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1.
Hosp Pharm ; 59(3): 318-323, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38764986

ABSTRACT

Purpose: To compare and evaluate 2 methods of inventory management in automated dispensing cabinets (ADCs). Methods: Ten profiled ADCs had 2 inventory management models implemented over 2 months. Implementation of the models on each ADC involved adjustment of par levels (desired accessible quantities of medication) and removal of medications not used in the past 90 days or more. The par levels of 5 ADCs were adjusted using a formula developed based on the economic order quantity model. The par levels of the other 5 ADCs were adjusted using a formula based on historical average daily usage. The study endpoints include stock out rate, vend:fill ratio, quantity of expired medications, and inventory carrying cost. Results: The total of number of medications stocked in the 10 ADCs was reduced from 3035 in a 2-month pre-implementation period to 2932 in a 2-month post-implementation period yielding a reduction of inventory carrying cost by $11 011. The mean stock out rate in both study groups increased and vend:fill ratio decreased after implementation. The quantity of expired medications increased in the modified economic order quantity formula inventory management model and decreased in the average daily usage inventory management model. Conclusion: The implementation of 2 inventory management models on ADCs had a negative impact on stock out rate and vend:fill ratio, a mixed impact on quantity of expired medications, and a positive impact on inventory carrying cost reduction.

2.
Pure Appl Chem ; 95(10)2023.
Article in English | MEDLINE | ID: mdl-37964805

ABSTRACT

The International Union of Pure and Applied Chemistry (IUPAC) has a long tradition of supporting the compilation of chemical data and their evaluation through direct projects, nomenclature and terminology work, and partnerships with international scientific bodies, government agencies and other organizations. The IUPAC Interdivisional Subcommittee on Critical Evaluation of Data (ISCED) has been established to provide guidance on issues related to the evaluation of chemical data. In this first report we define the general principles of the evaluation of scientific data and describe best practices and approaches to data evaluation in chemistry.

3.
J Proteome Res ; 21(11): 2664-2686, 2022 11 04.
Article in English | MEDLINE | ID: mdl-36181456

ABSTRACT

Protein turnover maintains the proteome's functional integrity. Here, protein turnover efficiency over time in wild-type Caenorhabditis elegans was assessed using inverse [15N]-pulse labeling up to 7 days after the egg-laying phase at 20 °C. Isotopic analysis of some abundant proteins was executed favoring data quality over quantity for mathematical modeling. Surprisingly, isotopic enrichment over time reached an upper limit showing an apparent cessation of protein renewal well before death, with protein fractions inaccessible to turnover ranging from 14 to 83%. For life span modulation, worms were raised at different temperatures after egg laying. Mathematical modeling of isotopic enrichment points either to a slowdown of protein turnover or to an increasing protein fraction resistant to turnover with time. Most notably, the estimated time points of protein turnover cessation from our mathematical model were highly correlated with the observed median life span. Thrashing and pumping rates over time were linearly correlated with isotopic enrichment, therefore linking protein/tracer intake to protein turnover rate and protein life span. If confirmed, life span extension is possible by optimizing protein turnover rate through modulating protein intake in C. elegans and possibly other organisms. While proteome maintenance benefits from a high protein turnover rate, protein turnover is fundamentally energy-intensive, where oxidative stress contributes to damage that it is supposed to repair.


Subject(s)
Caenorhabditis elegans Proteins , Caenorhabditis elegans , Animals , Caenorhabditis elegans/genetics , Caenorhabditis elegans Proteins/genetics , Caenorhabditis elegans Proteins/metabolism , Proteome/genetics , Proteome/metabolism , Longevity , Aging/metabolism , Eating
4.
Am J Clin Nutr ; 110(6): 1362-1369, 2019 12 01.
Article in English | MEDLINE | ID: mdl-31573611

