LC-MS/MS quantification of sulforaphane and indole-3-carbinol metabolites in human plasma and urine after dietary intake of selenium-fortified broccoli

This study aimed at developing a sensitive LC-MS/MS method for the quantification of sulforaphane (SFN) and indole-3-carbinol metabolites in plasma and urine after dietary intake of regular and selenium-fertilized broccoli using stable isotope dilution analysis. In a three-armed, placebo-controlled, randomized human intervention study with 76 healthy volunteers, 200 g of regular (485 μg of total glucosinolates and <0.01 μg of selenium per gram fresh weight) or selenium-fertilized broccoli (589 μg of total glucosinolates and 0.25 μg of selenium per gram fresh weight) was administered daily for 4 weeks. Glucoraphanin and glucobrassicin metabolites quantified in plasma and urine were SFN-glutathione, SFN-cysteine, SFN-cysteinylglycine, SFN-acetylcysteine, and indole-3-carboxaldehyde, indole-3-carboxylic acid, and ascorbigen, respectively. Dietary intake of selenium-fertilized broccoli increased serum selenium concentration analyzed by means of atomic absorption spectroscopy by up to 25% (p < 0.001), but affected neither glucosinolate concentrations in broccoli nor their metabolite concentrations in plasma and urine compared to regular broccoli.     

Bioavailability of pelargonidin-3-O-glucoside and its metabolites in humans following the ingestion of strawberries with and without cream

Plasma and urine were collected over a 24 h period after the consumption by humans of 200 g of strawberries, containing 222 micromol of pelargonidin-3- O-glucoside, with and without cream. The main metabolite, a pelargonidin- O-glucuronide, reached a peak plasma concentration ( C max) of 274 +/- 24 nmol/L after 1.1 +/- 0.4 h ( t max) when only strawberries were ingested. When the strawberries were eaten with cream, the C max was not statistically different but the t max at 2.4 +/- 0.5 h was delayed significantly ( p < 0.001). The pelargonidin- O-glucuronide, along with smaller quantities of other metabolites, was also excreted in urine in quantities corresponding to ca. 1% of anthocyanin intake. The quantities excreted over the 0-24 h collection period were not influenced significantly by cream. However, the 0-2 h excretion of anthocyanin metabolites was significantly lower when the strawberries were eaten with cream, whereas the reverse occurred during with the 5-8 h excretion period. In keeping with these observations, measurement of plasma paracetamol and breath hydrogen revealed that cream delayed gastric emptying and extended mouth to cecum transit time.     

Dietary intake and the insulin-like growth factor system: effects of migration in two related populations in India and Britain with markedly different dietary intake

BACKGROUND: The insulin-like growth factor (IGF) system is implicated in the pathogenesis of diabetes and cardiovascular disease. OBJECTIVE: We report the effects of total energy intake on the IGF system in two populations with markedly different dietary macronutrient intake and cardiovascular event rate. DESIGN, SUBJECTS AND SETTING: Dietary macronutrient intake was measured in a specific Gujarati migrant community in Sandwell, UK (n=205) compared with people still resident in the same villages of origin in India (n=246). Fasting IGF-I, IGF-binding protein (IGFBP)-1 and IGFBP-3, insulin and glucose (0 and 2-hour) were measured. RESULTS: Total energy and total fat intake were higher in UK migrants, as were IGFBP-3 and IGF-I (mean (95% confidence interval): 145.9 (138.1-153.6) vs. 100.9 (94.6-107.3) ng ml(-1); F=76.6, P<0.001). IGFBP-1 was lower in UK migrants (29.5 (25.9-33.0) vs. 56.5 (50.6-62.5) microg l(-1); F=48.4, P<0.001). At both sites, IGF-I correlated positively with total energy (Spearman's rho=0.45, P<0.001) and total fat (rho=0.44, P<0.001) as did IGFBP-3 with total energy (rho=0.21, P<0.05) and fat (rho=0.26, P<0.001). Conversely, in Indian Gujaratis, IGFBP-1 fell with increasing total energy (rho=-0.27, P<0.001) and fat intake (rho=-0.26, P<0.01) but not in UK Gujaratis. Multiple linear regression modelling showed that increasing quartiles of fat intake were associated with higher IGF-I (beta=0.42, P=0.007) independent of age, body mass index, plasma insulin, fatty acids and 2-hour glucose. CONCLUSION: In these genetically similar groups, migration to the UK and adoption of a different diet is associated with marked changes in the IGF system, suggesting that environmental factors profoundly modulate serum concentrations and actions of IGFs.     

