A new method for determining allowable daily intakes

The benchmark dose (BMD)4 approach is emerging as replacement to determination of the No Observed Adverse Effect Level (NOAEL) in noncancer risk assessment. This possibility raises the issue as to whether current study designs for endpoints such as developmental toxicity, optimized for detecting pair wise comparisons, could be improved for the purpose of calculating BMDs. In this paper, we examine various aspects of study design (number of dose groups, dose spacing, dose placement, and sample size per dose group) on BMDs for two endpoints of developmental toxicity (the incidence of abnormalities and of reduced fetal weight). Design performance was judged by the mean-squared error (reflective of the variance and bias) of the maximum likelihood estimate (MLE) from the log-logistic model of the 5% added risk level (the likely target risk for a benchmark calculation), as well as by the length of its 95% confidence interval (the lower value of which is the (BMD). We found that of the designs evaluated, the best results were obtained when two dose levels had response rates above the background level, one of which was near the ED05, were present. This situation is more likely to occur with more, rather than fewer dose levels per experiment. In this instance, there was virtually no advantage in increasing the sample size from 10 to 20 litters per dose group. If neither of the two dose groups with response rates above the background level was near the ED05, satisfactory results were also obtained, but the BMDs tended to be more conservative (i.e., lower). If only one dose level with a response rate above the background level was present, and it was near the ED05, reasonable results for the MLE and BMD were obtained, but here we observed benefits of larger dose group sizes. The poorest results were obtained when only a single group with an elevated response rate was present, and the response rate was much greater than the ED05. The results indicate that while the benchmark dose approach is readily applicable to the standard study designs and generally observed dose-responses in developmental assays, some minor design modifications would increase the accuracy and precision of the BMD.

The role of the benchmark dose in a regulatory context

Toxicological evaluation of sodium perfluorohexanoate

Increased cell proliferation increases the opportunity for transformations of normal cells to malignant cells via intermediate cells. Nongenotoxic cytotoxic carcinogens that increase cell proliferation rates to replace necrotic cells are likely to have a threshold dose for cytotoxicity below which necrosis and hence, carcinogenesis do not occur. Thus, low dose cancer risk estimates based upon nonthreshold, linear extrapolation are inappropriate for this situation. However, a threshold dose is questionable if a nongenotoxic carcinogen acts via a cell receptor. Also, a nongenotoxic carcinogen that increases the cell proliferation rate, via the cell division rate and/or cell removal rate by apoptosis, by augmenting an existing endogenous mechanism is not likely to have a threshold dose. Whether or not a threshold dose exists for nongenotoxic carcinogens, it is of interest to study the relationship between lifetime tumor incidence and the cell proliferation rate. The Moolgavkar-Venzon-Knudson biologically based stochastic two-stage clonal expansion model is used to describe a carcinogenic process. Because the variability in cell proliferation rates among animals often makes it impossible to detect changes of less than 20% in the rate, it is shown that small changes in the cell proliferation rate, that may be obscured by the background noise in rates, can produce large changes in the lifetime tumor incidence as calculated from the Moolgavkar-Venzon-Knudson model. That is, dose response curves for cell proliferation and tumor incidence do not necessarily mimic each other. This makes the use of no observed effect levels (NOELs) for cell proliferation rates often inadmissible for establishing acceptable daily intakes (ADIs) of nongenotoxic carcinogens. In those cases where low dose linearity is not likely, a potential alternative to a NOEL is a benchmark dose corresponding to a small increase in the cell proliferation rate, e.g., 1%, to which appropriate safety (uncertainty) factors can be applied to arrive at an ADI.

6-Mercaptopurine riboside (6-MPr) is used as a cytostatic chemotherapeutic. The teratogenic potential in rodents has been well known for several decades. In this study, the teratogenic risk of low doses of 6-MPr in NMRI mice was estimated based on a dose-response study. The effective doses corresponding to the incidences of 5%, 1%, and 0.1% were calculated using the probit and Weibull model. The evaluation was performed on the basis of both the fetus and the litter by evaluating the variable all gross structural abnormalities. From these experiments, benchmark doses were obtained which were used in low-dose risk assessment to define a reference dose. Depending on the biometrical model and the statistical unit used, values between 1.9 and 5.2 mg/kg (benchmark ED1) and 3.8 and 6.7 mg/kg (benchmark ED5) were obtained. These values were compared to the no observed adverse effect level (NOAEL) which was determined experimentally. The NOAEL was found to be 5 mg/kg, which is quite similar to the ED5 benchmark doses.

