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PHARMACEUTICAL STANDARDIZATION
Year : 2014  |  Volume : 35  |  Issue : 1  |  Page : 58-62  

Quality control evaluation of Keshamasi, Keshanjana and Keshamasi eye ointment


1 Department of Shalakya Tantra, Institute for Post Graduate Teaching and Research in Ayurveda, Gujarat Ayurved University, Jamnagar, India
2 Pharmaceutical Chemistry Laboratory, Institute for Post Graduate Teaching and Research in Ayurveda, Gujarat Ayurved University, Jamnagar, India
3 Analytical Development Laboratory, Zydus-Cadila, Ahmedabad, Gujarat, India
4 Marketing and Business Development, eVaidyaJi Wellness Pvt. Ltd., Jaipur, Rajashtan, India

Date of Web Publication29-Sep-2014

Correspondence Address:
Kartar Singh Dhiman
Professor and Head, Department of Shalakya, Tantra, IPGT and RA, Gujarat Ayurved University, Jamnagar - 361 008, Gujarat
India
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/0974-8520.141938

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   Abstract 

Background: Keshanjana (collyrium) is a well known Ayurvedic preparation prepared out of Keshamasi (ash prepared by scalp hairs) mixed with Goghrita (cow's ghee). This medicine is indicated for the treatment of Shushkakshipaka (dry eye syndrome) in the classical literature of Ayurveda; hence, it was under taken for standardization and clinical evaluation in an extra-mural research project from Central Council for Research in Ayurvedic Sciences, Department of AYUSH, New Delhi. Aim: To develop standard quality parameters for the Keshamasi, Keshanjana and Keshamasi ointment. Materials and Methods: Scalp hairs of male and females collected from saloons were converted to classical Masi Kalpana and mixed with cow ghee and petrolatum in the ratio of 1:5 to prepare the Keshanjana and Keshamasi ointment respectively. Standard Operation Procedure (SOP) were adopted and recorded accordingly. The raw material, furnished products and plain Goghrita were subjected for quality control parameters i.e., physico-chemical evaluation, anti-microbial study, particle size analysis, heavy metal analysis through inductive couple plasma spectroscopy with high performance thin layer liquid chromatography fingerprints. Results: Rancidity was negative in all the samples, indicating that the physico-chemical parameters are in acceptable range. Lead and zinc were present in most of the samples; while all samples are were free from microbial contamination. Conclusion: As no standards are available to compare the results of the current study, the observations cannot be compared. Thus the profile generated in the current study can be considered as standard to refer in future studies.

Keywords: Keshamasi , Keshanjana, ointment, quality control profile


How to cite this article:
Dhiman KS, Shukla VJ, Bhalodia NR, Sharma VR. Quality control evaluation of Keshamasi, Keshanjana and Keshamasi eye ointment. AYU 2014;35:58-62

How to cite this URL:
Dhiman KS, Shukla VJ, Bhalodia NR, Sharma VR. Quality control evaluation of Keshamasi, Keshanjana and Keshamasi eye ointment. AYU [serial online] 2014 [cited 2020 Sep 23];35:58-62. Available from: http://www.ayujournal.org/text.asp?2014/35/1/58/141938


   Introduction Top


Keshanjana (collyrium) is an Ayurvedic formulation prepared out of Keshamasi (ash prepared by scalp hairs) mixed with Goghrita (cow ghee). This particular preparation is indicated for treating Shushkashipaka (dry eye syndrome) in Vagbhatta Samhitas and classical Ayurvedic treatises. [1] Keshanjana has tried clinically as PG thesis work with control of artificial tears and better results were found in comparison to control group. [2] Encouraged by this outcome an EMR project on the "standardization and clinical evaluation of Keshanjana-an Ayurvedic formulation" has been granted by Central Council for Research in Ayurvedic Sciences, Department of AYUSH, New Delhi and in the present paper analytical/quality parameters for the standardization process are presented.

Keshmasi prepared by using Putapaka Vidhi (incineration method). Hairs were smeared with Goghrita (cow ghee) on glass slab. After proper mixing, mixture was equally divided in four parts and kept in four earthen saucers. Later, it was subjected for Putapaka in the conventional Puta (30 Angulas = 57 cm in length, breadth and height). After complete burning, it was allowed for self-cooling. Masi (ash) was collected from the inner surface of earthen saucers recovered from Puta after self-cooling and then collected Masi (ash) was packed in air tight bags and kept for further processing. Same method was adopted for the preparation of both male and female scalp hairs Keshamasi.

