Chronic Oral Toxicity Studies Research Articles

Safety Evaluation for Acute and Chronic Oral Toxicity of Maha Pigut Triphala Contains Three Medicinal Fruits in Sprague-Dawley Rats

Maha Pigut Triphala is the herbal mixture of three fruits consisting of T. bellirica, T. chebula, and E. officinalis also known as P. emblica. Humans regularly eat the fresh fruits of these plants on a daily basis. Maha Pigut Triphala is one of the widely known herbal medicinal formulas used in traditional Thai medicine. Besides studying pharmacological properties, attention should also be paid to the safety and toxicity studies of herbal medicines. The objective of the present study was to evaluate the acute and chronic oral toxicity of Maha Pigut Triphala (2:1:3) in Sprague-Dawley rats. A single dose of Maha Pigut Triphala at a concentration of 5000 mg/kg body weight was administered orally to female rats in the acute oral toxicity study. In the chronic oral toxicity study, male and female rats were treated with various concentrations of Maha Pigut Triphala (600, 1200, and 2400 mg/kg body weight) once daily for 270 consecutive days. The presence of abnormalities in the symptoms and behavior of the rats were observed and recorded throughout the experiment. Additionally, body weight, organ weight, and mortality were recorded. At the end of the study, blood samples were collected for hematological and blood chemistry analysis, while the internal organs were evaluated for gross pathological and histopathological changes. The acute oral toxicity study revealed no mortality and abnormal symptoms or behavior in Maha Pigut Triphala-treated rats. Moreover, gross pathological and histopathological findings did not reveal any abnormalities in the internal organs. In the chronic oral toxicity evaluation, although there were negligible changes in body weight, organ weight, and hematological and blood chemistry parameters in rats treated with Maha Pigut Triphala for 270 days, no behavioral or gross pathological and histopathological abnormalities were observed. Overall, the results of this study demonstrate that Maha Pigut Triphala (2:1:3) neither causes acute nor chronic oral toxicity in rats, proposing the safety of this herbal formula in animals prior to human trials and use.

Pre Clinical Safety Assessment of AYUSH Carctol S through Acute and Sub Chronic Oral Toxicity Study: Safety Assessment of AYUSH Carctol S

AYUSH Carctol S is a coded Ayurvedic compound formulation developed for treatment of cancer. The safety of the test drug was not established in the experimental animals yet, hence the present study was highlighted the same through acute and sub-chronic toxicity studies. The acute oral toxicity study of the test drug was performed in swiss albino mice with a dose of 2600 mg/kg. Similarly, the sub-chronic oral toxicity study was carried out in wistar rats with three different test doses i.e., 180 mg/kg, 900 mg/kg, and 1800 mg/kg. Cage side observations, feed intake, body weight, gross morphology, and necropsy findings of the animals of test drug-treated animals were normal and no mortality was reported in both either study. During the sub-chronic toxicity study, the hematological and biochemical parameters of AYUSH Carctol S-treated animals were found to be non-significant when compared to the control animals. Further, the histopathological analysis of major organs of AYUSH Carctol S 1800 mg/kg treated animals showed no major lesions and treatment-related changes. The test drug was considered to be nontoxic during the acute and sub-chronic oral toxicity studies. The LD50 of AYUSH Carctol S estimated during the acute toxicity was found as 2600 mg/kg and the sub-chronic safety dose i.e., NOAEL is up to 1800 mg/kg.