ABSTRACT

BACKGROUND: A highly soluble iron-casein complex has been developed for food fortification purposes with the aim to provide high iron bioavailability. OBJECTIVE: We aimed to determine the iron bioavailability of the iron-casein complex relative to that of ferrous sulfate (control) when given with whole milk in healthy young women. METHODS: A randomized comparator-controlled trial with a crossover design was conducted using the erythrocyte incorporation dual stable isotope (57Fe, 58Fe) technique. Iron absorption from the iron-casein complex was compared with that from ferrous sulfate in 21 healthy women aged 20-38 y with normal iron status. RESULTS: Fractional iron absorption (geometric mean; -SD, +SD) from the iron-casein complex (3.4%; 1.4%, 5.4%) and from ferrous sulfate (3.9%; 1.7%, 6.1%) were not statistically different (P > 0.05). The relative bioavailability value of the iron-casein complex to ferrous sulfate was determined to be 0.87 (-1 SD, +1 SD: -0.90, +2.64). CONCLUSIONS: The iron-casein complex has iron bioavailability comparable to that of ferrous sulfate in healthy young women. This trial was registered at www.anzctr.org.au as ACTRN12615000690550.


Subject(s)
Caseins/metabolism , Ferrous Compounds/metabolism , Food Additives/metabolism , Iron/metabolism , Milk/metabolism , Adult , Animals , Biological Availability , Female , Food, Fortified/analysis , Humans , Iron Isotopes/metabolism , Milk/chemistry , Young Adult
5.
Consult Pharm ; 32(7): 388-396, 2017 Jul 01.
Article in English | MEDLINE | ID: mdl-28701250

ABSTRACT

Americans have embraced a large number of diets in an attempt to manage obesity, improve quality of life, and address specific health problems. Among diets developed to address health problems, the ketogenic diet has had a long and variable history. Developed in the 1920s by a faith healer to help children with epilepsy, this diet induces a state that mimics carbohydrate starvation. As medications became available and effectively addressed seizures, the diet fell out of favor. During the last few decades, researchers and clinicians have learned that it can be useful in children and adults with refractory epilepsy and a variety of other conditions. Once again, pharmacists may encounter patients who are employing dietary management of serious health problems. This very high-fat diet almost eliminates carbohydrates from the patient's food selection. The result is the substitution of ketone bodies as a source of energy. Today's ketogenic diet has been modified with scientifically proven adjustments to increase palatability and help with adherence. Effective for some forms of epilepsy, the ketogenic diet also seems to have some utility in Alzheimer's disease, Parkinson's disease, and glaucoma, and many Americans are using it to lose weight. Consultant pharmacists may field questions about this diet, its potential to correct or alleviate health conditions, and its limitations. The article discusses the ketogenic diet's strengths, limitations, potential mechanisms, and use in a number of conditions with an emphasis on the elderly.


Subject(s)
Diet, Ketogenic , Alzheimer Disease/diet therapy , Animals , Consultants , Diet, Ketogenic/adverse effects , Epilepsy/diet therapy , Glaucoma/diet therapy , Humans , Parkinson Disease/diet therapy , Pharmacists
6.
J Nutr ; 144(11): 1703-9, 2014 Nov.
Article in English | MEDLINE | ID: mdl-25332469

ABSTRACT

BACKGROUND: Calcium inhibits and ascorbic acid (AA) enhances iron absorption from iron-fortified foods. Absorption efficiency depends on iron status, although the interaction is unclear. OBJECTIVE: We investigated the ability of AA to overcome calcium-induced inhibition of iron absorption in children differing in iron status. METHODS: The effect of calcium (0, 100, and 200 mg/test meal) on iron absorption in the absence and presence of AA (0, 42.5, and 85 mg/test meal) from a casein/whey-based drink fortified with ferrous sulfate was assessed in a series of randomized crossover studies both in iron-replete (IR) Indian schoolchildren and in children with iron deficiency anemia (IDA) (6-11 y; n = 14-16/group) by using stable isotopes. RESULTS: In the absence of calcium and AA, iron absorption from the casein/whey-based drink was 20% lower in IR children than in children with IDA. The addition of calcium reduced mean iron absorption by 18-27%, with the effect being stronger for high added calcium (P < 0.01). AA at a 2:1 or 4:1 molar ratio enhanced iron absorption by a factor of 2-4 and greatly overcompensated for the inhibitory effect of calcium on iron absorption in a dose-dependent manner (P < 0.001). The dose-response effect tended to be stronger (P < 0.1) in the IDA group, and iron status was of far less influence on iron absorption than the enhancing effect of AA. CONCLUSION: When adding AA to iron-fortified milk products, care should be taken not to provide absorbable iron in excess of needs.


