INTERNATIONAL PROGRAMME ON CHEMICAL SAFETY WORLD HEALTH ORGANIZATION SAFETY EVALUATION OF CERTAIN FOOD ADDITIVES AND CONTAMINANTS WHO FOOD ADDITIVES SERIES 40 Prepared by: The forty-ninth meeting of the Joint FAO/WHO Expert Committee on Food Additives (JECFA) World Health Organization, Geneva 1998 MICROCRYSTALLINE CELLULOSE First draft prepared by Dr J.B. Greig Department of Health, Skipton House, 80 London Road, London, SE1 6LW, UK 1. Explanation 2. Biological data 2.1 Biochemical aspects 2.1.1 Absorption, distribution and excretion 2.1.1.1 Rats 2.1.1.2 Humans 2.1.2 Presorption in animal species 2.2 Toxicological studies 2.2.1 Acute toxicity studies 2.2.2 Short-term toxicity studies 2.2.2.1 Rats 2.2.3 Long-term toxicity/carcinogenicity studies 2.2.3.1 Rats 2.2.4 Reproductive toxicity studies 2.2.4.1 Rats 2.2.5 Special studies on embryotoxicity and teratogenicity 2.2.5.1 Rats 2.2.6 Special studies on genotoxicity 2.2.7 Special studies on sensitization 2.2.8 Special studies on skin and eye irritation 2.2.9 Special studies on effects of cellulose fibre on tumour growth 2.3 Observations in humans 2.3.1 Toxicity consequent to substance abuse 2.3.2 Changes in gastrointestinal function and nutrient balance 3. Comments 4. Evaluation 5. References 1. EXPLANATION Microcrystalline cellulose was evaluated at the fifteenth, seventeenth and nineteenth meetings of the Committee (see Annex 1, references 26, 32 and 38). At the nineteenth meeting an ADI "not specified" was allocated. In the light of concern about possible persorption and consequential adverse effects of fine particles, the substance was re-evaluated at the present meeting. 2. BIOLOGICAL DATA 2.1 Biochemical aspects 2.1.1 Absorption, distribution and excretion 2.1.1.1 Rats Four rats were fed 14C-labelled microcrystalline cellulose at 10 or 20% of their diet. No evidence of degradation or digestion was noted. Faecal recoveries of radioactivity ranged from 96-104% and were complete for all labelled material. No radioactivity appeared in the urine (Baker, 1966). A study was specifically designed to investigate the possibility that persorption of microcrystalline cellulose might induce toxicological effects. Groups of male and female Sprague-Dawley CD rats (20 per group) from Charles River Laboratories were administered, by gavage, suspensions of a special fine particle-size microcrystalline cellulose (median particle size 6 µm). The rats were dosed orally daily for 90 consecutive days at a level of 5000 mg/kg bw per day by means of a 25% suspension in tap water. The animals were killed on study days 91-94 and necropsies were carried out under conditions that reduced the possibility of contamination of tissues with fine particulates. The birefringent microcrystalline cellulose particles were not detected in any organ or tissue, including gut-associated lymphoid tissue, liver, lung, spleen and brain. The size limit for detection of the particles was considered to be < 1 µm (Kotkoskie et al., 1996; FMC Corporation N.V., 1996). 2.1.1.2 Humans One human subject received 150 g of microcrystalline cellulose daily in two portions for a 15-day adaptation period. He then received 14C-labelled microcrystalline cellulose (47.6 µCi) in two portions on one day. Supplementation of the diet with unlabelled microcrystalline cellulose continued for 10 days. Twenty-four-hour faecal and urine collections were examined for radioactivity. No radioactivity appeared in the urine or in the expired CO2. All administered radioactivity (98.9 ± 3.0%) was recovered from the faeces within two days (Baker, 1968). Metabolism of a preparation of 14C-labelled cellulose by four volunteers has been shown to be increased by the consumption, for a period of 3 months, of an additional 7 g/per day of dietary fibre. In six subjects with an ileostomy, the cumulative excretion of 14CO2 was lower than in controls. In two constipated subjects metabolism appeared to be more extensive and occurred over a longer period (Walters et al., 1989). Examination of the stools of one male and one female patient given 30 g microcrystalline cellulose as dry flour or gel for 5´ weeks showed the presence of undegraded material of the same birefringence as the original microcrystalline cellulose administered. No significant effects on the human gastrointestinal tract were noted during the administration (Tusing et al., 1964). Most (87%) of the radiolabel associated with 131I-labelled alpha-cellulose fibres (retained by a sieve with pores of 1 mm diam) was excreted by 4 male and 4 female volunteers within 5 days of ingestion. Less than 2% of the faecal radiolabel was unbound; urinary excretion of unbound radio-iodine accounted for another 1.9% of the total dose (Carryer et al., 1982). Other studies have been carried out to demonstrate the relationship between persorbability and size and consistency of granules. Using quartz sand, the upper limit for persorbability was shown to be 150 µm. Starch granules must be structurally largely intact to possess the property of persorbability. Persorbed starch granules may be eliminated in the urine, pulmonary alveoli, peritoneal cavity, cerebrospinal fluid, via lactating milk and transplacentally (Volkheimer et al., 1968). In another study, dyed plant foods (oatmeal, creamed corn) were fed to human subjects, and blood and urine were examined for coloured fibres. Dyed fibres were shown to be present (Schreiber, 1974). Lycopodium spores and pollen grains have also been shown to be persorbed by humans (Linskens & Jorde, 1974). Mean intake of dietary microcrystalline cellulose in the USA has been estimated to range from 2.7 g/person per day (children 2 years of age) to 5.1 g/person per day (young adult males). For heavy consumer intake of microcrystalline cellulose (90th percentile) the values are 5.4 to 10.2 g/person per day for the same age groups (CanTox Inc., 1993). The mean intake of dietary microcrystalline cellulose in the United Kingdom has been estimated as 0.65 g/person per day. The highest mean intake, 0.90 g/person per day, was for children aged 10-11 (the youngest group for which data were available). For heavy consumer intake of microcrystalline cellulose (90th percentile) the values ranged from 1.13 g/person per day for adults age 16-24 to 1.83 g/person per day for males age 10-11 (Egan & Heimbach, 1994). 