Latest Studies 2019 – Pregnancy and Health | | Eannatto Delta Gold

Latest Studies 2019 – Pregnancy and Health

Female reproductive system has been observed to be constantly exposed to multiple deleterious factors which include nicotine exposure. Among the 3,500 chemical substances, nicotine being an addictive agent has been found to retard fetal development, cause low birth weight and delay parturition. Furthermore, administration of nicotine for five consecutive days in pregnant rats has the capability to alter the rate of embryo proliferation, delay implantation and even delay delivery process. Nicotine when present in higher concentrations in oocytes cultured in vitro, was observed to cause perturbation in the first and second meiotic division.

The mechanism by which the nicotine could damage tissues is related to its effects on gamete cells’ viability or change in the oviductal epithelial function. Earlier reports have shown that nicotine can act as free radicals. These free radicals can significantly damage the polyunsaturated fatty acids that are present in the cell membrane or as side – chains in certain chemical species. Moreover, administration of nicotine in rats has been found to lead to higher lipid peroxidation with subsequent decrease in antioxidant enzymes. Lipid peroxidation can lead to a variety of toxicological effects such as impaired mitochondrial functions and inhibition of antioxidant enzymes. The end product of this process can be measured by using malondialdehyde (MDA). To counter this condition, the effect of antioxidant such as vitamin E – Tocotrienol (Eannatto – DeltaGold) has been high – lighted. Vitamin E is known for its antioxidant properties since its discovery in 1922. Among all the components of the family of vitamin E, Tocotrienol (Eannatto – DeltaGold) is widely reported to have the highest biological activities. Recently, Gamma – Tocotrienol and Delta – Tocotrienol have provoked many researchers to explore its beneficial effects in various human diseases.

A population study on the hyper-cholesterolemic patients treated with palmvitee (tocotrienol-rich fraction of palm oil capsule) demonstrated a significant reduction in total cholesterol (10%), low-density cholesterol (13%) and Apo B (7%). The significance of vitamin E in male reproduction has been emphasized when its deficiency is found to cause testicular degeneration in rats. Evidences have therefore provided a new incentive to inspect the role of palm vitamin E in combating the radical – driven oxidative events.

Most research in the past 50 – 60 years has been focused on Tocopherols and 50% of all the research in last 30 years has been done on Tocotrienols in last 5 years. Half of the Tocotrienol research ever published has been published in last 10 years as shown in Fig. 1. Each day it is becoming increasingly understood that Tocotienols (especially Eannatto – DeltaGold) are the right form of Vitamin E. Well in excess of 100 studies and clinical trials have shown the surprising benefits of Tocotrienols – without any known side effects.

Fig. 1: In the graph, as you can see, R & D on Tocotrienol has increased exponentially over the years in all fields while research on Tocopherols has decreased. Whether it is cancer, Cardiovascular diseases (CVD), Diabetes, Anti – Oxidant activities or others, in all fields research on Tocotrienol has not only gained pace but quant as well.

Study – Role of Tocotrienol – rich palm Vitamin E on pregnancy and preim – plantation embryos in nicotine – treated rats.

The objective of this study was to investigate the possible protective profile of tocotrienol-rich palm vitamin E (TRF – PVE) on the embryo development and pregnancy outcome in nicotine – treated rats.

The experimental protocol was approved by the University Kebangsaan Malaysia Animal Care and Use Committee (UKMACUC) and was conducted at the Animal Biotechnology Laboratory, Department of Physiology of University Kebangsaan Malaysia (UKM). 24 fertile female Sprague Dawley rats weighing 160 – 240 g (of age 3 – 6 months) were obtained from UKM animal unit. Animals were kept in the animal house at 25 – 30 degree Celsius and had free access to rat chow and drinking water. Animals were divided into 4 groups of six rats in each. Fertile male rats were caged together for mating. Palm vitamin E with Tocotrienol rich fraction (Eannatto – DeltaGold) was a complimentary gift from Mr. Gapor Mat Top, Malaysian Palm Oil Board. PVE per 100 g contained: Alpha – Tocotrienol (17.3%), Gamma –Tocotrienol (21.6%), Delta – Tocotrienol (15.3%), Alpha – Tocopherol (18.9%) and palm olein (26.9%). was diluted with tocopherol-stripped corn oil (ICN USA) to obtain a desired concentration of 500 mg/kg of rat weight. The dispensing volume was 0.2 ml. Nicotine N-3876 obtained from SIGMA chemical company, USA was prepared weekly at a concentration of 20 mg/ml saline and stored away from light.