Subject(s)
Ascorbic Acid/pharmacokinetics , Beverages/analysis , Calcium/pharmacokinetics , Caseins/chemistry , Iron/metabolism , Milk Proteins/chemistry , Ascorbic Acid/chemistry , Child , Child Nutritional Physiological Phenomena , Cross-Over Studies , Dietary Supplements , Drug Interactions , Female , Humans , India , Iron/pharmacokinetics , Male , Whey Proteins
7.
J Food Sci ; 79(9): C1629-34, 2014 Sep.
Article in English | MEDLINE | ID: mdl-25124357

ABSTRACT

The iron storage protein ferritin is a potential vehicle to enhance the iron content of biofortified crops. With the aim of evaluating the potential of ferritin iron in plant breeding, we used species-specific isotope dilution mass spectrometry to quantify ferritin iron in bean varieties with a wide range of total iron content. Zinc, phytic acid, and polyphenols were also measured. Total iron concentration in 21 bean varieties ranged from 32 to 115 ppm and was positively correlated with concentrations of zinc (P = 0.001) and nonferritin bound iron (P < 0.001). Ferritin iron ranged from 13% to 35% of total iron and increased only slightly in high iron beans (P = 0.007). Concentrations of nonferritin bound iron and phytic acid were correlated (P = 0.001), although phytic acid:iron molar ratio decreased with increasing iron concentration (P = 0.003). Most iron in high iron beans was present as nonferritin bound iron, which confirms our earlier finding showing that ferritin iron in beans was lower than previously published. As the range of ferritin iron content in beans is relatively narrow, there is less opportunity for breeders to breed for high ferritin. The relevance of these findings to the extent of iron absorption depends on resolving the question of whether ferritin iron is absorbed or not to a greater extent than nonferritin bound iron.


Subject(s)
Crops, Agricultural/metabolism , Iron/metabolism , Phaseolus/metabolism , Seeds/metabolism , Breeding , Crops, Agricultural/chemistry , Ferritins/metabolism , Phytic Acid/metabolism , Plant Proteins/metabolism , Polyphenols/metabolism , Species Specificity , Zinc/metabolism
8.
Metallomics ; 6(9): 1709-17, 2014 Sep.
Article in English | MEDLINE | ID: mdl-25008269

ABSTRACT

Ferritin is a hollow sphere protein composed of 24 subunits that can store up to 4500 iron atoms in its inner cavity. It is mainly found in the liver and spleen but also in serum at trace levels. Serum ferritin is considered as the best single indicator in assessing body iron stores except liver or bone marrow biopsy. However, it is confounded by other disease conditions. Ferritin bound iron (FBI) and ferritin saturation have been suggested as more robust biomarkers. The current techniques for FBI determination are limited by low antibody specificity, low instrument sensitivity and possible analyte losses during sample preparation. The need for a highly sensitive and reliable method is widely recognized. Here we describe a novel technique to detect serum FBI using species-specific isotope dilution mass spectrometry (SS-IDMS). [(57)Fe]-ferritin was produced by biosynthesis and in vitro labeling with the (57)Fe spike in the form of [(57)Fe]-citrate after cell lysis and heat treatment. [(57)Fe]-ferritin for sample spiking was further purified by fast liquid protein chromatography. Serum ferritin and added [(57)Fe]-ferritin were separated from other iron species by ultrafiltration followed by isotopic analysis of FBI using negative thermal ionization mass spectrometry. Repeatability of our assay is 8% with an absolute detection limit of 18 ng FBI in the sample. As compared to other speciation techniques, SS-IDMS offers maximum control over sample losses and species conversion during analysis. The described technique may therefore serve as a reference technique for clinical applications of FBI as a new biomarker for assessing body iron status.