Safety profile of Naga Bhasma prepared by two classical methods

Abstract Background: Naga Bhasma (NB) (incinerated lead) has been indicated as a remedy for the treatment of a wide range of diseases including Prameha (diabetes), Kasa (cough), Raktapradara (menorrhagia), Shukradosha (defects in semen), etc., However, improperly prepared NB can cause harmful effects on the human body. In modern science also, lead consumption has been accepted to produce toxicity. Aim: The present study was undertaken to elucidate and compare the toxicity profile of two samples of NB prepared by two different methods to ascertain the safety aspects. Materials and methods: In the first method, NB was prepared by using Parada (mercury), Gandhaka (sulfur), and Nimbu Swarasa (juice of Citrus medica var acida of Watt). In the second method, NB was prepared by using Ashwattha-Twaka-Churna (Ficus religiosa L.), Manahshila (realgar) (Ariloha), and Nimbu Swarasa. In both methods, 30 calcination cycles were given. Charles foster albino rats were used for the pharmacological study. Acute oral toxicity study was carried out as per organization for economic cooperation and development (OECD) 425 and chronic oral toxicity study was carried out as per OECD 408 (90-day oral repeated dose toxicity study). Acute oral toxicity study was conducted at five dose levels, namely 100, 250, 500, 1000, and 2000 mg/kg body weight. Chronic toxicity was carried out at a dose level of 62.5 mg/kg body weight. Animals were sacrificed on the 91st day by cervical dislocation. Blood was collected from the jugular vein and sent for hematological and biochemical investigations were done. Organs were used for histopathological studies. The data were analyzed by unpaired t-test. Results: No mortality was observed in the acute toxicity study. Chronic toxicity study shows that both the test drugs do not produce significant toxicity, but both were not completely free from adverse effects at the dose level studied. Conclusion: The study shows that both the test drugs do not produce significant toxicity, but they are not completely free from adverse effects at the dose level studied.

OECD GUIDELINES FOR ACUTE ORAL TOXICITY STUDIES: AN OVERVIEW

The OECD Guidelines for testing chemicals provide a unique method for assessing the potential impact of chemicals on human health and the environment. These are divided into five sections. Oral toxicity studies are mentioned in the fourth section, i.e. Health Effects. This section cites the guidelines for acute, sub-acute, sub-chronic and chronic oral toxicity studies. Acute oral toxicity refers to those adverse effects of oral administration of a single dose or multiple doses given within 24 hours. OECD guidelines for Acute oral toxicity include 420 (Fixed Dose Procedure), 423 (Acute Toxic Class Method) and 425 (Up-and-Down procedure). OECD guidelines 420, 423 and 425 provide information on the hazardous properties and allow the substance to be ranked and classified according to the Globally Harmonised System (GHS) for the classification of chemicals which cause acute toxicity. In OECD Guideline 423, the method classifies the test substance as one of a series of toxicity classes defined by fixed LD50 cut-off values, while the OECD Guideline 425 method permits estimation of LD50 with a confidence interval. Based on these guidelines, a toxic dose of the drug is obtained. This study aims to compare and highlight the acute oral toxicity guidelines.

Safety evaluations of alternansucrase enzyme expressed in Escherichia coli shows no adverse effects invivo and invitro

Background Alternansucrase is a glucansucrase enzyme that mainly functions in synthesis of glucan-like polysaccharides and can be used for production of novel oligosaccharides and polysaccharides having low glycemic index and prebiotics properties which makes them ideal candidates as dietary fibers and food. From a regulatory perspective, it is necessary to prove the safety of alternansucrase before it can be used in any application as it is a novel enzyme and has not been historically used in food processing. Objectives Alternansucrase obtained from Escherichia coli was subjected to toxicological tests to determine its safety for use in various industrial applications. Design Toxicity studies were conducted at acute oral and repeated sub-chronic (14 days and 90 days) levels in rats following the OECD guidelines. The genotoxicity studies were conducted using the bacterial reverse mutation test as well as in vitro mammalian cell micronucleus test as per the OECD guidelines. Results Alternansucrase did not induce any clinical abnormalities or mortality in rats at the dose level of 2000 mg/kg of body weight in the acute oral toxicity test. A 90-day sub chronic repeated dose oral toxicity study with alternansucrase at and up to dose of 1000 mg TOS/kg of body weight did not show any treatment-related significant toxicological effects on body weight, food consumption, organ weights, hematological and clinical chemistry, or histopathology parameters. Alternansucrase was found to be non-mutagenic up to 5000 µg TOS/plate concentration in the bacterial reverse mutation test. Alternansucrase was determined to be non-clastogenic and non-aneugenic up to the test concentration of 1250 µg TOS/mL during the in vitro mammalian cell micronucleus test. Conclusion The lethal dose (LD50) based on this study is greater than 2000 mg/kg body weight, which falls into the category 5 criteria of Globally Harmonized System (GHS). No-Observed-Adverse-Effect-Level (NOAEL) was concluded to be greater than 1000 mg TOS/kg per day. The studies taken together substantiate the safety of alternansucrase enzyme in various food and associated industries. The present study paves a future for safe use of alternansucrase in varied industrial applications.

