“FORMULATION DEVELOPMENT AND STABILITY STUDIES OF VITAMIN D3: COMPARISION AMONG DIFFERENT PREPARATION METHODS”
DISSERTATION PROTOCOL
SUBMITTED TO
RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES
BANGALORE, KARNATAKA.
BY
NAVEEN KUMAR.J.C
I YEAR M.PHARM,
DEPARTMENT OF PHARMACEUTICS,
NARGUND COLLEGE OF PHARMACY,
BANGALORE-85. KARNATAKA.
(2009-2011)
RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES
BANGALORE, KARNATAKA
ANNEXURE-II
PROFORMA FOR REGISTRATION OF SUBJECTS FOR P.G. DISSERTATION
1. / Name of the candidate and address (in block letters) / NAVEEN KUMAR.J.CNARGUND COLLEGE OF PHARMACY,
DATTATREYANAGAR,
BANASHANKARI III STAGE,
BANGALORE-85.
2. /
Name of the institution
/ NARGUND COLLEGE OF PHARMACY,DATTATREYANAGAR,
BANASHANKARI III STAGE,
BANGALORE-85.
3. /
Course of study and subject
/MASTER OF PHARMACY IN PHARMACEUTICS
4. /Date of the admission
/ 20th June 20095. /
Title of the topic:
/ “FORMULATION DEVELOPMENT AND STABILITY STUDIES OF VITAMIN D3 : COMPARISION AMONG DIFFERENT PREPARATION METHODS”6.
7.
8. / BRIEF RESUME OF THE INTENDED WORK
6.1 NEED FOR THE STUDY
Vitamin D is also called the Antirachitic vitamin, as it counteracts rickets, breaking-down of the mineral substance in fully grown bones (Osteodistrophy), bone and joint deformations, growth disorders, spontaneous brittleness of bones and deficient eggshell strength.
The term vitamin D actually refers to a pair of biologically inactive precursors of a critical micronutrient. They are vitamin D3, also known as Cholecalciferol, and Vitamin D2 also known as Ergocalciferol. Cholecalciferol (D3) is produced in the skin by a photoreaction on exposure to Ultraviolet B light from the sun (wavelength 290 to 320nm). Erogcalciferol (D2) is produced in plants and enters the human diet through consumption of plant sources. D2 and D3 forms are inactive as such but once present in the circulation, enter the liver and kidneys where they are hydroxylated to form active forms of D2 and D3. D2 has shorter half life than D3.
Vitamin D3 plays an important part in human and animal nutrition. It regulates the absorption, metabolism and excretion of calcium and phosphorus from the intestine, and kidneys. Further it controls the incorporation of calcium and phosphorus in the skeleton.
It is common to this fat-soluble agent that, in its pure form, it can be handled only with difficulty or not at all, because it is oxidation-sensitive substance. Furthermore, a fine dispersion of the agent is advantageous for optimal absorbability and thus bioavailability. These substances are therefore often supplied in the form of emulsions or in the form of dry powders or as solution in physiologically tolerated oil or embedded in a fine dispersion in a protective colloid. All of these methods have common drawbacks such as rapid deformation and disintegration due to heat or physical force and also due to aqueous solution employed. Decreased content uniformity increased running cost and decreased productive are the other disadvantages. Recently soft capsules containing a vegetable oil solution of active vitamins D3 have also developed. However, soft capsules have their particular drawbacks such as that they are limited in their form and that the process of their preparation is complicated.
So an attempt will be made to produce an efficient formulation of granule or powder forms which will be stable through out its shelf life, dispersible in aqueous solutions and overcome the above mentioned drawbacks.
Powdering or granulating of vitamin D3 can be accomplished by any or all of the following methods:
O By Micro Encapsulation:
ü In this procedure the desired polymer is dissolved in a suitable organic solvent followed by the addition of the desired substances to be encapsulated. The substance is either dissolved or dispersed in the organic solvent. The resulting organic solution is dispersed in an aqueous phase to obtain oil in water emulsion where oily micro particles are dispersed in the aqueous phase. Upon complete removal of the solvent from the micro particles the microcapsules are formed. This method will be modified if required or any suitable method will be employed.
O By dry emulsion:
ü A process for preparing a dry powder composition containing vitamins D3 by homogenizing vitamin D3 in an aqueous solution containing one or more protective colloids for a time and under conditions effective to produce an emulsion containing vitamin D3, wherein said aqueous solution optionally contains one or more sugars or other additives and drying the mixture optionally in the presence of a coating material, to yield a dry powder. Modification of this method or any other suitable method will be employed.
O By spray drying:
ü Emulsifying the drug and oil at ordinary temperature together with soluble casein or a cellulose in an aqueous solution and spray-drying the emulsified liquid, so as to obtain powder or
ü After addition of the vitamins to a solvent and agitation, the solvent is removed and the residue is subjected to powdering. The powdering may be effected by spray drying or
ü Any other methods through which stable granules may be obtained will be employed.
6.2 Review of literature:
§ Vitamin D3 including its physical, chemical, biological properties and its pathological importance are discussed extensively in Martindale 34th edition [1].