2.1.2 Persorption in animal species Rats, pigs and dogs were used to study the persorption of microcrystalline cellulose. The animals were not fed for 12 hours prior to oral administration of the test compound. Rats, dogs and pigs were given 0.5, 140 and 200 g, respectively, of the test compound. Venous blood was taken from the animals 1-2 hours after administration of the test compound, and examined for particles. Persorbed particles were demonstrated in the blood of all three species. The average maximum diameter for persorbed particles was greater in rats than in dogs or pigs (Pahlke & Friedrich, 1974). Orally administered poly(DL-lactide-co-glycolide) microspheres of diameter 1-10 µm were specifically taken up into the Peyer's patch tissue of the gut of BALB/c mice. Those larger than or equal to 5 µm diameter remained in the Peyer's patches; those < 5 µm were translocated to the mesenteric lymph nodes and spleen (Eldridge et al., 1989). These microspheres degrade slowly and can be used for the delivery of antigens to the gut-associated lymphoid tissue of BALB/c mice, males and females, and the initiation of a secretory IgA response in other organs and tissues (Eldridge et al., 1990). The use of particles with differing sizes, i.e. styrene divinylbenzene microspheres (5.7 ± 1.5 µm, mean ± SE) and carbonized styrene divinylbenzene microspheres (15.8 ± 1.4 µm), indicated that smaller particles are more readily translocated to the lungs and mesenteric lymph nodes than are those of a larger size. Unlike earlier workers, the authors considered that the orally administered particles do not reach the peripheral blood but are transported to the lungs via the lymphatic system (LeFevre et al., 1980). Orally administered titanium dioxide particles of nominal size 500 nm are taken up by gut-associated lymphoid tissue of female Sprague-Dawley rats and are translocated to liver, spleen, lung and peritoneal tissues (Jani et al., 1994). The same research group has investigated the persorption of polystyrene microspheres with diameters of 50 nm to 3 µm administered by gavage for 10 days to female Sprague-Dawley rats at a dose of 1.25 mg/kg bw per day. The extent of persorption of the 50 and 100 nm particles was 34 and 26%, respectively; the uptake of larger particles was less than 14%. The 50 nm particles were detected in stomach, small intestine, colon, liver, spleen, kidney and blood. The 3.0 µm particles were detected only in the stomach, small intestine and colon (Jani et al., 1990). Fluorescent polystyrene microparticles of nominal diameters 0.15 and 1.0 µm were injected intraduodenally in the region of a Peyer's patch in male Wistar rats in which a cannula had been implanted in the mesenteric lymph duct. Uptake and detection in lymph was rapid for both sizes of particle, with the time of maximal uptake being 65 min after the start of the infusion (Jenkins et al., 1994). Sprague-Dawley rats, 10 per group, aged 25 days, 5 months or 15 months, were dosed with a suspension of fluorescent polystyrene microparticles of nominal diameter 2.247 µm for 5 days. There was no significant difference between age groups in the number of particles that were transferred into gut-associated lymphoid tissue (Simon et al., 1994) 2.2 Toxicological studies 2.2.1 Acute toxicity studies Acute toxicity studies of various cellulose preparations are summarized in Table 1. Table 1. Acute toxicity of microcrystalline cellulose in animals Species Sex Route LD50 Reference (mg/kg bw) Rat 1 M oral > 3160 Pallotta, 1959 Rat 2 M & F oral > 5000 Freeman, 1991a Rat 3 M & F oral > 5000 Freeman, 1996d Rat 1 M intraperitoneal > 3160 Pallotta, 1959 Rat 4 M & F dermal > 2000 Freeman, 1991b Rat 5 M & F dermal > 2000 Freeman, 1996e LC50 (mg/litre) Rat 6 M & F inhalation > 5.35 Signorin, 1996 1 Test material Cellan 300, five male rats per group. 2 Based on no deaths in 10 rats of each sex administered 5000 mg/kg of Avicel RCN-15. 3 Based on no deaths in 5 rats of each sex administered 5000 mg/kg of Avicel AC-815. 4 Based on no deaths in 5 rats of each sex treated with 2000 mg/kg of Avicel RCN-15. 5 Based on no deaths in 5 rats of each sex treated with 2000 mg/kg of Avicel AC-815. 6 Based on no deaths in 5 rats of each sex exposed to 5.35 mg/litre of Avicel AC-815. In the studies summarized in Table 1, there was no evidence of toxicity of microcrystalline cellulose preparations administered either orally or dermally to rats at doses of 5000 or 2000 mg/kg bw, respectively. The observations seen at necropsy in animals treated intraperitoneally with Cellan 300 at 3160 mg/kg bw are consistent with an irritant reaction caused by the presence of foreign material. An inhalation toxicity study showed only transient effects at a concentration of 5.35 mg/litre. Groups of five male Sprague-Dawley rats received a single oral dose, by stomach tube, of 10.0, 31.6, 100, 316, 1000 or 3160 mg/kg bw of a suspension of Cellan 300 (refined alpha-cellulose) in either distilled water or Mazola corn oil. The animals were observed for 7 days following administration. No differences were observed among the groups as regards the average body weight, appearance and behaviour compared to untreated rats. No observable gross pathology was revealed at autopsy in animals dosed with either suspension. Therefore, the acute oral LD50 was >3160 mg/kg (Pallotta, 1959). Similar single doses of refined alpha-cellulose were given i.p. in distilled water suspension to five male rats. During 7 days observation there were no abnormalities in the rats given 316 mg/kg bw or less. At 1000 and 3160 mg/kg bw inactivity, laboured respiration and ataxia were observed 10 min after administration and, at 3160 mg/kg bw, ptosis and sprawling of the limbs were observed. These animals appeared normal after 24 hours and for the remainder of the observation period. At sacrifice body weights were higher than normal and gross autopsy revealed adhesions between the liver, diaphragm and peritoneal wall and congestion of the kidneys. Masses resembling unabsorbed compound were also observed and these were found to a small extent in the mesentery of the animals administered 316 mg/kg bw. There were no deaths and therefore the acute i.p. LD50 was >3160 mg/kg bw (Pallotta, 1959). Ten male and ten female Sprague-Dawley rats fasted overnight were fed Avicel RCN-15 (a mixture of 85% microcrystalline cellulose with 15% guar gum) at a dose level of 5000 mg/kg bw mixed with parmesan cheese. Six of ten males and five of ten females consumed the mixture within 24 hours. After a 14-day period during which all rats gained weight normally they were killed. There were no gross lesions at necropsy. Under the specified conditions of administration the LD50 was >5000 mg/kg bw (Freeman, 1991a). An acute inhalation toxicity study using a preparation of Avicel AC-815 (composed of 85% microcrystalline cellulose and 15% calcium alginate) with mass median aerodynamic diameter of 8.48-8.61 µm (range of measures) was dispersed and delivered at a mean concentration of 5.35 mg/litre in a nose-only inhalation exposure chamber to 5 male and 5 female Crl:CDBR VAF Plus rats for a period of 4 hours. The rats were observed over the 14 days after removal from the chamber. The only signs of toxicity were on removal from the chamber and consisted of chromodacryorrhea, chromorhinorrhea and, in one male rat, decreased locomotion; these signs had resolved by the next day. After 14 days no gross lesions were observed at necropsy (Signorin, 1996). 