Evaluation of adaptogenic activity of Neurotip capsules by swimming endurance and chronic stress model

Stress is the disturbed homeostatic condition of the organism, and it is represented by non-specific response of the body to any demand imposed on it. Stress brings various changes in physiological condition of the organism, but various mechanism of the body will counteract to maintain homeostasis. Neurotip capsule is effective Ayurvedic treatment for stress related disorders. Neurotip capsules have unique combination of herbs like Jatamansi, Guduchi, Shankapushpi, Ashwagandha, and Probiotics. These herbs are known to sooth the mind, promote calmness and act as a stress buster. In present research study Neurotip formulation was screened for acute toxicity study and sub chronic oral toxicity study on adult albino mice. Animals were observed for behavioural changes, neurological changes and autonomic profile changes. There was no noticeable toxicity and death observed in animals when treated with highest dose of 5000mg/ kg of the formulation. Neurotip was tested on swimming endurance of mice. The results were compared with standard group treated with Geriforte. Test formulation was evaluated by chronic stress model in rats using Omeprazole as standard drug. Assessment of ulcer induction was done to check adaptogenic potential of Neurotip. Ulcer protection by test formulation was found to be 68.44% showing good antistress effects.

Chronic (52-week) oral toxicity study of herbal tea of Moringa stenopetala and Mentha spicata leaves formulation in Wistar albino rats

Background: Moringa stenopetala leaves have long been used to treat diabetes, hypertension, respiratory problems, and other diseases. The herbal formulation of Moringa stenopetala and Mentha spicata leaves was found to be more effective in lowering high blood pressure and blood sugar levels. Unlike its pharmacological properties, the long-term safety profile of this herbal formulation has not been investigated yet. Thus, this study investigated the long-term (chronic) oral toxicity of herbal tea of M. stenopetala and M. spicata leaves blended in rats. Methods: Wistar albino rats were randomly distributed into four groups (n = 10/sex/group), and then randomly assigned to a control group and three test groups. The control group (G I) received distilled water. The test groups (G II-IV) received 559.36, 1118.72, and 2237.44 mg/kg of herbal tea of M. stenopetala and M. spicata leaves blend respectively, for 360 consecutive days. During the treatment period, in-life parameters (mortality, clinical symptoms, body weight, and food intake) were evaluated. On the 361st day, hematological, serum biochemical, gross morphological, and histological parameters were investigated. Results: Throughout the 360-day treatment period, no herbal tea-related deaths, severe clinical symptoms, loss of body weight, or food intake were seen in any of the treated groups. Bodyweight, food consumption, organ weight, hematological, and serum biochemical findings showed no significant differences between the control and treated groups in both sexes. Macro-pathological and histopathological examinations of the major organs (liver, kidney, heart, pancreas, stomach, and spleen) revealed no herbal tea-related pathologic alterations. Conclusion: The findings indicate that long-term (360-days) oral administration of the herbal tea of M. stenopetala and M. spicata leaves blend is well tolerated by rats. Hence, it would be safe/low toxic up to a dose of 2237.44 mg/kg/day in chronic exposure.

Sub chronic oral toxicity study of Janma Ghunti Honey in Wistar rats

The present study was conducted to evaluate the safety profile of Janma Ghunti Honey in Wistar rats on oral administration for 28 consecutive days. Animals were randomized on the basis of body weight into 6 groups. Three groups received test item (TI) at three different dose levels (3 mL/k g, 6 mL/kg and 12 mL/kg body weight). One group served as high dose satellite reversal group (12 mL/kg). One group each served as the control and satellite control group. Animals were observed for clinical signs of toxicity and mortality at least once daily. Animals from satellite groups were observed for further 14 days without treatment to evaluate delayed occurrence or reversibility of any signs/toxicity. At the end of the study, animals were studied through clinical pathology and necropsy examination. No treatment related mortality was observed in any group. Except in female low dose and female high dose satellite groups, all the treated groups exhibited weight gain and no statistically significant alterations were observed. No TI related toxicity was found on hematological investigation, blood biochemistry parameters and absolute and relative organ weights among treated groups. 'No Observed Adverse Effect Level' of TI in male and female Wistar rats was found to be above 12 mL/kg body weight.

