|Year : 2017 | Volume
| Issue : 1 | Page : 1-7
Estimation of guggulsterone E and Z in the Guggul-based commercial formulations using high-performance thin-layer chromatography
Pramod Kumar Sairkar1, Anjana Sharma2, NP Shukla3
1 Centre of Excellence in Biotechnology, M. P. Council of Science and Technology, Bhopal, Madhya Pradesh, India
2 Department of Postgraduate Studies and Research in Biological Science, Bacteriology Laboratory, Rani Durgavati University, Jabalpur, Madhya Pradesh, India
3 Madhya Pradesh Pollution Control Board, Paryavaran Parisar, Bhopal, Madhya Pradesh, India
|Date of Web Publication||15-May-2017|
Pramod Kumar Sairkar
Centre of Excellence in Biotechnology, M.P. Council of Science and Technology, Vigyan Bhavan, Nehru Nagar, Bhopal, Madhya Pradesh
Source of Support: None, Conflict of Interest: None
| Abstract|| |
Background: Guggulsterone (GS) is a plant steroid and bioactive compound present in gum Guggul of Commiphora wightii. An Indian herbal medicine system “Ayurveda” has a long history of use of gum Guggul and plant extract of C. wightii as medicine for the treatment of various illnesses. Complex nature, low availability, and inconsistency of phytoconstituents make its analysis of difficult tasks. Aims: In this work, six different Guggul-based herbal formulations were examined for estimation of GS and their isomers (E and Z) through high-performance thin-layer chromatography technique. Materials and Methods: For that various concentrations of standard E-GS and Z-GS (50 ng–250 ng/spot) with samples (20 μg/spot) were applied on silica gel coated aluminum plate and developed with the mobile phase of toluene: ethyl acetate: formic acid: methanol (6:2:1:0.5). The scanning was performed at 254 nm wavelength and the absorbance (scan) spectrum of E-GS and Z-GS peak was generated at 200 nm–400 nm wavelength range. Results and Conclusions: Rf value and scan spectrum pattern of the samples reveal that they contain either one form of GS (E-GS, Z-GS) or both. The quantity of E-GS and Z-GS within the samples was ranged from 0.230 ± 0.0040–0.926 ± 0.0168% to 0.537 ± 0.0026–0.723 ± 0.0177%, respectively.
Keywords: Absorbance (scan) spectrum, Commiphora wightii, guggulsterone, herbal formulation, high-performance thin-layer chromatography
|How to cite this article:|
Sairkar PK, Sharma A, Shukla N P. Estimation of guggulsterone E and Z in the Guggul-based commercial formulations using high-performance thin-layer chromatography. J Pharm Bioall Sci 2017;9:1-7
|How to cite this URL:|
Sairkar PK, Sharma A, Shukla N P. Estimation of guggulsterone E and Z in the Guggul-based commercial formulations using high-performance thin-layer chromatography. J Pharm Bioall Sci [serial online] 2017 [cited 2017 Jul 23];9:1-7. Available from: http://www.jpbsonline.org/text.asp?2017/9/1/1/206225
| Introduction|| |
For the past few decades, compounds from natural sources have been gaining importance because of their vast chemical diversity, and many of them are being used as a medicine. Herbal-based medicines have traditional value in the curing of various diseases, and now it has become more popular due to its scientifically known pharmacological activity with fewer side effects. With the constant increase in the demand of herbal medicines/formulations worldwide, the safety and quality of herbal raw materials and their products have become a major concern for health authorities as well as pharmaceutical industries. The analysis of phytoconstituents is a difficult task because of their complex nature, usually low availability and inconsistency even within the same plant species. Due to the complex nature of combined/mixed herbal products in comparison to the chemical drug, it needs more attention and various analytical methods can be used for its quality assessment. The WHO has taken a keen interest in the quality control of herbal medicine and recommends the use of high-performance thin-layer chromatography (HPTLC) for the characterization and quality assurance of herbal medicines.