Biotransformation of vinclozolin in rat precision-cut liver slices: comparison with in vivo metabolic pattern

Vinclozolin is a dicarboxymide fungicide that presents antiandrogenic properties through its two hydrolysis products M1 and M2, which bind to the androgen receptor. Because of the lack of data on the biotransformation of vinclozolin, its metabolism was investigated in vitro in precision-cut rat liver slices and in vivo in male rat using [ (14)C]-vinclozolin. Incubations were performed using different concentrations of substrate, and the kinetics of formation of the major metabolites were studied. Three male Wistar rats were fed by gavage with [ (14)C]-VZ. Urine was collected for 24 h and analyzed by radio-HPLC for metabolic profiling. Metabolite identification was carried out on a LCQ ion trap mass spectrometer. In rat liver slices and in vivo, the major primary metabolite has been identified as 3',5'-dichloro-2,3,4-trihydroxy-2-methylbutyranilide (M5) and was mainly present as glucuronoconjugates. M5 is produced by dihydroxylation of the vinyl group of M2. Other metabolites have been identified as 3-(3,5-dichlorophenyl)-5-methyl-5-(1,2-dihydroxyethyl)-1,3-oxazolidine-2,4-dione (M4), a dihydroxylated metabolite of vinclozolin, which undergoes further conjugation to glucuronic acid, and 2-[[(3,5-dichlorophenyl)-carbamoyl]oxy]-2-methyl-3,4-dihydroxy-butanoic acid (M6), a dihydroxylated metabolite of M1.     

Healthy whole-grain choices for children and parents: a multi-component school-based pilot intervention

OBJECTIVE: The aim of the present study was to pilot-test a school-based intervention designed to increase consumption of whole grains by 4th and 5th grade children. DESIGN: This multi-component school-based pilot intervention utilised a quasi-experimental study design (intervention and comparison schools) that consisted of a five-lesson classroom curriculum based on Social Cognitive Theory, school cafeteria menu modifications to increase the availability of whole-grain foods and family-oriented activities. Meal observations of children estimated intake of whole grains at lunch. Children and parents completed questionnaires to assess changes in knowledge, availability, self-efficacy, usual food choice and role modelling. SETTING/SAMPLE: Parent/child pairs from two schools in the Minneapolis metropolitan area; 67 in the intervention and 83 in the comparison school. RESULTS: Whole-grain consumption at the lunch meal increased by 1 serving (P < 0.0001) and refined-grain consumption decreased by 1 serving for children in the intervention school compared with the comparison school post-intervention (P < 0.001). Whole-grain foods were more available in the lunches served to children in the intervention school compared with the comparison school post-intervention (P < 0.0001). The ability to identify whole-grain foods by children in both schools increased, with a trend towards a greater increase in the intervention school (P = 0.06). Parenting scores for scales for role modelling (P < 0.001) and enabling behaviours (P < 0.05) were significantly greater for parents in the intervention school compared with the comparison school post-intervention. CONCLUSIONS: The multi-component school-based programme implemented in the current study successfully increased the intake of whole-grain foods by children.     

Bioavailability and antioxidant effects of orange juice components in humans

Seven healthy females and six males consumed daily 256 mg of vitamin C, 271 mg of flavanones (mainly as glycosides), 6 mg of carotenoids (mainly xanthophylls and cryptoxanthins), and 0.16 mg of folate by incorporation of daily three times 236 mL of not from concentrate orange juice (OJ) into their habitual diet. At the end of 3 weeks, mean vitamin C, folate, carotenoid, and flavanone plasma concentrations increased significantly relative to baseline by 59% (p < 0.001), 46% (p = 0.018), and 22% (p < 0.001), and 8-fold (p = 0.045), respectively. Flavanones were excreted in urine 9-fold more at the end of the intervention (p = 0.01) but returned to baseline 2 days after study completion. After the 3 week intervention, plasma concentrations of vitamins A and E did not change. 8-Hydroxydeoxyguanosine in white blood cells declined by 16% (p = 0.38; n = 11), and in individuals with high baseline concentrations by 29% (p = 0.36; n = 7), respectively. Low-density lipoprotein (LDL)-/high-density lipoprotein (HDL)-cholesterol ratios decreased but cholesterol (HDL, LDL, total) and thiobarbituric acid reactive substance plasma concentrations did not change significantly. We conclude from this pilot study that OJ is an excellent food source to enhance circulating concentrations of valuable hydrophilic as well as lipophilic phytochemicals.     