Experimental study on effect of Naga Bhasma (lead calx) on hematological and biochemical parameters

Background: In recent decade, a lot of concerns has been raised regarding heavy metal content in Ayurveda medicines, especially Bhasma (metallic calx). Naga Bhasma (lead calx) is of the metallic preparation primarily indicated in treating chronic ailments such as Kamala (jaundice), Vatarakta (gout), Grdhrasi (sciatica), Kustha (skin diseases), and Prameha (diabetes). As being a compound of lead, its chronic toxicity study was planned to find out the effect of Naga Bhasma on serum biochemical and hematological parameters. Material and method: Naga Bhasma was prepared by Parada (mercury) and Gandhaka media (NBP) and Vasa Swarasa (juice of Adhatoda vasica Linn.) as herbal media (NBH) in seven Puta (incineration cycles). Chronic oral toxicity study was carried out as per Organization for Economic cooperation and Development (OECD) 408 (90-day oral repeated dose toxicity study). Wistar strain albino rats (Rattus norvegicus) were used for the experimentation. The study was conducted at three dosage levels, therapeutically effective dose (TED) of NBP and NBH was 45 mg/kg body weight of rat, TED × 5 and TED × 10. On 91st day, blood was collected by supraorbital puncture with the help of microcapillary tubes under mild ether anesthesia for estimation of serum biochemical and hematological parameters. Observation and results: Among hematological parameters, significant decrease was observed in MCV (Mean corpuscular volume), MCH (Mean corpuscular haemoglobin), and mean corpuscular hemoglobin concentration by NBH-treated group, whereas in biochemical parameters, both test drugs significantly increased BSL (Blood Sugar Level), SGOT (Serum Glutamic-Oxaloacetic Transaminase), and serum ALP (Alkaline Phosphatise) activity. Hematological and biochemical analysis indicates that both test drugs (prepared in seven incineration cycles) were not found completely safe, especially at higher dosage where the drug affected renal and hepatic functions. Conclusion: For the safety purpose, it is suggested that Naga Bhasma prepared in less number of incineration cycles may not be used for therapeutic purpose.

Combinative Approaches of Chemistry, Pharmacology and Toxicology for the Optimal Pharmaceutical Preparation of an Anti-arthritic Chinese Medicine Formulation QFJBT

ABSTRACT ObjectiveQing Fu Juan Bi Tang (QFJBT) is an anti-arthritic Chinese medicine formula consisting of five herbs: Aconiti Lateralis Radix Praeparata (Fu Zi, 附子), Sinomenii Caulis (Qing Feng Teng, 青风藤), Astragali Radix (Huang Qi, 黄芪), Paeoniae Radix Alba (Bai Shao, 白芍) and Moutan Cortex (Mu Dan Pi, 牡丹皮), which have well-established histories of use for treatment of rheumatic and arthritic diseases. We intended to establish the optimized and standardized pharmaceutical procedures and manufacturing processes for the pilot production of QFJBT to develop it as a novel botanical drug product for treatment of rheumatoid arthritis (RA). Methods The combinative approaches of chemical assessment, toxicological and pharmacological evaluation were explored to define the pharmaceutical preparation of QFJBT. Results The optimized and standardized pharmaceutical procedures and manufacturing processes for the pilot production of QFJBT were established in terms of greatest chemical contents of bioactive constituents, potent anti-inflammatory and antinociceptive activities, and favorable safety profile. Quality analysis of the pilot product of QFJBT by high-performance liquid chromatography (HPLC) demonstrated that the chromatographic fingerprint profiles of three batches of QFJBT were basically identical and the contents of four characteristic and bioactive markers were relatively consistent. General toxicological studies showed a favorable safety profile of QFJBT. The maximum tolerated single dose of QFJBT was determined in both sexes of rats to be 33.63 g/kg body weight which is equivalent to 346 times of clinical dose. In the chronic oral toxicity study, the results of laboratory investigation showed that QFJBT at doses of 3.89, 6.80 and 9.72 g/kg body weight (equivalent to 40, 70 and 100-fold clinical doses, respectively) caused no changes in all hematological parameters and blood biochemical parameters of rats. No mortality or specific toxic responses were observed in animals after three months of repeated dosing with QFJBT. Conclusion The optimized and standardized pharmaceutical and manufacturing processes for the production of QFJBT have been successfully screened and identified through established rigorous in-process controls.

