometry (GC-MS) in single ion monitoring (SIM) mode under repeatability conditions. Our results showed that for the 40% and 96% matrices, the ethanol and standard IS methods showed similar relative standard deviations (RSDs) variation of no more than 2% within the 250-1000 mg L-1 AA volatile compounds concentration range. For the 20% matrix as well as for the 250-5000 mg L-1 AA concentration range the resulting variation in calibration factors reaches 10% for both methods. As for the whole range of 20-96% alcohol by volume (ABV) and 250-5000 mg L-1 AA volatile concentration range, the resultant RRFs for the standard IS method (8% RSD) were more stable than those for the ethanol IS method (almost 40% RSD). Nonlinearity of the signal-to-amount dependence was also assessed with respect to the injection and detection processes.

Use of qNMR to determine HPLC relative response factors for botanical reference standards used in pharmacopeial monographs.

In modern botanical pharmacopeial monographs, one measurement of content is the quantitation of relevant constituents as marker compounds. The use of suitable reference standards (RSs) to quantify multiple compounds by HPLC is recommended in the U.S. Pharmacopeial (USP) botanical monographs. However, these substances may be expensive and difficult to develop into an RS. Surrogate RSs could be used instead of the actual constituents, provided that the relative response factors (RRFs) of each analyte to the selected surrogate RS are known. USP monographs of both Sichuan Lovage Rhizome and Dong Quai Root recognize Z-ligustilide as a major characteristic marker compound, making quantitation of Z-ligustilide and its analog(s) relevant for quality control. However, because Z-ligustilide is unstable, it is difficult to develop it into a quantitative RS. Instead, oxybenzone was selected as a surrogate external quantitative RS because of its similar chromatographic behavior to Z-ligustilide, its stability, and affordable cost. The RRF determination of Z-ligustilide to oxybenzone by the conventional HPLC procedure is challenging due to both the instability of the purified Z-ligustilide at ambient temperature and the difficulty of determining its purity. Therefore, a qNMR method was used to overcome these challenges as it enables to directly measure the mass ratio of Z-ligustilide to oxybenzone in the stock solution without the need for weighing and purity information. In the present study, RRF values of 1.01, 0.46, and 0.89 for Z-ligustilide, senkyunolide A, and ferulic acid relative to oxybenzone, respectively, were determined using the qNMR-based methodology.

A novel method for determining relative response factors using high-performance liquid chromatography with photodiode array detector and evaporative light scattering detection.

OBJECTIVES: Because the purity of the two impurities reference standard (RS) of cefathiamidine is not easy to obtain, HPLC-PDA-ELSD were used to determin the RRFs of cefathiamidine impurities. METHODS: Peak area correction was applied to calculate RRFs to eliminate the influence of different responses caused by the difference in pH between the two mobile phases of HPLC-PDA and HPLC-PDA-ELSD. The resulting RRF values have been verified by qNMR. CONCLUSION: The new calcution method described in this article provides a reliable research idea for determintion the RRFs by HPLC-PDA-ELSD, especially when the purity of RS is unknown and the mobile phase of HPLC-PDA and HPLC-PDA-ELSD have difference. This method can be mutually verified with qNMR to ensure the accuracy of RRFs. It is also promingsing replace determination RRF by qNMR becausing economical, simple and low cost.

Evaluation of a highly condensed SAPRC chemical mechanism and two emission inventories for ozone source apportionment and emission control strategy assessments in China.

The response of summertime O3 to changes in the nitrogen oxides (NOx) and volatile organic compounds (VOC) emissions, and contributions of different NOx and VOC sources to O3 in China are studied using a highly condensed photochemical mechanism in the Statewide Air Pollution Research Center (SAPRC) family (CS07A) and two popular anthropogenic emission inventories, the Multi-resolution Emission Inventory for China (MEIC) and Regional Emission inventory in ASia (REAS). Although CS07A predicts slightly lower O3 concentrations than the standard fix-parameter version of the SAPRC-11 mechanism, the two mechanisms predict almost identical relative responses to daily maximum 8-hour O3 (O3-8h) due to NOx and VOC emission reductions. A source-oriented version of the CS07A is applied to determine source contributions of NOx and VOCs to O3 using MEIC and REAS. The two inventories lead to similar model performance of O3, with MEIC predicting higher O3 in Beijing and Shanghai, especially on high O3 days. Source apportionment results show that industry and transportation are the top two contributors to non-background O3 for both inventories, followed by power and biogenic emissions. In general, the two inventories lead to similar source contribution estimations to O3 attributable to NOx. However, their estimations of relative contributions to VOC-related O3 differ for the industrial and transportation sectors. Differences in the source apportionment results are more significant in some urban areas, although both emissions capture the spatial variations in the source contributions. Our results suggest that future emission control policies should be assessed using multiple emission inventories, and the condensed CS07A is suitable for policy applications when a large number of simulations are needed.

