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ORIGINAL ARTICLE
Year : 2012  |  Volume : 3  |  Issue : 1  |  Page : 25-29  

Technological study of preparing gel from semi-solid extract of Cacalia hastata L.


1 Department of Pharmacy Management and Pharmaceutical Technology, School of Pharmacy, Health Sciences, University of Mongolia, Mongolia
2 Department of Clinical Pharmacy, School of Pharmaceutical Sciences, Universiti Sains, Malaysia

Date of Web Publication6-Mar-2012

Correspondence Address:
Syed Wasif Gillani
Lecturer/Researcher, Department of Clinical Pharmacy, School of Pharmaceutical Sciences, Universiti Sains Malaysia (USM), Penang 11800
Malaysia
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/2231-4040.93564

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   Abstract 

Most of the drugs which are used for wound healing are imported in Mongolia. It is required to develop drug formulation and increase local productions used for the treatment of wound healing. For the purpose of solving the above problems, we aimed to prepare new drug formulation from Cacalia hastata L. for the treatment of wound healing. Cacalia hastata L. is a medicinal plant, member of the family Asteraceae. Cacalia hastata L. is widely used for the Mongolian traditional medicine to treat wound healing, gastric ulcer, poisoning fever, liver fever, bile fever, oral cavity, and gynecological diseases. We prepared Cacalia gel from semi-solid extract of Cacalia hastata L. using various excipients such as gel former, solvent, neutralizer, antimicrobial preservative, and humectant. Gel formulation was standardized by such criteria, as the amount of biologically active compound, appearance (color, smell), pH, viscosity, and bacterial contamination. Stability testing of gel formulation was studied by long-term method. The quality of the Cacalia gel which was stored in room temperature, its appearance, viscosity, and amount of biological active compound were stable. The stability testing of the gel formulation from Cacalia hastata L. is continued.

Keywords: Cacalia hastata L. carbomer (Carbopol 934), excipients, gel, semi-solid extract


How to cite this article:
Jambaninj D, Syed Sulaiman S, Gillani SW, Davaasuren T S, Erdenetsetseg G, Dungerdorj D. Technological study of preparing gel from semi-solid extract of Cacalia hastata L. J Adv Pharm Technol Res 2012;3:25-9

How to cite this URL:
Jambaninj D, Syed Sulaiman S, Gillani SW, Davaasuren T S, Erdenetsetseg G, Dungerdorj D. Technological study of preparing gel from semi-solid extract of Cacalia hastata L. J Adv Pharm Technol Res [serial online] 2012 [cited 2020 May 29];3:25-9. Available from: http://www.japtr.org/text.asp?2012/3/1/25/93564


   Introduction Top


Nowadays, the vital problem of healthcare is to supply population with highly effective medicine with no adverse reaction. The world attitude is focused on the use of pure natural, herbal and animal preparation rather than synthetic preparation. [1]

Most of the drugs which are used for wound healing are imported in Mongolia. It is required to develop drug formulation and increase local production of drugs used for the treatment of wound healing.

Cacalia hastata L. is a medicinal plant of the family Asteraceae. It grows in botanical and geographical provinces such as Khangai, Khentii, Khuvsgul, and Mongol Daguur in Mongolia. [2],[3] Cacalia hastata L. has pyrrolizidine alkaloids: Hastacine up to 0.1%, platyphylline; total carotenoids up to 310 mg%, organic acid up to 13%, ascorbic acid, polysaccharide, flavonoids, phenolic acid, tannin, coumarin, and triterpenoids. [2],[3],[4],[5],[6],[7] Hastacine is alkaloid similar to platyphylline for its structure and has spasmolytic activity. [8] Flower and leaves of Cacalia hastata0 L. are widely used for the Mongolian traditional medicine to treat wound healing, gastric ulcer, poisoning fever, liver fever, bile fever, oral cavity, and gynecological diseases. It is also used to treat respiratory infection, cough, and antihemorrhagic for the Tibetan medicine named "King of hurt and break." [3],[4] Nowadays, extract of Cacalia hastata L. has been determined to have anti-inflammatory activity: Antibacterial, spasmolytic, antipyretic, gastroprotector, and antihemorrhagic. [9],[10]

For the purpose of solving the above problems, we aimed to prepare new drug formulation from Cacalia hastata L. for the treatment of wound healing.