HPTLC is preferred over other analytical method due to its several advantages such as automation, reproducible, fast, simple, many samples, as well as standards simultaneously, cost-effective, and hyphenation., The manufacturing of herbal product after proper quality assurance through HPTLC can enhance the efficiency and value of the herbal product.,
Guggulsterone (GS) (4,17 (20)-pregnadiene-3,16-dione) is a plant polyphenol (steroid) and bioactive compound present in gum Guggul of Commiphora wightii, (family Burseraceae; synonyms: Balsamodendron mukul or Commiphora mukul)., GS occurs in two isomeric forms, namely Z-GS and E-GS (Patil et al., 2015). GS act as antagonist ligands for the bile acid receptor, farnesoid X receptor, and as active ingredients responsible for the hypolipidemic activity., Both GS isomers were demonstrated to suppress lipopolysaccharide-induced inflammation by inhibiting IκB-α degradation and NF-κB activation., In addition, GS was reported to protect cardiac and neuronal damages in animal models. GS has medicinal properties such as anti-inflammatory,, hepatoprotective,, muscle relaxing, hypocholesterolemic and anti-obesity,,, antimycobacterial, antischistomal, larvicidal, and molluscicida.,,,,,, There are several gum Guggul-based standardized formulations are being used as cholesterol-lowering agents and also in the curing of joint pains., The GS is the main active ingredient of gum Guggul which has various pharmacological importance; therefore, it is essential to ensure that the high quality and right quantity of GS in the Guggul-based commercial formulations. In the present study, six different Guggul-based herbal formulations were examined for estimation of GS and their isomers (E and Z) through HPTLC technique.
| Materials and Methods|| |
Preparation of sample and standard
Gum Guggul-based six herbal formulations (Mf-A to Mf-F) of different manufacturers available in the Indian market were used. Tablets of these formulations were crushed and properly dissolved in methanol (10 μg/μl). The mixture was centrifuged at 10,000 rpm for 5 min, and the supernatant was used as a sample. Standard E-GS and Z-GS (Chromadex, USA) were prepared separately in methanol with concentrations of 50 ng/μl and 100 ng/μl.
Development of high-performance thin-layer chromatography plate
The samples were applied using Linomet 5 (CAMAG) as a form of 7 mm long bands on prewashed (with methanol) 20 cm × 10 cm, 200 μm layered silica gel 60 F254 coated aluminum HPTLC plate (Merck) using the stream of neutral gas (Nitrogen) at a constant rate of 150 nl/s. Since the standardization of E-GS and Z-GS estimation, two different amount standards (200 and 400 ng/spot) and samples (20 and 40 μg/spot) were applied to the plate, while in the estimation process, standards were applied in the concentration varied from 50 ng to 250 ng/spot and 20 μg/spot samples were applied in two replicates.
After sample application, plates were developed in CAMAG Automatic Developing Chamber 2 with the mobile phase of toluene: ethyl acetate: formic acid: methanol (6:2:1:0.5) solution. The developing chamber was saturated with mobile phase (20 ml) for 20 min, and 10 ml mobile phase was allowed to migrate up to 70 mm distance on the plate at the room temperature under controlled humid condition (60–80% with NaCl solution).
Estimation of E-guggulsterone and Z-guggulsterone
The developed plate was scanned in CAMAG TLC scanner 3 at 254 nm wavelength in deuterium lamp (D2) with 6.00 mm × 0.45 mm slit dimension, 10 mm/s scanning speed, and 100 μm/step data resolution. The scan spectrum of E-GS and Z-GS peak was generated through scanning at a wavelength range of 200 nm–400 nm with slit dimension of 6.00 mm × 0.10 mm.