Subject(s)
Ferritins/metabolism , Iron/blood , Isotope Labeling/methods , Mass Spectrometry/methods , Animals , Anions , Apoproteins/metabolism , Chromatography, High Pressure Liquid , Chromatography, Ion Exchange , Ferritins/blood , Ferritins/isolation & purification , Horses , Humans , Iron Isotopes , Native Polyacrylamide Gel Electrophoresis , Recombinant Proteins/isolation & purification , Recombinant Proteins/metabolism , Ultrafiltration
9.
Metallomics ; 6(8): 1417-26, 2014 Aug.
Article in English | MEDLINE | ID: mdl-24752826

ABSTRACT

Brain iron accumulation is supposed to play a central role in neurodegeneration by inducing oxidative stress. Currently it is unknown to which extent iron entering brain over lifetime exchanges with body iron or if uptake of iron is unidirectional without significant efflux from brain. To study brain iron dynamics in vivo, up to three stable isotope tracers were fed continuously with a standard rodent diet up to 5 months to healthy adult male Wistar rats (n = 8) in a staggered design. Brain iron uptake was found to be bi-directional but iron influx and efflux were unbalanced leading inevitably to brain iron accumulation over time. Brain iron turnover was found to be very low at a half-life of ca. 9 months for tracer iron entering brain. Observed tracer accumulation in brain iron can be extrapolated to an increase of brain iron by ca. 30% in the healthy rats from early adulthood to the end of their lives. In contrast to current beliefs that brain uptake of dietary iron is negligible during adulthood following short-term radiotracer studies, our long-term feeding experiments point to a possible role of the diet in brain iron accumulation and, subsequently, neurodegeneration.


Subject(s)
Brain/metabolism , Iron/metabolism , Animals , Isotope Labeling , Male , Models, Theoretical , Rats , Rats, Wistar
10.
Anal Chem ; 85(7): 3667-73, 2013 Apr 02.
Article in English | MEDLINE | ID: mdl-23419016

ABSTRACT

Subtle variations in the isotopic composition of elements carry unique information about physical and chemical processes in nature and are now exploited widely in diverse areas of research. Reliable measurement of natural isotope abundance variations is among the biggest challenges in inorganic mass spectrometry as they are highly sensitive to methodological bias. For decades, double spiking of the sample with a mix of two stable isotopes has been considered the reference technique for measuring such variations both by multicollector-inductively coupled plasma mass spectrometry (MC-ICPMS) and multicollector-thermal ionization mass spectrometry (MC-TIMS). However, this technique can only be applied to elements having at least four stable isotopes. Here we present a novel approach that requires measurement of three isotope signals only and which is more robust than the conventional double spiking technique. This became possible by gravimetric mixing of the sample with an isotopic spike in different proportions and by applying principles of isotope dilution for data analysis (GS-IDA). The potential and principle use of the technique is demonstrated for Mg in human urine using MC-TIMS for isotopic analysis. Mg is an element inaccessible to double spiking methods as it consists of three stable isotopes only and shows great potential for metabolically induced isotope effects waiting to be explored.


Subject(s)
Indicator Dilution Techniques , Magnesium/urine , Mass Spectrometry/methods , Humans , Isotopes/analysis , Isotopes/urine , Magnesium/analysis , Sensitivity and Specificity
11.
Metallomics ; 5(2): 167-73, 2013 Feb.
Article in English | MEDLINE | ID: mdl-23340610

ABSTRACT

Iron deposits in the brain are a common hallmark of Alzheimer's disease and Parkinson's disease. This has spurred the hypothesis that iron may play a functional role in the pathogenesis of neurodegenerative disorders through free radical damage. Previous short-term studies using radiotracers suggested that brain iron uptake is small as compared to other tissues in adult rodents. This has led to the assumption that brain iron uptake must also be marginal in humans after brain development is complete. In this study we applied a novel approach to determine directly the fraction of iron that was transferred over time from diet to brain and other organs in adult rats. A known amount of a stable iron isotope ((57)Fe) was fed with drinking water to adult rats over 4 months. Uptake of the tracer iron and final iron content in tissues were assessed by Negative Thermal Ionization Mass Spectrometry (NTI-MS). We found that only a very small amount of dietary iron entered the brain (0.000537 ± 0.000076%). This amount, however, is considerable relative to the total brain iron content (9.19 ± 0.71%), which was lower but comparable to percentage uptake in other tissues. Whereas it remains unclear whether excessive dietary iron intake is a risk factor in neurodegenerative diseases or whether high systemic iron correlates with iron deposits in the brain, our study suggests that uptake of dietary iron is much higher than previously thought. This finding challenges current beliefs and points to a possible role of iron nutrition in the pathogenesis of neurodegenerative disorders.