Carbomers are used in liquid or semisolid pharmaceutical formulation as rheology modifiers. It includes creams, gels, lotions, and ointments. In contrast, microorganisms grow well in unpreserved aqueous dispersions, and therefore an antimicrobial preservative such as 0.1% w/v chlorocresol, 0.18% w/v methylparaben - 0.02% propylparaben, or 0.1% w/v thimerosal should be added. The addition of certain antimicrobials, such as benzalkonium chloride or sodium benzoate, in high content rations (0.1% w/v), can cause cloudiness and a reduction in viscosity of carbomer dispersions. Carbomer aqueous gels are more viscous at pH 6 to 11. The viscosity is considerably reduced at pH values less than 3 or greater than 12, or in the presence of strong electrolytes. [11]

Carboxymethyl cellulose sodium is widely used in oral and topical pharmaceutical formulations, primarily for its viscosity-increasing properties. Higher concentrations, usually 3% to 6%, of medium-viscosity grade are used to produce gels; glycols are often included in such gels to prevent them drying out. [11]


   Materials and Methods Top


Herb of Cacalia hastata L. was collected as a raw material in Tuv Aimag province, Mongolia in July 2010 and was identified by prof E. Ganbold, Sc.D in biology (voucher number №00/28).

Methylparaben, propylparaben, and propylenglycol were gift samples from Moncream Co. ltd, Ulaanbaatar, Mongolia. Benzalkonium chloride was a gift sample from Monchemo Co. ltd, Ulaanbaatar, Mongolia. Carbomer (Carbopol 934P) and Sodium carboxymethyl cellulose were bought (Thianjin Well-Real Chemical Technology Co. ltd, China). All other chemicals were of analytical reagent grade.

The quantity of total alkaloids and total carotenoids are determined by spectrophotometric, tannin by Lowenthal permanganate titration method for standardization of gel and semi-solid extract.

Determination of Total Carotenoids

Accurately weighed 1 g gel was dissolved in 50 ml of water and shake well in 250 ml separation funnel. 50 ml of hexane was added and separated two times. Hexane solution was poured into 100 ml volumetric flask, diluted with hexane to volume. Absorbance of the hexane solution was determined at 450 nm. The total carotenoids content (mg%) was calculated by the formula.



D is the absorbance of the solution, m is weight of gel, and V is volume of solution. 2592 is specific absorbance of β- carotene in hexane, in 1 cm cuvette at 450 nm. [6]

Determination of Total Alkaloids

Accurately weighed 1 g gel was dissolved in 50 ml of NH 4 OH and shake at well in 250 ml volumetric separation funnel. 50 ml of chloroform was added and separated two times. Evaporate the chloroform layer to dryness, add 2% solution of hydrochloric acid then dissolve 50 ml methanol, sonicate to dissolve and filter. Methanolic solution was poured into 100 ml volumetric flask and diluted with methanol to volume. Absorbance of the solution was determined at 210 nm. The total alkaloids content (%) was calculated by the formula.



D is the absorbance of the solution, m is weight of gel, and V is volume of solution. 314.54 is specific absorbance of pyrrolizidine alkaloid presence of hastanecic acid in methanol, in 1 cm cuvette at 210.3 nm. [7]

Determination of Tannins

Samples were analyzed by adding 1 ml sample and 5 ml indigo carmine to the 500 ml flask and adding 200 ml water. This is titrated against the Potassium permanganate solution (N/40 or 0.005 M) until the royal blue fades to a light green. Then, it is titrated drop-wise until the lime green changes to yellow. This value is record as X ml.

A blank titration using 5 ml of indigo carmine alone in 200 ml water should also be carried out. The blank value should be 1 ml and should be recorded as Y ml.



Preparation of Sodium Carboxymethyl Cellulose Gel Bases

The sodium carboxymethyl cellulose gel bases were prepared by dispersing the sodium carboxymethyl cellulose of 6% and 10% w/w in half of total distilled water (20 ± 2°C). The solution of humectant was prepared in the remaining amount of water. The solution of humectants was added at the end of dispersion stage. The pH value of gel bases was measured using pH meter (Hanna 320, Germany) (n = 3). Viscosity of gel bases was measured using Brookfield DV- III Ultra programmable rheometer (Brookfield Engineering Laboratories. Inc., USA) (n = 3).

Preparation of Carbomer gel Bases

We prepared various carbomer gel bases with different ratio of gel former, antimicrobial preservative, neutralizer, and humectant. At first stage, the solution of antimicrobial preservative and neutralizer were prepared in half of total water. At second stage, carbomer (1% or 2%) was dispersed using mixer (Akira HM-202BSS) in the solution of antimicrobial preservative and neutralizer. At third stage, the humectants were added very slowly at the dispersion stage.

