DETERMINATION OF LEVELS OF GLUCOSAMINE HYDROCHLORIDE AND CHONDROITIN SULFATE IN MIXTURES IN TABLET AND CREAM FORMS USING HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY WITH FLUORESCENCE

  • Harmita Harmita Department of Pharmacy, Faculty of Pharmacy, Universitas Indonesia, Depok, Indonesia.
  • Catur Jatmika Department of Pharmacy, Faculty of Pharmacy, Universitas Indonesia, Depok, Indonesia.
  • Mufid Idan Nugraha Department of Pharmacy, Faculty of Pharmacy, Universitas Indonesia, Depok, Indonesia.

Abstract

Objective: Glucosamine hydrochloride (HCl) and chondroitin sulfate are glycosaminoglycan compounds and major structural components of bones
in the form of proteoglycans. These compounds maintain bone structure by stimulating the synthesis of synovial fluid and inhibiting the degradation
of joint cartilage, and they can be used for the treatment of osteoarthritis. The aim of this study was to identify a selective analytical method for
determining glucosamine HCl and chondroitin sulfate levels in tablet and cream forms.
Methods: After derivatization using ortho-phthalaldehyde and 2-mercaptoethanol, the samples were analyzed using high-performance liquid
chromatography (HPLC) with a fluorescence detector at an excitation wavelength of 335 nm and an emission wavelength of 445 nm. Deacetylation
using sodium hydroxide was required to break the acetyl group bond. The mobile phase used tetrahydrofuran 0.25% in water-acetonitrile (87:13)
with a flow rate of 1.5 ml/minute.
Results: The average levels of glucosamine HCl and chondroitin sulfate were 92.76% and 96.11% in tablets and 101.15% and 100.33% in creams,
which fulfilled the acceptance criteria.
Conclusions: Our validation method for glucosamine HCl and chondroitin sulfate met the acceptance criteria of accuracy, precision, selectivity, and
linearity.

Keywords: Chondroitin sulfate, Derivatization, Glucosamine hydrochloride, Fluorescence, High-performance liquid chromatography, Validation.

References

1. Tjahjono DH, Slamet IS, Sasanti TD. Analysis of glucosamine in
cream dosage form and difussion liquid by high performance liquid
chromatography. J Ion Exch 2007;18(4):1420-1.
2. Nagarajan P, Sathis KD, Somsubhra G, Harani A, Maheswaran N,
David B. Method development and validation for glucosamine and
chondroitin sulphate in soflet form by RP-HPLC. Res Rev J Pharm
Anal 2013;2:6-10.
3. Pratiwi AI. Diagnosis and Treatment Osteoarthritis, Thesis. Indonesia:
Universitas Lampung; 2015.
Table 13: Determination of glucosamine HCl levels in cream form
C (ppm) Standard area of glucosamine HCl Sample area of glucosamine HCl Measurable
concentration (ppm)
Measurable
concentration (%)
80 320059616 328605087 82.14 102.67
328510670 82.11 102.64
328473366 82.10 102.63
C: Concentration, HCL: Hydrochloride
Table 14: Determination of chondroitin sulfate levels in cream form
C (ppm) Standard area of chondroitin sulfate Sample area of chondroitin sulfate Measurable
concentration (ppm)
Measurable
concentration (%)
900 10382934 10431472 904.21 100.47
10400356 901.51 100.17
10419917 903.21 100.36
C: Concentration
4. Andriani S. Optimasi derivatisasi glukosamin hidroklorida dengan
9-fluorenilmetoksikarbonil klorida (FMOC-Cl) secara kromatografi
cair kinerja tinggi-fluoresensi. Depok: Universitas Indonesia; 2012.
5. Topping J. A review of methods available for the determination of
chondroitin sulphates in supplements. United Kingdom: Laboratory of
the Government Chemist Queens Road Teddington Middlesex; 2012.
6. David JI, Roman M, Zhou J, Hildreth J. Determination of chondroitin
sulphate content in raw materials and dietary supplements by high
performance liquid chromatography with ultraviolet detection after
enzimatic hydrolysis. J AOAC Int 2007;90(3):659-69.
7. Vaclavikova E, Frantisek K. Isotachophoretic determination of
glucosamine and chondroitin sulphate in dietary supplements. Czech J
Food Sci 2013;31:55-65.
8. Dionex Corporation. Determination of Glucosamine in Dietary
Supplements Using HPAE-PAD; Application Note 197, LPN 2001.
Sunnyvale, CA: Dionex Corporation; 2008.
9. Lee KS, Drescher DG. Derivatization of cysteine and cystine for
fluorescence amino acid analysis with the o-phthaldialdehyde/2-
mercaptoethanol reagent. J Biol Chem 1979;254(14):6248-51.
10. Rhee WM, Richard AB. Glycisaminoglycan-Synthetic Polymer
Conjugates. European Patent Apllication, Report; 1995.
11. Assocation of Official Analytical Chemist (AOAC). Guidelines
for Single Laboratory Validation of Chemical Methods for Dietary.
Available from: https://www.aoac.org/aoac_prod_imis/AOAC_Docs/
StandardsDevelopment/SLV_Guidelines_Dietary_Supplements.pdf.
12. Harmita H. Buku Ajar Analisis Fisikokimia. Depok: Universitas
Indonesia; 2006.
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How to Cite
Harmita, H., Jatmika, C., & Nugraha, M. (2017). DETERMINATION OF LEVELS OF GLUCOSAMINE HYDROCHLORIDE AND CHONDROITIN SULFATE IN MIXTURES IN TABLET AND CREAM FORMS USING HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY WITH FLUORESCENCE. International Journal of Applied Pharmaceutics, 9, 144-149. https://doi.org/10.22159/ijap.2017.v9s1.75_82
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Original Article(s)