International Journal of Advanced Scientific and Technical Research Issue 3 volume 2, March-April 2013

Available online on http://www.rspublication.com/ijst/index.html ISSN 2249-9954

Preparation and Evaluation of Stable Glucosamine sulphate Tablet

Formulation

Raju Merugu* (Registration No. J578600018),

Dr. M. Vijaya Bhaskara Reddy and Dr.Rajendra.G.Lala

VINAYAKA MISSION `S RESEARCH FOUNDATION DEEMED UNIVERSITY,

SALEM- TAMILNADU (INDIA)

Abstract
Pure Glucosamine is very "hygroscopic" and degrades (breaks down) rapidly when exposed to
moisture or air. To avoid this, Glucosamine needs to be bound to a stabilizer to be sold
commercially. The sulfate and the HCL forms are two of the most common "agents" that
Glucosamine is bound to ensure its stability. After Glucosamine is bound, it is stable and will not
degrade before it can get to the store shelf. Hence it is very difficult to prepare Glucosamine base
and instead find Glucosamine Sulfate or Glucosamine HCL.
Thus an attempt was made to stabilize the Glucosamine formulation by using the combination of
anti-oxidant, Desiccant to resolve the problems associated with the Glucosamine formulation .In
the present formulation studies were carried for the stability studies as ICH guidelines with Real
time and Accelerated stability studies and it was found to be stable during the study period of
stability for three months. All the formulas with single desiccant and single ant-oxidant did not
reduce the extent of the degradation. The tablets so prepared with conventional methods showed
good results physical evaluation parameters and chemical parameters such as Assay, and
Dissolution values. The granules prepared by using these ant-oxidants and desiccant, were good
in their flow properties. Assay and dissolution studies were conducted by the HPLC method.
However use of Ascorbic acid resulted in another problem that is it resulted in the spots on the
tablets.

Keywords: Stable glucosamine tablets,Oral solid formulation of glucosamine and Nutrceuticals

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International Journal of Advanced Scientific and Technical Research Issue 3 volume 2, March-April 2013
Available online on http://www.rspublication.com/ijst/index.html ISSN 2249-9954

Introduction:-

Glucosamine is a special formulation of several natural ingredients that have been found
to contribute to joint health. Glucosamine stimulates the body’s manufacture of collagen, the
protein portion of the fibrous substance that holds joints together and provides a shock-absorbing
cushion, as a person ages, the cartilage that cushions the joints often loses its ability to support
healthy cellular growth. In addition, the synovial fluid which lubricates these joints also
deteriorates. This condition, called osteoarthritis, often leads to rough bones that rub together and
cause distress with every twist or bend.( Nutraceuticals\ [Report] Global Nutraceuticals
Advances Market Research ; Remington the sciences and practice of pharmacy 20th edn )

Over time, the only option was to use nonsteroidal anti-inflammatory drugs (NSAIDs).
These preparations may cause side effects like slowing down the body’s rate of cartilage
replacement and actually destroying cartilage. Other side effects include: gastrointestinal tract
damage, hemorrhaging. However, modern science has discovered a nutrient that helps address
the dilemma of preserving and building healthy joint tissues and fluids. Glucosamine is a
necessary nutrient in the production of cartilage and synovial fluid. However, the body’s
production of it decreases as a person ages, thus inhibiting the new growth of cartilage destroyed
through wear and tear. (Deal CL, Moskowitz RW).
Pure Glucosamine is very "hygroscopic" and degrades (breaks down) rapidly when
exposed to moisture or air. To avoid this, Glucosamine needs to be bound to a stabilizer to be
sold commercially. The sulfate and the HCL forms are two of the most common "agents" that
Glucosamine is bound to ensure its stability. After Glucosamine is bound, it is stable and will not
degrade before it can get to the store shelf. Hence it is very difficult to prepare Glucosamine base
and instead find Glucosamine Sulfate or Glucosamine HCL. However some manufacturers add
up to 30% of a "1500 mg" mixture and replace it with plain table salt. This means you are only
taking 1050 mg of "actual" Glucosamine and it is legal because they put the ingredients on the
label and the product does contain 1,500 mg of Glucosamine sulfate (NaCl or 2KCL). It's just
that the NaCl and 2KCL that is added on is as filler. (www.glucosamine-osteoarthritis.org 05-04-
2006; McCarty MF, 1992) Thus there is a significant intake of salt. However, until now, it has
not been possible to eliminate several unfavorable properties of this compound, specifically, its

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International Journal of Advanced Scientific and Technical Research Issue 3 volume 2, March-April 2013
Available online on http://www.rspublication.com/ijst/index.html ISSN 2249-9954

highly hygroscopic nature and the facility with which its amino group oxidizes if not completely
saltified (www.glucosamine-osteoarthritis.org 05-04-2006, Reginster JY, 2001; Lisa R, Brian L).