The Keshanjana (collyrium prepared by ash of scalp hairs) prepared by using male and female Keshamasi (ash of scalp hairs) by adopting same method. During this process, Masi (ash) was transferred into a mixer grinder and sieved through 400#. Afterward, initially Keshamasi (ash of scalp hairs) and Goghrita was taken in the ratio of 1:2 and triturated for 6 hours. Then gradually 100 g of Goghrita was added and triturated again up to the formation of uniform mixture. After complete attrition, the Keshanjana (collyrium prepared by ash of scalp hairs) was weighed and transferred in to an air tight container for further processing. The complete process was repeated for preparation of ointments but instead of Goghrita, petrolatum was used as the base.

For the preparation of the Keshamasi, male and female hairs were taken as starting material. [3] This reference is indicated in the treatment of dry eye syndrome. Although no direct indication regarding the gender based nature of raw drug is indicated; however, here an attempt made to modify the Keshamasi into Keshanjana an Raskriya Anjana (ointment) from male and female hair. Modified preparations found more feasible with later for pharmacy scale as well as far as modishness of the finished product is concerned. [4]

Objectives of the study

Present research work provides physico-chemical parameters for its quality control of raw and finished product which includes its physico-chemical evaluation, anti-microbial study, particle size analysis, Heavy metal analysis through (Inductive Couple Plasma [ICP] spectroscopy) with high performance thin layer liquid chromatography (HPTLC) fingerprint of finished products. These qualitative parameters can be used for good manufacturing practices of the Drug preparation.


   Materials and Methods Top


Procurement of material

Finished product Keshamasi, Keshanjana and Keshamasi ointment prepared out of male and female scalp hair. All samples were subjected for the quality control process and coded as follows:

  • Keshamasi (prepared out from the male's scalp hair) - KMM
  • Keshamasi (prepared from female's scalp hair) - KMF
  • Keshanjana (prepared from male's scalp hair) - KSJM
  • Keshanjana (prepared from female's scalp hair) - KSJF
  • Ointment (prepared from male's scalp hair) - OM
  • Ointment (prepared from female's scalp hair) - OF
  • Goghrita - cow ghee
  • Keshamasi is considered as raw material in this study for preparation of Keshanjana and Keshamasi ointment.


Physico-chemical evaluation

In physical evaluation prescribed standardization parameters of ghee such as rancidity test, saponification value, acid value, iodine value, ester value, peroxide value, specific gravity, refractive index etc., studied as per standards. [5]

The analysis of raw materials included organoleptic parameters such as consistency, color, taste and odor, physico-chemical parameters such as loss on drying, total ash, acid-insoluble ash, water soluble ash, alcohol soluble extractive values, water soluble extractive values and petroleum ether soluble extractive values were also carried out. Particle size analysis has been done.

The analysis of finished products included the organoleptic parameters such as consistency, color, taste and odor, determination of pH, acid value, iodine value, saponification value, ester value, peroxide value, estimation of unsaponifiable matter. The determinations were performed in triplicate and results are expressed as mean ± standard deviation.

Microbiology study

In this study total microbial count, total bacterial count, total fungal count with pathogens- Escherichia More Details coli,  Salmonella More Details spp., Pseudomonas aeruginosa, Staphylococcus aureus in both aerobic and anaerobic conditions for KMM, KMF, KSJM, KSJF, OM and OF were carried out.

Particle size analysis

The KMM, KMF, KSJM, KSJF, OM and OF were analyzed in Symantec HELOS (H1004) instrument, by taking water as dispersion medium and measured at 0.5/4.5-875 μm range. It has three type Fourier lances: R1-0.1 μm to 35 μm, R2-0.5 μm to 175 μm, R3-0.5 μm to 875 μm. [6]

Heavy metal analysis

ICP spectroscopy study


The ICP analyses of the samples were carried out. All samples KSJM, KSJF, OM and OF were tested for the presence of heavy metals such as cadmium (Cd), lead (Pb), mercury (Hg), arsenic (As) and zinc (Zn). [7],[8] Microbiological study, particle size analysis and heavy meal analysis were carried out at SICART laboratory Vallabh Vidyanagar, Gujarat.

