“FWGE pulvis” is a powder consisting of an aqueous extract of fermented wheat germ, with the drying aids maltodextrin and silicon dioxide, standardized to contain approximately 200 µg/g of the natural constituent 2,6-dimethoxy-p-benzoquinone. The results of toxicological and clinical studies of this product demonstrate its safety for its intended use as a dietary supplement ingredient in the United States. Avemar pulvis has been used in Hungary since 1998 and is approved in that country, as well as in the Czech Republic, Bulgaria, and Romania, as a “medical nutriment for cancer patients.” Acute and subacute toxicity studies using rodents orally administered Avemar pulvis showed that dose levels (2000 to 3000 mg/kg body weight [bw]/day) exceeding the normal recommended oral dosage (8.5 g/day or 121 mg/kg bw/day for a 70-kg individual) by up to approximately 25-fold caused no adverse effects. 

The test substance showed no evidence of mutagenicity or genotoxicity in vitro or in vivo. Clinical studies using Avemar pulvis as a supplement to drug therapy in cancer patients at doses of 8.5 g/day not only showed no evidence of toxicity, but also showed a reduction in the side effects of chemotherapy. Overall, it was concluded that Avemar pulvis would not be expected to cause adverse effects under the conditions of its intended use as an ingredient in dietary supplements. The dietary supplement ingredient Avemar pulvis consists of an aqueous extract of the germ of wheat (Triticum vulgare) fermented with baker’s yeast (Saccharomyces cerevisiae) combined with the drying aids maltodextrin and silicon dioxide, and standardized to contain approximately 200 µg/g of the naturally occurring constituent 2,6-dimethoxy-p-benzoquinone (2,6-DMBQ). The process by which the 2,6-DMBQ content of fermented wheat germ extract is standardized was invented by Hungarian biochemist Mate Hidvegi in the early 1990s. It is produced by fermenting a fixed amount of food-grade wheat germ and water with a fixed amount of baker’s yeast in stainless steel fermentation vessels. The ratio of these ingredients is similar to those commonly used in the preparation of whole-wheat baked products. Fermentation proceeds with continuous stirring at controlled pH, temperature, and flow of filtered air for approximately 18 h. The fermentation product is decanted, separated, and fine-filtered to a cell-free solution, which is condensed under vacuum to a specified weight percentage. Food-grade maltodextrin and colloidal silicon dioxide are added and the mixture is spray-dried.

 The final product, FWGE pulvis, comprises 63.2% fermented wheat germ, 35.0% maltodextrin, and 1.8% colloidal silicon dioxide. The product is manufactured under pharmaceutical good manufacturing practices (GMP) and its consistent 2,6-DMBQ content has been confirmed by analysis of multiple lots of product (Tomoskozi-Farkas and Daood 2004; Boros, Nichelatti, and Shoenfeld 2005). Flavors and sweeteners may be added to Avemar pulvis and the product sold under various trade names, including Avemar and Av´e. Wheat germ has a long history of consumption as a food. The germ is part of the wheat kernel, along with the bran and the endosperm. Like the bran, the germ is removed in the processing of wheat to yield white flour, but it is retained in whole-wheat products. The germ constitutes approximately 2.5% (range 2% to 4%) of the total weight of the wheat kernel (Tsen 1985; Nichelatti and Hidvegi 2002). A loaf of whole-wheat bread containing about 2 cups of whole-wheat flour (∼270 g) includes about 7 g of wheat germ. Additionally, wheat germ is used as an ingredient in a wide variety of foods formulated to provide advantageous nutritional characteristics. Although it is likely that the average American’s current consumption of wheat germ is <1 g/day, it appears probable that many Americans— particularly those with a preference for whole grain foods—may consume >5 g/day of wheat germ. Those consuming whole wheat at the estimated 95th percentile of wheat consumption (600 g/day, based on data from the U.S. Department of Agriculture’s Economic Research Service [USDA/ERS 2005]) have a daily intake of wheat germ of approximately 15 g (2.5% of 600 g). 