2.2.2 Short-term toxicity studies 2.2.2.1 Rats Groups of four male rats were kept on diets containing 0.25, 2.5 or 25% of various edible celluloses for 3 months. No differences were observed among the groups with regard to growth and faecal output. Histopathology of the gastrointestinal tract revealed no treatment-related abnormalities (Frey et al., 1928). Three groups of five male rats received 0.5 or 10% microcrystalline cellulose in their diet for 8 weeks. Growth was comparable to controls but the 10% group showed slightly lower body weights. Haematology, serum chemistry and vitamin B1 levels in blood and faeces showed no differences from controls (Asahi Chemical Industry Co., 1966). Groups of five male weanling Sprague-Dawley rats received 0, 5, 10 or 20% of acid-washed cellulose in their diet during three consecutive nutrient balance trials over a period of 17 days. Absorption of magnesium and zinc were significantly lower in the animals that were receiving the 10 and 20% cellulose diets. Histopathology of the gastrointestinal tract revealed increased mitotic activity and the presence of increased numbers of neutrophils in crypt epithelial cells, particularly of the duodenum and jejunum (Gordon et al., 1983). A mixture of four types of Elceme (in the ratio of 1:1:1:1) was fed to groups of Wistar rats for 30 days at a dietary level of 50%, and for 90 days at a dietary level of 10% (Elceme is a microcrystalline cellulose, and the four types are identified by particle size, namely, 1-50 (powder), 1-100 (powder), 1-150 (fibrillar), 90-250 (granulate)). All test animals were observed for food intake and weight gain. For animals in the 10% group, urinalysis, haematological tests and serum biochemical tests were carried out at weeks 6 and 13 of the test. A complete autopsy including histopathology was carried out at the end of the study. Animals in the 50% group were subjected to a persorption test, on the last day of the study, by addition of a cellulose staining dye (Renal, Wine-red) to the food of the test animals at a level equivalent to 5% of the Elceme. The animals were sacrificed 24 hours after administration of the diet, and a careful histological examination was made of the gastrointestinal tract, spleen, liver, kidney and heart for stained particles. Animals in the 10% group gained significantly less weight than those in the control group; the marked decrease commenced in the third or fourth week of the study. Food intake was similar in test and control groups. Urinalysis, haematological values and biochemical values were similar for test and control group 1. At autopsy some of the rats on the test diet had distended stomachs, which often contained considerable amounts of the test diet. The absolute liver and kidney weights and the ratio of the weight of these organs to brain weight was increased in test animals when compared with control animals. No compound-related pathology was reported. Animals in the 50% group showed considerable less weight gain than control animals in spite of a marked increase in food consumption. No persorption of dyed fibres was observed (Ferch, 1973a,b). Randomly bred rats of both sexes were divided into groups that received a control diet or the control diet with 330 mg/kg microcrystalline cellulose for a period of 6 months. Six rats in each group were then killed, their organs were examined, and tissues were taken for histopathology. No effects of the treatment were observed (Yartsev et al., 1989). Groups of Crl:CD(R) BR/VAF/Plus rats (20/sex per group) were administered 0 (control), 25 000 or 50 000 mg/kg Avicel RCN-15 in the diet for 90 days. A few test animals were noted as having chromodacryorrhea/ chromorhinorrhea, but this was not considered to be biologically significant. In some early weeks the rats increased diet consumption, probably to allow for the increased dietary fibre content. Body weight gain was unaffected. During the study and at necropsy there was no evidence of treatment-related changes. Clinical chemistry, haematology and organ weights were unaffected by treatment. Histopathology of 34 organs or tissues, including gastrointestinal tract and gut-associated lymphoid tissue of the ileum, provided no evidence of toxicity of microcrystalline cellulose. The calculated daily consumption of microcrystalline cellulose was 3769 mg/kg bw per day for males and 4446 mg/kg bw per day for females. The author noted that the NOEL exceeded 50 000 mg/kg diet (Freeman, 1992a). Groups of Sprague-Dawley CD rats (20 rats/dose per sex) from Charles River Laboratories were administered 0 (control), 25 000 or 50 000 mg/kg Avicel CL-611 in the diet for 90 days. (Avicel CL-611 or Avicel(R) Cellulose Gel is composed of 85% microcrystalline cellulose and 15% sodium carboxymethyl cellulose). There were no differences in weight gain of the males; a body weight gain decrement in females was attributed to a decreased caloric intake. No adverse effects attributable to the treatment were observed. At necropsy organ weights of the test groups were normal other than changes to adrenals of males receiving 50 000 mg/kg and to absolute brain and kidney weights in females receiving 25 000 mg/kg, but these were not attributed to the treatment. Histopathology of 36 organs or tissues from the control and high-dose groups, including gastrointestinal tract and gut-associated lymphoid tissue of the ileum, provided no evidence of toxicity of the microcrystalline cellulose. The mean nominal consumptions, averaged over weekly periods, of Avicel CL-611 by males and females of the top-dose groups ranged from 2768 to 5577 and 3673 to 6045 mg/kg bw per day, respectively (Freeman, 1994a). Microcrystalline cellulose (Avicel) was used as a positive control in a short-term toxicity study (approximately 13 weeks) of Cellulon, a cellulose fibre. Sprague-Dawley Crl:CB (SD) BR rats, 20 rats/sex per group, received a diet containing 0, 5 or 10% of the appropriate fibre ad libitum. Animals were checked daily, and body weights and food consumption were monitored weekly. Haematology (10 parameters) and clinical chemistry (14 parameters) were performed on blood samples taken from 10 rats/sex per group. All animals were necropsied, and gross observations and the weights of liver, testes with epididymes, adrenals and kidneys were recorded. Histological examination was carried out on tissue sections from control and high-dose groups. Food consumption was increased in the groups fed cellulose fibre, although there were no differences in body weight between the fibre-fed and control groups. This effect was attributed to the altered nutritional value of the diet. From the haematology and clinical chemistry there was only one significant difference of a microcrystalline cellulose group from the control value; this was in the group of female rats fed 5% microcrystalline cellulose in which there was an elevation of the haematocrit. There was no evidence of a dose response. Study of the necropsy results and the histological observations indicated that there was no evidence of any treatment-related effects of microcrystalline cellulose during the 13-week feeding study in rats at either 5 or 10% in the diet (Schmitt et al., 1991). Groups of Sprague-Dawley (CD) rats (20 rats/dose per sex) from Charles River Laboratories were administered, by gavage, suspensions of a special, fine particle size, microcrystalline cellulose (median particle size 6 µm). The dose levels were 0 (control), 500, 2500 or 5000 mg/kg per day as a 25% suspension in tap water. Dosing was performed daily for 90 consecutive days. No treatment-related deaths occurred during the study and the only treatment-related clinical sign (pale faeces) was not attributed to toxicity. There were no toxicologically significant effects in treated animals with respect to body weight, absolute and relative organ weights (5 organs weighed), food consumption, clinical chemistry measurements, haematology measurements or opthalmoscopic examinations. In animals that had received 5000 mg/kg per day there were no treatment-related lesions detected histopathologically (in 36 tissues including gut-associated lymphoid tissue, liver, lung, spleen and brain) nor was there any macroscopic or microscopic finding of microemboli or granulomatous inflammatory lesions (Kotkoskie et al., 1996). 