A liquid chromatographic (LC) method was validated for the determination of total vitamin B6 in i... more A liquid chromatographic (LC) method was validated for the determination of total vitamin B6 in infant formula. Total vitamin B6 was quantified by converting the phosphorylated and free vitamers into pyridoxine. Pyridoxine was determined by ion pair reversed-phase LC with fluorescence detection. The method was subjected to an AOAC collaborative study involving a factory-manufactured, milk- and soy-based infant formula. Each was spiked at 3 concentrations in the range of 0-1 microg/g and sent as blind duplicate to participant laboratories. Nine laboratories returned valid data which were statistically analyzed for outliers and precision parameters. The repeatability relative standard deviation (RSD(r)) ranges were 2.0-4.0 and 3.5-5.9% for fortified milk- and soy-based formulas, respectively. The reproducibility relative standard deviation (RSD(R)) ranges were 8.2-8.4 and 6.7-11.2% for fortified milk- and soy-based formulas, respectively. HORRAT values ranged from 0.42 to 0.53, indica...
Determination of total fluoroacetic acid and its salts in all forms of infant formula is evaluate... more Determination of total fluoroacetic acid and its salts in all forms of infant formula is evaluated. Infant formula includes breast-milk substitute specially manufactured to satisfy, by itself, the nutritional requirements of infants during the first months of life up to the introduction of appropriate complementary feeding, made from any combination of milk, soy, rice, whey, hydrolyzed protein, starch, and amino acids, with and without intact protein. Suitable methods will include blank check samples, and check standards at the lowest point and midrange point of the analytical range.
Abstract Infant formulas are strictly regulated and rigorously tested for compliance. Recently, n... more Abstract Infant formulas are strictly regulated and rigorously tested for compliance. Recently, new official analytical methods/standards have been established for nutrient analyses in these product categories through the Stakeholder Panel on Infant Formula and Adult Nutritionals (SPIFAN), governed by AOAC INTERNATIONAL. Many of these methods have been adopted or are in the process of being adopted as reference methods by Codex Alimentarius. The purpose of this paper is to assess the ability of these cutting-edge analytical methods to deliver acceptable results in the context of established regulatory limits for nutrients in food standards and regulations. For this evaluation, the analytical method variability is considered as one of the three main sources of overall process variability, which also includes variation in raw materials/ingredients and the manufacturing process. The process capability (Cp) is a concept for determining the overall process variability relative to specification limits for a parameter in the final product. Based on this principle an analytical method capability (Cm) was defined and calculated for SPIFAN methods. Global regulatory requirements were evaluated including minimum and maximum limits and tolerances from the declared label values. Compared to these requirements, analytical methods for vitamins A, B12, D and folic acid are of particular concern in relation to the requirements in China, some EU member states, Pakistan, Russia, Singapore, South Africa and Thailand. For a product with a manufacturing target at the midpoint of the regulatory range for these nutrients, the probability of obtaining an analytical result outside the regulatory requirements due to analytical variation alone can be as high as 19%. This does not consider variability caused by the production process and/or raw materials/ingredients. These currently used analytical methods are state-of-the-art and represent the latest developments in technology. However, this work demonstrates that continuous method improvements for the nutrients identified must be pursued. In addition, this work supports a risk management approach that takes into consideration analytical method capability when establishing regulatory limits for nutrients in infant formulas. Ongoing efforts towards harmonization of regulatory requirements across global markets will facilitate evaluation of regulatory compliance in infant formulas.
Background: Monochloropropanediol (MCPD) and its fatty acid esters as well as glycidyl fatty acid... more Background: Monochloropropanediol (MCPD) and its fatty acid esters as well as glycidyl fatty acid esters are substances generated during oil refining or food processing. The free form released during digestion, 3-MCPD and glycidol, have shown adverse effects in animal studies. Objective: So far, the available analytical methods have not been validated in a collaborative study for infant and adult nutritional formulas. This manuscript describes a single-laboratory validation method in view of a future multilaboratory validation trial. Methods: The method described is for the direct determination of 2- and 3-MCPD and indirect determination of 2- and 3-MCPD esters and glycidyl esters in infant and adult/pediatric nutritional formulas by GC coupled to MS. Results: The analytical range was found to be 4–2000 μg/kg powder formula and 0.7–333 μg/kg liquid formula for fatty acid esters of MCPD and glycidol, and 2.5–750 μg/kg samples for free MCPD. The recovery rates were within 91–124% for ...