§ Marc S. Micozzi and Bethesda Maryland in their review article “Confronting the Worldwide Epidemic of Vitamin D Deficiency” reported the physiological importance of vitamin D and metabolism to its active forms. They explained the requirement of vitamin D in various disease state and normal daily intake [2].
§ Stephen Allen Christensen in his article “Is Vitamin D3 Better than D2?” has explained why vitamin D3 is more stable than D2 and clearly pointed the need of Vitamin D3[3].
§ Hähnlein, Wolfgang, Hansen, Morten Mohr, Olesen, Jes Elenius et al. have clearly reported the need for the stable formulation. They also demonstrated dry powder formulation with much emphasis on particle size, homogeneous fine dispersion, and stability concluding its importance for optimal absorbability and bioavailability [4].
§ Masaaki Nakahara, Akihiko Kurosaki, Hiroshi Otake, Hitoshi Kanai, Hiroshi Yokokawa, Kiyoshi Kumabe et al. have clearly demonstrated the conventional methods for powdering oil-soluble substance mentioning their common drawbacks. They discovered that elution of the same to heat, pressure, heat, water, or the like, can be prevented by allowing the same to adsorb on a calcium component in an aqueous solution in the presence of surfactant[5].
§ Gregory Paul Dittmar Andrew Irvine Sokolik in their patent application title “Vitamin D content uniformity in pharmaceutical dosage forms” have come up with new dosage forms of vitamin D3 and calcium carbonate having improved content uniformity. The improvements are realized through modifications to the formulation, the raw material specifications, and the process of manufacture [6].
§ Vladimir Babtsov, Kiryat Shmona, Yury Shapiro, Givat Shmuel, Emma Kvitnitsky, Kiryat Shmona in their patent article “Methods of Microencapsulation” have used excipients like Polymethyl Methaacrylate, cellulose acetate, polyvinyl alcohol, sodium Lauryl acetate etc in their preparation involving Microencapsulations of water insoluble drugs [7].
§ Hähnlein, Wolfgang, Hansen, Morten Mohr, Olesen, Jes Elenius et al. have prepared a dry emulsion preparation of vitamin D3 by homogenizing vitamin D3 in an aqueous solution containing one or more protective colloids for a time and under conditions effective to produce an emulsion containing vitamin D3. The said aqueous solution optionally contains one or more sugars or other additives and drying the mixture optionally in the presence of a coating material, to yield a dry powder[8].
§ Yajima and Mizuo have demonstrated the powdering of vitamin D3 by spray drying a method that involves addition of the vitamins to a solvent and agitation. The solvent is removed and the residue is subjected to powdering. The powdering may be effected by spray drying, drying in vacuum, freeze-drying and drying in drum or other known drying methods. They concluded that when a diluted acid is used as a solvent, the pH should preferably be adjusted to 3 to 4. On the contrary, when diluted ammonia is used, the pH should preferably adjusted to 10-11. Subsequently, vitamins are added. [9].
§ Makino, Yuji, Suzuki, Yoshiki, have introduced a pharmaceutical solid preparation of active form of vitamin D3 of improved stability which comprises of active form of vitamin D3 dispersed in an excipient readily soluble in organic solvent and a basic substance. They also examined the stability and ‘residual percentage of active vitamin D 3’ of the prepared specimen comparing with the control (standard vitamin D3) [10].
§ Ishii, Kuniaki, Toriumi, Yumiko Itai, Shigeru Hayashi, Hidefumi ,Nemoto, Masami have prepared solid pharmaceutical preparations containing a vitamin D3 derivative. The composition of this preparation consisted of excipients like Mannitol, sugar, Hydroxypropyl cellulose, and a binder polyvinyl Pyrrolidone[11].
§ Moroi, Masami, Yokoyama, Toshio, Iwasa and Akira have demonstrated the method, for the preparation of a stable dosage-form of active vitamin D 3, which comprises an active vitamin D3 and a stabilizer selected from Polyvinylacetal diethylaminoacetate and Hydroxy-propylcellulose adding a pharmaceutically-acceptable carrier to the resultant mixture [12].
§ Nemoto, Kaoru, Igusa, Kazuo, Ogasawara, Toshichika in their patent article “Stabilized active forms of vitamin D3” demonstrated the preparation containing an active form of vitamin D3 which is stabilized by incorporation of an amino acid. They concluded amino acids that do not contain a sulfur atom or an acid amino group in its structure stabilized the vitamin D3[13].
§ John M. Ballard , Limin Zhu, Eric D. Nelson, Randal A. Seburg conducted LC-UV profile of a thermally stressed vitamin D3 tablet and concluded that four major degradants formed are identified as the Octanoate and Decanoate esters of D3 and pre-vitamin D3.This observation reinforces the need to be aware of potential interactions when designing formulations of apparently inert excipients and active pharmaceutical ingredients. They also concluded that even minor drug-excipient reactivity can be significant in the long-term stability of pharmaceutical products due to the stringent quality standards to which pharmaceutical formulations are held[14].