Triethanolamine was used as a neutralizer and the pH of the gel systems was adjusted by per cent of 1.1%, 1.15%, 1.2%, 1.25%, and 1.3% w/w. As a conserver were used 0.18% methylparaben-0.02% propylparaben, 0.18% methylparaben, 0.02% propylparaben, and 0.01% benzalkonium chloride. As a humectant were used 1% propylene glycol, 1% polyethylene glycol-400, and 10%glycerol. The pH value of gel bases was measured using pH meter (Hanna 320, Germany) (n = 3). Viscosity of gel bases was measured using Brookfield DV-III Ultra programmable rheometer (Brookfield Engineering Laboratories. Inc., USA) (n = 3).

Formulation of Cacalia Gel from Cacalia Hastata Semi-solid Extract

Cacalia thick extract was added by 5% at the end of the dispersion stage. The gels had specific smell, dark brown to white brown color. The pH value of gels were measured using pH meter (Hanna 320, Germany) (n = 3). Viscosities of gels were measured using Brookfield DV-III Ultra programmable rheometer (Brookfield Engineering Laboratories. Inc., USA) (n = 3).

The quality of the gel formulation was investigated comparatively by its appearance (color, smell), pH, viscosity, and bacterial contamination.

Stability testing of gel formulation from Cacalia hastata L. was studied by long-term method.

Bacterial and mould contamination was defined according to MNS-5189-2002, MNS-5190-2002, MNS -5193-2002, and MNS -5194-2002. [12],[13],[14],[15]

Tests for significant differences between means were performed by Student's t-test or one-way ANOVA using the software SPSS 16. Differences were considered significant at P<0.05 and P<0.02 levels.


   Results Top


For choosing of appropriate gel former, gels were prepared by 1% and 2% carbomer, 6% and 10% sodium carboxymethyl cellulose. The result is shown in [Table 1].
Table 1: Quality of Cacalia gel with different concentration and various gel former

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Gels with carbomer gel base were prepared with various concentration triethanolamine and determined quality. The result is shown in [Table 2].
Table 2: Quality of Cacalia gel with different concentration triethanolamine

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For the determination of the quality of gel dependence on the antimicrobial preservative, we prepared gels using methylparaben-0.18%, propylparaben-0.02%, benzalkonium chloride-0.01%, combined methylparaben-0.18%, and propylparaben-0.02%. The result is shown in [Table 3].
Table 3: Microbial contamination of Cacalia gel with various antimicrobials

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Moisturizing effect of Cacalia gel was studied using various humectants such as propylene glycol, polyethylene glycol-400, and glycerol. The result is shown in [Table 4].
Table 4: Quality of Cacalia gel with various humectant

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Stability testing of gel formulation from Cacalia hastata L. was studied by long-term method. The quality of the gel was tested in each terms freshly prepared, after 2 weeks, 30 days, 2 months, 3 months, and 4 months. The results are shown in the [Table 5] and [Table 6].
Table 5: Viscosity of Cacalia gel at various storage condition (n = 3)

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Table 6: Amount of biological active compounds of Cacalia gel at various storage condition (n = 5)

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   Discussion Top


We prepared semi-solid extract by previous investigation. For preparing the semi-solid extract from leaves of Cacalia hastata L., it has been chosen by extraction method as a percolation, as the solvents by 70% ethanol three times at 20 ± 2°C and by distilled water two times at 96 ± 2°C. 70% ethanol 1:6 and distilled water 1:3 ratio was used as raw material with solvents. After that, it was evaporated to have 25% of moisture by vacuum vapor. Thick extract from Cacalia hastata L. has dark brown color, specific smell, and bitter taste. It should have not less than 25% of moisture and total alkaloids 0.36%, carotenoids 245 mg%, and tannins 13.6%. Thick extract had no heavy metals and bacterial contamination.

Cacalia gel was prepared with 5% semi-solid extract . Quantity of total alkaloids and total carotenoids, the appearance was not changed when using 1.2% triethanolamine when pH = 7.02 ± 0.077, and, Cacalia gel containing 1% propylene glycol had good moisturizing effect and did not change viscosity. These results were similar to the reference on the Handbook of pharmaceutical excipients. [4] Furthermore, Cacalia gel base composition is similar to Indian researcher's gel composition. They prepared herbal gel containing Clerodendron infortunatum leaves extract with 1% carbomer, 0.2% methylparaben 0.1% propylparaben, 5% propylene glycol, and 1.2% triethanolamine. [16]

The quality of the gel from Cacalia hastata L., which was stored in room temperature, its appearance, viscosity, and amount of biological active compound were comparative stable. The stability testing of the gel formulation from Cacalia hastata L. is continued.