Validity of protein, retinol, folic acid and n-3 fatty acid intakes estimated from the food-frequency questionnaire used in the Danish National Birth Cohort

OBJECTIVE: To validate intakes of protein, folic acid, retinol and n-3 fatty acids estimated from a food-frequency questionnaire in week 25 of pregnancy (FFQ-25). DESIGN: Validation was done against a 7-day weighed food diary (FD) and biomarkers of the nutrients in gestation week 32-38. SUBJECTS AND SETTING: The FFQ-25 to be validated was used in the Danish National Birth Cohort comprising 101 042 pregnant Danish women, of whom 88 participated in the present validation study. RESULTS: Estimated intakes of protein, retinol and folic acid did not differ significantly between the two dietary methods, but intake of n-3 fatty acids was one third larger when estimated from the FFQ-25. The intakes estimated from the two dietary methods were all significantly correlated, ranging from 0.20 for retinol intake to 0.57 for folic acid intake. Sensitivities of being correctly classified into low and high quintiles were between 0.22 and 0.77, and specificities were between 0.62 and 0.89. Urinary protein content did not correlate significantly with protein estimated from the FFQ (r = 0.17, P > 0.05), but did with intake estimated from the FD (r = 0.56, P < 0.0001). Erythrocyte folate correlated significantly with the estimated total intake from the FFQ (r = 0.55, P < 0.0001) and the FD (r = 0.52, P < 0.0001). No correlations with plasma retinol were found. Erythrocyte eicosapentaenoic acid (C20:5n-3) correlated significantly with n-3 fatty acids intake estimated from both the FFQ-25 (r = 0.37, P < 0.001) and the FD (r = 0.62, P < 0.0001). CONCLUSION: The FFQ-25 gives reasonable valid estimates of protein, retinol and folic acid intakes, but seems to overestimate intake of n-3 fatty acids.     

Absorption, disposition, metabolism, and excretion of [3-(14)C]caffeic acid in rats

Male Sprague-Dawley rats ingested 140 × 10(6) dpm of [3-(14)C]trans-caffeic acid, and over the ensuing 72 h period, body tissues, plasma, urine, and feces were collected and the overall levels of radioactivity determined. Where sufficient radioactivity had accumulated, samples were analyzed by HPLC with online radioactivity and tandem mass spectrometric detection. Nine labeled compounds were identified, the substrate and its cis isomer, 3'-O- and 4'-O-sulfates and glucuronides of caffeic acid, 4'-O-sulfates and glucuronides of ferulic acid, and isoferulic acid-4'-O-sulfate. Four unidentified metabolites were also detected. After passing down the gastrointestinal tract, the majority of the radiolabeled metabolites were excreted in urine with minimal accumulation in plasma. Only relatively small amounts of an unidentified (14)C-labeled metabolite were expelled in feces. There was little or no accumulation of radioactivity in body tissues, including the brain. The overall recovery of radioactivity 72 h after ingestion of [3-(14)C]caffeic acid was ～80% of intake.     

Increased salicylate concentrations in urine of human volunteers after consumption of cranberry juice

The aim of this study was to assess whether regular consumption of cranberry juice results in elevations in urinary salicylate concentrations in persons not taking salicylate drugs. Two groups of healthy female subjects (11/group) matched for age, weight, and height consumed 250 mL of either cranberry juice or a placebo solution three times a day (i.e., 750 mL/day) for 2 weeks. At weekly intervals, salicylic acid and salicyluric acid (the major urinary metabolite of salicylic acid) concentrations were determined in urine by HPLC with electrochemical detection. Concentrations of salicylic acid in plasma were also determined. Consumption of cranberry juice was associated with a marked increase (p < 0.001) of salicyluric and salicylic acids in urine within 1 week of the intervention. After 2 weeks, there was also a small but significant (p < 0.05) increase in salicylic acid in plasma. The regular consumption of cranberry juice results in the increased absorption of salicylic acid, an anti-inflammatory compound that may benefit health.     