A detection method for relative response of a detector matrix / 中华放射医学与防护杂志

Objective:To test the relative response of a detector matrix used for measuring of the dose distribution in radiotherapy in order to ensure the accuracy and reliability of dose distribution measurement result.Methods:The two adjacent detectors can be irradiated at the same radiation dose by translating the detector matrix. The relative response of each detector to the reference detector can be obtained by combining the recursive algorithm. At the same time, the measurement data at different test steps are corrected to the reference step by setting the reference step, which correct the influence on the relative response result caused by the variation of the radiation condition and the detector response.Results:Based on the actual test of a 32 × 32 detector matrix, the relative response of each detector of the detected detector matrix to the reference detector varied from 0.896 to 1.077, with the expanded uncertainty of the relative response result being 0.8% ( k=2). Conclusions:On the premise of no known distribution of a radiation field, the relative response relationship of each detector of a detector matrix can be obtained by this method accurately and expediently, which provides a basic method for the performance evaluation of the detector matrix. The same idea can also be used to determine the relative response relationship of other detector matrices for different measurement purposes.

Determination of vacuum ultraviolet detector response factors by hyphenation with two-dimensional comprehensive gas chromatography with flame ionization detection.

Two-dimensional comprehensive gas chromatography is an established technique, employed for the characterization of complex samples. Broadband vacuum ultraviolet absorption spectroscopy detection has recently attracted a lot of attention as it is a universal detection technique characterized by good selectivity but also ease of use and amenability to coupling with two-dimensional comprehensive gas chromatography. Vacuum ultraviolet spectroscopy is particularly interesting due to the possibility of performing spectral decomposition for species that coelute in gas chromatography analysis. This detector has quantitative capabilities, however not all species absorb vacuum ultraviolet radiation the same. Unfortunately, vacuum ultraviolet relative response factors for compounds are not always available. Methods to rapidly measure vacuum ultraviolet relative response factors and generate a large database that would allow calibration free quantitative analysis of complex mixtures are therefore of great interest. In this work, a universal methodology that permits rapid measurement of vacuum ultraviolet relative response factors is reported. It involves flow modulated two-dimensional comprehensive gas chromatography with dual vacuum ultraviolet and flame ionization detection. In this set-up, flame ionization detection is employed as a quantitative reference allowing to scale vacuum ultraviolet responses of investigated compounds. This approach was validated by flow measurements and by comparing relative response factors obtained for model compounds with literature data.

Delineating the extra-virgin olive oil aroma blueprint by multiple headspace solid phase microextraction and differential-flow modulated comprehensive two-dimensional gas chromatography.

