Absorption, Distribution and Excretion
/ Milk / Non-nutritive sweeteners (NNS), including saccharin, sucralose, aspartame, and acesulfame potassium, are commonly consumed in the general population. Except for saccharin, all other sweeteners are considered safe for use during pregnancy and lactation. Sucralose (Splenda) currently holds a large share of the NNS market and is frequently used in combination with acesulfame potassium in various foods and beverages. To date, saccharin has been reported as the only NNS detected in breast milk after maternal consumption, while information on other NNSs remains unclear. This study collected breast milk samples from 20 lactating volunteers whose NNS intake was not related to their daily intake. Saccharin, sucralose, and acesulfame potassium were detected in the breast milk samples of 65% of the participants, while aspartame was not detected. These data suggest that lactating infants frequently ingest non-nutritive sweeteners (NNS), thus requiring prospective clinical studies to determine the clinical significance of early NNS exposure through breast milk. /Acesulfame potassium/
In rats and dogs, a single oral dose of 10 mg (14)C-acesulfame potassium/kg body weight resulted in rapid absorption. Peak plasma concentrations of 0.75 ± 0.2 g/mL were observed in rats 0.5 hours after administration, and in dogs 6.56 ± 2.08 g/mL 1–1.5 hours after administration. 82–100% of the dose was excreted in rat urine and 85–100% in dog urine; 97–100% of the total radioactivity was excreted in the feces of both animals, with a total recovery rate close to 100%. No accumulation was observed in rats after 10 mg/kg daily for 10 consecutive days. Three days after administration, the drug concentration in the liver was 0.4 nmol/g, and in other tissues <0.2 nmol/g. Seven days after administration, the drug concentration in all tested tissues in dogs was <0.2 nmol/g. /Acesulfame K/
Male rats were pre-fed a diet containing 3% acesulfame K for 7 days, and then administered 250 mg of acesulfame K by gavage, which contained 14C-acesulfame K (9.6 x 10⁸ dpm). Animals were sacrificed 8 hours later, and livers and spleens were removed; DNA and chromatin proteins were isolated from these organs. No radioactivity was detected in any DNA samples. Low levels of radioactivity (8–11 dpm/mg protein) were present in the chromatin proteins, which were believed to be due to non-covalent interactions with the unlabeled acesulfame K. /Acesulfame K/
Male and female rats pre-fed unlabeled acesulfame K at a dose of 300 mg/kg feed for 60 days were administered a single oral dose of approximately 15 mg (14)C-acesulfame K/kg body weight. Control group animals that did not undergo pretreatment were also fed the same dose of (14)C-acesulfame K. 95.1–98.2% of the dose was recovered from urine and cage flushing fluid in all animals, and 0.95–2.86% from feces. The overall recovery rate was 96.3–99.2%. Radioactive material was rapidly excreted, exhibiting a biphasic kinetic profile; 92.6–96.8% of the dose was excreted within 24 hours. …No significant differences were observed between sexes or between the control group and animals pretreated with acesulfame potassium for 60 days in terms of excretion pathway or rate. /Acesulfame Potassium/
For more complete data on the absorption, distribution, and excretion of acesulfame potassium (8 types), please visit the HSDB records page.

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Metabolism/Metabolites
This study examined the metabolism of acesulfame potassium in the urine and feces of rats and dogs that received a single oral dose of 10 mg/kg body weight, and in the urine and bile of pigs that received an oral dose of 5 mg/kg body weight. The analytical methods used (thin-layer chromatography, mass spectrometry, and isotope dilution) detected only the original substance in these samples. In the rat urine extract used in the above studies, only one peak was found after thin-layer chromatography separation, which was identical to that of acesulfame potassium. No metabolites were detected in either the control group or the acesulfame potassium pretreatment group. Similarly, no metabolites were detected in animals pretreated with 1% acesulfame potassium for 7 days. /Acesulfame Potassium/
The metabolism of acesulfame potassium in serum and urine was studied after a single administration of 30 mg/person to human volunteers. Only the original drug was detected in all samples. /Acesulfame potassium/