2.2.3 Long-term toxicity/carcinogenicity studies 2.2.3.1 Rats Three groups of 50 male and 50 female rats received in their diet for 72 weeks either 30% ordinary cellulose or dry microcrystalline cellulose or micro-crystalline cellulose gel. Appearance and behaviour was comparable in all groups. No adverse effects were noted. The body weights of males given microcrystalline cellulose gel were higher than those of the controls. Food efficiency, survival and haematology were comparable in all groups. The liver and kidney weights of males receiving microcrystalline cellulose gel were higher than the controls. Gross and histopathology showed some dystrophic calcification of renal tubules in females on microcrystalline cellulose but all other organs appeared unremarkable. Tumour incidence did not differ between the groups (Hazleton Labs, 1963). The Committtee was aware of a study in which a microcrystalline cellulose preparation, of which 90% of the particles had a diameter < 20 µm, was fed to male and female rats at 0 (control), 30, 100 or 200 g/kg diet. The high mortality during the course of the study, the evidence of confounding infection, the limited number of animals for which there was histopathological examination, and the absence of details of the first year of feeding do not provide adequate reassurance as to the ability of this study to detect other than gross effects (Lewerenz et al., 1981). A review article makes reference to what may be the earlier, first year portion of the above study; however, inadequate referencing did not permit a firm conclusion (Steege et al., 1980). 2.2.4 Reproductive toxicity studies 2.2.4.1 Rats Groups of eight male and 16 female rats were used to produce P, F1a, F1b, F2 and F3 generations after having been fed on diets containing 30% microcrystalline cellulose flour or gel or ordinary cellulose as a control. The presence in the diet of such an amount of non-nutritious material, which contributed no calories, had an adverse effect on reproduction. Fertility and numbers of live pups were relatively depressed, and lactation performances in all three generations, as well as survival and the physical condition of the pups, were unsatisfactory throughout the study. The new-born pups appeared smaller, weak and showed evidence of disturbed motor coordination. Liver weights were increased in the group receiving microcrystalline cellulose gel in all generations but other organ weights showed no consistent patterns. At autopsy female rats of all generations showed kidney changes comprising pitting, occasional enlargement and zonation of the cortex. Other organs showed no consistent changes. No teratological deformities were seen (Hazleton Labs, 1964). 2.2.5 Special studies on embryotoxicity and teratogenicity 2.2.5.1 Rats Seventy-two rats (Sprague-Dawley CD) divided into eight groups were fed a mixture of four types of Elceme in the ratio of 1:1:1:1 in the diet at a level of 0, 2.5, 5 or 10% for 10 days, between days 6 and 15 of pregnancy. Rats of four test groups were killed on day 21 of pregnancy and the following parameters studied: number of fetuses and resorption sites, litter size and average weight of rats, average weight of fetuses and average backbone length. Fetuses were also examined for soft tissue or skeletal defects. The remaining groups were allowed to bear young, which were maintained to weaning (21 days). The following parameters were studied: litter size, weight of pups at days 7 and 21, and there was a histological study of the offspring. Although there is some suggestion that administration of dietary Elceme resulted in a dose-dependent increase in resorption sites, as well as a change in sex ratio, and possible defects such as opaque crystalline lenses, the data has not been presented in a manner that permits a meaningful interpretation. However, the author concluded that Elceme is non-teratogenic (Ferch, 1973a,b). Groups of 25 presumed pregnant Crl:CD(R) BR VAF/Plus rats were administered 0 (control), 25 000 or 50 000 mg Avicel RCN-15/kg diet (equal to 2.1 and 4.5 g/kg bw per day, respectively) ad libitum on days 6 to 15 of gestation. Animals received basal diet at all other times. In the group receiving 50 000 mg/kg the food consumption on days 6 to 15 was significantly higher than that of controls, probably because of the increased fibre content. On day 20 of gestation the dams were killed by carbon dioxide inhalation and the following parameters studied: number and distribution of implantation sites, early and late resorptions, live and dead fetuses and corpora lutea. External, visceral and skeletal examinations of the fetuses were also performed. There was no evidence of any adverse effects of the test material on either the dams or the fetuses. Due to a protocol error fetal sex was not recorded (Freeman, 1992b). Groups of 25 presumed pregnant Charles River Sprague-Dawley CD rats were administered 0 (control), 25 000 or 50 000 mg Avicel CL-611/kg (equal to 2.2 and 4.6 g/kg bw per day, respectively) diet ad libitum on days 6 to 15 of gestation. Animals received basal diet at all other times. In the test groups the food consumption on days 6 to 15 was significantly higher than for controls, probably because of the increased fibre content. The parameters studied and examinations performed were the same as in the study of Freeman (1992b). There was no evidence of any effects of the Avicel treatment on the fetuses, and there was no evidence of a change of sex ratio in the pups or of eye defects. Under the conditions of the study, the maternal and fetal NOEL was > 50 000 mg/kg diet (equal to 4.6 g/kg bw per day) (Freeman, 1994b). 2.2.6 Special studies on genotoxicity Various microcrystalline cellulose preparations have been tested for genotoxicity in several different assay systems. The results, all of which were negative, are summarized in Table 2. Table 2. Results of genotoxicity assays of microcrystalline cellulose Test system Test cells Concentration Results Reference Reverse mutation 1,2 Salmonella typhimurium 50-5000 µg/plate negative Batt, 1992 TA98, TA100, TA1535, TA1537, TA1538 Reverse mutation 1,3 Salmonella typhimurium 10-5000 µg/plate negative Lawlor, 1996 TA98, TA100, TA1535, TA1537, TA1538 Escherichia coli 10-5000 µg/plate negative WP2uvrA Forward mutation 1,2 Mouse lymphoma 100-1000 µg/ml negative Cifone, 1992 L5178Y cells, TK locus Forward mutation 1,4 Mouse lymphoma 125-1000 µg/ml negative Cifone, 1994 L5178Y cells, TK locus UDS with confirmatory Rat liver primary cell 10-1000 µg/ml inactive McKeon, 1992 assay 2 cultures In vivo mammalian Bone marrow polychromatic 5000 mg/kg bw, negative Murli, 1992 micronucleus assay 2,5 erythrocytes of ICR mice oral In vivo mammalian Bone marrow polychromatic 5000 mg/kg bw, negative Murli, 1994a micronucleus assay 6 erythrocytes of CD-1 oral (ICR) mice In vivo mammalian Bone marrow polychromatic 5000 mg/kg bw, negative Murli, 1994b micronucleus assay 4 erythrocytes of CD-1 oral (ICR) mice Table 2. Continued... 