Comprehensive two-dimensional gas chromatography with parallel mass spectrometry and flame ionization detection (GC × GC-MS/FID) enables effective chromatographic fingerprinting of complex samples by comprehensively mapping untargeted and targeted components. Moreover, the complementary characteristics of MS and FID open the possibility of performing multi-target quantitative profiling with great accuracy. If this synergy is applied to the complex volatile fraction of food, sample preparation is crucial and requires appropriate methodologies capable of providing true quantitative results. In this study, untargeted/targeted (UT) fingerprinting of extra-virgin olive oil volatile fractions is combined with accurate quantitative profiling by multiple headspace solid phase microextraction (MHS-SPME). External calibration on fifteen pre-selected analytes and FID predicted relative response factors (RRFs) enable the accurate quantification of forty-two analytes in total, including key-aroma compounds, potent odorants, and olive oil geographical markers. Results confirm good performances of comprehensive UT fingerprinting in developing classification models for geographical origin discrimination, while quantification by MHS-SPME provides accurate results and guarantees data referability and results transferability over years. Moreover, by this approach the extent of internal standardization procedure inaccuracy, largely adopted in food volatiles profiling, is measured. Internal standardization yielded an average relative error of 208 % for the fifteen calibrated compounds, with an overestimation of + 538% for (E)-2-hexenal, the most abundant yet informative volatile of olive oil, and a -89% and -80% for (E)-2-octenal and (E)-2-nonenal respectively, analytes with a lower HS distribution constant. Compared to existing methods based on 1D-GC, the current procedure offers better separation power and chromatographic resolution that greatly improve method specificity and selectivity and results in lower LODs and LOQs, high calibration performances (i.e., R2 and residual distribution), and wider linear range of responses. As an artificial intelligence smelling machine, the MHS-SPME-GC × GC-MS/FID method is here adopted to delineate extra-virgin olive oil aroma blueprints; an objective tool with great flexibility and reliability that can improve the quality and information power of each analytical run.

Assay of active pharmaceutical ingredients in drug products based on relative response factors: Instrumentation insights and practical considerations.

Relative Response Factors (RRFs) can be used for quantitation of one compound against another and it is widely used for Impurity analysis of pharmaceutical products; however, the application in potency assay is limited. Through an extensive study shown in this paper, it can be concluded that using the "RRF methodology" for potency assay is much more challenging compared to impurity analysis, due to the much tighter criteria required for potency analysis. The effects of instrument settings, which are rarely discussed or recognized in current HPLC analytical method development and quality release testing, are discussed. These factors impact the RRF just as much as other commonly recognized HPLC parameters. The effects of UV detector settings, i.e. Slit Width, Step Width, Band Width, and Data Collection Module, have been explored. This phenomenon has been demonstrated using three compounds to observe the impact of their quantitation due to the significant RRF variations. Finally, principles to reduce RRF variations have been discussed, and practical considerations of RRF application to method development and method transfer are provided.

Ampicillin sodium: Isolation, identification and synthesis of the last unknown impurity after 60 years of clinical use.

Ampicillin, discovered in 1958, was the first broad spectrum semisynthetic penicillin introduced into the market. Despite its wide use not all the impurities have been identified to date. Herein, the last unknown impurity present in commercially available medicines was isolated and identified. This impurity that accounts up to 0.8 in area % by HPLC (EP 10.0) in the Reference Listed Drugs (RLD) was characterized and identified to be the 16-keto penicillin G. The structure was confirmed by comparison with a chemically synthesized sample. The determination of the Relative Response Factor (RRF) of the impurity respect to the parent drug allowed to recalculate the real amount that is consistently below the reporting threshold.

Quantification of cranberry proanthocyanidins by normal-phase high-performance liquid chromatography using relative response factors.

INTRODUCTION: American cranberries (Vaccinium macrocarpon) contain primarily A-type proanthocyanidins (PACs), which have been shown to prevent urinary tract infection. Currently, the accurate quantification of cranberry PACs is still lacking. OBJECTIVE: A normal-phase high-performance liquid chromatography (NP-HPLC) method using relative response factors was developed and validated to quantify cranberry PAC oligomers and polymers. MATERIALS AND METHODS: PAC oligomers with degree of polymerisation (DP) 3-9 and total polymers were isolated from the cranberry juice concentrate. Characterisation of the isolated PAC oligomers was performed by ultra-performance liquid chromatography-high resolution mass spectrometry. The relative response factors of oligomers from DP 2-9 and total polymers were determined against procyanidin A2. Method validation was conducted to assess limit of detection, limit of quantification, the linearity and working range, precision and accuracy. In addition, quantifications of PACs by NP-HPLC using relative response factors and two other commonly used methods were conducted in three cranberry food products. RESULTS: Cranberries PACs oligomers contained both A-type and B-type linkage, with epicatechin and epigallocatechin as basic units. Method validation results suggested this method is reliable and reproducible. Quantifications of PACs by NP-HPLC using relative response factors yielded higher values than that by the other two methods. CONCLUSION: A NP-HPLC method using the relative response factors was developed and validated. This method provides a more accurate approach in determining cranberry PACs. It can be used to quantify individual oligomers from DP 2-9, total polymers and total PACs in cranberries and cranberry products.