Safety assessment of vitacoxib: 180-day chronic oral toxicity studies

Vitacoxib, a selective COX-2 inhibitor, is approved for the relief of pain and inflammation associated with orthopedic surgery and osteoarthritis in dogs. In the current study, a chronic toxicity research was performed to evaluate the safety of vitacoxib in male and female rats for long-term. Vitacoxib was dosed orally to groups of rats for 180 days at 1.2, 6, 30 mg/kg bw/day by gavage. The chronic study oral administration of vitacoxib did not show observational or toxicological effects on the body or organ weights, food consumption, hematology and biochemistry at dose 6 mg/kg bw. However, vitacoxib (30 mg/kg) showed minor alterations to histopathology of liver, kidney and stomach related to treatment. These results provide further indication that vitacoxib is safe and well-tolerated in rats after 180 days of daily oral administration at 6 mg/kg bw and the NOAEL for both sexes was 6 mg/kg bw for 180 consecutive days.

Evaluation of the acute and chronic toxicity of the jiangu capsules.

Saussureae involucratae Flos is the primary component of the jiangu capsule, which also consists of 12 types of natural medicine. It is a traditional Chinese medicine used to treat a number of conditions including arthralgia, cold and sore muscles, stiff limbs, inconvenient flexion and activity limitation caused by different types of osteoarthritis. The present study was performed to assess the safety of the jiangu capsule following acute and chronic administration in mice and rats. In the acute study, mice were treated with the jiangu capsule orally at 16 g/kg/day, the highest dose, for 14 days. In the chronic oral toxicity study, rats were treated with 2, 4 and 8 g/kg/day of the jiangu capsule for 6 months. The general condition, body weight changes, food and water consumption, hematological and biochemical parameters, urinalysis, systematic anatomy and organ weights of the rats were monitored and histopathological examination was performed at the end of 3 and 6 months' daily administration, and following 14 days of withdrawal. There were no signs of toxicity in the general condition and body weight of the mice was not significantly affected by the treatment. Furthermore, necropsy findings for the animals in the acute and chronic toxicity studies demonstrated that no mortalities had occurred. The results of the current study indicate that treatment with the jiangu capsule did not induce mortality or any detectable abnormalities in mice and rats.

Acute and sub chronic oral toxicity study of Antangin Fit in rats and its immunostimulatory activity

Acute and sub chronic oral toxicity study of Antangin Fit in rats and its immunostimulatory activity