The presence of E-GS and Z-GS was assessed only in the samples of which scan spectrum was similar to standard E-GS and Z-GS. The quantity of E-GS and Z-GS was estimated through the calibration curve between standard E-GS and Z-GS concentration and their peak area using WinCATs Planner Chromatography manager software (CAMAG) Switzerland Excel (Microsoft Office 10) USA. The E-GS and Z-GS content (percentage) in the samples were calculated using following formula:
| Results|| |
Presence of E-guggulsterone and Z-guggulsterone in the sample
Standard E-GS and Z-GS had 0.43 and 0.48 Rf value. According to these Rf values, E-GS was present in all the samples except sample Mf-E, while Z-GS was present in all samples. The Rf of E-GS on the sample Mf-A, Mf-B, Mf-C, Mf-D, and Mf-F were ranged from 0.43 to 0.44, and the Rf of the Z-GS in all samples were ranged from 0.48 to 0.52 [Table 1] and [Figure 1] and [Figure 2]. The pattern of scan spectra and λmax of all the samples (λmax 249–250 nm) were almost similar to the standard E-GS (λmax of 250 nm) except sample A (λmax of 213 nm), similarly the scan spectra and λmax of all the samples (λmax of 251 and 257) and standard Z-GS (λmax of 250) had nearly same except sample Mf-F (λmax of 235) [Table 1] and [Figure 3] and [Figure 4].
|Table 1: Rf and λmax of E-guggulsterone and Z-guggulsterone in the standard and sample|
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|Figure 1: Peak of guggulsterone E; (a) in the standard; (b) in the sample|
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|Figure 2: Peak of guggulsterone Z; (a) in the standard; (b) in the sample|
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Quantification of guggulsterone
In the estimation of E-GS and Z-GS, the respective linear calibration curves were plotted with five different concentrations of standard E-GS and Z-GS (50–200 ng) and their peak area. The correlation coefficient (r2) of E-GS and Z-GS calibration curve was 0.9979 and 0.9991, while the residual standard deviation of the standard point (sdv) was 3.142 and 2.1873, respectively. All the samples contain GS, either in the form of E, Z, or both. Sample Mf-A and sample Mf-E had only Z-GS with 0.590 ± 0.0205 and 0.723 ± 0.0177%, respectively, while sample F had only 0.926 ± 0.0168% GS in the form of E trans isomer. Both forms of GS were present in the sample Mf-B, Mf-C, and Mf-D with 0.382 ± 0.0025, 0.230 ± 0.0040, and 0.240 ± 0.0206% E-GS and 0.573 ± 0.0028, 0.537 ± 0.0026, and 0.245 ± 0.0034% Z-GS, respectively [Table 2] and [Figure 5] and [Figure 6].
| Discussion|| |
Traditional medicinal plants are being used in curing the diseases, particularly in remote parts of the developing countries having few health facilities. However, herbal-based medicine is now becoming popular in the developed countries  might because medicinal plants are the best source to obtain a variety of drugs due to its various advantages over other drugs. The use of herbal medicine in the form of formulations/extract would have certain drawbacks because the inconsistent amount of main constituents in the raw material, therefore, raw materials should be used in the drug formulation after proper investigation of their properties, safety, and efficacy. Based on traditional knowledge of herbal medicines, its scientific validation using standardized techniques may be a safe path of reverse pharmacology approach. This may play a vital role in drug discovery, development, and therapeutics.