1 With and without rat liver S9 metabolic activation 2 Test material: Avicel RCN-15 3 Test material: Avicel AC-815 4 Test material: Avicel CL-611 5 Test material: Avicel RCN-15 6 Test material: Avicel PH101 Pharmaceutical In the reverse mutation assays the microcrystalline cellulose formulations produced a heavy precipitate on the plate at the highest concentration. Solubility also affected the forward mutation assays and it was not possible to include concentrations of the test material that were cytotoxic. In the in vivo mammalian micronucleus assays it is improbable that there was appreciable persorption of the test materials, and, therefore, there was little exposure of the bone marrow cells. In the test in which Avicel RCN-15 was used it was administered admixed with the diet of male and female ICR mice. Only mice that had consumed all the diet within 10 hours were retained in the study and were killed after 24, 48 or 72 hours. Because one group of control mice had 0 micronuclei per 1000 polychromatic erythrocytes, the comparison with the test group was statistically significant. This was not considered to be a valid observation. There is no evidence that microcrystalline cellulose is genotoxic. 2.2.7 Special studies on sensitization Avicel RCN-15 was determined to be non-sensitizing when topically applied to ten male and ten female Hartley guinea-pigs (Freeman, 1991e). Avicel AC-815 was determined to be non-sensitizing when topically applied to ten male Hartley guinea-pigs (Freeman, 1996c). 2.2.8 Special studies on skin and eye irritation Avicel RCN-15 was judged to be minimally irritating after instillation into the eyes of four male and two female New Zealand White rabbits (Freeman, 1991c). Avicel AC-815 was judged to be minimally irritating after instillation into the eyes of four male and two female New Zealand White rabbits (Freeman, 1996a). Avicel RCN-15 was judged to be non-irritating after a 4-hour occlusive contact with the skin of three male and three female New Zealand White rabbits (Freeman, 1991d). Avicel AC-815 was judged to be non-irritating after a 4-hour occlusive contact with the skin of three male and three female New Zealand White rabbits (Freeman, 1996b). 2.2.9 Special studies on effects of cellulose fibre on tumour growth The effect of artifical diets containing varied concentrations of either wheat bran or pure cellulose fibre on the induction of mammary tumours by N-nitrosomethylurea (i.v., 40 mg/kg) was studied in female F344 rats. The wheat bran diet appeared to possess anti-promotion properties that pure cellulose lacked. The concentrations of serum estrogens, urinary estrogens and faecal estrogens did not vary in a consistent, statistically significant manner (Cohen et al., 1996). The effect of a high-fibre diet containing 45 000 mg/kg Avicel PH- 105 on the development of colon tumours was investigated in male Wistar rats that were injected with 1,2-dimethylhydrazine dihydrochloride (25 mg/kg, s.c., once weekly for 16 weeks). The test and control diets were administered for 2 weeks prior to the first injection of the carcinogen. There was a reduction in the number of animals bearing colon tumours and a statistically significant reduction in the number of colon tumours/rat in the high-fibre dietary group. However, for small bowel tumours and tumours of the ear canal there was no significant difference between the dietary groups Freeman et al., 1978). A later study by the same authors demonstrated that there was no significant effect of increasing the level of cellulose in the diet to 90 000 mg/kg (Freeman et al., 1980). 2.3 Observations in humans 2.3.1 Toxicity consequent to substance abuse Intravenous abuse of drugs available in tablet form has led to the detection of excipients, e.g., talc, magnesium stearate or microcrystalline cellulose, in the tissues of a series of 33 fatality cases of intravenous drug addicts. Microcrystalline cellulose (21 cases) and talc (31 cases) were detected most frequently and, in some cases, were associated with granulomatous lesions (Kringsholm & Christoffersen, 1987). In one case intravenous abuse of the drug pentazocine, possibly for longer than six months, led to a fatal pulmonary granulomatosis (Zeltner et al., 1982). In other cases of pentazocine abuse, in addition to pulmonary granulomas there were granulomas at the injection site and in the right ventricular endocardium. The principal vascular lesion was thrombosis (Tomashefski et al., 1981). 2.3.2 Changes in gastrointestinal function and nutrient balance A number of clinical studies using refined cellulose as roughage in the human diet for the treatment of constipation showed no deleterious effects. Groups of 18 children received regular amounts of edible cellulose instead of normal cereal for three months. The only effect noted was an increase in bowel movements but no diarrhoea or other gastrointestinal disturbances were seen (Frey et al., 1928). Eight male and eight female volunteers supplemented their normal diet with 30 g microcrystalline cellulose per day as either dry powder or gel (15% aqueous) for 6 weeks followed by 2 weeks without supplementation. No adverse findings were reported regarding acceptance or body weight but most subjects complained of fullness and mild constipation. Haematology was normal in all subjects. Biochemical blood values showed no differences between treatment and control periods, nor was there evidence of liver or kidney function disturbance. Urinalysis produced normal findings. The faecal flora remained unchanged. The cellulose content of faeces increase five to eight times during the test period. Microscopy revealed the presence of microcrystalline cellulose (Hazleton Labs, 1962). In another study, eight healthy males received 30 g microcrystalline cellulose daily as supplement to their diet for 15 days. D-xylose absorption varied between pretest, test and post-test periods, being lower during microcrystalline cellulose ingestion. The absorption of 131I-triolein was unaffected by microcrystalline cellulose ingestion. No change was noted in the faecal flora nor was there any significant effect on blood chemistry during ingestion of microcrystalline cellulose. Examination of urine, blood and faecal levels of vitamin B1 during microcrystalline cellulose ingestion showed no difference from control periods (Asahi Chemical Industry Co., 1966). Twelve men consumed diets containing fibres from various sources for periods of 4 weeks. There was no significant difference between values of serum cholesterol, triglyceride and free fatty acid levels measured after consumption of the basal diet, compared with the values measured after consumption of a diet containing cellulose fibres (90% cellulose, 10% hemicellulose; James River Corp., Berlin, New Hampshire, USA). There were no significant differences in plasma VLDL and HDL cholesterol or in the ratio of HDL/VLDL+LDL cholesterol. However, the increase in plasma LDL cholesterol after the cellulose diet was significant (Behall et al., 1984). A similar study in a group of four men and six women could detect no effect of a diet containing added alpha-cellulose (15 g daily) on serum total cholesterol, triglycerides, HDL cholesterol and the ratio of HDL to total cholesterol. The cellulose was well tolerated (Hillman et al., 1985). A double-blind cross-over trial of the effects of guar gum and microcrystalline cellulose on metabolic control and serum lipids in 22 Type 2 diabetic patients has been carried out. The fibre preparations were given at 15 g/day for a 2-week period and then at 5 g/day for the remaining 10-week period of each treatment phase. There was no effect of the microcrystalline cellulose diet on fasting blood glucose level, glycosylated haemoglobin, serum HDL-cholesterol, serum triglycerides, serum zinc or ferritin, or urinary magnesium excretion (Niemi et al., 1988). The effect of various dietary fibres, including microcrystalline cellulose (40 g), on the uptake of vitamin A (approximately sixty times the daily requirement) from a test meal was investigated in 11 female subjects aged 19 to 22. All the dietary fibres significantly increased the absorption of the vitamin A over a period of 9 hours (Kasper et al., 1979). A study of apparent mineral balance in a group of eleven men revealed that there was no significant effect of cellulose, added to the diet at 7.5 g per 1000 kcal for 4 weeks, on the mineral balance of calcium, magnesium, manganese, iron, copper or zinc. However, in this report the source of the cellulose fibre was not specified (Behall et al., 1987). The addition of nutritional grade cellulose (21 g) to the daily diet of healthy adolescent girls resulted in reduction of the serum calcium, phosphorus and iron concentrations. The authors suggested that high-fibre diets may not be advisable (Godara et al., 1981). A study of only three men on a low-fibre diet claimed changes in mineral balance consequent on the consumption of additional cellulose fibre, 10 g of Whatman No. 3 filter paper daily, in the diet (Ismail-Beigi et al., 1977). Microcrystalline cellulose (5 g) did not appear to inhibit the uptake of iron in women who were neither pregnant nor lactating (Gillooly et al., 1984). A group of twenty women, aged 27-48, who were given 20 g packs of alpha-cellulose to be consumed daily for three months, were included in a study of the effect of indole-3-carbinol on estrogen metabolite ratios. Because the control group and the group fed indole-3-carbinol received capsules, the cellulose group could not be blinded; in addition, an unspecified number of subjects in this group dropped out as they found that the cellulose suspension was unpalatable. However, the authors suggest that the estrogen metabolite ratio in the high- fibre group was not different from that in the control group (Bradlow et al., 1994). 3. COMMENTS Persorption of microcrystalline cellulose was reported in various species, which included rats, in early studies. A recent study in which a special fine particle size preparation of microcrystalline cellulose (median diameter of particles 6 µm) was administered orally to rats (5 g/kg bw per day) for 90 days has failed to confirm the earlier observations. In this study precautions were taken to ensure that, at autopsy, there was no cross-contamination of the tissues with fine particulate matter. In various parenteral studies of the acute toxicity of microcrystalline cellulose in animals there have been signs consistent with a tissue response to foreign particles. Similarly, microcrystalline cellulose has been associated with the formation of granulomas in human lung when it has been injected intravenously during drug abuse. No such lesions have been described as a consequence of oral ingestion of microcrystalline cellulose by rats or humans. In 90-day toxicity tests during which microcrystalline cellulose was administered to rats in the diet at concentrations of 2.5 to 50%, increased consumption of food to compensate for the content of this material was observed. Although this may have some effects on mineral absorption there was, in general, no compound-related systemic toxicity. The NOEL exceeded 50 g/kg diet, at which dose level the mean intakes of microcrystalline cellulose by male and female rats were 3.8 and 4.4 g/kg bw per day, respectively. A two-year feeding study of microcrystalline cellulose in rats was brought to the attention of the Committee. Despite a lack of evidence of toxic effects, the Committee considered that the execution and reporting of the study were not adequate to identify a NOEL. In vitro and in vivo genotoxicity studies were negative. In a three-generation reproductive toxicity study in rats that had been reviewed by an earlier Committee, there were some effects of using 30% microcrystalline cellulose in the diet; these had been considered to be a consequence of the quantity of material reducing the energy density of the diet. However, in recent embryotoxicity and teratogenicity studies in rats there was no evidence of compound-related effects at dietary levels up to 50 g of microcrystalline cellulose per kg diet (equal to 4.6 g/kg bw per day), given on days 6 to 15 of pregnancy. In some human studies there have been reports of alterations to gastrointestinal function following ingestion of microcrystalline cellulose. The changes do not appear to be related to systemic toxicity. 4. EVALUATION The Committee concluded that the toxicological data from humans and animals provided no evidence that the ingestion of microcrystalline cellulose can cause toxic effects in humans when used in foods according to good manufacturing practice. It is recognized that small particles of other materials may be persorbed and that the extent of persorption is greater with sub-micrometre particles. Despite the absence of any demonstrated persorption of microcrystalline cellulose in the recent study in rats, the Committee, as a precautionary measure, revised the specifications for microcrystalline cellulose at the present meeting to limit the content of particles less than 5 µm in diameter. The Committee retained the ADI "not specified" for microcrystalline cellulose conforming to these specifications. 