Glucosamine and Methylsulfonylmethane Tablets
DEFINITION
Glucosamine and Methylsulfonylmethane Tablets are prepared from either Glucosamine Hydrochloride, Glucosamine Sulfate Sodium Chloride, Glucosamine Sulfate Potassium Chloride, or a mixture of any of them, with Methylsulfonylmethane. Tablets contain NLT 90.0% and NMT 120.0% of the labeled amount of glucosamine (C6H13NO5) and NLT 90.0% and NMT 110.0% of the labeled amount of methylsulfonylmethane (C2H6O2S).
IDENTIFICATION
• A. Presence of Glucosamine:
The retention times of the major peaks of the Sample solution correspond to those of the Standard solution, as obtained in the Content of Glucosamine.
• B. Presence of Methylsulfonylmethane:
The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Content of Methylsulfonylmethane.
STRENGTH
• Content of Glucosamine
Diluent:
Transfer 29 µL of acetic acid and 5 mL of acetonitrile to a 100-mL volumetric flask containing 50 mL of water, and dilute with water to volume.
Borate buffer:
0.2 M (76.3 g/L of sodium borate in water) adjusted with hydrochloric acid TS to a pH of 9.5. [Note—Buffer must be stored at room temperature. It must be warmed to dissolve if crystallization occurs. ]
Acetate buffer:
6.80 g/L of sodium acetate trihydrate in water adjusted with dilute acetic acid to a pH of 5.9
Derivatizing reagent:
In a 14-mL polypropylene culture tube dissolve 50 mg of o-phthalaldehyde in 1.25 mL of anhydrous methanol. Add 50 µL of 3-mercaptopropionic acid and 11.2 mL of Borate buffer, and mix gently. Allow to stand in the dark for 30 min before use. [Note—Reagent strength is maintained by adding 10 µL of 3-mercaptopropionic acid every 2 days. Storage should be in the dark, at room temperature, and can be used for NMT 2 weeks. ]
Mobile phase:
Methanol and Acetate buffer (1:9)
Standard solution:
1.0 mg/mL of USP Glucosamine Hydrochloride RS in water. Allow to stand at room temperature for 1 h.
Sample solution:
Transfer an equivalent to 25 mg of glucosamine from NLT 20 Tablets, finely powdered, to a 25-mL volumetric flask, and dilute with Diluent to volume. Mix on a vortex mixer to suspend the powder in solution. Sonicate in a 65
water bath for 20 min. Remove from the bath, stir for 5 min with the aid of a magnetic stirrer, and centrifuge.
water bath for 20 min. Remove from the bath, stir for 5 min with the aid of a magnetic stirrer, and centrifuge.
Chromatographic system
Mode:
LC
Detector:
UV 340 nm
Column:
3.0-mm × 5-cm; packing L1
Flow rate:
1 mL/min
Injection size:
10 µL
System suitability
Samples:
Five individual aliquots of the Standard solution derivatized as directed for Analysis. Each derivatized aliquot is injected only once.
[Note—The relative retention times for the 
-anomer and the 
-anomer are 1.0 and 1.8, respectively. The retention time for the -anomer is NLT 4 min. ]
-anomer and the -anomer are 1.0 and 1.8, respectively. The retention time for the -anomer is NLT 4 min. ]
Suitability requirements
Relative standard deviation:
NMT 2.0% for five replicate injections
Analysis
Samples:
Standard solution and Sample solution
Transfer 100 µL of the Derivatizing reagent and 100 µL of the Standard solution or the Sample solution to a vial containing 400 µL of Borate buffer. Allow the derivatization to proceed for 1 min. Inject the derivatized solutions immediately after the derivatization reaction.
Calculate the percentage of the labeled amount of glucosamine (C6H13NO5) in the portion of Tablets taken:
Result = (rU/rS) × (CS/CU) × (Mr1/Mr2) × 100
| rU | = | = peak response of the -anomer from the derivatized Sample solution |
| rS | = | = peak response of the -anomer from the derivatized Standard solution |
| CS | = | = concentration of USP Glucosamine Hydrochloride RS in the Standard solution (mg/mL) |
| CU | = | = nominal concentration of glucosamine in the Sample solution (mg/mL) |
| Mr1 | = | = molecular weight of glucosamine, 179.17 |
| Mr2 | = | = molecular weight of glucosamine hydrochloride, 215.63 |
Acceptance criteria:
90.0%–120.0% of the labeled claim
• Content of Methylsulfonylmethane
Diluent:
Transfer 950 mL of methanol to a 1-L volumetric flask. Add 0.60 mL of diethylene glycol methyl ether, and dilute with methanol to volume.