Guggulipid extract has been used in Ayurveda medicine to treat various diseases such as dyslipidemia, inflammation, and arthritis and also have cholesterol-lowering properties and thyroid-stimulating activity. The cis and trans GS (E-GS and Z-GS) are the main ingredients in guggulipid and preliminary studies indicate that the medicinal value of guggulipid extract is due to these isomers of GS.,,, Looking the necessity of quantitative estimation of GS in herbal medicine, six Ayurveda formulations were undertaken using HPLTC because HPTLC was found to be specific, reproducible, and precise for the estimation of E-GS and Z-GS., This estimation process would reveal the presence of either both the isomers of GS or only one isomer of GS in the observed herbal formulations, which subsequently leads the authentication of proper pharmaceutical doses of GS.,
For HPTLC method development, the suitable stationary and mobile phase is the primary requirement. Silica gel 60 F254-coated aluminum plate as a stationary phase used in more than 80% of HPTLC analysis for the quantification of compound. In context with mobile phase, various combinations of organic solvents were used for GS estimation like light petroleum: ethyl acetate (3:1), n-hexane: chloroform: ethyl acetate: methanol in (10:3:3:1), toluene: acetone with the ratio of 9:1,, and 9.3:0.7 and petroleum ether: ethyl acetate: methanol (6:2:0.5). In addition to these combinations of solvent toluene: ethyl acetate: formic acid: methanol (6:2:1:0.5) solution is preferred for quantitative estimation of GS in the herbal formulations.
Both the isomers of GS absorb the same wavelength of light, which was 250 nm in the present study. Thus, for the quantitative estimation of GS isomers, it is necessary that they should be discriminated through their Rf. The used mobile phase properly separates these isomers with Rf value of 0.35 and 0.42 for E-GS and Z-GS, respectively. However, the presence of compound within the samples is also determined by the pattern of the absorbance spectrum (scan spectrum). The absorbance spectrum of the sample has to be similar to the standard of the compound analyzed and the purity of the absorbance spectrum band was checked by overlaying both the absorbance spectrum band at start, middle, and end position. Using these criteria, GS was found in the all six samples, while the both forms of GS were present only in four samples.
The correlation coefficient (r2) near to 0.999 and sdv lesser than 5 is better for the quantification of the compound through the linear calibration curve. The regression analysis data for the calibration plots for E-GS and Z-GS showed a linear relationship with a correlation coefficient (r2) of 0.9979 and 0.9991 and sdv of 3.142 and 2.187, respectively, showed that the quantity of GS within the samples would be precise and accurate. The content of E-GS and Z-GS was estimated in the ayurvedic medicine, which reveals that in the sample, the content of E-GS form was varied from 0.230 ± 0.0040 to 0.926 ± 0.0168%, while the content of Z-GS form varied from 0.245 ± 0.0034 to 0.723 ± 0.0177%, which was near about to earlier studies upon the herbal formulations such as Rakta Dosh Nashak vati  and Jivitprada vati.
High-performance liquid chromatography (HPLC) was used by many researchers for estimation of E-GS and Z-GS in blood serums, commercial formulations, plant materials, and health supplements; reversed-phase liquid chromatographic method was used for the estimation of E-GS and Z-GS in rate blood plasma  and human blood serum.,, Reversed-phase HPLC and HPLC methods successfully used for the estimation of GSs E and Z in many commercial formulations, i.e., in nine commercial formulations available in the US market, in Guggul formulations, in nanoemulsion formulation and Habb-e-Khardal Unani tablet, in a polyherbal formulation containing C. wightii extracts, in polyherbal formulation, Yogaraja Guggul tablets, and in Ariflex tablet formulation. This method was also used for the estimation of E-GS and Z-GSs in plant materials,, as well as in vitro generated plant callus.,
The authors wish to thank Prof. Pramod K. Verma, Director General, MPCST and Dr. R.K. Singh, Resource Scientist and Group Head, Advance Research Instrumentation Facility, MPCST for the providing laboratory facilities.
Financial support and sponsorship
Conflicts of interest
There are no conflicts of interest.
| References|| |
Hutchings MR, Athanasiadou S, Kyriazakis I, Gordon IJ. Can animals use foraging behaviour to combat parasites? Proc Nutr Soc 2003;62:361-70.
Liang YZ, Xie P, Chan K. Quality control of herbal medicines. J Chromatogr B Analyt Technol Biomed Life Sci 2004;812:53-70.
World Health Organization. Guidelines on Good Manufacturing Practices (GMP) for Herbal Medicines. Geneva: World Health Organization; 2008.