Toxicity studies on Harpagophytum procumbens (Devil's claw) capsules in mice

Acute (24 h) and chronic (90 days) oral toxicity studies on Harpagohytum procumbens (Devil’s claw) capsules manufactured by Boiron (France), were carried out. Male and female mice were used as experimental model. Acute dosages were: 0.5, 1.0 and 3 g/kg while chronic dosage was 100 mg/kg per day of the capsule contents. All morphological, biochemical, haematological and spermatogenic changes, in addition to mortality, body weight changes and any change in vital organs were recorded. Histopathological investigations were done on vital organs. During acute toxicity experiment, mice treated with higher dose were found to have reduced locomotor activity as compared to the control animals. Biochemical studies showed reduction in blood glucose level of mice in the treatment groups as compared to the control. During chronic toxicity studies, both male and female mice in the treatment groups gained statistically significant weight which was similar and comparable to respective control groups. The water intake increased in the treatment as well as the control groups. One male animal was found to develop forelimb inflammation and snout alopecia during chronic toxicity studies. All other animals were normal and comparable to the control animals. There was no mortality of statistical significance observed in any group. Biochemical studies revealed a significant decrease in blood sugar levels and uric acid level of Devil’s claw treatment groups. A slight increase in the aspartate aminotransferase (AST) levels was noticed in the treatment groups as compared to the control groups. However, hematological parameters remained comparable to the control. At the end of the treatment, the visceral condition and the vital organs of animals were found to be normal and comparable to the control. The results were substantiated by histopathological studies. The male treatment group was subjected to sperm abnormality test to assess any mutagenic potential of Devil’s claw prolonged treatment. Devil’s claw capsules treatment group, after chronic treatment, showed no spermatotoxic effect. All the observations recorded revealed that Devil’s claw treatment in the given dose levels, poses low toxicity. The findings of present study might be used in designing future preliminary and preclinical studies on Devil claws capsules.

Safety Assessment of a Hydroethanolic Extract of Caralluma fimbriata

This toxicological assessment evaluated the safety of a hydroethanolic extract prepared from Caralluma fimbriata (CFE), a dietary supplement marketed worldwide as an appetite suppressant. Studies included 2 in vitro genotoxicity assays, a repeated dose oral toxicity study, and a developmental study in rats. No evidence of in vitro mutagenicity or clastogenicity surfaced in the in vitro studies at concentrations up to 5000 μg of extract/plate (Ames test) or 5000 μg of extract/mL (chromosomal aberration test). No deaths or treatment-related toxicity were seen in the 6-month chronic oral toxicity study in Sprague-Dawley rats conducted at 3 doses (100, 300, and 1000 mg/kg body weight (bw)/d). The no observed effect level for CFE in this study was considered to be 1000 mg/kg bw/d. A prenatal developmental toxicity study conducted at 3 doses (250, 500, and 1000 mg/kg bw/d) in female Sprague-Dawley rats resulted in no treatment-related external, visceral, or skeletal fetal abnormalities, and no treatment-related maternal or pregnancy alterations were seen at and up to the maximum dose tested. CFE was not associated with any toxicity or adverse events.

Acute and 30-day oral toxicity studies of administered carnosic acid

PurposeIncreasing interest in carnosic acid (CA) is due to its pharmacological properties. The aim of this study was to evaluate the acute and 30-day oral toxicity of CA. MethodsThe acute oral toxicity study in Kuming mice design followed the OECD-guidelines 423, and a 30-day chronic oral toxicity study in Wistar rats based on the enhanced OECD test guideline 407 were performed. ResultsThe oral lethal dose (LD50) for mice was 7100mg/kg of body weight in the acute toxicity study. The histopathological changes were observed in the heart, liver and kidney for the survival mice treated with a single dose CA. For the sub chronic toxicity study, CA administered for 30days produced slightly reductions in the weight gain pattern, which did not reach the significant level when compared with the control values. With respect to serum biochemistry test, decreased total serum protein levels, but conversely increased aspartate aminotransferase (AST) levels were detected in the high-dose and moderate-dose groups. Histopathologically, light pathological changes were observed in the heart, liver, and kidney of rats treated with the high-dose CA. ConclusionThe present work suggests that a short-term oral administration of CA has a relatively low toxicity profile.

Safety Evaluation of a Proprietary Food-Grade, Dried Fermentate Preparation of Saccharomyces cerevisiae

A safety evaluation was performed for EpiCor, a product produced by a proprietary fermentation process using Saccharomyces cerevisiae. Studies included the following assays: bacterial reverse mutation, mouse lymphoma cell mutagenicity, mitogenicity assay in human peripheral lymphocytes, and a cytochrome P450 ([CYP] CYP1A2 and CYP3A4) induction assessment as well as 14-day acute, 90-day subchronic, and 1-year chronic oral toxicity studies in rats. No evidence of genotoxicity or mitogenicity was seen in any of the in vitro or in vivo studies. The CYP assessment showed no interactions or inductions. No toxic clinical symptoms or histopathological lesions were observed in the acute, subchronic, or chronic oral toxicity studies in the rat. Results of the studies performed indicate that EpiCor does not possess genotoxic activity and has a low order of toxicity that is well tolerated when administered orally. The no observable adverse effect level (NOAEL) was 1500 mg/kg body weight (bw)/d for the 90-day study and 800 mg/kg bw/d for the 1 year study, for the highest doses tested.