5. REFERENCES Asahi Chemical Industry Co. (1966) Effect of ingestion of avicel-contained foods on living organisms. Unpublished report from Yoshitoshi Internal Seminar (Submitted to WHO by Asahi Chemical Industry Co., Ltd). Baker, E.M. (1966) Microcrystalline cellulose: oral administration - Rats. Unpublished report from Fitzsimmons General Hospital (Submitted to WHO by FMC Corporation). Baker, E.M. (1968) Microcrystalline cellulose: oral administration - Humans. Unpublished report from Fitzsimmons General Hospital (Submitted to WHO by FMC Corporation). Batt, K.J. (1992) Avicel RCN-15 - Salmonella/mammalian microsome plate incorporation assay (Ames test). Unpublished report No. I91-1214 from FMC Corporation Toxicology Laboratory, Princeton, New Jersey, USA (Submitted to WHO by FMC Europe N.V.). Behall, K.M., Lee, K.H., & Moser, P.B. (1984) Blood lipids and lipoproteins in adult men fed four refined fibers. Am. J. Clin. Nutr., 39: 209-214. Behall, K.M., Scholfield, D.J., Lee, K., Powell, A.S., & Moser, P.B. (1987) Mineral balance in adult men: effect of four refined fibers. Am. J. Clin. Nutr., 46: 307-314. Bradlow, H.L., Michnovicz, J.J., Halper, M., Miller, D.G., Wong, G.Y.C., & Osborne, M.P. (1994) Long-term response of women to indole-3-carbinol or a high fiber diet. Cancer Epidemiol. Biomarkers Prev, 3: 591-593. CanTox Inc. (1993) Estimated consumption of microcrystalline cellulose and sodium carboxymethylcellulose from current and proposed food uses of Avicel cellulose gel. Unpublished report dated December 1993, prepared by CanTox Inc. for FMC Corporation (Submitted to WHO by FMC Europe N.V.). Carryer, P.W., Brown, M.L., Malagelada, J.-R., Carlson, G.L., & McCall, J.T. (1982) Quantification of the fate of dietary fiber in humans by a newly developed radiolabeled fiber marker. Gastroenterology, 82: 1389-1394. Cifone, M.A. (1992) Mutagenicity test on Avicel RCN-15 in the L5178Y TK+/- mouse lymphoma forward mutation assay with an independent repeat. Unpublished report by Hazleton Washington Inc., Vienna, Virginia, USA (FMC Study No. 191-1230) (Submitted to WHO by FMC Europe N.V. Cifone, M.A. (1994) Mutagenicity test on Avicel CL-611, E329N in the L5178Y TK+/- mouse lymphoma forward mutation assay with a confirmatory assay. Unpublished report by Hazleton Washington Inc., Vienna, Virginia, USA (FMC Study No. 194-1834) (Submitted to WHO by FMC Europe N.V.) Cohen, L.A., Zhao, Z., Zang, E., & Rivenson, A. (1996) Dose-response effects of dietary fiber on NMU-induced mammary tumorigenesis, estrogen levels and estrogen excretion in female rats. Carcinogenesis, 17: 45-52. Egan, S.K. & Heimbach, J.T. (1994) Microcrystalline cellulose, MCC, E460(i). Part four: Exposure data - Estimated intake of MCC in the United Kingdom. Unpublished report dated April 11, 1994, prepared by TAS, Inc., Washington, DC, USA for FMC Corporation (Submitted to WHO by FMC Europe N.V.). Eldridge, J.H., Gilley, R.M., Staas, J.K., Moldoveanu, Z., Meulbroek, J.A., & Tice, T.R. (1989) Biodegradable microspheres: vaccine delivery system for oral immunization. Curr. Top. Microbiol. Imunol., 146: 59-66. Eldridge, J.H., Hammond, C.J., Meulbroek, J.A., Staas, J.K., Gilley, R.M. & Tice, T.R. (1990) Controlled vaccine release in the gut-associated lymphoid tissues: I. Orally administered biodegradable microspheres target the Peyer's patches. J. Control. Release, 11: 205-214. Ferch, H. (1973a) Innocuity of Elceme (R). Part I. Pharm. Ind., 35(9): 578-583. Ferch, H. (1973b) Innocuity of Elceme (R). Part II. Pharm. Ind., 35(9): 658-661. FMC Corporation N.V. (1996) Microcrystalline cellulose. MCC, Ins 460 (i). Technical and Scientific Dossier. Unpublished report from FMC Europe N.V., Brussels, Belgium, dated November 1996 (Submitted to WHO by FMC Europe N.V.). Freeman, C. (1991a) Avicel RCN-15. Acute oral toxicity study in rats. Unpublished report No. I91-1217 from FMC Corporation Toxicology Laboratory, Princeton, New Jersey, USA (Submitted to WHO by FMC Europe N.V.). Freeman, C. (1991b) Avicel RCN-15. Acute dermal toxicity study in rats. Unpublished report No. I91-1219 from FMC Corporation Toxicology Laboratory, Princeton, New Jersey, USA (Submitted to WHO by FMC Europe N.V.). Freeman, C. (1991c) Avicel RCN-15. Primary eye irritation study in rabbits. Unpublished report No. I91-1218 from FMC Corporation Toxicology Laboratory, Princeton, New Jersey, USA (Submitted to WHO by FMC Europe N.V.). Freeman, C. (1991d) Avicel RCN-15. Primary skin irritation study in rabbits. Unpublished report No. I91-1220 from FMC Corporation Toxicology Laboratory, Princeton, New Jersey, USA (Submitted to WHO by FMC Europe N.V.). Freeman, C. (1991e) Avicel RCN-15. Skin sensitisation study in guinea pigs. Unpublished report No. I91-1216 from FMC Corporation Toxicology Laboratory, Princeton, New Jersey, USA (Submitted to WHO by FMC Europe N.V.). Freeman, C. (1992a) Avicel RCN-15. Ninety-day feeding study in rats. Unpublished report No. I91-1202 from FMC Corporation Toxicology Laboratory, Princeton, New Jersey, USA (Submitted to WHO by FMC Europe N.V.). Freeman, C. (1992b) Avicel RCN-15. Teratology study in rats (dietary). Unpublished report No. I91-1213 from FMC Corporation Toxicology Laboratory, Princeton, New Jersey, USA (Submitted to WHO by FMC Europe N.V.). Freeman, C. (1994a) Avicel CL-611. Ninety-day feeding study in rats. Unpublished report No. I92-1711 from FMC Corporation Toxicology Laboratory, Princeton, New Jersey, USA (Submitted to WHO by FMC Europe N.V.). Freeman, C. (1994b) Avicel CL-611. Teratology study in rats (dietary). Unpublished report No. I92-1712 from FMC Corporation Toxicology Laboratory, Princeton, New Jersey, USA (Submitted to WHO by FMC Europe N.V.). Freeman, C. (1996a) Avicel AC-815. Primary eye irritation study in rabbits. Unpublished report No. I95-2042 from FMC Corporation Toxicology Laboratory, Princeton, New Jersey, USA (Submitted to WHO by FMC Europe N.V.). Freeman, C. (1996b) Avicel AC-815. Primary skin irritation study in rabbits. Unpublished report No. I95-2043 from FMC Corporation Toxicology Laboratory, Princeton, New Jersey, USA (Submitted to WHO by FMC Europe N.V.). Freeman, C. (1996c) Avicel AC-815. Skin sensitization study in guinea pigs. Unpublished report No. I95-2044 from FMC Corporation Toxicology Laboratory, Princeton, New Jersey, USA (Submitted to WHO by FMC Europe N.V.). Freeman, C. (1996d) Avicel AC-815. Acute oral toxicity study in rats. Unpublished report No. I95-2040 from FMC Corporation Toxicology Laboratory, Princeton, New Jersey, USA (Submitted to WHO by FMC Europe N.V.). Freeman, C. (1996e) Avicel AC-815. Acute dermal toxicity study in rats. Unpublished report No. I95-2041 from FMC Corporation Toxicology Laboratory, Princeton, New Jersey, USA (Submitted to WHO by FMC Europe N.V.). Freeman, H.J., Spiller, G.A., & Kim, Y.S. (1978) A double-blind study on the effect of purified cellulose dietary fiber on 1,2- dimethylhydrazine-induced rat colonic neoplasia. Cancer Res., 38: 2912-2917. Freeman, H.J., Spiller, G.A., & Kim, Y.S. (1980) A double-blind study on the effects of differing purified cellulose and pectin fiber diets on 1,2-dimethylhydrazine-induced rat colonic neoplasia. Cancer Res., 40: 2661-2665. Frey, J.W., Harding, E.R., & Helmbold, T.R. (1928) Dietetic investigations of edible pure cellulose. Med. J. Rec., 127: 585-589. Gillooly, M., Bothwell, T.H., Charlton, R.W., Torrance, J.D., Bezwoda, W.R., MacPhail, A.P., Derman, D.P., Novelli, L., Morrall, P., & Mayet, F. (1984) Factors affecting the absorption of iron from cereals. Br. J. Nutr., 51: 37-46. Godara, R., Kaur, A.P., & Bhat, C.M. (1981) Effect of cellulose incorporation in a low fiber diet on fecal excretion and serum levels of calcium, phosphorus, and iron in adolescent girls. Am. J. Clin. Nutr., 34: 1083-1086. Gordon, D.T., Besch-Williford, C., & Ellersieck, M.R. (1983) The action of cellulose on the