Article pubs.acs.org/JAFC
Exposure Assessment to Ochratoxin A in Chinese Wine Qi Ding Zhong,*,† Guo Hui Li,†,§ Dao Bing Wang,† Yi Shao,# Jing Guang Li,# Zheng He Xiong,† and Yong Ning Wu*,# †
China National Institute of Food and Fermentation Industries, Beijing, China 100015 Key Laboratory of Analytical Science of Hebei Province, Key Laboratory of Medical Chemistry and Molecular Diagnosis, Ministry of Education, and College of Chemistry and Environmental Science, Hebei University, Baoding, China 071002 # Key Laboratory of Food Safety Risk Assessment, Ministry of Health, China National Centre for Food Safety Risk Assessment, Beijing, China 100021 §
ABSTRACT: A rapid, sensitive, reproducible, and inexpensive method of high-performance liquid chromatography with fluorescence detection after an anion-exchange solid-phase extraction cleanup step for the analysis of ochratoxin A (OTA) in Chinese wine was developed. The average recovery rate and the average RSD of recovery were 97.47% and about 4%. The relative standard deviations of both the interday and intraday precision were 6.7 and 12.6%, respectively. The limits of detection and quantitation were determined to be 0.01 and 0.03 μg/L, respectively. A total of 223 samples from the major wine-producing areas of China were analyzed for OTA. OTA was detected at levels of 0.01−0.98 μg/L. The mean was 0.15 μg/L. Then, participants as representative inhabitants were invited to answer the designed questionnaire about the quantity and frequency of wine consumption. All data were simulated by the point evaluation for the risk assessment of OTA contamination from wine. Those results indicated that daily intake (DI) of OTA for the average adult consumer varies between 0.86 and 1.08 ng/kg bw per week, which was lower than all the reference standards. However, the DI value (4.38−5.54 ng/kg bw per week) in the high percentile (97.5) was slightly above 5% PTWI (100 ng kg−1 week−1) of the JECFA. In conclusion, OTA exposure from Chinese wine has no risk of harm. This research will provide the scientific basis for determining the maximum limit of OTA content in Chinese wine. KEYWORDS: exposure assessment, ochratoxin A, wine, Chinese
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INTRODUCTION Ochratoxin is a derivative resulting from the combination of isocoumarin with β-phenylalanine and exists in four major types, ochratoxins A, B, C, and D. Ochratoxin A (OTA) is produced by toxigenic mold species (Penicillium verrucosum, Aspergillus ochraceus, Aspergillus niger, and Aspergillus carbonarius) that occur in human food and animal feed. OTA commonly contaminates several human food types, including cereals, cacao, coffee, fruits, peanuts, cotton seed, corn, and rice.1,2 OTA has also been detected in beverages such as beer, wine, and grape juice.3,4 An evaluation by the European Commission showed that OTA was ingested mainly from grain (50%) and wine (13%). Once the juice of grapes becomes infected with Penicillium, Aspergillus, carbon black Aspergillus, or other mucedine species, OTA is produced under favorable conditions and ultimately infiltrates the wine.5−11 OTA has been classified as a possible human carcinogen (group 2B) by the International Agency for Research on Cancer on the basis of experimental studies demonstrating evidence for OTA carcinogenicity in animals.12 Results of toxicology experiments of OTA have revealed its renal toxicity, hepatotoxicity, teratogenicity, mutagenicity, carcinogenicity, and immunotoxicity properties. Generally, risk assessment is the systematic characterization of the potential adverse effects resulting from exposure to hazardous agents. Exposure of a certain population to OTA can be quantitatively assessed by analyzing the exposure data of the contaminants in foods coupled with a daily food intake survey data. There are many © 2014 American Chemical Society
research studies on the assessment and exposure assessment of OTA in different food commodities.1,13−17 Nevertheless, in contrast to other countries, evidence from human studies to evaluate the risk of OTA at current dietary exposures in China is still inadequate. Up to now, no studies have been reported to clearly identify the conclusive risk of OTA from the diet to human health in China, especially on wine because the dietary habits in China are completely different from those in most other countries, in addition to the large geographic area and different environmental conditions, that is, temperature and humidity. It is a critical issue to carry out a risk assessment to reveal the realistic probabilities of adverse effects associated with OTA to humans in China. Wine is the second contributor (13%) of OTA at EU dietary exposure followed by cereals (50%).18 Assessment of dietary intake of ochratoxin A has been done by the population of EU member states.19 In China, the results have are different, as found by Su et al.20 and Han et al.;11 their research shows that wine is the second contributor (0.5%) of OTA at EU dietary exposure after by cereals (90%), in the same order. On the basis of genetic toxicity and renal toxicity data obtained from laboratory animals, Canadian authorities proposed a tolerable daily intake (TDI) of 1.2−5.7 ng/kg Received: Revised: Accepted: Published: 8908
February 12, 2014 July 27, 2014 July 31, 2014 July 31, 2014 dx.doi.org/10.1021/jf500713x | J. Agric. Food Chem. 2014, 62, 8908−8913
Journal of Agricultural and Food Chemistry
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body weight (bw) per day. The toxin was also evaluated by the Scientific Committee on Food, which proposed that exposure should be below 5 ng/kg (bw) per day.21 The Joint FAO/ WHO Expert Committee for Food Additives (JECFA) reconfirmed the provisional tolerable weekly intake (PTWI) of OTA as 100 ng kg−1 week−1.22 At present, the International Office of Vine and Wine (OIV) is considering the application of a limit of 2 μg/kg OTA for wine;23 however, there is no regulation or limit standards with respect to OTA in wine from China currently. In 2013, Zhang et al.24 analyzed a total of 119 wines, which were obtained from four major regions around China from 1999 to 2009; the OTA levels ranged from