   Conclusion Top


Appropriate composition of gel formulation for Cacalia hastata L. was developed. The gel which used 1% carbomer as a gel former, 1.65% triethanolamine as a neutralizer, 0.18% methylparaben-0.02% propylparaben as an antimicrobial preservative, and 1% propylene glycol as a humectant completely satisfied the quality criteria. The gel of Cacalia hastata L., which was stored at room temperature (25°C ± 2°C/60% RH ± 5% RH), was stable.

 
   References Top

1.Jambaninj D. Technological study of preparing children's drug formulation from Bergeniacrassifolia. In: Fritisch L, editor. Thesis of dissertation for M.Sc in pharmacy. Ulaanbaatar: Health Sciences University of Mongolia; 2004. p. 32.  Back to cited text no. 1
    
2.Khaidav TS, Altanchimeg B, Varlamova TS. Medicinal plants of Mongolian medicine. 2nd ed. Ulaanbaatar, Mongolia: 1985. p. 137-8.  Back to cited text no. 2
    
3.Ligaa U. Medicinal plants of Mongolia used in western and eastern medicine. Ulaanbaatar, Mongolia: 2005. p. 222-3  Back to cited text no. 3
    
4.Balhaev MI. Study of preparations of Cacalia hastata L. Bulletin VSNTs so RAMN 2005;13:199-200.  Back to cited text no. 4
    
5.Olennikov DN, Tankhaeva LM, Nikolaeva GG, Tsyrenzhapov AV, Nikolaev SM, Chekhirova GV. Biologically active substances from Cacalia hastata leaves: Carbonhydrates from leaves and their hypoglycemic activity. Chem Nat Compd 2004;4:21-3.  Back to cited text no. 5
    
6.Olennikov DN, Potanina OG, Tankhaeva LM, Nikolaeva GG. Pharmacognostic characteristic leaves of Cacalia hastata L. Chem Plant Raw Mater 2004;3:43-52.  Back to cited text no. 6
    
7.Hayashi K, Natorigawa A, Mitsuhashi H. Integerrimine from Cacalia hastata L. subsp. Orientalis Kitamura. Pharm Soc Japan 1972;20:201-2.  Back to cited text no. 7
    
8.Àltanchimeg D. Chemical composition of pyrrolizidine alkaloids of some species Mongolia. Thesis of dissertation for Ph.D in chemical science. Ulaanbaatar: Mongolian Academy of Chemistry and Chemical Technology; 2001. p. 32.  Back to cited text no. 8
    
9.Ayushieva SR, RazuvaevaIa G, Olennikov DN, Lonshakova KS. Phytofilm Khastaplen use in case of experimental parodontitismodelling.; Available from: http://www.ncbi.nlm.gov [Last Accessed on 2011 Dec 9].  Back to cited text no. 9
    
10.Volodiya TS. Gastroprotector activity of herbal preparations. Thesis of dissertation for Sc.D in veterinary medicine. Ulaanbaatar; Mongolia: 2002. p. 10-21.  Back to cited text no. 10
    
11.Rowe RC, Editor. Handbook of pharmaceutical excipients. 6 th ed. London and Chicago: Pharmaceutical Press: 2009.  Back to cited text no. 11
    
12.Basic indices of microbiological requirements, method for sampling. Mongolian National Standard. Dosage form of Drugs, raw materials. MNS-5189-2002. Ulaanbaatar: Center of Standardization and Measurement of Mongolia; 2002. p. 1-7.  Back to cited text no. 12
    
13.Preparation of samples for microbiological analysis. Mongolian National Standard. Dosage form of Drugs, raw materials. MNS- 5190-2002. Ulaanbaatar: Center of Standardization and Measurement of Mongolia; 2002. p. 1-6.  Back to cited text no. 13
    
14.Method for definition of total number of bacterium. Mongolian National Standard. Dosage form of Drugs, raw materials. MNS- 5193-2002. Ulaanbaatar: Center of Standardization and Measurement of Mongolia; 2002. p. 1-3.  Back to cited text no. 14
    
15.Method for definition of mould and fungus. Mongolian National Standard. Dosage form of Drugs, raw materials. MNS-5194-2002. Ulaanbaatar: Center of Standardization and Measurement of Mongolia; 2002. p.1-5.  Back to cited text no. 15
    
16.S, Haldar P, Pramanik G. Formulation and evaluation of herbal gel containing Clerodendroninfornatum leaves extract. Int J Pharm Tech Res 2011;3:140-3.  Back to cited text no. 16
    



 
 
    Tables

  [Table 1], [Table 2], [Table 3], [Table 4], [Table 5], [Table 6]



 

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