Rapid liquid chromatography tandem mass spectrometry assay to quantify plasma (-)-epicatechin metabolites after ingestion of a standard portion of cocoa beverage in humans

A rapid liquid chromatography electrospray ionization tandem mass spectrometry with negative ion detection method was developed and validated to determine cocoa flavonoid metabolites in human plasma and urine after the intake of a standard portion of a cocoa beverage. A chromatographic run time of only 9 min provided clear separation of all metabolites and internal standards. Samples were analyzed in a product-ion scan of m/z 289, 369, and 465 to identify the metabolites and in multiple reaction monitoring acquisition mode to quantify (-)-epicatechin ((-)-Ec) (289/ 245), (-)-epicatechin-glucuronide ((-)-EcG) (465/289), and (-)-epicatechin-sulfate ((-)-EcS) (369/289). One (-)-Ec-G and three (-)-Ec-S were identified and confirmed in urine as the major metabolites, and one (-)-Ec-G was the only metabolite present in plasma volunteers (n = 5) at a mean concentration of 625.7 +/- 198.3 nmol/L at 2 h after consumption of a cocoa beverage containing 54.4 mg of (-)-Ec.     

Homocysteine and cobalamin status in German vegans

OBJECTIVE: The study aimed to evaluate the homocysteine and cobalamin status of German vegans and determine whether the intake of very small amounts of foods of animal origin can improve this status. DESIGN AND SETTING: Cross-sectional study, Germany. SUBJECTS: The dietary and nutritional intakes of 131 vegans (73 women, 58 men; age range: 20.2-82.1 years) were evaluated using a general questionnaire and two food-frequency questionnaires. RESULTS: The prevalence of inadequate cobalamin status in volunteers of the German Vegan Study was 28.2%, and that of hyperhomocysteinaemia, 38.1%. Moderate vegans were affected to a lesser extent than were strict vegans. Duration of veganism and cobalamin concentration were inversely correlated (Spearman's r=-0.175, P=0.047). Folate concentration and erythrocyte aspartic acid aminotransferase activity were not correlated with plasma homocysteine concentration, but duration of veganism correlated positively with homocysteine concentration (Spearman's r=0.319, P<0.001). Cobalamin and homocysteine concentrations were inversely correlated (when controlling for duration of veganism; r=-0.602, P<0.001). CONCLUSION: Cobalamin status needs to be improved in order to minimise the risk of hyperhomocysteinaemia.     

Tissue depletion of amoxicillin and its major metabolites in pigs: influence of the administration route and the simultaneous dosage of clavulanic acid

A residue depletion study of amoxicillin (AMO) and its major metabolites, amoxicilloic acid (AMA) and amoxicillin diketopiperazine-2',5'-dione, was performed after a single oral (p.o.) and intravenous (i.v.) administration of amoxicillin (20 mg kg (-1)) and amoxicillin/clavulanic acid (20 and 5 mg kg (-1)) to pigs. Animals were slaughtered 12, 36, 48, 60, 72, and 84 h after dosing. Tissue samples were analyzed using liquid chromatography-tandem mass spectrometry. Kidney samples contained high concentrations of amoxicilloic acid metabolite, which depleted much slower from tissues than amoxicillin, both after p.o. (t1/2AMO = 4.5 h vs t1/2AMA = 8 h) and i.v. (t1/2AMO = 4 h vs t1/2AMA = 8 h) administration. Moreover, after oral administration, significantly higher amoxicilloic acid concentrations were measured in liver and kidney than after i.v. administration. The coadministration of amoxicillin with clavulanic acid provoked no significant differences in amoxicilloic acid tissue concentrations as compared to an amoxicillin dosing. The prolonged presence of residues of amoxicilloic acid in edible tissues can play an important role in food safety, because the compound could give rise to a possible health risk, although it is not included in the maximum residue limit legislation.     

Metabolite profiling based on lipophilic compounds for quality assessment of perilla (Perilla frutescens) cultivars

Lipophilic compounds from Korean perilla ( Perilla frutescens ) seeds were characterized to determine the diversity among their phytochemicals and to analyze relationships between their contents. Twenty-four metabolites consisting of policosanol, phytosterol, tocopherol, and fatty acids were identified. The metabolite profiles were subjected to data mining processes, including principal component analysis (PCA), partial least-squares discriminate analysis (PLS-DA), and Pearson's correlation analysis. PLS-DA could distinguish between all cultivars except between Daesil and Daeyeup cultivars. Linolenic acid contents were positively correlated with β-sitosterol (r = 0.8367, P < 0.0001) and γ-tocopherol contents (r = 0. 7201, P < 0.001) among all perilla grains. The Daesil and Daeyeup cultivars appear to be good candidates for future breeding programs because they have simultaneously high linolenic acid, phytosterol, and tocopherol levels. These results demonstrate the use of metabolite profiling as a tool for assessing the quality of food.     