In addition to the wheat germ consumed in whole-wheat products, many Americans may ingest wheat germ marketed as a food supplement. Milled wheat germ has been sold for many years for incorporation into baked products, addition to breakfast cereals, use as a meat filler, and other applications. Naturally occurring 2,6-DMBQ has been identified in numerous plant families in addition to wheat, including several species that are consumed as food (e.g., Compositae [ettuce, endive, artichoke, chicory, sunflower, safflower, etc.], Leguminosae [soy, beans, peas, lentils, etc.], Proteaceae [macadamia], Gramineae [cereal grains], Ericaceae [blueberry, huckleberry, cranberry, wintergreen]) (Handa et al. 1983). Given that some of these plant families are widely consumed, frequently in large quantities, it is likely that humans have considerably greater exposure to 2,6-DMBQ in their diets than would result from consumption of wheat germ alone. Uses in Food Avemar pulvis has been used in Hungary since 1998 as a dietary supplement and was approved in 2002 as a “medical nutriment for cancer patients. It is classified by the European Union (EU) as a “dietary food for special medical purposes,” a category of foods specifically processed or formulated and intended for the dietary management of patients and to be used under medical supervision, and is sold with this designation in the Czech Republic, Bulgaria, and Romania. It is also sold as a dietary supplement in Italy, Serbia, Switzerland, Cyprus, Russia, Israel, Austria, Slovakia, South Korea, Taiwan, Japan, Hong Kong, Australia, and New Zealand.

 In the United States, FWGE pulvis is intended to be added to dietary supplements at a level of 8.5 g/dose for an average adult. For a 70-kg individual, this is equivalent to an intake of 121 mg Avemar pulvis/kg body weight (bw)/day. Because individuals weighing more than 90 kg (200 pounds) may increase the amount consumed up to a limit of 2 doses, it is anticipated that their intake of Avemar pulvis, in mg/kg bw/day, will generally be in the same range. The highest intake would be for a 90-kg individual consuming 2 doses: 189 mg Avemar pulvis/kg bw/day. Because the 2,6-DMBQ content of Avemar pulvis is 200 µg/g, an 8.5-g dose of Avemar pulvis contains 1.7 mg 2,6- DMBQ. This intake of 2,6-DMBQ could be supplied by eating 17 g of wheat germ (Tomoskozi-Farkas and Daood 2004) or about 700 g of whole wheat bread (Posner 1985), respectively. TOXICOLOGICAL STUDIES Although wheat germ is a commonly consumed food with no known adverse effects, the effects of FWGE pulvis, a product containing 63% fermented wheat germ, were investigated in various studies, mostly unpublished, using rats or mice (Hidvegi et al. 1998, 1999; Csik´o, Semjen, and Ratz 1999; Lehel, Semjen, and Ratz 1999; Zalatnai et al. 2001; Szende et al. 2004). The experimental protocols were approved by the Hungarian Ethical Committee for Preventing Abuse and Torture of Animals. As described below, there were no instances of any adverse reactions. Acute Studies Two acute oral toxicity studies were conducted by the same laboratory using mice and rats. 

These studies, as well as a third acute oral toxicity study conducted by a different laboratory, were performed according to Good Laboratory Practice (GLP) guidelines and following the Guidelines for the Testing of Chemicals(No. 401: “Acute Oral Toxicity”) promulgated by the Organization for Economic Cooperation and Development (OECD). Groups of 6-week-old CD-1 mice (10/sex/group) were administered by gavage a single dose of 0 (vehicle control) or 2000 mg/kg bw of Avemar pulvis in distilled water and observed for up to 14 days (Lehel, Semjen, and Ratz 1999). Animals were housed 5 per cage and were given rodent diet and tap water ad libitum; intakes were not recorded. Body weights were recorded on the day of arrival, day of randomization, prior to treatment, and 24 h post dosing. Because some weight loss (not statistically significant) was noted among some animals in both the experimental and control groups at the 24-h interval (although there were no differences in the group mean weights), mice were weighed daily thereafter until an increase in body weight was noted. Animals also were weighed on days 7 and 14 post dosing. The animals were observed twice daily for any adverse clinical signs; after the observation period, the mice were killed for a postmortem examination. No deaths or abnormal clinical signs were reported. There were no statistically significant differences in body weight between control and treated mice. Pathological examination showed no macroscopic lesions. 