HPTLC study

For HPTLC investigation, the chromatogram of unsaponifiable matter of cow ghee, raw materials and formulated finished products obtained. A Camag HPTLC equipped with a sample applicator Linomat V, twin trough plate development chamber, thin layer liquid chromatography Scanner III, Reprostar and Wincats 4.02, integration software (Switzerland) were used. [9]

Experimental condition

  • Stationary phase: Precoated silica gel Gf 254.
  • Mobile phase:Hexane:diethyl ether:acetic acid (7:3:0.5).
  • Spray reagents: Anisaldehyde sulphuric acid.



   Observations and Results Top


Cow ghee shows specific gravity value of 0.9097 at 40°C and refractive index value of 1.4620 at 40°C, which are preferable under as given in application programming interfaces (Ayurvedic Pharmacopoeia of India) references for cow ghee samples [Table 1].
Table 1: Results of physico‑chemical parameters for cow's ghee

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Organoleptic characters of raw materials both the KMM and KMF show black color, disagreeable in odor, bitter in taste and show fine powder texture [Table 2].
Table 2: Organoleptic characters of raw materials

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Organoleptic characters of finished products show that both KSJM and KSJF show black color, disagreeable in odor, smooth in touch and shows black oily appearance. While both OM and OF shows black color, disagreeable in odor, smooth in touch and shows black paste like appearance [Table 3].
Table 3: Organoleptic characters of finished products

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Physico-chemical characters of KMM show more percentage of acid insoluble ash in comparison to KMF while KMF shows more percentage of loss on drying and ash value in comparison to KMM. Both raw materials show nearly 7 pH values. KMM shows more alcohol soluble and petroleum Ether soluble extractive values than KMF. KMF shows more water soluble extractive values than KMM [Table 4].
Table 4: Results of physico‑chemical parameters for Kesha masi samples

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Physico-chemical parameters of cow's ghee and finished products (KSJM and KSJF) were performed, where KSJM shows more acid value, saponification value, iodine value and ester value in comparison to KSJF. While KSJF shows more peroxide value in comparison to KSJM. OM shows more acid value, saponification value, iodine value and ester value in comparison to OF and OF shows more peroxide value than OM to similar Keshanjana drugs. Cow ghee shows all values, which are preferable under as given in Ayurvedic pharmacopoeia of India references for cow's ghee samples. Cow ghee shows negative result to rancidity test. Hence, it is considerable for analysis [Table 5].
Table 5: Results of physico‑chemical parameters for cow ghee and finished products

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Microbial study

Determinations of microorganism of the samples were carried out and the result of the same has been presented. Cow ghee shows all the results of total microbial count and pathogens in aerobic and anaerobic culture are under as limit as per World Health Organization (WHO) safety guideline [8] [Table 6].
Table 6: Results of microbial study of cow ghee in aerobic and anaerobic culture

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Determination of microorganism and the bacteriological examination of the samples were carried out and it is found that finished products of OM, OF, KSJM and KSJF show total microbial count and pathogens in culture are under the limit mentioned in WHO safety guideline [8] [Table 7].
Table 7: Results of microbial study

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Particle size analysis

The distribution curve of particle seemed near to skewness as observed in KMM is 60 μm to 300 μm and in KMF is 60 μm to 400 μm range. The operational confirmation and force may be modified during method validation stage [Table 8].
Table 8: Observations of particle size distribution test

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ICP spectroscopy study

ICP analysis show the presence of lead in all the samples, OM and OF shows lead in prescribed limits. While KSJM and KSJF show lead above prescribed limits. Zinc is present in OM, OF and KSJM while it is absent in KSJF. In OM zinc is present more than prescribed limits while in OF and KSJM shows zinc in prescribed limits. Arsenic is present in KSJF above the prescribed limits. Cadmium and mercury are not detected in any samples. So, finished products are free from these elements [Table 9].
Table 9: ICP analysis of samples