AOAC SMPRs describe the minimum recommended performance characteristics to be used during the eva... more AOAC SMPRs describe the minimum recommended performance characteristics to be used during the evaluation of a method. The evaluation may be an on-site verification, a singlelaboratory validation, or a multi-site collaborative study. SMPRs are written and adopted by AOAC stakeholder panels composed of representatives from the industry, regulatory organizations, contract laboratories, test kit manufacturers, and academic institutions. AOAC SMPRs are used by AOAC expert review panels in their evaluation of validation study data for methods being considered for Performance Tested MethodsSM or AOAC Official Methods of AnalysisSM, and can be used as acceptance criteria for verification at user laboratories.
A brief overview of the main analytical approaches and practices to determine food authenticity i... more A brief overview of the main analytical approaches and practices to determine food authenticity is presented, addressing, as well, food supply chain and future requirements to more effectively mitigate food fraud. Food companies are introducing procedures and mechanisms that allow them to identify vulnerabilities in their food supply chain under the umbrella of a food fraud prevention management system. A key step and first line of defense is thorough supply chain mapping and full transparency, assessing the likelihood of fraudsters to penetrate the chain at any point. More vulnerable chains, such as those where ingredients and/or raw materials are purchased through traders or auctions, may require a higher degree of sampling, testing, and surveillance. Access to analytical tools is therefore pivotal, requiring continuous development and possibly sophistication in identifying chemical markers, data acquisition, and modeling. Significant progress in portable technologies is evident a...
Water-soluble vitamins (WSVs) are a group of organic compounds which are essential micronutrients... more Water-soluble vitamins (WSVs) are a group of organic compounds which are essential micronutrients. WSVs could be divided between the B complex group and vitamin C (l-ascorbic acid). Within the B complex group, eight vitamins are recognized: vitamins B1 (thiamin), B2 (riboflavin), B3 (niacin or niacinamide), B5 (pantothenic acid), B6 (pyridoxine, pyridoxal, or pyridoxamine), B7 (biotin), B9 (folic acid), and B12 (various cobalamins). This paper reviews the new methods for the analysis of these vitamins, with a focus on infant formula and adult nutritionals.
Determination of monofluoroacetate in powdered nutritional products by derivatization with 2-nitr... more Determination of monofluoroacetate in powdered nutritional products by derivatization with 2-nitrophenylhydrazine and LC-MS/MS IS discussed. Samples are prepared by dilution in water followed by protein precipitation with acetonitrile. An aliquot of the sample extract is derivatized with 2-nitrophenylhydrazine (2-NPH) in the presence of 1-ethyl-3-(-3-dimethylaminopropyl) carbodiimide hydrochloride (EDC). This reaction achieves the coupling of the carboxyl group of monofluoroacetate to the amino group of 2-NPH with the formation of an amide bond. Samples are processed through a solid-phase extraction (SPE) step in order to exchange.
Methods under consideration as part of the AOAC Stakeholder Panel on Infant Formula and Adult Nut... more Methods under consideration as part of the AOAC Stakeholder Panel on Infant Formula and Adult Nutritionals process are to be evaluated against a set of Standard Method Performance Requirements(SM) (SMPRs) via peer review by an expert review panel (ERP). A validation protocol and a checklist have been developed to assist the ERP to evaluate experimental data and to compare multiple candidate methods for each nutrient. Method performance against validation parameters mandated in the SMPRs as well as additional criteria are to be scored, with the method selected by the ERP proceeding to multilaboratory study prior to Final Action approval. These methods are intended to be used by the infant formula industry for the purposes of dispute resolution.