Subject(s)
Brain/metabolism , Iron Isotopes/metabolism , Animals , Iron Isotopes/blood , Male , Mass Spectrometry , Rats , Rats, Wistar
12.
J Biol Inorg Chem ; 18(1): 1-7, 2013 Jan.
Article in English | MEDLINE | ID: mdl-23053533

ABSTRACT

Persistent impairments in the regulation of intestinal iron absorption result in iron deficiency or iron accumulation in the long term. Diagnosis remains difficult unless pathological symptoms develop as iron absorption varies strongly between meals and days. Variations in the natural iron isotopic composition of whole blood have recently been suggested as a novel parameter to assess long-term differences in intestinal absorption efficiency between individuals. In this study, baseline blood samples collected in two previous conventional iron absorption studies in Swiss and Thai women using stable isotope tracers were reanalyzed by multicollector inductively coupled plasma mass spectrometry. The natural iron isotopic compositions obtained were compared with fractional absorption from the test meals observed in these earlier trials. Correlations of natural blood iron isotopic composition and fractional absorption from the test meals were found to be highly significant in both cohorts (for Swiss women, r = 0.40, P = 0.01, n = 38; for Thai women, r = 0.57, P < 0.01, n = 24), with the blood of both ethnicities clearly differing in iron isotopic composition (P < 0.001). Combining the findings of this study and those of recent animal and human studies confirms that blood iron isotopic patterns may serve as a novel compound biomarker of iron metabolism to assess impairments in regulation of intestinal iron absorption in individuals or population groups.


Subject(s)
Intestinal Absorption , Iron Isotopes/blood , Iron, Dietary/metabolism , Female , Humans , Time Factors , Young Adult
13.
Eur J Nutr ; 52(4): 1361-8, 2013 Jun.
Article in English | MEDLINE | ID: mdl-22956195

ABSTRACT

PURPOSE: The main purpose of this study was to establish bioavailability data in humans for the new (Fe) fortification compound ferrous ammonium phosphate (FAP), which was specially developed for fortification of difficult-to-fortify foods where soluble Fe compounds cannot be used due to their negative impact on product stability. METHODS: A double-blind, randomized clinical trial with cross-over design was conducted to obtain bioavailability data for FAP in humans. In this trial, Fe absorption from FAP-fortified full-cream milk powder was compared to that from ferric pyrophosphate (FPP) and ferrous sulfate. Fe absorption was determined in 38 young women using the erythrocyte incorporation dual stable isotope technique (57Fe, 58Fe). RESULTS: Geometric mean Fe absorption from ferrous sulfate, FAP and FPP was 10.4, 7.4 and 3.3 %, respectively. Fe from FAP was significantly better absorbed from milk than Fe from FPP (p < 0.0001). Fe absorption from FAP was significantly lower than Fe absorption from ferrous sulfate, which was used as water-soluble reference compound (p = 0.0002). Absorption ratios of FAP and FPP relative to ferrous sulfate as a measure of relative bioavailability were 0.71 and 0.32, respectively. CONCLUSIONS: The results of the present studies show that replacing FPP with FAP in full-cream milk could significantly improve iron bioavailability.


Subject(s)
Beverages , Dairy Products , Ferrous Compounds/metabolism , Food, Fortified , Iron, Dietary/administration & dosage , Phosphates/metabolism , Adult , Cross-Over Studies , Diphosphates/chemistry , Diphosphates/metabolism , Double-Blind Method , Erythrocytes/metabolism , Female , Ferrous Compounds/chemistry , Food, Preserved , Humans , Intestinal Absorption , Iron/chemistry , Iron/metabolism , Iron Isotopes , Iron, Dietary/metabolism , Nutritive Value , Phosphates/chemistry , Solubility , Young Adult
14.
Anal Bioanal Chem ; 402(7): 2463-9, 2012 Mar.
Article in English | MEDLINE | ID: mdl-22287047