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Biological half-life
/After/ Male and female rats were fed a single oral dose of approximately 15 mg (14)C-acesulfame potassium/kg body weight to unlabeled acesulfame potassium for 60 days prior at a feed level of 300 mg/kg; the half-life of the rapid phase was 4–4.5 hours, and the half-life of the slow phase (<0.5% of the dose) was 109–257 hours. /Acesulfame potassium/
After a single intravenous injection of 10 mg (14C)-acesulfame potassium/kg body weight in rats, the radioactive material was excreted quantitatively in the urine, with a plasma half-life of 0.23 hours. /Acesulfame potassium/
After a single oral dose of approximately 10.6 mg/kg body weight of (14)-acesulfame potassium in lactating rats, the biological half-life in milk (5.6 hours) and blood (4 hours) was similar. /Acesulfame K/
The purpose of this study was to determine the levels of acesulfame K (AcK) in the plasma and urine of C57BL mice. Male and female animals were administered the drug orally (by gavage) and intravenously at a dose of 10 mg/kg, respectively, and plasma and urine samples were collected within 24 hours post-administration. Plasma samples were analyzed using high-performance liquid chromatography (HPLC)...urine samples were analyzed using a different HPLC method...saccharin was used as an internal standard in both methods, and UV absorption was detected at 230 nm. Following intravenous administration, plasma acetylcholine (AcK) concentrations decreased rapidly and linearly within 120 minutes, with a second AcK peak at 240 minutes. The half-life was estimated to be 11–15 minutes. Following oral administration, plasma AcK concentrations peaked within 45 minutes and decreased rapidly. Plasma AcK concentrations were below the detection limit at 480 minutes post-administration. A second peak was also observed after oral administration, suggesting enterohepatic circulation. Following intravenous and oral administration, 45% (males) and 70% (females) of the dose are excreted in the urine within 24 hours. Based on urinary data, the oral bioavailability is estimated to be 90-100%.

Toxicity Summary
Identification and Uses: Acesulfame potassium (AS) is a solid. It is used as an artificial sweetener in food and also in cosmetics. Human Studies: A case study has been reported in which an individual with a known allergy to sulfites and sulfonamides developed a hypersensitivity reaction after ingesting taurine and the non-nutritive sweetener acesulfame potassium at levels exceeding the threshold, compounds that typically do not cause allergic reactions. In vitro studies suggest that long-term intake of acesulfame potassium may accelerate atherosclerosis and aging by impairing the function and structure of apolipoprotein AI (apoA-I) and high-density lipoprotein (HDL). Animal Studies: Acesulfame potassium was non-irritating in a primary skin irritation test in rabbits. Acesulfame potassium did not show an antigenic effect; only guinea pigs sensitized with bovine serum albumin (BSA) showed allergic reactions. Acesulfame potassium was not carcinogenic in mice and rats. A multigenerational rat study fed male and female rats (two litters per generation) diets supplemented with 0%, 0.3%, 1.0%, and 3.0% acesulfame potassium, respectively, for three consecutive generations. Slightly decreased growth rates were observed in the highest-dose groups of the F0 and F1 generations, and in the medium-dose group of the F0 generation. Teratogenicity studies showed no adverse effects on appearance, food consumption, maternal necropsy, organ weight, or litter size; no visceral or skeletal abnormalities were observed associated with this treatment. Acetylsulfame potassium was negative in both in vivo and in vitro genotoxicity studies, including Ames assays in Salmonella TA98, TA100, TA15325, TA1537, and TA1538 strains at concentrations of 0–100 mg/plate and 4–5000 μg/plate, respectively, and in Escherichia coli WP2uvrA strain at concentrations of 4–5000 μg/plate. Ecotoxicity Studies: Acetylsulfamate is listed as an emerging pollutant due to its environmental persistence and widespread presence in the environment. Increased toxicity of acesulfamate was observed in the livers of goldfish exposed to UV radiation. Embryotoxicity studies showed that low concentrations (g/L) of acesulfame potassium metabolites had significant adverse effects on tail detachment, heart rate, hatching rate, and survival rate during fish embryonic development.

