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 Table of Contents  
ORIGINAL ARTICLE
Year : 2022  |  Volume : 7  |  Issue : 1  |  Page : 47-54

Comparative shelf-life study of Raw Guggulu (Commiphora wightii oleo-gum resin) and Shodhita Guggulu (cow urine processed C. wightii oleo-gum resin)


1 Department of Ayurveda, Central Ayurveda Research Institute, Jhansi, Uttar Pradesh 284003, India
2 Department of Chemistry, Central Ayurveda Research Institute, Jhansi, Uttar Pradesh 284003, India
3 Department of Microbiology, Central Ayurveda Research Institute, Jhansi, Uttar Pradesh 284003, India
4 Department of Chemistry, Central Council for Research in Ayurvedic Sciences, New Delhi, India
5 Department of Ayurveda, Central Council for Research in Ayurvedic Sciences, New Delhi, India

Date of Submission26-Jan-2022
Date of Decision03-Feb-2022
Date of Acceptance06-Feb-2022
Date of Web Publication07-Jul-2022

Correspondence Address:
Dr. Vijay Kumar
Department of Chemistry, Central Ayurveda Research Institute, Jhansi, Uttar Pradesh 284003
India
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/jdras.JDRAS_14_22

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  Abstract 

BACKGROUND: Shelf-life is an important aspect of raw as well as finished drugs. Recently, shelf-life parameters have been included in the Ayurvedic Pharmacopeia of India. The objective of this article is to evaluate and compare the long-term shelf-life study of Raw Guggulu (RG) (C. wightii oleo-gum resin) and Shodhita Guggulu (SG) (cow urine processed C. wightii oleo-gum resin). MATERIALS AND METHODS: A stability chamber with environmental conditions 30°C ± 2°C/60% ± 5% RH was used for long-term shelf-life study of RG and SG. Physico-chemical parameters such as loss on drying, pH, total ash, acid-insoluble ash, water extractive value, and alcohol extractive value were tested at regular frequency (0th, 3rd, 6th, 9th, and 12th month). RESULTS: Significant changes have been observed w.r.t. analysis of physico-chemical parameters. Physico-chemical parameters revealed that the shelf-life of RG ranged from 42 to 66 months and shelf-life of SG ranged from 39 to 62 months. CONCLUSION: The average shelf-life of RG and SG was found out to be 55.16 and 48.16 months, respectively. These studies may help to understand the shelf-life of various ayurvedic formulations of RG and SG.

Keywords: API, Commiphora wightii, resin, shelf-life, Shodhita Guggulu


How to cite this article:
Charde V, Jagtap C, Kumar V, Kushwaha V, Grewal J, Mishra SK, Shakya SK, Soni H, Singh G, Babu G, Singh A, Singh R, Khanduri S, Sharma BS, Srikanth N. Comparative shelf-life study of Raw Guggulu (Commiphora wightii oleo-gum resin) and Shodhita Guggulu (cow urine processed C. wightii oleo-gum resin). J Drug Res Ayurvedic Sci 2022;7:47-54

How to cite this URL:
Charde V, Jagtap C, Kumar V, Kushwaha V, Grewal J, Mishra SK, Shakya SK, Soni H, Singh G, Babu G, Singh A, Singh R, Khanduri S, Sharma BS, Srikanth N. Comparative shelf-life study of Raw Guggulu (Commiphora wightii oleo-gum resin) and Shodhita Guggulu (cow urine processed C. wightii oleo-gum resin). J Drug Res Ayurvedic Sci [serial online] 2022 [cited 2022 Dec 7];7:47-54. Available from: http://www.jdrasccras.com/text.asp?2022/7/1/47/350050