These unfavorable properties give rise to difficulties and limitations the practical use of
Glucosamine sulphate. For example, oral forms, such as tablets or capsules, require anti-
oxidants, such as sodium hyposulphite, to be present in their formulations, which substances,
although blocking the oxidation of the amino group, leave the problem of the hygroscopic nature
of the sulphate unsolved; this necessitates the preparation of these forms in environments with a
relative humidity not greater than 30%, with results which are even then unsatisfactory and with
a stability lifetime which is practically insufficient for their use. Similar comments apply to
rectal forms (suppositories) which, even if kept under dry, refrigerated conditions, degrade with
considerable rapidity. (United States Patent, Patent Number 4,642,340, European Patent, Patent
Number 444000B1, Daniel O. C, Domenic M.D., Reda J, 2006)

Materials and Methods.

Materials:
Glucosamine sulfate was obtained as gift samples from Chaitianya Biological Pvt. Ltd- (India).
Aerosol 200(Degussa Germany) ,Croscarmellose sodium (Ac-Di-Sol,FMC Corp), Lactose
Anhydrous (DMV), Stearic acid (Taurus Chemicals, Hyderabad),
Kesilughur (Alpha chemie), Ascorbic acid (Roche), Pregelatinized starch (Colorcon Asia Pvt.
Ltd), Alpha Tocopheryl acetate Concetrate - Powder Form (Vitamin E-Roche) and Sodium carbonate
(SD fine Chemical -Mumbai India) were procured from commercial sources.All other
ingredients and reagents used were of analytical grade and used as such.

Methods:
Preparation of tablets
All materials were simply dry-blended with a minimum amount of lubricant and compressed by
using a CADMACH 27-station compression machine. A set of 17 x7.5mm capsule shape
punches and die were used for the compression and compression force was set at five tons.
Compressed tablets were film coated by using auto coater and film suspension used was opadry
white supplied by Colorcon Asia Pvt Ltd.

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International Journal of Advanced Scientific and Technical Research Issue 3 volume 2, March-April 2013
Available online on http://www.rspublication.com/ijst/index.html ISSN 2249-9954

Table 1. Various Glucosamine sulphate Tablet 500 mg Formulations

Sl Ingredients
No Formulations (mg / tablet)
A B C D E F G
Glucosamine sulphate
1 500 500 500 500 500 500 500

2 Pregelatinized starch 38 38 38 38 38 38 38
3 Aerosil 200
3.0 3.0 3.0 3.0 3.0 3.0 3.0
4 Lactose Anhydrous
100 100 100 100 100 100 100
Ac-di-sol
20.0 20.0 20.0 20.0 20.0 20.0 20.0
5
Stearic acid
6.0 6.0 6.0 6.0 6.0 6.0 6.00
6
Butylated hydroxy
7 - - - - - 0.2 0.2
anisole
8 Calcium carbonate
- - 10.0 - 10.0 - -
9 Sodium bicarbonate
- - - 10.0 - 10.0 10.0
Butyl hydroxytoluene
- - - - - - 0.2
10
Vitamin E
- - - - - - 10.0
11
Sodium meta
12 3.0 - - - - - -
bisulphate
13 Ascorbic acid
- 20.0 - - - - -
14 Sodium chloride
- - 3.0 - - - -
Kesilughur
- - - 20.0 - - 50.0
15
Total weight of core tablet 670 687 680 697 677 677.2 737.4
Opadry white
16 15 15 15 15 15 15 15
Total weight of
685 702 695 712 692 692.2 752.4
Coated tablet

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International Journal of Advanced Scientific and Technical Research Issue 3 volume 2, March-April 2013
Available online on http://www.rspublication.com/ijst/index.html ISSN 2249-9954

