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Human liver s9 fractions

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Human liver S9 fractions are subcellular liver preparations containing a mix of metabolic enzymes and cofactors. They are used for in vitro studies of drug metabolism and toxicity.

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Lab products found in correlation

Sulfatase, by Merck Group (2 mentions) Sodium phosphate monobasic monohydrate, by Merck Group (2 mentions) Glucose 6 phosphate dehydrogenase, by Merck Group (2 mentions) D glucose 6 phosphate disodium salt hydrate, by Merck Group (2 mentions) Mgcl2, by Merck Group (2 mentions) β glucuronidase, by Merck Group (2 mentions) Sodium phosphate dibasic, by Merck Group (2 mentions) Acetic acid, by Merck Group (2 mentions) β nicotinamide adenine dinucleotide phosphate sodium salt hydrate, by Merck Group (1 mentions)

3 protocols using human liver s9 fractions

1

Prodrug Metabolism Screening Using Human Liver S9

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Example 79

Method H: In Vitro Metabolic Conversion Assay Using Human Liver S9 Fractions

A representative sampling of prodrug compounds was incubated with Human Liver S9 Fractions (Life Technologies Corporation) at 10 μM in duplicates. Incubations were carried out at 37° C. in a shaker water bath. Samples were taken at 0 and 60 minutes. Negative controls (without S9 fractions) and positive controls (7-ethoxycoumarin and 7-hydroxycoumarin) were run concurrently with the test articles in duplicates. All samples were analyzed using LC/MS/MS; depletion of the parent compounds and the formation of gemcitabine were monitored, though the latter was not quantitatively monitored. The test results are tabulated in Table 1.

TABLE 1
% of ParentFormation of
Test SampleCompoundRemainingGemcitabine
131%✓
268%✓
3100%✓
4110%✓
51313% ✓
6140%✓
7580%✓
8591%✓
9622%✓

After incubation with Human Liver S9 Fractions for 60 minutes, depletion of each test compound was observed along with formation of gemcitabine as a metabolite.

US09890189B2. Gemcitabine prodrugs and uses thereof (2018-02-13). BoYen Therapeutics, Inc. [TW]. Inventors: Laurence I. Wu [TW].
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2

In vitro metabolism of reactive oxygen species

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The chemicals and reagents used in our experiment were obtained from the following sources. PIO (purity = 97%) standard was obtained from Toronto Research Chemicals (North York, Ontario, Canada). ROS labeled with deuterium (on the benzene ring; ROS-D4, purity = 96%) standard was obtained from BDG Synthesis Limited (Wellington, New Zealand). Human liver S9 fractions (20 mg/mL protein base) were obtained from Thermo Fisher Scientific (Runcorn, United Kingdom). ROS (purity ≥98%) standard, glucose-6-phosphate dehydrogenase (activity: 225 units/mg), MgCl2 (purity ≥98%), sodium phosphate monobasic monohydrate (purity ≥98%), sulfatase (activity: 11 units/mL), D-glucose 6-phosphate disodium salt hydrate (purity ≥98%), β-glucuronidase (activity >85,000 units/mL), β-nicotinamide adenine dinucleotide phosphate sodium salt hydrate (NADP, purity ≥98%), acetic acid (purity ≥99%), and sodium phosphate dibasic (purity ≥99%) were obtained from Sigma-Aldrich (St. Louis, MO, United States).
Lin I.S., Chuang C.Y, & Shih C.L. (2023). Dose-response technique combined with stable isotope tracing for drug metabolite profiling by using high-resolution mass spectrometry. Frontiers in Pharmacology, 14, 1293540.
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3

Assay for Glucose-6-Phosphate Dehydrogenase Activity

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Glucose-6-phosphate dehydrogenase (activity: 225 units/mg), MgCl2 (purity: ≥98%), d-glucose 6-phosphate disodium salt hydrate (purity: ≥98%), sulfatase (activity: 11 units/mL), β-glucuronidase (activity: >85000 units/mL), sodium phosphate monobasic monohydrate (purity: ≥98%), β-nicotinamide adenine dinucleotide phosphate sodium salt hydrate (NADP, purity: ≥98%), sodium phosphate dibasic (purity: ≥99%), and acetic acid (purity: ≥99%) were obtained from Sigma–Aldrich (St. Louis, MO, USA). PIO (purity = 97%), deuterium-labeled rosiglitazone (purity = 98%), and deuterium-labeled (on the benzene ring) PIO (D4-PIO, purity = 97%) were obtained from Toronto Research Chemicals (Toronto, Canada). Human liver S9 fractions (20 mg/mL protein base) were obtained from Thermo Fisher Scientific (Runcorn, UK).
Ting H.H., Chiou Y.S., Chang T.Y., Lin G.Y., Li P.J, & Shih C.L. (2023). Development of a metabolomics-based data analysis approach for identifying drug metabolites based on high-resolution mass spectrometry. Journal of Food and Drug Analysis, 31(1), 152-164.
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