  Introduction Top


Commiphora wightii (Arn.) Bhandari belongs to the plant family Burseraceae. It produces a fragrant oily resin known as Guggulu. Guggulu, in its raw form “Raw Guggulu” and purified form “Shodhita Guggulu,” has been used in various Ayurvedic and traditional medicines. Raw Guggulu (RG) is in demand worldwide including Europe, Africa, and America because of its significant medicinal values.[1],[2] In fact, the demand for Guggulu has been more than its production. Due to overexploitation, the World Conservation Union listed C. wightii (Arn.) Bhandari in IUCN Red list of threatened plants.[1],[2] In recent years, the Indian government and National Medicinal Plants Board (NMPB) have done a tremendous job for the cultivation of C. wightii (Arn.) Bhandari. Guggulu is an important Ayurvedic drug used in various herbal formulations.[3]Guggulu has been used against obesity, hyperlipidemia, inflammation, cholesterol, and cardiovascular diseases. Ketosteroids (E-guggulsterone and Z-guggulsterone) are considered as major phytochemicals responsible for its biological activities.[2],[3],[4],[5],[6],[7],[8],[9],[10]

In classical literatures, it has been reported that potency of Guggulu increases with time.[3] Shelf-life is termed as “Virya,” which means potency of any drug.[11] Shelf-life and expiry date are two different concepts, but these terms are interrelated to each other. Expiry of any drug depends on the shelf-life of the product.[11] Shelf-life concept has been in practice in classical literatures since 12th century AD.[11] The International Conference on Harmonisation (ICH) has issued the shelf-life guidelines to maintain the safety regulations.[3],[12],[13],[14],[15],[16] Recently, in 2016, the Indian government amended Drugs and Cosmetics (Amendment) Rules, 1940 to meet the international standards, and shelf-life of various formulations has been introduced in the Ayurvedic Pharmacopeia of India (API).[17] Basic parameters that are included in the API for shelf-life studies are testing of physico-chemical parameters and biological testing. Finding the shelf-life of traditional or herbal drugs is a tedious work. Shelf-life of a polyherbal formulation [tablet (vatti), powder (churna), etc.] with three or more ingredients depends on various factors. Even a single contaminated ingredient (contamination like presence of microbial load or aflatoxins or pesticides or heavy metals) can spoil the whole formulation. While finding the shelf-life of herbal drugs, microbial contamination (microbial load or aflatoxins) is a major concern. Environmental parameters such as temperature and humidity, storage, handling, transportation, etc. also play a vital role in the shelf-life of any product.[18],[19],[20],[21]

There is no published report on the shelf-life study of RG and Shodhita Guggulu (SG), till date. The aim of the present study is to evaluate the comparative long-term shelf-life study of RG and SG.

Materials and Methods

The authentic material of RG was collected from Rajasthan, India by the Botanist of Regional Ayurveda Research Institute, Jaipur. SG was prepared in the in-house pharmacy of Central Ayurveda Research Institute, Jhansi as per the procedure mentioned under Ayurvedic Formulary of India (AFI). Standards of metal ions lead (Pb), cadmium (Cd), arsenic (As), and mercury (Hg) of analytical grade (AR) and purity >98% were purchased from Merck Ltd India. Agar (MacConkey, Bismuth sulfite, Mannitol Salt, Soyabean Casein, etc.) and standard chloramphenicol of AR grade were purchased from HiMedia. Microorganisms with MTCC numbers were purchased from CSIR—Institute of Microbial Technology Chandigarh, India.

Long-term shelf-life studies

Well-dried RG and SG samples were packed in seal packs in December 2020 and kept in food grade HDPE containers. The containers were placed in the stability chamber for long-term shelf-life studies at 30°C ± 2°C/60% ± 5% RH.[13] A control sample was kept in the deep freezer at 0°C.

Quality control parameters assessment

Quality control parameters such as loss on drying (LOD), pH, total ash (TA), acid-insoluble ash (AIA), water extractive value (WEV), and ethanol extractive value (AEV) were performed at regular frequency (0th, 3rd, 6th, 9th, and 12th month) as per the protocols mentioned under the API.[22],[23],[24]

Kinetic analysis

The results of quality control parameters were fitted for the first-order kinetic model. By plotting log Ct vs. time (t), a straight line was observed:





For 10% degradation time, t0.9 (at Ct=0.9C0) was calculated from the first-order kinetic model. The time required for 10% decomposition is as:





Here, rate constant (kobs) is the slope obtained from the linear plot of log Ct vs. time (t).