§ Semih OTLES, Yasar HISIL have clearly demonstrated the determination of vitamin D3 by high pressure liquid chromatography (HPLC). They estimated the quantity of vitamin D3 in hen egg and the recovery study of vitamin D3. They finally concluded that HPLC method is rapid simple, sensitive, reproducible and very efficient technique for the determination of vitamin D3 in hen eggs [15].
6.3 Objectives of the study :
The aim of the present work is to formulate and evaluate the stable vitamin D3 formulation and comparison of different methods.
· It is therefore an objective of this study is to provide a composition for solid pharmaceutical preparations of active vitamins D 3 wherein the stability of said active vitamins D 3 is remarkably improved.
· It is another objective of this invention is to provide a process for the preparation of a composition for solid pharmaceutical preparations of active vitamins D 3. It should be simple and efficient method whereby a composition uniform in size distribution and homogeneous in active vitamins D 3 content can be obtained.
· Further, this invention is to provide solid pharmaceutical preparations of active vitamins D3, wherein the stability of said active vitamins D 3 to light, heat, oxidation, etc. is remarkably improved.
· It is yet another objective of this invention is to provide a composition of active vitamins D3 which is extremely suited as a composition for making solid preparations such as tablets, powders, granules, etc. of active vitamins D 3 given as such or added to milk powder, rice flour, soft drinks etc.
· The other objective is to prepare in compatible with taste masking, to retain drug properties and to improve flow property of powders.
· Finally in-vitro analysis of different formulations produced using standard analytical tools like HPLC, UV, etc.,
MATERIALS AND METHOD
Materials:
1. Active forms of vitamin D3: wherein the active form of vitamin D3 is at least one selected from the group consisting of 1α-hydroxycholecalciferol, 1α, 25-dihydroxycholecalciferol,1α,24-dihydroxycholecalciferol, 1α,24,25- Trihydroxycholecalciferol, 1α-hydroxy-24-oxocholecalciferol, 1α, 25-dihydroxy-24-oxo-cholecalciferol, 1α, 25-dihydroxy-cholecalciferol-26,23-lactone, 1α, 25-dihydroxy-cholecalciferol-26,23-peroxylactone, 26,26,26,27,27,27-hexafluoro-1α,25-dihydroxy-cholecalciferol, 25-hydroxycholecalciferol, 24-hydroxycholecalciferol, 24-oxocholecalciferol, 24,25-dihydroxycholecalciferol, 25-hydroxy-24-oxo-cholecalciferol, 25-hydroxycholecalciferol-26,23-lactone, and 25-hydroxycholecalciferol-26,23-peroxylactone.
2. Excipient readily soluble in an organic solvent is at least one selected from the group consisting of polyvinyl Pyrrolidone, Hydroxypropylcellulose, Hydroxypropyl methylcellulose, methylcellulose, cholesterol, β-Sitosterol, Campesterol, and Deoxycholic acid etc.
3. Excipient which is slightly soluble in an organic solvent crystalline cellulose, starch, casein, cyclodextrin, lactose, Hydroxypropyl starch, dextrin, gelatin, etc.,
4. Organic solvents: methanol, ethanol, and Propanol; solvents of Haloganated hydrocarbons such as dichloromethane and chloroform; and such ethereal solvents as diethyl ether, etc.,
5. other excipients: D-Mannitol or white sugar, starch, dextrin, Tragacanth, gelatin, PVP, HPC, PVA, calcium carbonate etc,
6. Coloring agents, such as dyestuffs of tar source, Sunset Yellow,
7. Antioxidants - butyl Hydroxytoluene (BHT), Propyl Gellate, butyl Hydroxyanisol (BHA), Lecithine, α-Tocopherol, hydroquinone, ascorbic acid, Octyl Gallate, and Dodecyl GallateAs Corrigents, citric acid, Fumaric acid, menthol, and citrus perfume.
8. Surfactants: sodium Dodecyl sulfate and Sodiumlauryl sulfate,etc .
9. Calcium components like calcium Carbonate,Calcium citrate etc
10. Thickening and stabilizing agents like Xanthum Gum,Guar gum, locust bean Gum, etc.
11. Reducing sugars like glucose, fructose, lactose, maltose etc
12. Amino acids such as Alanine, Valine, Proline, phenylalanine, tryptophan, Leucine, Isoleucine, Glycine and serine, and basic amino acids such as lysine, Arginine and Histidine.
METHOD:
Some of the commonly used methods which are to be employed for the formulation of the stable Vitamin D3 preparations are:
1. Spray drying method.
2. A method using hardening of oil.
3. Encapsulation method.
4. Hydrogenation method.
5. Dry emulsion method.
6. Clathration with bile acids, cyclodextrin.
7. Complexation with sterols.
8. Dispersion with polyvinyl Pyrolidone. etc
7.1 Sources of data
1. Library: Nargund College of pharmacy.
2. e-library: Nargund college of pharmacy.
3. Official books (IP, BP, USP, and EP).
4. RGUHS Library, Bangalore.
5. International and national Pharmaceutical abstracts etc
Journals & articles :
1. Journal of Pharmaceutical and Biomedical Analysis.