A Validated Method for Cholesterol Determination in Turkey Meat Products Using Relative Response Factors.

The objective of this study was to develop a precise and accurate method to quantify cholesterol in turkey meat products using relative response factors, based on a modification of a previously published method for plant sterols determination. Validation was performed using neat solutions to determine linearity, precision, and accuracy. The method was linear in the concentration range considered (1-20 µg/mL, r2 ≥ 0.991). Precision and accuracy were within the acceptability guidelines of the U.S. Food and Drug Administration (FDA) for method validation (<20% relative standard deviation (RSD) at the lower limit of quantification (LLOQ) and <15% RSD for other standards). Turkey meat was spiked with cholesterol at two levels (low = 3 µg/mL and high = 18 µg/mL), either before or after saponification, to establish the recovery and matrix effects. Recovery ranged from 94% to 105%, with a mean value of 105% at the low spike level and 95% at the high spike level. No significant matrix effects were found (90% to 112% recovery). This method is reliable for the quantification of cholesterol in turkey meat products in the range 0.4-8 mg/g.

[Chemical profiling for bile acid derivatives in yak bile].

Bile acids( BAs),the major constituents of bile,are also known to be potential biomarkers of various diseases,especially liver disease. The systematic analysis of BAs is believed to be of great importance towards the clarification of the effective material basis for bile-type medicines,and the diagnosis and therapy of related diseases as well. As a part of systematic study on bile-type medicine ongoing in our group,this study lays emphasis on the isomer discrimination,and the improvement of analytical method of BAs. Further,this method was subsequently applied to elucidate in depth the chemical profile of BAs in yak bile. Regarding isomer discrimination for BAs,we constructed relative response-collision energy curves( RRCECs) by high performance liquid chromatographyion trap-time of flight-mass spectrometry( HPLC-IT-TOF-MS) in combination with high performance liquid chromatography-triple quadrupole-linear ion trap mass spectrometry( HPLC-Qtrap-MS). As a result,both the optimum collision energy( OCE) and CE_(50) exhibited great correlations with structural characteristics,thus enabling the isomer distinguishing,such as unconjugated BAs,glycine-conjugated BAs,and taurine-conjugated BAs. According to information provided by mass spectrometry,the comparison of OCE and CE_(50),retention time matching,combined with reference substances and database retrieval,a total of 30 bile acid derivatives were observed and identified in yak bile. The newly developed method could serve as a feasible tool for the in-depth characterization of BAs in bile and biological samples.

Relative efficiency of Gafchromic EBT3 and MD-V3 films exposed to low-energy photons and its influence on the energy dependence.

Energy-dependence of Gafchromic films exposed to low-energy photons has been reported to be a function of absorbed-dose. However, these studies are based on a relative-response, R, which considers the absorbed-dose in water and not within the film sensitive-volume. This work investigated the relative-efficiency, REfilm, (ratio of absorbed-dose required to produce the same net optical density (netOD) by 60Co gamma and by x-ray) of Gafchromic EBT3 and MD-V3 films exposed to five x-ray beams from 20 kV to 160 kV and 60Co gamma rays. A factor that accounts for the energy-dependence, fx,Q,med, based on REfilm, phantom-material and depth at which the films are placed during irradiation was used to remove the influence of absorbed dose. Values of REfilm indicated that the absorbed dose from 60Co gamma rays needs to be 4 and 3 times larger than those from 20 kV x-rays to produce the same netOD within the EBT3 and MD-V3 sensitive volumes, respectively. Thus, saturation could help explain why Gafchromic films show under-response to very low doses from low-energy photon beams, regardless of film model. Furthermore, REfilm, was found to be nearly independent of netOD and colour-channels. Consequently, fx,Q,med is independent of the absorbed dose and colour-channels. In contrast, besides the variation with the photon energy, fx,Q,med varied with film model, depth and phantom material used during the irradiation. Thus, the results suggest that fx,Q,med is a more reliable wide-ranging parameter for evaluating the degree of energy-dependence of the film rather than the relative-response method commonly considered.