Evaluation of Iron Bioavailability From Iron Chlorophyllin
Condition: Iron Deficiency
MeSH terms: Anemia, Iron-Deficiency; Iron Deficiencies
Study type: Interventional

Summary: 
      The overall objective of this study is to evaluate the iron absorption from iron
      chlorophyllin. Iron deficiency is a public health problem in both developing and
      industrialized countries. There are several approaches to combat iron deficiency. Most
      supplements in the present day, to address the problem of iron deficiency, are in the form of
      iron salts, known as ferrous salts, especially ferrous sulfate. However, we can only usually
      absorb about 20% of the total iron content in ferrous sulfate. The common strategy of food
      supplement companies is to increase the amount of iron in the supplements to compensate for
      the low absorption rate. However, this often causes gastrointestinal side effects. In the
      present study, we would like to measure the iron bioavailability from sodium iron
      chlorophyllin, which made up from ferrous salts and chlorophyllin and where we hypothesize
      that it is absorbed via a different pathway than ferrous sulfate. Via this mechanism, we
      further hypothesize that sodium iron chlorophyllin will therefore have an enhanced
      bioavailability and more favorable side effect profile than ferrous sulfate and other iron
      salts.
    

Detailed description: 
      New approaches to treat iron deficiency include developing novel iron compounds with possible
      iron absorption routes that increase iron bioavailability and reduce gastrointestinal side
      effects. Heme iron is considered to be highly bioavailable (10-20%) and less affected by meal
      composition than non-heme iron. Heme iron is iron (Fe2+) that is bound to the iron
      protoporphyrin IX prosthetic groups of proteins, mainly hemoglobin and myoglobin, which are
      present in animal tissue. Heme is released from hemoglobin during digestion so that it can be
      taken up by the duodenal enterocytes. The intact iron porphyrin is transported across the
      brush border membrane by the Heme Carrier Protein 1 (HCP1). Once inside the cell, the iron is
      released, and it is then likely to enter the low molecular weight pool of iron. The use of
      heme analogues from vegetable origin could provide an alternative iron source of potentially
      high bioavailability.

      Sodium iron chlorophyllin (SIC) is a water-soluble semisynthetic chlorophyll derivative where
      the magnesium in the porphyrin ring has been substituted by iron. It is a greenish brown
      pigment that dissolves in water, alcohols, and chloroform, but not ethers. SIC is known to
      have a superior processing stability to chlorophyll and is commonly used as food colorings in
      Asian countries including in Korea and Japan, for foods that do not contain meat or fish.
      Magnesium Chlorophyllin has an E number of E140 and is approved as a food additive in Europe.
      Currently sodium iron chlorophyllin is present in the Designated Additives List in Japan with
      a number 261. Designated additives are those designated by the Minister of Health, Labour and
      Welfare as substances that are unlikely to harm human health based on Article 10 of the Food
      Sanitation Act.

      Given the porphyrin ring heme-like structure of SIC, it may be an alternative delivery route
      for iron, suitable also for vegetarians and vegans, yet there are few studies investigating
      the use of SIC for this purpose.

      A recent study by Ding et al. 2019 suggests a positive effect of SIC on hemoglobin (Hb)
      values in iron-deficiency anemia (IDA) in children and adults after 1 month of treatment.
      They assume that SIC is taken up by the heme carrier protein. The researchers used
      shengxuening tablets, also known as SXN, which are based on chlorophyll/porphyrin structures
      extracted from silkworms in which the Mg is replaced by the Fe and then coadjutants are
      added. This study provided no data on the absorption pathway of the SXN compound. Thus, it
      remains unclear if it was taken up by the heme transporter or whether it was digested and the
      Fe released in the gut lumen to join the non heme Fe pool, and thereby improved Hb in the IDA
      subjects. The authors also describe Xray diffraction assays to assess if the chlorophyllin
      porphyrin ring contained Fe and not Mg, however these data are not reported.