A liquid chromatographic (LC) method was validated for the determination of total vitamin B6 in i... more A liquid chromatographic (LC) method was validated for the determination of total vitamin B6 in infant formula. Total vitamin B6 was quantified by converting the phosphorylated and free vitamers into pyridoxine. Pyridoxine was determined by ion pair reversed-phase LC with fluorescence detection. The method was subjected to an AOAC collaborative study involving a factory-manufactured, milk- and soy-based infant formula. Each was spiked at 3 concentrations in the range of 0-1 microg/g and sent as blind duplicate to participant laboratories. Nine laboratories returned valid data which were statistically analyzed for outliers and precision parameters. The repeatability relative standard deviation (RSD(r)) ranges were 2.0-4.0 and 3.5-5.9% for fortified milk- and soy-based formulas, respectively. The reproducibility relative standard deviation (RSD(R)) ranges were 8.2-8.4 and 6.7-11.2% for fortified milk- and soy-based formulas, respectively. HORRAT values ranged from 0.42 to 0.53, indica...
Determination of total fluoroacetic acid and its salts in all forms of infant formula is evaluate... more Determination of total fluoroacetic acid and its salts in all forms of infant formula is evaluated. Infant formula includes breast-milk substitute specially manufactured to satisfy, by itself, the nutritional requirements of infants during the first months of life up to the introduction of appropriate complementary feeding, made from any combination of milk, soy, rice, whey, hydrolyzed protein, starch, and amino acids, with and without intact protein. Suitable methods will include blank check samples, and check standards at the lowest point and midrange point of the analytical range.
Abstract Infant formulas are strictly regulated and rigorously tested for compliance. Recently, n... more Abstract Infant formulas are strictly regulated and rigorously tested for compliance. Recently, new official analytical methods/standards have been established for nutrient analyses in these product categories through the Stakeholder Panel on Infant Formula and Adult Nutritionals (SPIFAN), governed by AOAC INTERNATIONAL. Many of these methods have been adopted or are in the process of being adopted as reference methods by Codex Alimentarius. The purpose of this paper is to assess the ability of these cutting-edge analytical methods to deliver acceptable results in the context of established regulatory limits for nutrients in food standards and regulations. For this evaluation, the analytical method variability is considered as one of the three main sources of overall process variability, which also includes variation in raw materials/ingredients and the manufacturing process. The process capability (Cp) is a concept for determining the overall process variability relative to specification limits for a parameter in the final product. Based on this principle an analytical method capability (Cm) was defined and calculated for SPIFAN methods. Global regulatory requirements were evaluated including minimum and maximum limits and tolerances from the declared label values. Compared to these requirements, analytical methods for vitamins A, B12, D and folic acid are of particular concern in relation to the requirements in China, some EU member states, Pakistan, Russia, Singapore, South Africa and Thailand. For a product with a manufacturing target at the midpoint of the regulatory range for these nutrients, the probability of obtaining an analytical result outside the regulatory requirements due to analytical variation alone can be as high as 19%. This does not consider variability caused by the production process and/or raw materials/ingredients. These currently used analytical methods are state-of-the-art and represent the latest developments in technology. However, this work demonstrates that continuous method improvements for the nutrients identified must be pursued. In addition, this work supports a risk management approach that takes into consideration analytical method capability when establishing regulatory limits for nutrients in infant formulas. Ongoing efforts towards harmonization of regulatory requirements across global markets will facilitate evaluation of regulatory compliance in infant formulas.
Background: Monochloropropanediol (MCPD) and its fatty acid esters as well as glycidyl fatty acid... more Background: Monochloropropanediol (MCPD) and its fatty acid esters as well as glycidyl fatty acid esters are substances generated during oil refining or food processing. The free form released during digestion, 3-MCPD and glycidol, have shown adverse effects in animal studies. Objective: So far, the available analytical methods have not been validated in a collaborative study for infant and adult nutritional formulas. This manuscript describes a single-laboratory validation method in view of a future multilaboratory validation trial. Methods: The method described is for the direct determination of 2- and 3-MCPD and indirect determination of 2- and 3-MCPD esters and glycidyl esters in infant and adult/pediatric nutritional formulas by GC coupled to MS. Results: The analytical range was found to be 4–2000 μg/kg powder formula and 0.7–333 μg/kg liquid formula for fatty acid esters of MCPD and glycidol, and 2.5–750 μg/kg samples for free MCPD. The recovery rates were within 91–124% for ...