Safety and toxicological parameter analysis

Heavy metal analysis

The heavy metals such as Pb, Cd, Hg, and As were analyzed by using the inductively coupled plasma-optical emission spectrometry (ICP-OES) instrument (Agilent 5800, USA). The samples were prepared by using an automatic microwave sample digestor (Microwave 130000 Multiwave GO Plus 50 Hz).

Microbial load and specific pathogens

Microbial assay like microbial load and specific pathogens (Aspergillus niger,  Escherichia More Details coli, Pseudomonas aeruginosa,  Salmonella More Details typhi, and Staphylococcus aureus) were analyzed as per the protocol, according to the limits mentioned in the API.

Pesticides residues analysis and aflatoxin analysis

The analysis of aflatoxins (B1, B2, G1, and G2) and pesticide’s residues (organochlorine, organophosphate, and pyrethroid pesticides) was done by outsourcing testing. The gas chromatography-tandem mass spectrometry-based QuEChERS (quick, easy, cheap, effective, rugged, and safe) procedure was used to analyze the pesticides residues.[25] The quantification of aflatoxins was done by using the high-performance liquid chromatography method.[20]


  Results Top


The results of long-term shelf-life study of RG and SG are mentioned in [Table 1]. In RG, after 12 months, pH, LOD, WEV, AEV, TA, and AIA changed by 4.90%, 4.72%, 4.15%, 5.12%, 6.99%, and 7.19%, respectively, as shown in [Table 1] and [Figure 1]. While in the case of SG, after 12 months, a change of 6.43%, 5.96%, 4.54%, 6.56%, 7.39%, and 7.97% was noticed for pH, LOD, WEV, AEV, TA, and AIA parameters, respectively. After 12 months, significant changes (at P < 0.001) in all the parameters were observed when compared with the initial day (0th month) results.
Table 1: Shelf-life data of RG and SG

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Figure 1: Percent change in parameters with time (months) for shelf-life data of RG and SG. LOD: loss on drying, WEV: water extractive value, EEV: ethanol extractive value, TA: total ash, AIA: acid-insoluble ash

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Safety parameters for both RG and SG were analyzed and mentioned in [Table 2] and [Table 3]. The microbial load (cfu/g) with respect to the total bacterial count (TBC) and total fungal count (TFC) was found to be increased when compared with the initial day (0th month) results [Table 2]. The observed TBC values for RG and SG increased from 65 to 321 cfu/g and from 55 to 267 cfu/g, respectively. Similarly, TFC values for RG and SG also increased from 23 to 122 cfu/g and from 62 to 166 cfu/g, respectively. Specific pathogens, viz. A. niger, E. coli, Salmonella, S. aureus, and P. aeruginosa, were not observed in RG and SG samples throughout the period from 0th to 12th month. The above-mentioned results were quite within the prescribed limits of API as well as WHO. The observed results for aflatoxins B1, B2, G1, and G2 were well within the tolerance limits, as none of the aflatoxins was detected during the whole shelf-life analysis period [Table 2]. Heavy metal and pesticides residues were examined at initial day (0th month) time interval, and no pesticide residue and heavy metal was detected [Table 4]. There are many published reports on heavy metal toxicity and pesticides contaminations of herbs or raw drugs.[20],[21],[25],[26] In the present study, RG was collected from natural source and it could be a reason for the absence of heavy metal toxicity and pesticides residues.
Table 2: Microbial load and test for specific pathogens in Raw Guggulu (RG) and Shodhita Guggulu (SG)

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Table 3: Pesticide residues and heavy metals analysis of Raw Guggulu (RG) and Shodhita Guggulu (SG)

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Table 4: Kinetic study data of Raw Guggulu (RG) and Shodhita Guggulu (SG)