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HPTLC study

In the chromatogram of unsaponifiable matter of cow ghee and formulated finished products; R f values of spots were noted under different conditions. Then all tracks were scanned under 254 nm and 366 nm and also post chromatographic derivation through spray methanolic sulphuric acid and R f are mentioned in the table. Different tracks are represented at the wavelength of 254 nm and 366 nm [Table 10] and [Figure 1].
Figure 1: 3D image of all tracks at 366 nm

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Table 10: Rf values of unsaponifiable matter of cow's ghee, raw materials and finished products

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   Discussion Top


The parameters studied such as saponification value, iodine value, acid value, ester value, peroxide value, specific gravity, refractive index can be considered as standard parameters. Rancidity is an important qualitative parameter to check the quality of ghee sample. Since Bhaishajya of ghee always categorized in to three level of heat Mridu, Madhyama and Khara in short Ghrita preparations when prepared is heated as suggested in the drug regime in case of KSJ no other drug added in preparation. During the process of preparation, the analytical values of raw ghee are expected to undergo alterations. Hence, a detailed analytical study was carried out. Further to rule out the influence of extraneous factors on the expression of pharmacological activity and for safety concerns WHO quality control criteria included heavy metal. Hence in this study, the test samples were analyzed for the presence of heavy meals also.

The analysis of raw materials included organoleptic parameters such as consistency, color, taste and odor, physico-chemical parameters such as loss on drying, total ash, acid-insoluble ash, water soluble ash, alcohol soluble extractive values, water soluble extractive values and petroleum ether soluble extractive values were also carried out. Particle size analysis has done to check the uniformity of raw materials.

The analysis of finished products included the organoleptic parameters such as consistency, color, taste and odor, determination of pH, acid value, iodine value, saponification value, ester value, peroxide value, estimation of unsaponifiable matter. In addition the HPTLC fingerprinting was also carried out, to check the presence of more unsaponifiable matter and separation of more components in the sample with HPTLC fingerprinting. All samples were tested for the presence of heavy metals such as mercury, lead, cadmium, zinc and arsenic. Microbial study has also done to check the absence of pathogens, which are harmful in finished products.

All these qualitative parameters can be taken for good manufacturing of these preparations for APIs and other standard reference.

 
   References Top

1.Vagbhata, Astanga Sangrha uttara tantra 20/2. edited by Prasad SS, 2 nd ed. Chaukhamba Sanskrit Series, Varanasi, 2008; 727.  Back to cited text no. 1
    
2.Vardhan P, Dhiman KS, Sharma SK. A clinical study on the effect of Keshanjana and Parisheka in the management of Sushkakshipaka w.s.r. to Dry Eye Syndrome. Shimla: HPU; 2005.  Back to cited text no. 2
    
3.Vagbhata, Astanga Hridayam, Uttar Tantra, 16/32, edited by Shashtri HS, Chaukhambha Sura Bharati Prakashana, Varanasi, 2002; 832.  Back to cited text no. 3
    
4.Dhiman KS, Vyas M, Prajapati PK. Standard operational procedures of Keshanjana. Int J Ayu Med 2012;3:152-5.  Back to cited text no. 4
    
5.Anonymous. Manual of Methods of Analysis of Foods Oils and Fats. New Delhi: Directorate General of Health Services Ministry of Health and Family Welfare, Government of India; 2005.  Back to cited text no. 5
    
6.Chatwal GR, Anand SK. Instrumental Methods of Chemical Analysis. 5 th revised and reprint ed. Mumbai: Himalaya Publishing House; 2005. p. 317.  Back to cited text no. 6
    
7.Dwivedi SK, Dey S. Medicinal herbs: A potential source of toxic metal exposure for man and animals in India. Arch Environ Health 2002;57:229-31.  Back to cited text no. 7
    
8.Anonymous. Quality Control Method for Herbal Materials. Switzerland: WHO Library Cataloguing in Publication Data; 1998. pp. 67-75.  Back to cited text no. 8
    
9.Anonymous. Planner Chromatography, Modern Thin Layer Chromatography. Switzerland, London, New York: Springer, Heidleberg, Dordbrecht, 1999. p. 2-16.  Back to cited text no. 9
    


    Figures

  [Figure 1]
 
 
    Tables

  [Table 1], [Table 2], [Table 3], [Table 4], [Table 5], [Table 6], [Table 7], [Table 8], [Table 9], [Table 10]



 

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