AOAC SMPRs describe the minimum recommended performance characteristics to be used during the eva... more AOAC SMPRs describe the minimum recommended performance characteristics to be used during the evaluation of a method. The evaluation may be an on-site verification, a singlelaboratory validation, or a multi-site collaborative study. SMPRs are written and adopted by AOAC stakeholder panels composed of representatives from the industry, regulatory organizations, contract laboratories, test kit manufacturers, and academic institutions. AOAC SMPRs are used by AOAC expert review panels in their evaluation of validation study data for methods being considered for Performance Tested MethodsSM or AOAC Official Methods of AnalysisSM, and can be used as acceptance criteria for verification at user laboratories.
A brief overview of the main analytical approaches and practices to determine food authenticity i... more A brief overview of the main analytical approaches and practices to determine food authenticity is presented, addressing, as well, food supply chain and future requirements to more effectively mitigate food fraud. Food companies are introducing procedures and mechanisms that allow them to identify vulnerabilities in their food supply chain under the umbrella of a food fraud prevention management system. A key step and first line of defense is thorough supply chain mapping and full transparency, assessing the likelihood of fraudsters to penetrate the chain at any point. More vulnerable chains, such as those where ingredients and/or raw materials are purchased through traders or auctions, may require a higher degree of sampling, testing, and surveillance. Access to analytical tools is therefore pivotal, requiring continuous development and possibly sophistication in identifying chemical markers, data acquisition, and modeling. Significant progress in portable technologies is evident a...
Water-soluble vitamins (WSVs) are a group of organic compounds which are essential micronutrients... more Water-soluble vitamins (WSVs) are a group of organic compounds which are essential micronutrients. WSVs could be divided between the B complex group and vitamin C (l-ascorbic acid). Within the B complex group, eight vitamins are recognized: vitamins B1 (thiamin), B2 (riboflavin), B3 (niacin or niacinamide), B5 (pantothenic acid), B6 (pyridoxine, pyridoxal, or pyridoxamine), B7 (biotin), B9 (folic acid), and B12 (various cobalamins). This paper reviews the new methods for the analysis of these vitamins, with a focus on infant formula and adult nutritionals.
Determination of monofluoroacetate in powdered nutritional products by derivatization with 2-nitr... more Determination of monofluoroacetate in powdered nutritional products by derivatization with 2-nitrophenylhydrazine and LC-MS/MS IS discussed. Samples are prepared by dilution in water followed by protein precipitation with acetonitrile. An aliquot of the sample extract is derivatized with 2-nitrophenylhydrazine (2-NPH) in the presence of 1-ethyl-3-(-3-dimethylaminopropyl) carbodiimide hydrochloride (EDC). This reaction achieves the coupling of the carboxyl group of monofluoroacetate to the amino group of 2-NPH with the formation of an amide bond. Samples are processed through a solid-phase extraction (SPE) step in order to exchange.
Methods under consideration as part of the AOAC Stakeholder Panel on Infant Formula and Adult Nut... more Methods under consideration as part of the AOAC Stakeholder Panel on Infant Formula and Adult Nutritionals process are to be evaluated against a set of Standard Method Performance Requirements(SM) (SMPRs) via peer review by an expert review panel (ERP). A validation protocol and a checklist have been developed to assist the ERP to evaluate experimental data and to compare multiple candidate methods for each nutrient. Method performance against validation parameters mandated in the SMPRs as well as additional criteria are to be scored, with the method selected by the ERP proceeding to multilaboratory study prior to Final Action approval. These methods are intended to be used by the infant formula industry for the purposes of dispute resolution.
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