Oral toxicological studies of black soybean (Glycine max) hull extract: Acute studies in rats and mice, and chronic studies in mice

Black soybean (Glycine max) has been used for traditional medicine and food in Asian countries, but safety of its hull has not been studied. We conducted acute and chronic oral toxicity studies. For the acute study, an extract of black soybean hull (BE; 2.5g/kg body weight) was administered singly by intragastric intubation to Sprague–Dawley rats and C57BL/6 mice. There was no death or significant decrease in body weight in rats and mice, and the oral LD50 of BE was >2.5g/kg body weight. In the chronic study, BE was administered at dietary levels of 0% (control), 2.0%, and 5.0% to male and female C57BL/6 mice for 26weeks. No mortality or toxicologically significant clinical changes were observed through the experimental period. Although body weights, as well as abdominal fat, blood levels of triglyceride and total cholesterol in 5.0% males were significantly lower than that in control and 2.0% groups, these changes were considered not to be adverse. Hematology and histopathological observation revealed no toxicologically significant changes. The no-observed adverse-effect-level of BE was estimated to be 5.0% in the diet (5074.1mg/kg body weight/day for males and 7617.9mg/kg body weight/day for females).

Safety assessment of AGPC as a food ingredient

α-Glycerylphosphorylcholine (AGPC) is a semi-synthetic derivative of lecithin. Following oral administration, it is converted to phosphatidylcholine, a metabolically active form of choline that is able to reach cholinergic synaptic endings where it increases acetylcholine synthesis and release. A series of studies were conducted to demonstrate the safety of AGPC. The oral LD50 was equal to or greater than 10,000mg/kg in rats and mice. Deaths were preceded by convulsions in some animals. Dosing of dogs with up to 3000mg/kg AGPC resulted only in reduced activity. Sub-chronic and chronic oral toxicity studies in rats (up to 1000mg/kg/day) and beagles (up to 300mg/kg/day) produced symptomology primarily consisting of reduced activity; slight decreases in food consumption and body weight gain; and slight reduction in liver weight, paralleled by significant decreases in plasma triglycerides, bilirubin, and alkaline phosphatase. There were no histopathological correlates. The in vivo and in vitro assays clearly indicated that AGPC was devoid of mutagenic activity. Based on these results, AGPC is not genotoxic in vitro or in vivo, exhibits low acute oral toxicity and, has an oral NOAEL of 150mg/kgbw/day following 26weeks oral exposure.

One-year chronic oral toxicity with combined reproduction toxicity study of a novel probiotic, Bacillus coagulans, as a food ingredient

Some strains of Bacillus coagulans can survive extremes of heat, stomach acid and bile acids, to which commonly consumed probiotics are susceptible. A toxicological safety assessment was published in 2009 on a proprietary preparation of B. coagulans – GanedenBC30™ – a novel probiotic. It was concluded that GanedenBC30™ is safe for chronic human consumption based upon scientific procedures, supported by a safe history of use (Endres et al., 2009).A one-year chronic oral toxicity study combined with a one-generation reproduction study was conducted to further investigate safety of long-term consumption. The one-year study of GanedenBC30™ administered to male and female HsdBrlHan: Wistar rats in their diet showed no signs of toxicity at the highest dose tested. The conclusion from the reproduction toxicity study is that administration of GanedenBC30™ in the diet caused no signs of toxicity in the parental generation (male or female) nor the F1 offspring. Using the lowest NOEL of 1948mg/kg concluded at the end of the 1-year feeding study, a 100-fold safety factor, a test article concentration of 6.88×1010CFU (colony forming units) per gram, and an average 70kg human, it is determined that GanedenBC30™ is safe for chronic consumption at up to 9.38×1010CFUs per day.