      Miret et al. (2010) studied the stability of heme-analogous SIC absorption using the Caco-2
      cell model. The SIC remained stable and only 5% of the Fe from the compound was released at
      pH 2 to 4. In-house dissolution studies conducted at the Human Nutrition Laboratory (HNL)
      show only a 2% release of Fe from the compound at similar pH levels. Using solid phase
      extraction at a higher pH of 7, similar to that in the duodenum, we observed a 5% Fe loss,
      which may correspond to free iron or iron liberated from any other binding sites on the
      chlorophyll structure.

      Toyoda et al. (2014) performed a toxicity study of SIC performed in male and female rats with
      oral administration of SIC in their diet at concentrations of 0%, 0.2%, 1.0%, and 5.0% for 13
      weeks. No abnormal clinical signs, no mortality and no abnormal hematological changes were
      observed in any of the groups during the experiment. Based on the histopathology of the
      parotid glands, the no-observed-adverse-effect level (NOAEL) of SIC in this study was
      estimated to be 1.0% (609 mg SIC/kg bodyweight (bw)/day for males and 678 mg/kg bw/day for
      females). In the present study, we will use 6 mg elemental iron, in 100 mg SIC, 0.002% of the
      equivalent dose administered by Toyoda et al.

      SIC has been produced at the HNL with a 75% incorporation of iron into the chlorophyll. Our
      protocol is based on a Unilever patent that is no longer active, with in-house adaptation.
      The SIC has a neutral taste and will be presented as a dark green liquid, being the solution
      of SIC in water.

      The goal of this project is to develop a novel iron compound that can be used as a food
      supplement. The iron content in our SIC will be limited to 6 mg per dose, which therefore
      does not exceed the 14 mg daily iron dose stipulated as the legal maximum in Switzerland for
      food supplements. SIC is already used widely as a food colorant and is safe for human
      consumption.

      We propose to test our SIC against a known comparator, ferrous sulfate, to ascertain whether
      SIC is an effective iron delivery compound. If SIC, as hypothesized, behaves in a similar way
      to heme, then commonly used enhancers of iron absorption effective on ferrous sulfate, such
      as ascorbic acid, will not have an effect on the absorption of iron from SIC
    

Intervention name(s): SIC; SIC + AA combined; FeSO4; FeSO4 + AA combined; EP + FeSO4 combined; EP + FeSO4 + AA combined
Intervention type(s): Dietary Supplement; Dietary Supplement; Dietary Supplement; Dietary Supplement; Other; Other
Intervention description(s): Sodium Iron Chlorophyllin, whose bioavailability is to be studied; Sodium Iron Chlorophyllin and ascorbic acid. The ascorbic acid should not have an effect on Sodium Iron Chlorophyllin; Ferrous sulfate serves as a positive control, whose iron bioavailability is known; Ferrous sulfate serves as a positive control and the addition of ascorbic acid, further enhances its bioavailability; Chlorophyllin with an empty porphyrin ring, given along with FeSO4 to study if there is an incorporation of Fe into the porphyrin ring as it passes the gastric system; Chlorophyllin with an empty porphyrin ring, given along with FeSO4 to study if there is an incorporation of Fe into the porphyrin ring as it passes the gastric system. Ascorbic acid is given along with the intervention to see if there is any difference in the fractional iron absorption when compared to EP+ FeSO4
Intervention MeSH terms: Fesoterodine
Allocation: Randomized
Intervention model: Crossover Assignment
Intervention model description: The study will be a single-center, prospective cross-over trial in which each subject receives all test conditions. During the screening, about 2 weeks before the start of the study, women will be assessed for eligibility. Finally, 55 eligible women will be invited to participate.
Each subject will complete six iron absorption studies in which they will receive supplemental iron doses of 6 mg iron.The experimental phase will last for 37 days. The labeled iron doses will be administered on days 1, 3, 5, 19, 21 and 23. Since only three isotopes are used in the study, after day 5, there will be a wait period of 14 days to allow for incorporation of the labeled iron into erythrocytes. On day 19, 21 and 23 the other three test doses are administered. Randomization of the interventional products listed above will be set up as such, to ensure that participants do not receive the same isotopes in a given week. On day 1, day 19 and day 37, a venous blood sample will be collected.
Masking: None (Open Label)
Primary purpose: Treatment
Keywords: iron deficiency; Iron chlorophyllin