The LD50 of FWGE pulvis in male and female mice in this study was >2000 mg/kg bw. In a similar study, groups of 5-week-old Wistar rats (10/sex/group) were administered by gavage a single dose of 0 (vehicle control) or 2000 mg/kg bw of FWGE pulvis in distilled water and observed for up to 14 days (Csik´o, Semjen, and Ratz 1999). The rats were treated in the same manner as the mice in the previous study and the same parameters were examined as those in the mouse study. No deaths or abnormal clinical signs were reported. There were no statistically significant differences in body weight between control and treated rats. Pathological examination showed no macroscopic lesions. The LD50 of FWGE pulvis in male and female rats in this study was >2000 mg/kg bw. A third acute oral toxicity study was conducted using Wistar rats (IPCM 1999). Groups of male and female rats were administered a single gavage dose of 0 (control) or 5000 mg/kg bw of FWGE pulvis in distilled water. There were no differences between the test animals and controls in the body weights of the animals, and no cardiovascular, respiratory, neurological, or other adverse effects were reported. Based on the absence of adverse effects, the oral LD50 of Avemar pulvis in male and female rats in this study was >5000 mg/kg bw. Subacute Studies A 28-day study with a 14-day recovery period was conducted under GLP using rats (Csik´o et al. 2000). The study design met the criteria outlined in OECD Guideline 407 (“Repeated Dose 28-Day Oral Toxicity Study in Rodents”) and was conducted as a limit test. Groups of 4.5-week-old Wistar BR rats (20/sex/group) were given by gavage 0 (vehicle control) or 2000 mg/kg bw/day of Avemar pulvis in 1% methylcellulose for 28 days.

 Half of the animals (10/sex/group) were followed for a 14-day recovery period. The animals were housed 2 per cage and were given rodent diet and tap water ad libitum. Body weights were recorded on the day of arrival, day of randomization, prior to treatment, on days 7, 14, 21, and 28 post dosing, and on days 7 and 14 of the recovery period. The rats were observed twice daily for any adverse clinical signs or mortality during the treatment and recovery periods. Feed consumption (g/day) was recorded weekly throughout the entire study and water intake (g/day) was recorded daily during weeks 1 and 4 of the treatment period and during week 2 of the recovery period. Prior to termination, fasting blood samples were taken from the femoral vein for hematological and clinical chemistry evaluations. Hematological parameters included red blood cell count, white blood cell count, hemoglobin, hematocrit, mean corpuscular volume, mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), platelet count, prothrombin time, and differential leucocyte count. Clinical chemistries included glucose, total protein, albumin, total bilirubin, urea nitrogen, creatinine, aspartate transaminase (AST), alanine transaminase (ALT), alkaline phosphatase, cholesterol, triacylglycerols, Na, K, Ca, Cl, and P. Urine samples also were taken for urinalysis by test strip (Hemoket/GPH5) at the end of the treatment and recovery periods.