Biotransformation of vinclozolin by the fungus Cunninghamella elegans

This study investigated the biotransformation of the dicarboximide fungicide vinclozolin [3-(3,5-dichlorophenyl)-5-methyl-5-vinyl-1,3-oxazolidine-2,4-dione] by the fungus Cunninghamella elegans. Experiments with phenyl-[U-ring-14C]vinclozolin showed that after 96 h incubation, 93% had been transformed to four major metabolites. Metabolites were separated by HPLC and characterized by mass and NMR spectroscopy. Biotransformation occurred predominantly on the oxazolidine-2,4-dione portion of vinclozolin. The metabolites were identified as the 3R- and 3S- isomers of 3',5'-dichloro-2,3,4-trihydroxy-2-methylbutyranilide, N-(2-hydroxy-2-methyl-1-oxobuten-3-yl)-3,5-dichlorophenyl-1-carbamic acid, and 3',5'-dichloro-2-hydroxy-2-methylbut-3-enanilide. The enanilide compound has been reported previously as a plant and mammalian metabolite and is implicated to contain antiandrogenic activity. The 3R- and 3S- isomers of 3',5'-dichloro-2,3,4-trihydroxy-2-methylbutyranilide are novel metabolites.     

Impact of promotion of mango and liver as sources of vitamin A for young children: a pilot study in Burkina Faso

OBJECTIVE: To assess the effectiveness of a behaviour change approach, with or without financial support, in improving vitamin A (VA) intake and serum retinol concentration through mango and liver consumption by children. DESIGN: A parallel design (no control area) was used to assess changes in VA intake and serum retinol over a 15-week period. SETTING AND SUBJECTS: A pilot study was implemented in the Department of Kokologho, a rural area in central west Burkina Faso. One hundred and fifty children aged 2-3 years were randomly selected and assigned to two treatment groups: PA$$ (promotional activities and financial support) and PA (promotional activities). RESULTS: The intervention significantly increased (P < 0.001) total VA intake by 56% in PA$$ and by 50% in PA. VA intake from liver increased significantly (P < 0.001) from 12.7 +/- 23.5 to 155.3 +/- 56.3 microg retinol activity equivalents (RAE) in PA$$ and from 21.6 +/- 29.7 to 135.3 +/- 44.9 microg RAE in PA. Changes in VA intake from liver were significantly higher (P = 0.004) in PA$$ compared with PA. Mean serum retinol concentration increased significantly by 26% (P < 0.001) in PA$$ and 30% (P < 0.001) in PA. Changes in serum retinol concentration (0.13 micromol l(-1) in PA$$ vs. 0.17 micromol l(-1) and in PA) did not differ significantly (P = 0.455) between groups over the intervention. CONCLUSION: Promotional activities on mango and liver intake effectively increased VA intake and serum retinol concentrations. Although an additional beneficial effect of financial support on liver intake was observed, this did not translate into a further increase in serum retinol concentration.     

Sorghum bran in the diet dose dependently increased the excretion of catechins and microbial-derived phenolic acids in female rats

Sorghum bran is concentrated with procyanidins (predominately polymers), which may be beneficial for health in humans; however, the bioavailability of procyanidins is not well-understood. Female Sprague-Dawley rats were fed an AIN93G diet containing 0, 5, 10, 20, or 40% Hi-tannin sorghum bran (n = 5-7 for each group) for 50 days. Sorghum bran contained 23.3 mg/g of procyanidins. The urinary excretions of catechin, epicatechin, methylated catechins, and phenolic acids were analyzed using liquid chromatography-tandem mass spectrometry. Sorghum bran dose dependently increased the urinary excretion of catechin (0-2.2 nmol/day) and 3'-O-methylcatechin (0-9.5 nmol/day). Their serum concentrations also increased with dose (range of 0-14 nM for 3'-O-methylcatechin). Among the 14 phenolic acids analyzed, 3,4-dihydroxybenzoic acid, 3-methoxy-4-hydroxybenzoic acid, and 4-hydroxyphenylacetic acid dominated in the serum (1.8-8 micromol/L). In the urine, 3-methoxy-4-hydroxyphenylacetic acid, 3-hydroxyphenylacetic acid, and 3-hydroxyphenylpropionic acid dominated and their excretion increased significantly with the level of sorghum bran in the diet. The summed phenolic acid excretion was 0.8 micromol/day in the control group and increased to 23 micromol/day for 40% sorghum bran group. The hippuric acid excretion ranged from 2.2 to 16.2 micromol/day and peaked in the 10% sorghum bran group. On the basis of chromic oxide, a nonabsorbable marker, total procyanidins and polymers disappeared progressively, and significant degradation occurred in the cecum and colon. Catechins and procyanidins in sorghum were bioavailable; however, bacteria-derived phenolic acids were the predominant metabolites of procyanidins. Procyanidins degraded in the gastrointestinal tract. Depolymerization was not observed.     