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Impact During Pregnancy and Lactation
◉ Overview of Drug Use During Lactation
Various concentrations of acesulfame potassium were detected in the breast milk of lactating mothers who consumed artificially sweetened beverages and sweetener packets within the past 24 hours. Even some mothers who did not consume artificial sweeteners had trace amounts of acesulfame potassium in their breast milk. However, their intake is unlikely to exceed the infant's daily acceptable intake. Consuming sugar-free beverages containing low-calorie sweeteners may increase the risk of vomiting in breastfed infants. Some authors suggest that breastfeeding women should limit their intake of non-nutritive sweeteners because their effects on breastfed infants are unclear.
◉ Effects on Breastfed Infants
A cross-sectional survey assessed the dietary history of US mothers between 11 and 15 weeks after their infants' birth. The survey was used to estimate the amount of sugar-free soft drinks and fruit juices consumed by these mothers. Results showed no statistically significant difference between infant weight or z-score and the intake of low-calorie sweeteners. However, infants who consumed low-calorie sweeteners once a week or less had a significantly higher risk of vomiting than infants who did not consume such sweeteners. Increased intake was not associated with vomiting. The effects of specific sweeteners could not be assessed.
◉ Effects on Lactation and Breast Milk
No relevant published information was found as of the revision date.

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Interactions
The ecotoxicity of heavy metals largely depends on their speciation, which is influenced by other coexisting substances with chelating capabilities. This study investigated the toxic effects of Cd(2+) and Cu(2+) on Scenedesmus obliquus in the presence of two artificial sweeteners—acesulfame K (ACE) and sucralose (SUC)—by comparing the specific cell growth rate μ and pulse amplitude modulation (PAM) parameters (maximum photosystem II photochemical efficiency Fv/Fm, actual photochemical efficiency Yield, and non-photochemical quenching NPQ) over 96 hours. Simultaneously, the bioaccumulation of metals in algal cells in the presence of activated sludge (ASs) was determined. The presence of ACE promoted the growth of Scenedesmus obliquus and increased its bioaccumulation of Cd²⁺, while the effect of SUC was not significant. Furthermore, the EC50 values of Cd²⁺ on the growth of Scenedesmus obliquus increased from 0.42 mg/L to 0.54 mg/L and 0.48 mg/L, respectively, after the addition of 1.0 mg/L ACE and SUC. For Cu²⁺, the EC50 values increased from 0.13 mg/L to 0.17 mg/L and 0.15 mg/L, respectively, after the addition of 1.0 mg/L ACE and SUC. In conclusion, both activated sludge types can reduce the toxicity of metals to algae, with ACE being more effective than SUC. Although PAM parameters are less sensitive than cell-specific growth rates, they can reveal the mechanisms of metal toxicity at the subcellular level. This study provides the first evidence of the potential impact of aspartame on the ecotoxicity of heavy metals. Swiss albino male mice were exposed by gavage to a mixture of aspartame (3.5, 35, 350 mg/kg body weight) and acesulfame potassium (1.5, 15, 150 mg/kg body weight). Bone marrow cells were isolated from the femur, and chromosomal aberrations were analyzed. Statistical analysis showed that the combination of aspartame and acesulfame potassium did not have significant genotoxicity. /Acesulfame potassium/
/Authors/...The ability of zinc sulfate (5, 25, 50 mM) to inhibit the sweetness of 12 chemically distinct sweeteners, all of which had an intensity matching that of 300 mM sucrose [800 mM glucose, 475 mM fructose, 3.25 mM aspartame, 3.5 mM saccharin, 12 mM sodium cyclohexylsulfamate, 14 mM acesulfame potassium, 1.04 M sorbitol, 0.629 mM sucralose, 0.375 mM neohesperidin dihydrochalcone (NHDC), 1.5 mM steviol glycosides, and 0.0163 mM thomatose]. Zinc sulfate inhibited the sweetness of most compounds in a concentration-dependent manner, with a peak inhibition rate of 80% at a concentration of 50 mM. Interestingly, zinc sulfate never inhibited the sweetness of sodium cyclohexylsulfamate. This suggests that sodium cyclohexylsulfamate may act on a different sweetness mechanism than other sweeteners (except for saccharin), which are inhibited at all concentrations of zinc sulfate. We hypothesize that this group of compounds may act on a single receptor or multiple receptors that are equally inhibited by zinc sulfate at all concentrations. This study aimed to determine the effect of repeated presentation of the same sweet stimulus on sweetness intensity ratings. The sweet stimuli tested in this study were binary and ternary mixtures of 14 sweeteners with significantly different chemical structures. A trained sensory evaluation team tasted each given mixture in four bites, 30 seconds apart, and assessed its sweetness intensity. The intensity of each component in binary sweetener combinations was anchored to 5% sucrose, while the intensity of each sweetener in ternary mixtures was anchored to 3% sucrose… Each submixture was also evaluated (e.g., acesulfame potassium-acesulfame potassium). The main finding of this study is that mixtures of two or three different sweeteners, after four repeated sippings, exhibited a smaller decrease in sweetness intensity than single sweeteners of the same sweetness level. Furthermore, ternary mixtures were often slightly more effective than binary mixtures in mitigating the effects of repeated exposure to specific sweet stimuli. These findings suggest that mixed sweeteners can reduce the decrease in sweetness intensity after repeated exposure to sweet stimuli. Human bitterness is mediated by G protein-coupled receptors of the hTAS2R family. Using a high-throughput screening method, this study discovered a novel bitterness receptor antagonist (GIV3727) that inhibits the activation of hTAS2R31 (formerly hTAS2R44) by saccharin and acesulfame potassium (two common artificial sweeteners). Pharmacological analysis indicates that GIV3727 is likely an ortho-irreversible antagonist of hTAS2R31. Surprisingly, the study also found that this compound can inhibit five other hTAS2R receptors, including the closely related hTAS2R43. Molecular modeling and site-directed mutagenesis studies have shown that two residues in helix 7 are crucial for the antagonistic activity of hTAS2R31 and hTAS2R43. In human sensory studies, GIV3727 significantly reduced the bitterness of both sulfonamide sweeteners, indicating that the hTAS2R antagonist is active in vivo.