intestinal mucosa and element absorption by the rat. J. Nutr., 113: 2545-2556. Hazleton Labs (1962) Microcrystalline cellulose; oral administration - Human. Unpublished report from Hazleton Labs, Inc. (Submitted to WHO by FMC Corporation). Hazleton Labs (1963) Long-term nutritional balance study - Rats. Unpublished report from Hazleton Labs, Inc. (Submitted to WHO by FMC Corporation). Hazleton Labs (1964) Microcrystalline cellulose: reproduction study - Rats. Unpublished report from Hazleton Labs, Inc. (Submitted to WHO by FMC Corporation). Hillman, L.C., Peters, S.G., Fisher, C.A., & Pomare, E.W. (1985) The effects of the fiber components pectin, cellulose and lignin on serum cholesterol levels. Am. J. Clin. Nutr., 42: 207-213. Ismail-Beigi, F., Reinhold, J.G., Faraji, B., & Abadi, P. (1977) Effects of cellulose added to diets of low and high fiber content upon the metabolism of calcium, magnesium, zinc and phosphorus by man. J. Nutr., 107: 510-518. Jani, P., Halbert, G.W., Langridge, J., & Florence, A.T. (1990) Nanoparticle uptake by the rat gastrointestinal mucosa: quantitation and particle size dependency. J. Pharm. Pharmacol., 42: 821-826. Jani, P.U., McCarthy, D.E., & Florence, A.T. (1994) Titanium dioxide rutile particle uptake from the rat GI tract and translocation to systemic organs after oral administration. Int. J. Pharm., 105: 157-168. Jenkins, P.G., Howard, K.A., Blackhall, N.W., Thomas, N.W., Davis, S.S., & O'Hagan, D.T. (1994) The quantitation of the absorption of microparticles into the intestinal lymph of Wistar rats. Int. J. Pharm., 102: 261-266. Kasper, H., Rabast, U., Fassl, H., & Fehle, F. (1979) The effect of dietary fiber on the postprandial serum vitamin A concentration in man. Am. J. Clin. Nutr., 32: 1847-1849. Kotkoskie, L.A., Butt, M.T., Selinger, E., Freeman, C., & Weiner, M.L. (1996). Qualitative investigation of uptake of fine particle size microcrystalline cellulose following oral administration in rats. J. Anat., 189: 531-535. Kringsholm, B. & Christoffersen, P. (1987) The nature and the occurrence of birefringent material in different organs in fatal drug addiction. Forensic Sci. Int., 34: 53-62. Lawlor, T.E. (1996) Mutagenicity test with Avicel AC-815 in the Salmonella-Escherichia coli/mammalian microsome reverse mutation assay with a confirmatory assay. Unpublished report by Corning Hazleton Inc., Vienna, Virginia, USA (FMC Study No. I95-2047) (Submitted to WHO by FMC Europe N.V.). LeFevre, M.E., Hancock, D.C., & Joel, D.D. (1980) Intestinal barrier to large particulates in mice. J. Toxicol. Environ. Health, 6: 691. Lewerenz, H.J., Bleyl, D.W.R., & Plass, R. (1981) Report on investigations in the second test year of continuous administration of microcrystalline cellulose into rats with their feed. Translation (and German original) of an unpublished report from the Academy of Sciences of the German Democratic Republic, Research Center for Molecular Biology and Medicine, Central Institute for Nutrition, Potsdam-Rehbrücke (Submitted to WHO by FMC Europe N.V.). Linskens, H.F. & Jorde, W. (1974) Persorption of lycopodium spores and pollen grains, Naturwissenschaften, 61: 275-276. McKeon, M.E. (1992). Genotoxicity test on Avicel RCN-15 in the assay for unscheduled DNA synthesis in rat liver primary cell cultures with a confirmatory assay. Unpublished report by Hazleton Washington Inc., Kensington, Maryland, USA (FMC Study No. I91-1229) (Submitted to WHO by FMC Europe N.V.). Murli, H. (1992) Mutagenicity test on Avicel RCN-15 in vivo mammalian micronucleus assay. Unpublished report by Hazleton Washington Inc., Kensington, Maryland, USA ( FMC Study No. I91-1228) (Submitted to WHO by FMC Europe N.V.). Murli, H. (1994a) Mutagenicity test on Avicel pH101 Pharmaceutical in an in vivo mouse micronucleus assay. Unpublished report by Hazleton Washington, Inc., Vienna, Virginia, USA (FMC Study No. I94-1837) (Submitted to WHO by FMC Europe N.V.). Murli, H. (1994b) Mutagenicity test on Avicel CL-611 in an in vivo mouse micronucleus assay. Unpublished report by Hazleton Washington, Inc., Vienna, Virginia, USA (FMC Study No. I94-1835) (Submitted to WHO by FMC Europe N.V.). Niemi, M.K., Keinänen-Kiukaanniemi, S.M., & Salmela, P.I. (1988) Long-term effects of guar gum and microcrystalline cellulose on glycaemic control and serum lipids in Type 2 diabetes. Eur. J. Clin. Pharmacol., 34: 427-429. Pahlke, G. & Friedrich, R. (1974) Persorption of microcrystalline cellulose, Naturwissenschaften, 61: 35. Pallotta, A.J. (1959) Acute oral administration - Rats; and acute intraperitoneal administration - Rats, of microcrystalline cellulose. Unpublished report from Hazleton Labs, Inc. (Submitted to WHO by FMC Corporation). Schmitt, D.F., Frankos, V.H., Westland, J., & Zoetis, T. (1991) Toxicologic evaluation of Cellulon fiber; genotoxicity, pyrogenicity, acute and subchronic toxicity. J. Am. Coll. Toxicol., 10: 541-554. Schreiber, G. (1974) Ingested dyed cellulose in the blood and urine of man. Arch. Environ. Health, 29: 39. Signorin, J. (1996) Avicel AC-815. Acute inhalation study in rats. Unpublished report No. I95-2045 by FMC Corporation Toxicology Laboratory, Princeton, New Jersey, USA (Submitted to WHO by FMC Europe N.V.). Simon, L., Shine, G., & Dayan, A.D. (1994) Effect of animal age on the uptake of large particulates across the epithelium of the rat small intestine. Int. J. Exp. Pathol., 75: 369-373. Steege, H., Lewerenz, H.J., Philipp, B., & George, J. (1980) Characterization of cellulose powders with special attention to the physiological aspects. International Dissolving Pulp Conference, German Democratic Republic, 5, 169-183. Tomashefski, J.F., Hirsch, C.S., & Jolly, P.N. (1981) Microcrystalline cellulose pulmonary embolism and granulomatosis. A complication of illicit intravenous injections of pentazocine tablets. Arch. Pathol. Lab. Med., 105: 89-93. Tusing, T.W., Paynter, O.E., & Battista, O.A. (1964) Birefringence of plant fibrous cellulose and microcrystalline cellulose in human stools freezer-stored immediately after evacuation. Agric. Food Chem., 12(3): 284-287. Volkheimer, G., Schultz, F.H., Lehmann, H., Aurich, I., Hubner, R., Hubner, M., Hallmayer, A., Munch, H., Opperman, H., & Strauch, S. (1968) Primary portal transport of persorbed starch granules from the intestinal wall. Med. Exp., 18: 103-108 Walters, M.P., Kelleher, J., Findlay, J.M., & Srinivasan, S.T. (1989) Preparation and characterisation of a [14C]cellulose suitable for human metabolic studies. Br. J. Nutr., 62: 121-129. Yartsev, N.M., Ivanova, V.S., Altymyshev, A.A., Sarybayeva, R.I., & Vasil'kova, T.V. (1989) Anatomical and histological state of rats given microcrystalline cellulose in long-term experiments. Izvestiya AN Kirgizskoi SSR, 3: 63-65. Zeltner, T.B., Nussbaumer, U., Rudin, O., & Zimmermann, A. (1982) Unusual pulmonary vascular lesions after intravenous injections of microcrystalline cellulose. A complication of pentazocine tablet abuse. Virchows Arch. [Pathol. Anat.], 395: 207-216.
See Also: Toxicological Abbreviations Microcrystalline cellulose (WHO Food Additives Series 1) Microcrystalline cellulose (WHO Food Additives Series 5) MICROCRYSTALLINE CELLULOSE (JECFA Evaluation)