ABSTRACT

Despite the importance of stating the measurement uncertainty in chemical analysis, concepts are still not widely applied by the broader scientific community. The Guide to the expression of uncertainty in measurement approves the use of both the partial derivative approach and the Monte Carlo approach. There are two limitations to the partial derivative approach. Firstly, it involves the computation of first-order derivatives of each component of the output quantity. This requires some mathematical skills and can be tedious if the mathematical model is complex. Secondly, it is not able to predict the probability distribution of the output quantity accurately if the input quantities are not normally distributed. Knowledge of the probability distribution is essential to determine the coverage interval. The Monte Carlo approach performs random sampling from probability distributions of the input quantities; hence, there is no need to compute first-order derivatives. In addition, it gives the probability density function of the output quantity as the end result, from which the coverage interval can be determined. Here we demonstrate how the Monte Carlo approach can be easily implemented to estimate measurement uncertainty using a standard spreadsheet software program such as Microsoft Excel. It is our aim to provide the analytical community with a tool to estimate measurement uncertainty using software that is already widely available and that is so simple to apply that it can even be used by students with basic computer skills and minimal mathematical knowledge.


Subject(s)
Uncertainty , Monte Carlo Method , Probability , Software
15.
J Biol Inorg Chem ; 17(2): 301-9, 2012 Feb.
Article in English | MEDLINE | ID: mdl-22002846

ABSTRACT

We recently showed in an animal model that iron isotopic composition varies substantially between different organs. For instance, iron in ferritin-rich organs--such as the major storage tissues liver, spleen, and bone marrow--contain a larger fraction of the heavy iron isotopes compared with other tissues, including blood. As a consequence, partitioning of body iron into red blood cells and storage compartments should be reflected in the isotopic pattern of blood iron. To confirm this hypothesis, we monitored blood iron isotope patterns in iron-overloaded subjects undergoing phlebotomy treatment by multicollector inductively coupled plasma mass spectrometry. We found that bloodletting and consequential replacement of lost blood iron by storage iron led to a substantial increase of the heavy isotope fraction in the blood. The progress of iron depletion therapy and blood loss was quantitatively traceable by isotopic shifts of as much as +1‰ in δ((56)Fe). These results show that--together with iron absorption efficiency--partitioning of iron between blood and iron storage tissues is an important determinant of blood iron isotopic patterns, which could make blood iron isotopic composition the first composite measure of iron metabolism in humans.


Subject(s)
Iron/blood , Iron/metabolism , Adult , Female , Humans , Iron Isotopes/blood , Iron Isotopes/metabolism , Male , Mass Spectrometry , Middle Aged , Phlebotomy
16.
J Nutr ; 140(11): 1977-82, 2010 Nov.
Article in English | MEDLINE | ID: mdl-20861210

ABSTRACT

Low iron absorption from common beans might contribute to iron deficiency in countries where beans are a staple food. High levels of phytic acid (PA) and polyphenols (PP) inhibit iron absorption; however, the effect of bean PP on iron absorption in humans has not been demonstrated and, with respect to variety selection, the relative importance of PP and PA is unclear. To evaluate the influence of bean PP relative to PA on iron absorption in humans, 6 stable iron isotope absorption studies were conducted in women (16 or 17 per study). Bean PP (20, 50, and 200 mg) were added in studies 1-3 as red bean hulls to a bread meal. Studies 4- 6 investigated the influence on iron absorption of PP removal and dephytinization of whole red bean porridge and PP removal from dephytinized porridge. Iron absorption was lowered by 14% with 50 mg PP (P < 0.05) and by 45% with 200 mg PP (P < 0.001). The mean iron absorption from whole bean porridge was 2.5%. PP and PA removal increased absorption 2.6-fold (P < 0.001) and removal of PP from dephytinized porridge doubled absorption (P < 0.001). Between-study comparisons indicated that dephytinization did not increase iron absorption in the presence of PP, but in their absence, absorption increased 3.4-fold (P < 0.001). These data suggest that in countries where beans are a staple food, PP and PA concentrations should be considered when selecting bean varieties for human consumption. Lowering only one inhibitor will have a modest influence on iron absorption.