Standard solution:
0.4 mg/mL of USP Methylsulfonylmethane RS in Diluent. Sonicate at 50 for 1 min, and allow to cool to room temperature.
for 1 min, and allow to cool to room temperature.
Sample solution:
Finely powder NLT 20 Tablets. Dissolve a portion of the finely powdered material, equivalent to 1 Tablet, in Diluent, and sonicate for 15 min at 50. Allow to cool to room temperature, dilute with Diluent to volume, and mix. Quantitatively dilute with Diluent to obtain a final concentration of 0.4 mg/mL of methylsulfonylmethane. Transfer 1 mL of the suspension to a 1.5-mL microcentrifuge tube, and centrifuge for 20 s. Use the supernatant.
. Allow to cool to room temperature, dilute with Diluent to volume, and mix. Quantitatively dilute with Diluent to obtain a final concentration of 0.4 mg/mL of methylsulfonylmethane. Transfer 1 mL of the suspension to a 1.5-mL microcentrifuge tube, and centrifuge for 20 s. Use the supernatant.
Chromatographic system
Mode:
GC
Detector:
Flame ionization
Column:
0.53-mm × 30-m capillary; 5-µm phase G2 coating
Temperature
Column:
120
Injector:
250
Detector:
250
Carrier gas:
Helium
Flow rate:
5 mL/min
Injection size:
1 µL
Injector type:
Split ratio, 2:1
System suitability
Sample:
Standard solution
Suitability requirements
Relative standard deviation:
NMT 2.0% for the peak response ratio of methylsulfonylmethane to diethylene glycol methyl ether from replicate injections
Analysis
Samples:
Standard solution and Sample solution
Calculate the percentage of the labeled amount of methylsulfonylmethane (C2H6O2S) in the portion of Tablets taken:
Result = (RU/RS) × (CS/CU) × 100
| RU | = | = peak response ratio of methylsulfonylmethane to diethylene glycol methyl ether from the Sample solution |
| RS | = | = peak response ratio of methylsulfonylmethane to diethylene glycol methyl ether from the Standard solution |
| CS | = | = concentration of USP Methylsulfonylmethane RS in the Standard solution (mg/mL) |
| CU | = | = nominal concentration of methylsulfonylmethane in the Sample solution (mg/mL) |
Acceptance criteria:
90.0%–110.0% of the label claim
PERFORMANCE TESTS
• Disintegration and Dissolution 
2040
:
Meet the requirements for Dissolution
2040:
Meet the requirements for Dissolution
Medium:
Water; 900 mL
Apparatus 2:
75 rpm
Time:
60 min
Determine the percentage of glucosamine dissolved as follows.
Standard solution:
Prepare as directed in the test for Content of Glucosamine. Dilute with a suitable quantity of water, if necessary.
Sample solution:
Use the solution under test.
Borate buffer, Acetate buffer, Derivatizing reagent, Mobile phase, Chromatographic system, and Analysis:
Proceed as directed in the test for Content of Glucosamine.
Calculate the percentage of the labeled amount of glucosamine (C6H13NO5) dissolved:
Result = (rU/rS) × (CS × V/L) × (Mr1/Mr2) × 100
| rU | = | = peak area obtained from the derivatized Sample solution |
| rS | = | = peak area obtained from the derivatized Standard solution |
| CS | = | = concentration of USP Glucosamine Hydrochloride RS in the Standard solution (mg/mL) |
| V | = | = volume of Medium, 900 mL |
| L | = | = label claim of glucosamine (mg/Tablet) |
| Mr1 | = | = molecular weight of glucosamine, 179.17 |
| Mr2 | = | = molecular weight of glucosamine hydrochloride, 215.63 |
Tolerances:
NLT 75% of the labeled amount of glucosamine (C6H13NO5) is dissolved.
• Weight Variation 2091:
Meet the requirements
2091:
Meet the requirements
ADDITIONAL REQUIREMENTS
• Packaging and Storage:
Preserve in tight, light-resistant containers.
• Labeling:
The label indicates the types of glucosamine salts contained in the article.
• USP Reference Standards 11
11
USP Glucosamine Hydrochloride RS 
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USP Methylsulfonylmethane RS
Dimethyl sulfone.
C2H6O2S 94.13
Dimethyl sulfone.
C2H6O2S 94.13
Auxiliary Information—
Please check for your question in the FAQs before contacting USP.
| Topic/Question | Contact | Expert Committee |
|---|---|---|
| Monograph | Natalia Davydova
Scientific Liaison 1-301-816-8328 |
(DS2010) Monographs - Dietary Supplements |
| Reference Standards | RS Technical Services 1-301-816-8129 rstech@usp.org |
USP35–NF30 Page 1338
Pharmacopeial Forum: Volume No. 32(4) Page 1137