In vitro release study

Glucosamine sulphate release was determined with six tablets per formulation using a USP Type
II dissolution assembly (Hansen SR-8 Plus, Chatsworth, CA, and Erweka DT-70, Milford, CT)
in deionized water with a paddle speed of 50 rpm and bath temperature of 37.0+0.5°C. Samples
were collected interval of 15 minutes, 30 minutes and 45 minutes. Each samples were collected
were replaced with fresh equal volume of dissolution media. Six replicates were tested for each
batch of the tablet formulations. Dissolution samples were filtered and this apparatus was
equipped for filtered fraction collection diluted to appropriate concentrations for drug analysis.
Each filtered sample was analyzed by HPLC method using Waters Alliance 2695 Seperations
module; 2996 PDA detector was used with BDS Hypersil C8 column (250mm x 4.6mm; 5 μm
particle size)
Formulae for the calculation of Assay values

AT WS 200 P (100 - L/W)

------x---------x----------x----------x---------------x Average wt.
AS 50 Wt 100 100

Where, AT' is area of test solution. 'AS' of area of standard solution. 'WS' is weight of standard
"WT" is weight of test solution. 'P' is purity of standard. 'L/W' is LOD or water content.
Tablet hardness and friability
Hardness of finished tablets was determined using a VanKel VK200 hardness tester (Cary, NC).
The values reported here are the averages of 10 determinations. Friability was determined using
a VanKel single drum friabilator (Cary, NC) following USP method guidelines: 10 tablets were
weighed, rotated in the drum for 100 revolutions, removed and re-weighed. Change in mass was
calculated and reported as percentage lost.

RESULTS AND DISCUSSION

All tablet formulations possessed satisfactory tableting properties such as good mixing
granulating characteristics, flowability and compressibility. The manufacturing process was
simple and satisfactory with conventional granulation, mixing and coating procedures. No
complex process was required for production of Tablet of Glucosamine sulphate.
All the tablets were not show much change in the physical as well as chemical evaluation
parameter.

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International Journal of Advanced Scientific and Technical Research Issue 3 volume 2, March-April 2013
Available online on http://www.rspublication.com/ijst/index.html ISSN 2249-9954

Table : 2
Physical parameters of the tablets during stability studies at 40 0C+2 o C/75+5% RH
Name of Hardness in kg/cm2 Friability in percentage %w/w
Formulation Initial 1st 2nd 3rd Initial 1st 2nd 3rd
Month Month Month Month Month Month
Formulation A 5.0 4.8 4.5 4.2 0.1 0.3 0.4 0.5
Formulation B 5.2 4.8 4.5 4.2 0.1 0.2 0.25 0.3
Formulation C 5.5 5.0 4.8 4.6 0.1 0.3 0.42 0.53
Formulation D 5.8 5.2 5.0 4.6 0.1 0.25 0.3 0.35
Formulation E 5.5 5.0 4.9 4.8 0.2 0.3 0.32 0.38
Formulation F 5.8 4.9 4.2 4.0 0.2 0.25 0.3 0.35
Formulation G 6.0 6.0 6.0 6.0 0.1 0.2 0.25 0.25

percentage assay at 25/60% RH

101
percentage Assay

100 Formulation A
99 Formulation B
98 Formulation C
97 Formulation D
96 Formulation E
95 Formulation F
0 1 2 3 Formulation G
study period in months

Fig :1 Graph of Percentage Assay Vs Stability study period at 25oC+2/60%+5 RH

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International Journal of Advanced Scientific and Technical Research Issue 3 volume 2, March-April 2013
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Dissolution profile at 25/60%RH

percentage of dissolution

100 Formulation A

Formulation B

Formulation C

Formulation D
50 Formulation E

Formulation F

Formulation G

0
0 0.5 1 1.5 2 2.5 3
Period of study in months

Fig:2- Graph of Percentage Dissolution Vs Stability study period at 25oC+2/60%+5 RH

Percentage assay at 40 oC/75%RH

120

100 Formulation A

Formulation B
percentage assay

80
Formulation C

60 Formulation D

Formulation E
40
Formulation F

20 Formulation G

0
0 1 2 3 4 5
study period in months

Fig : 3 Graph of Percentage Assay Vs Stability study period at 40 oC+2/75%+5 RH

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International Journal of Advanced Scientific and Technical Research Issue 3 volume 2, March-April 2013
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Dissolution values at 40/75%RH

120

100
Formulation A

Formulation B
dissolution values

80

Formulation C
60
Formulation D

40 Formulation E

Formulation F
20

Formulation G

0
0 1 2 3 4 5

study period in months

Fig:4 Graph of Percentage dissolution Vs Stability study period at 40oC+2/75%+5 RH

