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Evidence that glyphosate, the active ingredient in Roundup, is not present in human milk Michelle K McGuire, Kimberly Lackey, Janae Carrothers, Bahman Shafii, William Price, and Mark A McGuire School of Biological Sciences and Paul G. Allen School for Global Animal Health, Washington State University, Pullman, WA, USA; Statistical Programs, College of Agricultural and Life Sciences, University of Idaho, Moscow, ID, USA; Department of Animal and Veterinary Science, University of Idaho, Moscow, ID, USA Abstract Conclusions Background and Hypothesis Subjects and Methods Results, cont. Glyphosate [N-(phosphonomethyl) glycine], the active ingredient in Roundup, is used to control broadleaf weeds and grasses in row crops, ornamental plants, lawns, and forest plantings. Despite publication of numerous studies showing that use of glyphosate as directed poses no risk to humans, there continues to be public concern. Recently, an on-line publication of a small survey conducted by an advocacy group claimed that 3 out of 10 human milk samples they tested had high levels of glyphosate (76-166 g/mL), and some urine samples contained glyphosate levels 10-times higher than those found in Europe. This non-peer-reviewed publication has generated some attention and concern. However, the ELISA- based assay used in this study was neither validated nor optimized for human milk, a complex biological matrix notoriously difficult to accurately analyze. In response, after receiving Institutional Review Board approval and consent from subjects, we collected milk and urine samples from 41 healthy lactating women living in the Moscow, ID-Pullman, WA area a highly-productive agricultural region of the US Pacific Northwest where glyphosate is used routinely in farming practices. Questionnaires documenting potential glyphosate exposure from the environment, activities, and foods were also completed. Milk and urine samples were analyzed for glyphosate and aminomethylphosphonic acid (AMPA, a glyphosate metabolite) using highly sensitive LC-MS/MS methods validated for and optimized to each sample matrix. Limits of detection (LOD) and quantification (LOQ) for glyphosate in milk were 2.0 and 10.0 g/L, respectively; those for urine were 0.02 and 0.10 g/L, respectively. LOD and LOQ for AMPA in milk were 2.0 and 10.0 g/L, respectively; those for urine were 0.03 and 0.10 g/L, respectively. Ten of the women reported living on or directly adjacent to a farm or ranch; 23 of the women described their personal diet as conventional; and 5 had personally mixed or applied glyphosate sometime in the past. Contrary to the advocacy groups report, our assay detected neither glyphosate nor AMPA in any milk sample. Glyphosate concentrations of 8 and 3 of the urine samples were