 Rats dying during the study and those terminated at end of the treatment and recovery periods underwent necropsy, including gross pathology and histopathological examination of the lymph nodes, mammary glands, salivary glands, sternebrae, femur (including marrow), pituitary, thymus, trachea, lungs, heart, thyroid, esophagus, stomach, small intestine, colon, liver, gall bladder, pancreas, spleen, kidneys, adrenals, bladder, prostate, testes, ovaries, uterus, brain, eyes, and spinal cord. Organ weights were recorded. Results were statistically analyzed using the t test. No adverse clinical signs were reported with the exception of one treated female rat which showed mild lethargy with decreased motor activity and piloerection 1 day prior to death on the first day of the recovery period. Two other rats from the Avemar pulvis–treated group died during the treatment period. Pathological examination revealed these deaths to be gavage errors rather than inherent toxicity of the test compound. No statistically significant differences were observed in body weight, feed consumption, water intake, urinalysis, pathological examinations, organ weights, or histopathology between control and treated rats. Hematological parameters were similar between control and treated rats, with the exception of slight but statistically significant increases in hemoglobin content, MCH, and MCHC in treated females at the end of the 2-week recovery period. 

These small changes were considered not to be related to the test compound and of no biological importance. Clinical chemistry values remained within normal ranges, with the exception that control males had statistically significantly higher AST (124.70 ± 25.56 U/L versus 83.70 ± 16.04 U/L) and ALT (65.23 ± 7.57 U/L versus 54.73 ± 6.07 U/L) activities compared to treated males after 28 days of treatment, but these were considered to be due to moderate hemolysis of samples. Further information on the extent of sample hemolysis was not provided in the study report. Na, K, and Cl ions were significantly elevated in control animals; however, the differences were small and not considered biologically important. Based on these findings, the no observed adverse effect level (NOAEL) in this oral subacute study was the tested dose of 2000 mg/kg bw/day of Avemar pulvis. Subchronic Studies Groups of F344 rats and C57BL/6 mice (13/group) were administered 0 or 3000 mg/kg bw/day of Avemar pulvis by gavage for up to 77 days (FIP 1997). Although this study was not conducted under GLP or in accordance with OECD guidelines, histological samples were extracted and prepared under Registry of Industrial Toxicology Animal-Data (RITA) guidelines (Bahnemann et al. 1995). In addition to Avemar pulvis, 900 mg vitamin C/kg bw/day was also administered to the animals by gavage. Animals were observed for any abnormal clinical signs and body weights were recorded on treatment days 1, 4, 8, 11, 17, 22, 29, 40, 52, 61, and 74 for rats and days 1, 7, 13, 18, 32, 39, 48, 61, 67, and 77 for mice. On day 77, all rats and mice were killed using anesthesia and underwent necropsy. No hematology or clinical chemistry was performed. 

Organ and tissue samples were taken from 3 randomly selected animals per species in both the experimental and control groups and prepared using hematoxylin and eosin staining for histopathological examination by a board-certified pathologist in a blinded setting. Organs (heart, lung, thymus spleen, liver, kidneys, and testicles) were removed and weighed for both species from the experimental groups only; organs from the control groups were not weighed. There were no statistically significant differences in body weight between treated animals and their corresponding controls. No pathological changes were noted in either rats or mice. Based on these findings, the NOAEL in this oral subchronic study was the tested dose of 3000 mg/kg bw/day of Avemar pulvis. Mutagenicity/Genotoxicity Studies Avemar pulvis In a study of the ability of Avemar pulvis to inhibit the development of colonic tumors, 100 4-week-old inbred male F344 rats were randomized into four groups which were treated in the following manner over a 32-week period: (A) untreated control (n = 10); (B) animals (n = 48) received three subcutaneous injections (1 week apart) of 15 mg azoxymethane/kg bw starting at week 2 to induce colon carcinogenesis; (C) animals (n = 32) were gavaged daily with 3000 mg Avemar pulvis/kg bw for the entire 32-week period and received three subcutaneous injections (1 week apart) of 15 mg azoxymethane/kg bw starting at week 2; and (D) animals (n = 10) were gavaged daily with 3000 mg Avemar pulvis/kg bw for the entire 32-week period (Zalatnai et al. 2001). Rats from all four groups were killed by exsanguination after 32 weeks.