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Non-human toxicity values
Oral LD50 in rats: 7431 mg/kg /acesulfame potassium/
Intraperitoneal LD50 in rats: 2243 mg/kg /acesulfame potassium/

[1]. BioCong WN, et al. Long-term artificial sweetener acesulfame potassium treatment alters neurometabolic functions in C57BL/6J mice. PLoS One. 2013 Aug 7;8(8):e70257.chemical Assay Reagents

Acetylsulfamic acid potassium is an aminosulfonate with the structure 1,2,3-oxathiazine-4(3H)-one-2,2-dioxide, with a methyl group substituted at the 6-position. It is an exogenous substance, an environmental pollutant, and a sweetener. Acetylsulfamic acid potassium belongs to the categories of aminosulfonates, organic nitrogen heterocyclic compounds, oxygen heterocyclic compounds, and organic heteromonocyclic compounds.

C4H5NO4S

163.15

162.993

33665-90-6

Acesulfame potassium;55589-62-3

36573

Needles from benzene or chloroform

1.7±0.1 g/cm3

332.7±25.0 °C at 760 mmHg

123.2 °C
MP: 225 °C /Acesulfame potatssium/
123 - 123.5 °C

155.0±23.2 °C

0.0±1.6 mmHg at 25°C

1.609

-0.31

1

4

0

10

283

0

[K+].CC1=CC(=O)N=S(=O)([O-])O1

YGCFIWIQZPHFLU-UHFFFAOYSA-N

InChI=1S/C4H5NO4S/c1-3-2-4(6)5-10(7,8)9-3/h2H,1H3,(H,5,6)

6-methyl-2,2-dioxooxathiazin-4-one

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples

Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.

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Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)
*Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution.
Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)
Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil)
Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals).

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Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)]
*Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.
Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline)
Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline)
Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline)
Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline
Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil)
Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)

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Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium)
Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose
Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals).

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Oral Formulation 3: Dissolved in PEG400
Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose
Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose
Oral Formulation 6: Mixing with food powders

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Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.

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 (Please use freshly prepared in vivo formulations for optimal results.)