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International Journal of Advanced Scientific and Technical Research Issue 3 volume 2, March-April 2013
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Stability of Glucosamine sulphate oral dosage forms:

The stability data obtained from the three month study showed that the formulae G was stable
with respect to colour change and Assay values (as shown in the photographs)

Fig 5 : Colour changes of the tablets containing Glucosamine sulphate

I: Initial colour of tablets II: Colour of tablet after 3 months at
400C+2/75%+5 RH. A, B, C, D, E, F,and G are the formulations and colour
of the tablets of respective batches at 400C+2/75%+5 RH after 3 months

Conclusions
Trials conducted using combinations of various antioxidants and the results of are given as
follows
1. Formulation A containing only one anti oxidant that is sodium meta bisulphate did not
give any significant help in reduction in the colour change thus formulation A was not
suitable.
2. Formulation B containing ascorbic acid an anti oxidant has given rise other problems
like spotting was observed it may be due reduction of ascorbic acid
3. Formulation C containing sodium chloride salt, calcium carbonate a desiccant did not
solve the problem of the degradation of the Glucosamine sulphate

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International Journal of Advanced Scientific and Technical Research Issue 3 volume 2, March-April 2013
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4. Formulation D containing Kesilughur a desiccant and sodium bicarbonate salt also did
not help in the achieving the stability of the Glucosamine sulphate tablet.
5. Formulation E containing calcium carbonate a desiccant was added but it also did not
serve the purpose.
6. Formulation F containing Butylated hydroxy anisole a stronger anti-oxidant and a salt of
Sodium bicarbonate was added but it did not produce satisfactory results.
Thus by above all the trials we conclude that single anti-oxidants, single desiccants will not help
in achieving a stable formulation of Glucosamine sulphate, hence an attempt was made by using
a combination of the anti-oxidants and desiccants which was as per the formula G.
7. Formulation G containing Butylated hydroxy anisole, Sodium bicarbonate, Butyl
hydroxy toluene, Vitamin E and Kesilughur has shown the best results with respect to the
physical appearance and Assay values.
8. The stability studies conducted during three months shows formulae stable yet to confirm
this stability studies has to be continued for more days.
9. Glucosamine sulphate are highly instable in higher humidity which confirmed by
the above studies hence the final packing of the Glucosamine or Glucosamine sulphate
containing products should be packed or stored in a moisture proof /resistant containers
like glass or HDPE. Aluminum packing have shown to be less effective in preventing the
breakdown of the Glucosamine sulphate
Thus the formulation of the Glucosamine sulphate can be effectively formulated by using a
combination of the anti-oxidants and desiccants which will in solving the problems associated
with Glucosamine sulphate such as the hygroscopicity and oxidation of the amino groups.

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International Journal of Advanced Scientific and Technical Research Issue 3 volume 2, March-April 2013
Available online on http://www.rspublication.com/ijst/index.html ISSN 2249-9954

List of references
1. On line artcle:-Nutraceuticals\ [Report] Global Nutraceuticals Advances Market
Research, Trends, Analysis.htm 05-04-2006.
2. www.glucosamine-osteoarthritis.org 05-04-2006
3. United States Patent, Patent Number 4,642,340 Rotta Research Laboratorium S.p.A.,
Milan, Italy
4. European Patent, Patent Number 444000B1 Rotta Research Laboratorium S.p.A.,
Milan, Italy.
5. Daniel O. C, Domenic M.D., Reda J. the new England journal of medicine Volume
354:795-808 February 23, 2006 Number 8
6. Reginster JY, Deroisy R, Rovati LC, et al. Longterm effects of glucosamine sulphate on
osteoarthritis progression: a randomised, placebo- controlled clinical trial. Lancet
2001;357 (9252):251-6.
7. McCarty MF. The neglect of glucosamine as a treatment for osteoarthritis-a personal
perspective. Med Hypotheses 1994;42 (5):323-7.
8. Deal CL, Moskowitz RW. Nutraceuticals as therapeutic agents in osteoarthritis. The role
of glucosamine, chondroitin sulfate, and collagen Arthritis Foundation,1330 West
Peachtree Street, Atlanta GA 30309 bulletin on the rheumatic diseases a publication of
the arthritis foundation vol. 50 no. 7
9. Remington the sciences and practice of pharmacy 20th edn
10. Lisa R, Brian L School of pharmacy and pharmaceutical sciences, England UK 15-26 pp.

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