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Kinetic results of the shelf-life study are mentioned in [Table 4] and [Figure 2]. Various kinetic models were examined and finally the first-order kinetic model was applied to the data to obtain a linear plot of log Ct vs. time (t) with regression coefficient r2 >0.95 [Figure 2]. The shelf-life period w.r.t. 10% degradation was calculated. The shelf-life period of RG w.r.t. LOD, pH, WEV, AEV, TA, and AIA parameters was 62, 62, 66, 55, 44, and 42 months, respectively. Similarly, the shelf-life period of SG w.r.t. LOD, pH, WEV, AEV, TA, and AIA parameters was 53, 48, 62, 46, 41, and 39 months, respectively.
Figure 2: Linearity plot log (Ct) vs. time (t in months) for shelf-life data of RG and SG. LOD: loss on drying, WEV: water extractive value, EEV: ethanol extractive value, TA: total ash, AIA: acid-insoluble ash

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  Discussion Top


Recently, work on the shelf-life study of herbal drugs has been reported by quite a few researchers. This highlights the keenness of world community on the shelf-life study of herbal drugs. Few shelf-life reports of ayurvedic drugs such as churna, avaleha, ghrita, kwath churna, and herbo-minerals have been reported.[23],[24],[27],[28],[29],[30],[31],[32],[33],[34],[35] It has been observed that herbal or polyherbal drugs contain a wide class of phytochemicals and act as good inhibitors for microbes and aflatoxins.[23],[24],[27],[28],[29],[30],[31],[32],[33] For the easy understanding of shelf-life, the terms such as “true shelf life,” “estimated shelf life,” “supported shelf life,” “maximum shelf life,” and “labeled shelf life” have been well mentioned in the literature.[35] Few researchers have experimentally claimed that shelf-life and expiry are even 10 years more than the labeled shelf-life and expiry.[36] This shows that potency of the drug does not depend on a single parameter or single phytochemical. The average shelf-life of RG and SG was found out to be 55.16 and 48.16 months, respectively. A relatively smaller shelf-life of SG when compared with RG could be due to experimentations on SG in controlled conditions by using water and cow urine. Usage of water leads to an increase in the moisture content and high microbial load, which leads to a low shelf-life period. It is documented that the recommended media for the Guggulu shodana are gomutra (cow urine), triphala kasaya (decoction of triphala), vasapatra kasaya (decoction of Adhatoda vasica leaves), vasapatra savrasa (aqueous extract of Adhatoda vasica leaves), dugdha (milk), and water.[37]Shodhana or purification is used to remove the foreign matters and to reduce toxicity.[37] There are a few reports available in literatures in which Guggulu shodhana is well mentioned but none of them has studied the effect of purification on shelf-life period of Guggulu. It is the first study in which comparative shelf-life study of RG and SG has been established and reported. To obtain homogeneous results and to enhance the global acceptance of herbal drugs, it is important to follow the same methodology as prescribed by the ICH[38] and the same has been followed by the Government of India. For the better understanding of shelf-life of herbal drugs, the shelf-life of raw drug and its intermediate is an importance aspect, which needs to be covered by the researchers while performing their shelf-life study experiments.


  Conclusions Top


This is the first kinetics-based shelf-life study in which ayurvedic raw drug and its intermediate were assessed. Overall, the first-order kinetic model was fitted to the shelf-life experiments of RG and SG. Good linearity was maintained with r2 value more than 0.95. The average shelf-life of RG and SG was established to be 55.16 and 48.16 months, respectively. These studies may help to understand the shelf-life of various ayurvedic formulations of RG and SG as well the mechanism for their shelf-life period.

Acknowledgments

DG-CCRAS is highly acknowledged for funding and facilities at CARI, Jhansi, Uttar Pradesh, India. CARI, Patiala and RARI, Jaipur are highly acknowledged for supply of Guggulu samples.

Financial support and sponsorship

CCRAS, Ministry of Ayush, New Delhi.

Conflicts of interest

There are no conflicts of interest.

Ethics approval and consent to participate

Not applicable.





 
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    Figures

  [Figure 1], [Figure 2]
 
 
    Tables

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



 

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