Subject(s)
Diet , Fabaceae/chemistry , Flavonoids/administration & dosage , Intestinal Absorption , Iron, Dietary/metabolism , Phenols/administration & dosage , Phytic Acid/administration & dosage , Seeds/chemistry , Adolescent , Adult , Bread/analysis , Cross-Over Studies , Female , Flavonoids/analysis , Food Handling/methods , Humans , Iron Isotopes , Middle Aged , Phenols/analysis , Phytic Acid/analysis , Polyphenols , Young Adult
17.
Anal Chem ; 81(17): 7368-72, 2009 Sep 01.
Article in English | MEDLINE | ID: mdl-19653660

ABSTRACT

Ferritin is nature's predominant iron storage protein. The molecule consists of a hollow protein shell composed of 24 subunits which is capable of storing up to 4500 iron atoms per molecule. Recently, this protein has been identified as a target molecule for increasing iron content in plant staple foods in order to combat dietary iron deficiency, a major public health problem in developing countries. Here, we present a novel technique for quantification of ferritin-bound iron in edible plant seeds using species-specific isotope dilution mass spectrometry (IDMS) by means of a biosynthetically produced (57)Fe-labeled ferritin spike and negative thermal ionization mass spectrometry (NTIMS). Native plant ferritin and added spike ferritin were extracted in 20 mM Tris buffer (pH 7.4) and separated by anion exchange chromatography (DEAE Sepharose), followed by isotopic analysis by thermal ionization mass spectrometry. The chosen IDMS approach was critically evaluated by assessing the (i) efficiency of analyte extraction, (ii) identical behavior of spike and analyte, and (iii) potential iron isotope exchange with natural iron. Repeatabilities that can be achieved are on the order of <5% RSD for quintuplicate analyses at an absolute detection limit of 60 ng of ferritin-bound iron for plant seeds. Studies in six different legumes revealed ferritin-iron contents ranging from 15% of total iron in red kidney beans up to 69% in lentils.


Subject(s)
Ferritins/analysis , Iron/analysis , Mass Spectrometry/methods , Plants, Edible/chemistry , Seeds/chemistry , Ferritins/isolation & purification , Iron Isotopes/analysis
18.
Scand J Clin Lab Invest ; 69(2): 190-7, 2009.
Article in English | MEDLINE | ID: mdl-18942019

ABSTRACT

OBJECTIVE: The aim of this study was to develop and validate a high-pressure liquid chromatography (HPLC) method for assessing vitamin D status as 25-hydroxyvitamin D(2) (S-25OHD(2)) and 25-hydroxyvitamin D(3) (S-25OHD(3)) in serum. MATERIAL AND METHODS: We assessed the within- and between-subject variation of vitamin D status in serum samples from four different dietary intervention studies in which subjects (n = 92) were supplemented with different doses of vitamin D(3) (5-12 microg/day) and for different durations (4-20 months). RESULTS: The HPLC method was applicable for 4.0-200 nmol S-25OHD/L, while the within-day and between-days variations were 3.8 % and 5.7 %, respectively. There was a concentration-dependent difference between results obtained by a commercial radioimmunoassay and results from the HPLC method of -5 to 20 nmol 25OHD/L in the range 10-100 nmol 25OHD/L. The between-subject variation estimated in each of the four human intervention studies did not differ significantly (p = 0.55). Hence, the pooled standard deviation was 15.3 nmol 25OHD(3)/L. In the studies with 6-8 samplings during 7-20 months of supplementation, the within-subject variation was 3.9-7.2 nmol 25OHD(3)/L, while vitamin D status was in the range 47-120 nmol/L. CONCLUSIONS: The validated HPLC method was applied in samples from human intervention studies in which subjects were supplemented with vitamin D(3). The estimated standard deviation between and within subjects is useful in the forthcoming decision on setting limits for optimal vitamin D status.