Whole-grain intake as a marker of healthy body weight and adiposity

OBJECTIVE: To review evidence relating to the consumption of whole grains and healthy body weight (BW). DESIGN: Systematic review and analysis of observational studies reporting whole-grain consumption and measures of BW and adiposity, including the effect on macronutrient intakes and lifestyle factors. SETTING: Medline and other databases were searched for the period 1990 to 2006 to produce a full reference list; observational studies were retained for further analysis if they included an appropriate control group and reported whole-grain intake and body mass index (BMI) or a measure of adiposity. SUBJECTS: Fifteen trials were identified which included data from 119 829 male and female subjects aged 13 years and over. RESULTS: The combined and weighted mean difference in BMI from 15 studies representing 20 treatment groups (n = 119 829) using a random-effects model was 0.630 kg/m2 lower when high versus low whole-grain intake was compared, P < 0.0001 (95 % confidence interval (CI) 0.460, 0.800 kg/m2). In high consumers, adiposity assessed as waist circumference was reduced by 2.7 (95 % CI 0.2, 5.2) cm, P = 0.03 (six data sets, n = 4178) or as waist:hip ratio by 0.023 (95 % CI 0.016, 0.030), P < 0.0001 (four data sets, n = 20 147). Higher intake of whole grains led to increased dietary fibre intake (9 g, P < 0.01), while total and saturated fat intakes decreased by 11 g and 3.9 g, respectively. CONCLUSION: A higher intake of whole grains (about three servings per day) was associated with lower BMI and central adiposity. In addition, people who consume more whole grains are likely to have a healthier lifestyle as fewer of them smoke, they exercise more frequently and they tend to have lower fat and higher fibre intakes.     

植物乳杆菌对苏尼特羊肠道菌群、血浆代谢物及肉品质的影响

为了研究植物乳杆菌对苏尼特羊肠道菌群、血浆代谢物和肉品质的影响及其作用机理,选取三月龄苏尼特羊为试验对象,随机分为2组:对照组(C组,基础日粮)和植物乳杆菌组(R组,基础日粮、活菌数为3×1010cfu/g植物乳杆菌),进行为期90 d的饲喂试验.屠宰后取其肠道内容物及背最长肌,利用高通量测序技术、代谢组学液相-色谱联...     

Preparative-scale synthesis of two metabolites isolated from soil treated with zoxium fungicide and kerb herbicide.

The preparation in multigram scale of two metabolites 3-(3,5-dichloro-4-methyl-benzoylamino)-2-hydroxy-3-methyl-pentanoic acid and 3-(3,5-dichloro-benzoylamino)-3-methyl-2-oxo-butyric acid isolated from soil treated with either Zoxium fungicide or Kerb herbicide was efficiently accomplished using a common 5-step synthetic process starting from easily available raw materials.     

Comparison of Lead Extraction and Detection Procedures for Six Canadian Soils

The effects of acid digestion procedures and instrumentation on extracted lead (Pb) concentrations from several soils, including a mildly Pb-contaminated soil, were determined using a two-factor factorial experiment. The two factors were (i) digestion procedure [seven levels: U.S. EPA, AOAC (dry ashing), nitric acid (NA), three aqua regia procedures (AR1, AR2, and AR3), and hydrofluoric acid (HF)] and (ii) instrumentation [two levels: atomic absorption spectrometry (AAS) and inductively coupled plasma (ICP) spectroscopy]. The greatest Pb recovery was obtained when soils were digested with HF and analyzed by AAS. The results suggest that the AOAC procedure (a standard procedure for recovery of soil nutrients and trace elements in Canadian laboratories) and EPA procedure (a standard procedure in American laboratories) may underestimate Pb concentrations in some Atlantic Canadian soil types. The AAS procedure is more accurate than ICP for determining Pb concentrations in soil with a history of PbHAsO₄, at least for Atlantic Canadian soils.