Effects of PVE on pregnancy outcome in rats treated with nicotine:

Pregnant females were treated daily from day 1 pc until term. Animals of Group A (control) were given 0.5 ml 0.9% sodium chloride and 0.2 ml tocopherol – stripped corn oil orally. Animals of Group B were given nicotine [5 mg/kg of body weight sodium chloride] concurrently with 0.2 ml tocopherol – stripped corn oil orally. Group C was also given nicotine [5 mg/kg of body weight sodium chloride] and PVE at 60 mg/kg of body weight orally. Group D was given 0.5 ml 0.9% sodium chloride and PVE at a dose of 60 mg/kg of body weight orally. Vaginal smears were taken daily and subjected for Schorr’s staining. Sperm – positive vaginal smear was counted as day 0 of pregnancy. Upon delivery, anthropometric measurements were performed on each pup.

Effects of PVE on embryo development in rats treated with nicotine:

The distribution of animals groups was the same as described in the previous experiment. Immature female rats were around 9 – 12 weeks old and weighed between 120 – 140g. Treatments were given and administered for 30 days consecutively. Animals were super – ovulated by an intra – peritoneal injection of 150 IU/kg of body weight of PMSG and 48 hours later by another injection of sodium chloride of hCG (75 IU/kg of body weight). Animals were then mated with fertile males and sacrificed at 48 hours post mating. The embryos were flushed from the fallopian tubes and examined under a dissecting microscope. Venous blood samples were taken from the orbital sinus after the pregnant rats had delivered. Plasma levels of malondialdehyde (MDA) were measured as an indicator of lipid peroxidation. Also, vitamin E in the plasma was also measured. Cotinine level in urine was investigated using a spectrophotometer according to the published protocol. Values are given as a median and the comparisons were analyzed by non-parametric Kruskal – Wallis analysis of variance on rank. Probability levels <0.05 were taken as statistical significance.


Nicotine treatment until term reduced the rate of pregnancy outcome to 33.3% as shown in Table 1. The pregnant (2/6) had longer duration of gestation (median 23.5 days) compared to controls (median 21.7 days). The harmful effect of nicotine as viewed by the lowest number of embryos survived in Group B which was 26 out of the 183 which was around 14.21% as shown in Table 3. The extended length of gestation in the nicotine – primed rats could possibly be due to delayed embryo cleavage in which 61.54% of retrieved embryos remained un-cleaved. The excretion of cotinine as the metabolite of nicotine in the urine in Group B animals was found to be significantly higher compared to the controls (p<0.05) as shown in Table 2. The MDA levels were similarly recorded to be higher in Group B animals (P<0.05) as again shown in Table 2.

Table 1: The % of pregnancy, gestation period and postnatal development of pups following treatment throughout the pregnancy. n = 6 in each groups.
Table 2: Biochemical analysis of plasma MDA and vitamin E with urinary cotinine. All values are expressed in median.
Table 3: Number and % of oocytes and embryo derived from all the groups.

Supplementation of palm vitamin E – Tocotrienol rich fraction in Group C rats was observed to improve the rate of pregnancy to 83.3% with the gestation length of median 22 days as shown in Table 1. The number of retrieved embryos accounted for 47 out of 183 which is around 25.68%. Algthough, 76.56% of them were found to be in a 2 – cell and 4 – cell stage as shown in Table 3. The benefit of palm vitamin E – Tocotrienol rich fraction supplementation was found to be more obvious in Group D animals, 100% of them became pregnant with the highest number of retrieved embryos (57 out of 183). Moreover, all the embryos obtained in this group were fertilized and cleaved. 90% of them were found to be in 2 – cell and 4 – cell embryos and seven percent had reached 8 – cell and 16 – cell stage.