Use of benchmark dose and meta-analysis to determine the most sensitive endpoint for risk assessment for dimethoate

Objectives: The aim of this study was to determine the no observed adverse effect level (NOAEL), the lowest observed adverse effect level (LOAEL) and the benchmark dose low (BMDL) of cadmium exposure by re-evaluation of the dose–response relationship between cumulative cadmium exposure and renal tubular damage reported previously. Methods: The participants were workers (326 men and 114 women) employed for at least three months between 1931 and 1982. Blood cadmium (Cd-B) and air cadmium (Cd-A) were collected at regular intervals with urinary β2-microglobulin as the tubular effect marker. Cumulative Cd-A and Cd-B were estimated by multiplying concentration and working period. The BMDL was calculated using Benchmark Dose Software (version 3.1.2). The benchmark response (BMR) was set at 5% or 10%. Results: By logistic regression, the NOAEL of mean cumulative Cd-B was 7122 months nmol/L. The LOAEL of cumulative Cd-A and least-squares cumulative Cd-B was 691 yrs μg/m3 and 8586 months nmol/L, respectively. Among various models for dose–response relationships, a probit model was adopted as the best fitting model. The obtained BMDLs of cumulative Cd-A were 272.3 yrs µg/m3 (BMR5%) and 707.5 yrs µg/m3 (BMR10%). The BMDLs of mean cumulative Cd-B were 3967.2 months nmol/L (BMR5%) and 7798.1 months nmol/L (BMR10%). The BMDLs of least-squares cumulative Cd-B were 3588.6 months nmol/L (BMR5%) and 8616.3 months nmol/L (BMR10%). Assuming a working period of 40 years, the BMDLs for BMR10% corresponded to 17.7 µg/m3 (Cd-A) and 1.8~2.0 µg/L (Cd-B). Discussion: This study provides new valuable information to enhance the reliability of limit values and thereby make a significant contribution to preventing the health effects of Cd in exposed workers.

Detection of glyphosate residues in companion animal feeds

Panax notoginseng is a traditional valuable Chinese medicine plant that can be used for the treatment of cardiovascular diseases,inflammation,trauma,and hemorrhage. In recent years,the heavy metal pollution problem of P.notoginseng has attracted widely attention,however studies on this regard are limited. In this study,the heavy metal pollution of P. notoginseng and soil in Wenshan( Yunnan,China) was investigated; the bioaccumulation and transfer of heavy metal in P. notoginseng was revealed; and the associated health risk was evaluated. Concentrations of Cd,Cr and Cu in soil samples fell in the ranges of 0. 13—1. 87 mg /kg,71. 35—273. 73 mg /kg and 13. 96—86. 16 mg /kg,respectively.Cd concentrations in 75%,Cr concentrations in 38%,and Cu concentrations in 50% of the soil samples exceeded the Environmental Quality Standard( GB15618—1995),indicating that serious pollution existed in P. notoginseng planting area. Shapiro-Wilk test revealed that concentrations of soil Cd,Cr and Cr were not normally distributed( The P values of Cd,Cr and Cu were 0. 004,0. 02 and 0. 03,respectively,which were lower than 1),indicating that human activities had caused skewed probability distribution. Nemerow index analysis indicated that the magnitude of pollution followed the order of,from high to low,Qiubei,Wenshan,Yanshan and Maguan county. The concentrations of Cd,Cr and Pb in P. notoginseng were 0. 23—5. 50 mg /kg,0—30. 57 mg /kg and 0. 12—27. 45 mg /kg,respectively. and approximately81% —100%,75% —100%,and 25% —63% of the herb samples exceeded the Green Standards of Medicinal Plants and Preparations for Foreign Trade and Economy( WM /T2—2004) or NSF International Draft Standard( 173—2001).Bioaccumulation factors( BCF) of Cd,Cr,Cu and Pb were 1. 51—4. 01,0. 01—0. 11,0. 13—0. 37 and 0. 11—0. 37,respectively in different part of P. notoginseng. Transfer factors( TF) of Cd,Cr,Cu and Pb were 1. 23,0. 41,1. 14 and3. 61,respectively in P. notoginseng. Different parts of P. notoginseng accounted for 13. 46% —46. 40%,8. 67% —24.67% and 1. 90% —14. 40%,respectively,of the ingestion rates with ADI( Allowable Daily Intake). The correlation coefficient between heavy metal concentration in soil and the percent of main root ingestion rates with ADI were 0. 841**( Cd),0. 789**( Cr) and 0.753**( Cu)( The double-asterisk is used to represent P 0.01),respectively. This means that the health risk of Cd,Cr and Cu in the main root of the herb increased with the increasing metal concentrations. In conclusion,heavy metal pollution of soil and P. notoginseng in the planting area of Wenshan should not be neglected,and effective measures should be adopted to reduce heavy metal accumulation in the herb and human health risks.