Exposure Assessment to Ochratoxin A in Chinese Wine Qi Ding Zhong,*,† Guo Hui Li,†,§ Dao Bing Wang,† Yi Shao,# Jing Guang Li,# Zheng He Xiong,† and Yong Ning Wu*,# †
China National Institute of Food and Fermentation Industries, Beijing, China 100015 Key Laboratory of Analytical Science of Hebei Province, Key Laboratory of Medical Chemistry and Molecular Diagnosis, Ministry of Education, and College of Chemistry and Environmental Science, Hebei University, Baoding, China 071002 # Key Laboratory of Food Safety Risk Assessment, Ministry of Health, China National Centre for Food Safety Risk Assessment, Beijing, China 100021 §
ABSTRACT: A rapid, sensitive, reproducible, and inexpensive method of high-performance liquid chromatography with fluorescence detection after an anion-exchange solid-phase extraction cleanup step for the analysis of ochratoxin A (OTA) in Chinese wine was developed. The average recovery rate and the average RSD of recovery were 97.47% and about 4%. The relative standard deviations of both the interday and intraday precision were 6.7 and 12.6%, respectively. The limits of detection and quantitation were determined to be 0.01 and 0.03 μg/L, respectively. A total of 223 samples from the major wine-producing areas of China were analyzed for OTA. OTA was detected at levels of 0.01−0.98 μg/L. The mean was 0.15 μg/L. Then, participants as representative inhabitants were invited to answer the designed questionnaire about the quantity and frequency of wine consumption. All data were simulated by the point evaluation for the risk assessment of OTA contamination from wine. Those results indicated that daily intake (DI) of OTA for the average adult consumer varies between 0.86 and 1.08 ng/kg bw per week, which was lower than all the reference standards. However, the DI value (4.38−5.54 ng/kg bw per week) in the high percentile (97.5) was slightly above 5% PTWI (100 ng kg−1 week−1) of the JECFA. In conclusion, OTA exposure from Chinese wine has no risk of harm. This research will provide the scientific basis for determining the maximum limit of OTA content in Chinese wine. KEYWORDS: exposure assessment, ochratoxin A, wine, Chinese
■
INTRODUCTION Ochratoxin is a derivative resulting from the combination of isocoumarin with β-phenylalanine and exists in four major types, ochratoxins A, B, C, and D. Ochratoxin A (OTA) is produced by toxigenic mold species (Penicillium verrucosum, Aspergillus ochraceus, Aspergillus niger, and Aspergillus carbonarius) that occur in human food and animal feed. OTA commonly contaminates several human food types, including cereals, cacao, coffee, fruits, peanuts, cotton seed, corn, and rice.1,2 OTA has also been detected in beverages such as beer, wine, and grape juice.3,4 An evaluation by the European Commission showed that OTA was ingested mainly from grain (50%) and wine (13%). Once the juice of grapes becomes infected with Penicillium, Aspergillus, carbon black Aspergillus, or other mucedine species, OTA is produced under favorable conditions and ultimately infiltrates the wine.5−11 OTA has been classified as a possible human carcinogen (group 2B) by the International Agency for Research on Cancer on the basis of experimental studies demonstrating evidence for OTA carcinogenicity in animals.12 Results of toxicology experiments of OTA have revealed its renal toxicity, hepatotoxicity, teratogenicity, mutagenicity, carcinogenicity, and immunotoxicity properties. Generally, risk assessment is the systematic characterization of the potential adverse effects resulting from exposure to hazardous agents. Exposure of a certain population to OTA can be quantitatively assessed by analyzing the exposure data of the contaminants in foods coupled with a daily food intake survey data. There are many © 2014 American Chemical Society
research studies on the assessment and exposure assessment of OTA in different food commodities.1,13−17 Nevertheless, in contrast to other countries, evidence from human studies to evaluate the risk of OTA at current dietary exposures in China is still inadequate. Up to now, no studies have been reported to clearly identify the conclusive risk of OTA from the diet to human health in China, especially on wine because the dietary habits in China are completely different from those in most other countries, in addition to the large geographic area and different environmental conditions, that is, temperature and humidity. It is a critical issue to carry out a risk assessment to reveal the realistic probabilities of adverse effects associated with OTA to humans in China. Wine is the second contributor (13%) of OTA at EU dietary exposure followed by cereals (50%).18 Assessment of dietary intake of ochratoxin A has been done by the population of EU member states.19 In China, the results have are different, as found by Su et al.20 and Han et al.;11 their research shows that wine is the second contributor (0.5%) of OTA at EU dietary exposure after by cereals (90%), in the same order. On the basis of genetic toxicity and renal toxicity data obtained from laboratory animals, Canadian authorities proposed a tolerable daily intake (TDI) of 1.2−5.7 ng/kg Received: Revised: Accepted: Published: 8908
February 12, 2014 July 27, 2014 July 31, 2014 July 31, 2014 dx.doi.org/10.1021/jf500713x | J. Agric. Food Chem. 2014, 62, 8908−8913
Journal of Agricultural and Food Chemistry
■
body weight (bw) per day. The toxin was also evaluated by the Scientific Committee on Food, which proposed that exposure should be below 5 ng/kg (bw) per day.21 The Joint FAO/ WHO Expert Committee for Food Additives (JECFA) reconfirmed the provisional tolerable weekly intake (PTWI) of OTA as 100 ng kg−1 week−1.22 At present, the International Office of Vine and Wine (OIV) is considering the application of a limit of 2 μg/kg OTA for wine;23 however, there is no regulation or limit standards with respect to OTA in wine from China currently. In 2013, Zhang et al.24 analyzed a total of 119 wines, which were obtained from four major regions around China from 1999 to 2009; the OTA levels ranged from