Subject(s)
25-Hydroxyvitamin D 2/blood , Calcifediol/administration & dosage , Calcifediol/blood , Chromatography, High Pressure Liquid/methods , Humans , Middle Aged , Reproducibility of Results , Sensitivity and Specificity
19.
Am J Clin Nutr ; 87(4): 881-6, 2008 Apr.
Article in English | MEDLINE | ID: mdl-18400710

ABSTRACT

BACKGROUND: Ascorbic acid (AA) enhances and tea inhibits iron absorption. It is unclear whether iron status influences the magnitude of this effect. OBJECTIVE: We evaluated the influence of the iron status of young women on iron absorption from a rice meal with or without added tea or AA. DESIGN: Two stable-isotope iron absorption studies were made in 2 groups of 10 subjects with iron deficiency anemia (IDA) and 10 subjects who were iron replete (control subjects). In study 1, the reference rice meal was fed alone or with 1 or 2 cups of black tea. In study 2, the reference meal was fed alone or with AA (molar ratio to iron, 2:1 or 4:1). Iron absorption was measured by the erythrocyte incorporation of (57)Fe and (58)Fe labels at 14 d. RESULTS: Mean fractional iron absorption from the reference rice meal was approximately 2.5 times as great in the IDA group as in the control group (P < 0.05). The consumption of 1 or 2 cups of tea decreased iron absorption in the control subjects by 49% (P < 0.05) or 66% (P < 0.01), respectively, and in the IDA group by 59% or 67% (P < 0.001 for both), respectively. AA (molar ratio to iron, 2:1 or 4:1) increased iron absorption by 270% or 343%, respectively, in control subjects and by 291% or 350%, respectively, in subjects with IDA (P < 0.001). CONCLUSIONS: The inhibitory effect of tea and the enhancing effect of AA on iron absorption were similar in the 2 groups. Overall differences in iron absorption in the 2 groups, however, continued to be dictated by iron status.


Subject(s)
Anemia, Iron-Deficiency/metabolism , Ascorbic Acid/pharmacology , Iron, Dietary/pharmacokinetics , Nutritional Status , Tea/chemistry , Adolescent , Adult , Anemia, Iron-Deficiency/blood , Anemia, Iron-Deficiency/diet therapy , Biological Availability , Case-Control Studies , Cross-Over Studies , Erythrocytes/chemistry , Female , Humans , Intestinal Absorption/drug effects , Iron Isotopes
20.
J Nutr ; 138(5): 878-84, 2008 May.
Article in English | MEDLINE | ID: mdl-18424595

ABSTRACT

Biofortification of staple foods with iron in the form of ferritin-iron is a promising approach to fighting iron-deficiency anemia in developing countries. However, contradictory results regarding iron bioavailability to humans from ferritin are not yet fully clarified. Furthermore, the question has been raised whether ferritin can potentially survive gastric passage intact and be absorbed via a ferritin-specific uptake mechanism. We studied changes of ferritin-iron and protein during cooking and in vitro gastric digestion. Water soluble, native ferritin-iron, measured in different legumes, represented 18% (soybeans) up to maximally 42% (peas) of total seed iron. Ferritin-iron was no longer detectable after boiling the legumes for 50 min in excess water. When the same cooking treatment was applied to recombinant bean ferritin propagated in Escherichia coli, some ferritin-iron remained measurable. During in vitro gastric digestion of recombinant bean ferritin and red kidney bean extract, ferritin-iron was fully released from the protein and dissolved at pH 2. Stability tests at varying pH at 37 degrees C showed that the release of ferritin-iron starts at pH 5 and is complete at pH 2. We concluded that ferritin-iron is efficiently released from the ferritin molecule during cooking and at gastric pH and that it should be absorbed as efficiently as all other nonheme iron in food.


Subject(s)
Digestion , Ferritins/metabolism , Hot Temperature , Iron/metabolism , Animals , Biological Availability , Drug Stability , Edetic Acid/pharmacology , Escherichia coli/genetics , Fabaceae/chemistry , Ferritins/genetics , Food, Fortified , Gastric Acid/metabolism , Gastric Mucosa/enzymology , Gene Expression , Hydrogen-Ion Concentration , In Vitro Techniques , Iron/analysis , Kinetics , Pepsin A/metabolism , Phaseolus/chemistry , Recombinant Proteins/metabolism , Swine
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