  • Antioxidants, especially Tocotrienol was observed to exhibit anti-oxidant properties by lowering inflammation and oxidative stress as shown in Fig. 2.
Fig. 2: In the study conducted by Dr. Qureshi, he saw that at 250 mg of Tocotrienols, the endogenous anti-oxidant, TAS (represented with grey colour) increased, while the C-reactive protein (CRP) dropped by 40%, oxidized fat (MDA) dropped by 34% and Total Anti-oxidant increased by 22%.
  • Cytokines associated with cardiovascular diseases were down-regulated by Tocotrienols according to the study done at the University of Missouri – Kansas City.
  • MicroRNAs which are usually down-regulated in hyper-cholesterolemic individuals are up-regulated by Tocotrienol treatment as per the study.
  • Vitamin E (Tocotrienol) acts as a lipid-soluble antioxidants protecting cells of the body from damage by harmful free radicals.
  • Lipid peroxides in blood vessels and blood pressure have been observed to be lowered by Tocotrienols which improves total anti-oxidant status thus preventing hypertension.
  • Oxidative Stress has been observed to be prevented by the synergistic act of Tocotrienol. Oxidative Stress is an imbalance in the amount of freer radicals circulating in the body and the availability of antioxidants to counter the free radicals.
  • Cholesterol reduction is a significant feature of Tocotrienols by suppression of HMGR (3-hydroxy3methyl-glutaryl-CoA reductase) the enzyme/protein responsible for the body’s cholesterol production.
  • Cancer stem cell death has been observed by the action of Tocotrienols especially Delta – Tocotrienols (DeltaGold – Eannatto). Even after chemotherapies, radiation and surgeries, there are stem cells of those cancerous tissues left revolving in your body which can lead to your cancer coming back. Henceforth, their death is very necessary and Tocotrienols have been observed to kill cancer stem cells.
Fig. 4: About 1% of cancer cells are Cancer Stem Cells (CSC) which keep circulating in your body even after nailing the cancer through chemo. It has been observed that Gamma – Tocotrienol and Delta – Tocotrienol, both specifically target CSC.


  • Typically a 200 mg dose of Tocotrienol has been found to be quite suitable but dosages up to 900 mg have also been found to be effective without any adverse effects.

Why Tocotrienol and Not Tocopherol?

  • Tocopherol, the enemy of Tocotrienol: In the studies, Tocopherol was observed to interfere in the functioning of Tocotrienol! Tocopherol has been observed to attenuate cancer inhibition, inhibits absorption, reduces adipose storage, and compromises cholesterol and triglyceride reduction.
  • Tocotrienol, the protector of State: Tocotrienol has more mobility than Tocopherol due to its small structure so it can cover a larger area targeting more number of colon cancer cells.
  • Small structure and less molecular weight: The higher anti-oxidant activity of Tocotrienols is due to their small structure and less molecular weight which assist in their integration of the cell, unlike Tocopherols.
Fig. 5: In the first graph, it can be observed that Tocotrienol uptake is maximum when Alpha – Tocopherol concentration is 0 but as the concentration of Alpha – Tocopherol increases, the concentration of uptake of Tocotrienol decreases. While, in the 2nd graph it can be observed that as the concentration of Alpha – Tocopherol increases in the dosage of Tocotrienol, the apoptosis induction property of Tocotrienol was suffered and cancer kill was compromised.
  • Absorption: As compared to Tocopherols, Tocotrienols absorb better in the body and Tocopherols have been observed to prevent absorption of Tocotrienols.
Fig. 6: The 2nd pie chart represents Palm Tocotrienol rich fraction with 32% Alpha – Tocopherol which was given to people and when the Alpha – Tocopherol was removed then it was represented by the 1st pie chart with 0.3% Alpha – Tocopherol which was then given to people. In the graph, the hollow bar represents the Tocotrienol with Tocopherol which reduced the concentrations of Alpha, Gamma and Delta Tocotrienol in the body but when Tocopherol was removed from the dosage (Solid grey bars in graph), the concentrations of Alpha, Gamma and Delta – Tocotrienol significantly increased.


  • Tocotrienols: Latest Cancer Research in Vitamin E by Barrie Tan, Ph.D., and Anne M.Trias, MS.
  • Tocotrienols: The Promising Analogues of Vitamin E for Cancer Therapeutics                         

  • Pregnancy and Birthing Online Resource Center

  • Role of Tocotrienol – rich palm Vitamin E on pregnancy and preim – plantation embryos in nicotine – treated rats by Norfilza M Mokhtar, Mohd Hamim Rajikins and Zaiton Zakaria.

  • WHO: e-Library of Evidence for Nutrition Actions (eLENA)

  • Weisberg E, Smoking and reproductive health, Clin Reprod Fertil 1985; 3: 175-186.
  • Ashakumary L, Vijayammal PL. Effect of nicotine on antioxidant defence mechanisms in rats fed a high-fat diet, Pharmacology 1996; 52: 153-158.
  • Gutteridge JM, Halliwell B. Free radicals and antioxi-dants in the year 2000. A historical look to the future, Ann N Y Acad Sci 2000; 899: 136-147.
  • Sen CK, Khanna S, Roy S. Tocotrienol: the natural vitamin E to defend the nervous system?, Ann N Y Acad Sci 2004; 1031: 127-142.

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