Environmental Chemicals: Evaluating Low-Dose Effects

Determination of a site-specific reference dose for methylmercury for fish-eating populations.

The Scientific Committee (SC) reconfirms that the benchmark dose (BMD) approach is a scientifically more advanced method compared to the no‐observed‐adverse‐effect‐level (NOAEL) approach for deriving a Reference Point (RP). The major change compared to the previous Guidance (EFSA SC, 2017) concerns the Section 2.5, in which a change from the frequentist to the Bayesian paradigm is recommended. In the former, uncertainty about the unknown parameters is measured by confidence and significance levels, interpreted and calibrated under hypothetical repetition, while probability distributions are attached to the unknown parameters in the Bayesian approach, and the notion of probability is extended to reflect uncertainty of knowledge. In addition, the Bayesian approach can mimic a learning process and reflects the accumulation of knowledge over time. Model averaging is again recommended as the preferred method for estimating the BMD and calculating its credible interval. The set of default models to be used for BMD analysis has been reviewed and amended so that there is now a single set of models for quantal and continuous data. The flow chart guiding the reader step‐by‐step when performing a BMD analysis has also been updated, and a chapter comparing the frequentist to the Bayesian paradigm inserted. Also, when using Bayesian BMD modelling, the lower bound (BMDL) is to be considered as potential RP, and the upper bound (BMDU) is needed for establishing the BMDU/BMDL ratio reflecting the uncertainty in the BMD estimate. This updated guidance does not call for a general re‐evaluation of previous assessments where the NOAEL approach or the BMD approach as described in the 2009 or 2017 Guidance was used, in particular when the exposure is clearly lower (e.g. more than one order of magnitude) than the health‐based guidance value. Finally, the SC firmly reiterates to reconsider test guidelines given the wide application of the BMD approach.

Although benchmark-dose methodology has existed for more than 20 years, benchmark doses (BMDs) still have not fully supplanted the no-observed-adverse-effect level (NOAEL) and lowest-observed-adverse-effect level (LOAEL) as points of departure from the experimental dose–response range for setting acceptable exposure levels of toxic substances. Among the issues involved in replacing the NOAEL (LOAEL) with a BMD are (1) which added risk level(s) above background risk should be targeted as benchmark responses (BMRs), (2) whether to apply the BMD methodology to both carcinogenic and noncarcinogenic toxic effects, and (3) how to model continuous health effects that aren’t observed in a natural risk-based context like dichotomous health effects. This paper addresses these issues and recommends specific BMDs to replace the NOAEL and LOAEL.

We present a critical assessment of the benchmark dose (BMD) method introduced by Crump as an alternative method for setting a characteristic dose level for toxicant risk assessment. The no-observed-adverse-effect-level (NOAEL) method has been criticized because it does not use all of the data and because the characteristic dose level obtained depends on the dose levels and the statistical precision (sample sizes) of the study design. Defining the BMD in terms of a confidence bound on a point estimate results in a characteristic dose that also varies with the statistical precision and still depends on the study dose levels. Indiscriminate choice of benchmark response level may result in a BMD that reflects little about the dose-response behavior available from using all of the data. Another concern is that the definition of the BMD for the quantal response case is different for the continuous response case. Specifically, defining the BMD for continuous data using a ratio of increased effect divided by the background response results in an arbitrary dependence on the natural background for the endpoint being studied, making comparison among endpoints less meaningful and standards more arbitrary. We define a modified benchmark dose as a point estimate using the ratio of increased effect divided by the full adverse response range which enables consistent placement of the benchmark response level and provides a BMD with a more consistent relationship to the dose-response curve shape.

Derived no-effect level (DNEL)