6-hydroxy Chlorzoxazone (Synonyms: NSC 24955) |
Catalog No.GC42580
One-Click Copy Product Info
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6-hydroxy Chlorzoxazone is the primary metabolite of Chlorzoxazone in vivo. 6-hydroxy Chlorzoxazone is mainly formed through the hepatic cytochrome P450 enzyme system and is commonly used as a probe in biochemical research to quantitatively evaluate CYP2E1 activity.
Products are for research use only. Not for human use. We do not sell to patients.
Cas No.: 1750-45-4
Sample solution is provided at 25 µL, 10mM.
6-hydroxy Chlorzoxazone is the primary metabolite of Chlorzoxazone in vivo. 6-hydroxy Chlorzoxazone is mainly formed through the hepatic cytochrome P450 enzyme system and is commonly used as a probe in biochemical research to quantitatively evaluate CYP2E1 activity. 6-hydroxy Chlorzoxazone is also involved in studies of hepatic xenobiotic metabolism, pharmacokinetics, and toxicology to elucidate metabolic pathways and drug interaction mechanisms[1-2].
References:
[1] Dreisbach AW, Ferencz N, Hopkins NE, et al. Urinary excretion of 6-hydroxychlorzoxazone as an index of CYP2E1 activity. Clin Pharmacol Ther. 1995 Nov;58(5):498-505.
[2] O'Shea D, Davis SN, Kim RB, et al. Effect of fasting and obesity in humans on the 6-hydroxylation of chlorzoxazone: a putative probe of CYP2E1 activity. Clin Pharmacol Ther. 1994 Oct;56(4):359-67.
Determination of CYP2E1 Enzyme Activity Using 6-hydroxy Chlorzoxazone
This protocol is adapted from research data and provided for reference only. Adjustments may be necessary based on specific experimental requirements.
1. Reagent and Material Preparation
(1) Standard stock solution: Dissolve 6-hydroxy Chlorzoxazone in methanol to prepare a 1mg/mL master stock solution, and store at -20°C.
(2) Incubation system: Contains 0.25mg/mL human liver microsomal protein, 50μM Chlorzoxazone, 1.15% KCl, 0.2M phosphate buffer (pH=7.4), and 5mM NADPH.
(3) Stop solution: Water/acetonitrile/formic acid (42:55:3, v/v), containing the internal standard Chlorpropamide.
2. Chromatographic and Mass Spectrometry Conditions
(1) Chromatographic column: Waters Acquity UPLC BEH C18 (50mm × 2.1mm, 1.7μm) with a VanGuard pre-column (5mm × 2.1mm, 1.7μm).
(2) Mobile phase: Phase A is 0.1% formic acid in water, Phase B is 0.1% formic acid in acetonitrile; gradient elution (5% B → 80% B, 4min), flow rate 0.4mL/min, column temperature 35°C.
3. Incubation and Sample Processing: Take 50μL of microsomal suspension and mix with 50μL of Chlorzoxazone. Add buffer and KCl, and pre-incubate for 3 minutes (37°C). Initiate the reaction by adding NADPH, incubate for 15 minutes, then add 25μL of stop solution to terminate the reaction. After cooling on ice, centrifuge (20,000×g, 10min, 4°C), and collect the supernatant for injection analysis.
Precautions:
(1) The total analysis time of this method is 5.25 minutes, making it suitable for high-throughput detection. 6-hydroxy Chlorzoxazone must be detected in negative ion mode and analyzed in two separate injections along with other metabolites.
(2) For your safety and health, please wear a lab coat and disposable gloves during operation.
References:
[1] De Bock L, Boussery K, Colin P, et al. Development and validation of a fast and sensitive UPLC-MS/MS method for the quantification of six probe metabolites for the in vitro determination of cytochrome P450 activity. Talanta. 2012 Jan 30;89:209-16.
| Cas No. | 1750-45-4 | SDF | |
| Synonyms | NSC 24955 | ||
| Chemical Name | 5-chloro-6-hydroxy-2(3H)-benzoxazolone | ||
| Canonical SMILES | O=c1[nH]c2cc(Cl)c(O)cc2o1 | ||
| Formula | C7H4ClNO3 | M.Wt | 185.6 |
| Solubility | Slightly soluble in DMSO, Slightly soluble in Methanol | Storage | Store at -20°C |
| General tips | Please select the appropriate solvent to prepare the stock solution according to the
solubility of the product in different solvents; once the solution is prepared, please store it in
separate packages to avoid product failure caused by repeated freezing and thawing.Storage method
and period of the stock solution: When stored at -80°C, please use it within 6 months; when stored
at -20°C, please use it within 1 month. To increase solubility, heat the tube to 37°C and then oscillate in an ultrasonic bath for some time. |
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| Shipping Condition | Evaluation sample solution: shipped with blue ice. All other sizes available: with RT, or with Blue Ice upon request. | ||
| Prepare stock solution | |||
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1 mg | 5 mg | 10 mg |
| 1 mM | 5.3879 mL | 26.9397 mL | 53.8793 mL |
| 5 mM | 1.0776 mL | 5.3879 mL | 10.7759 mL |
| 10 mM | 538.8 μL | 2.694 mL | 5.3879 mL |
Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)
g
μL
Step 2: Enter the in vivo formulation (This is only the calculator, not formulation. Please contact us first if there is no in vivo formulation at the solubility Section.)
Calculation results:
Working concentration: mg/ml;
Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid concentration mg/mL, Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug. )
Method for preparing in vivo formulation: Take μL DMSO master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL saline, mix and clarify.
Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.
Note: 1. Please make sure the liquid is clear before adding the next solvent.
2. Be sure to add the solvent(s) in order. You must ensure that the solution obtained, in the previous addition, is a clear solution before proceeding to add the next solvent. Physical methods such as vortex, ultrasound or hot water bath can be used to aid dissolving.
3. All of the above co-solvents are available for purchase on the GlpBio website.
Quality Control & SDS
- View current batch:
- Purity: >97.00% Appearance: A solid
- COA (Certificate of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
Average Rating: 5 (Based on Reviews and 28 reference(s) in Google Scholar.)