Jaiswal Y, Tatke P, Gabhe S, Vaidya A. Rapid high performance thin layer chromatographic method for quantitation of catechin from extracts of cashew leaves – A short report. Pol J Food Nutr Sci 2013;63:49-54.
Mahady GB, Fong HH, Farnsworth NR. Botanical Dietary Supplements: Quality, Safety and Efficacy. Lisse, the Netherlands: Swets and Zeitlinger Publishers; 2001. p. 283-9.
Mohammad A, Bhawani SA, Sharma S. Analysis of herbal products by thin-layer chromatography: A review. Int J Pharma Bio Sci 2010;1:1-50.
Shen T, Lou HX. Bioactive constituents of myrrh and frankincense, two simultaneously prescribed gum resins in Chinese traditional medicine. Chem Biodivers 2008;5:540-53.
Urizar NL, Moore DD. Gugulipid: A natural cholesterol-lowering agent. Annu Rev Nutr 2003;23:303-13.
Urizar NL, Liverman AB, Dodds DT, Silva FV, Ordentlich P, Yan Y, et al.
A natural product that lowers cholesterol as an antagonist ligand for FXR. Science 2002;296:1703-6.
Wu J, Xia C, Meier J, Li S, Hu X, Lala DS. The hypolipidemic natural product guggulsterone acts as an antagonist of the bile acid receptor. Mol Endocrinol 2002;16:1590-7.
Meselhy MR. Inhibition of LPS-induced NO production by the oleogum resin of Commiphora wightii
and its constituents. Phytochemistry 2003;62:213-8.
Shishodia S, Aggarwal BB. Guggulsterone inhibits NF-kappaB and IkappaBalpha kinase activation, suppresses expression of anti-apoptotic gene products, and enhances apoptosis. J Biol Chem 2004;279:47148-58.
Kaul S, Kapoor NK. Reversal of changes of lipid peroxide, xanthine oxidase and superoxide dismutase by cardio-protective drugs in isoproterenol induced myocardial necrosis in rats. Indian J Exp Biol 1989;27:625-7.
Gujral ML, Sareen K, Tangri KK, Amma MK, Roy AK. Antiarthritic and anti-inflammatory activity of gum guggul (Balsamodendron mukul
Hook). Indian J Physiol Pharmacol 1960;4:267-73.
Francis JA, Raja SN, Nair MG. Bioactive terpenoids and guggulusteroids from Commiphora mukul
gum resin of potential anti-inflammatory interest. Chem Biodivers 2004;1:1842-53.
Al-Howiriny TA, Al-Sohaibani MO, Al-Said MS, Al-Yahya MA, El-Tahir KH, Rafatullah S. Hepatoprotective properties of Commiphora opobalsamum
(“Balessan”), a traditional medicinal plant of Saudi Arabia. Drugs Exp Clin Res 2004;30:213-20.
Al-Howiriny T, Al-Sohaibani M, Al-Said M, Al-Yahya M, El-Tahir K, Rafatullah S. Effect of Commiphora opobalsamum
(L.) Engl. (Balessan) on experimental gastric ulcers and secretion in rats. J Ethnopharmacol 2005;98:287-94.
Allam AF, el-Sayad MH, Khalil SS. Laboratory assessment of the molluscicidal activity of Commiphora molmol
(Myrrh) on Biomphalaria alexandrina
, Bulinus truncatus
and Lymnaea cailliaudi
. J Egypt Soc Parasitol 2001;31:683-90.
Tripathi SN, Sastri VV, Satyavati GV. Experimental and clinical studies on the effects of guggul (C. mukul
) in hyperlipidemia and thrombosis. J Res Indian Med 1968;2:10.
Satyavati GV, Dwarakanath C, Tripathi SN. Experimental studies on the hypocholesterolemic effect of Commiphora mukul
. Engl. (guggul). Indian J Med Res 1969;57:1950-62.
Bhatt AD, Dalal DG, Shah SJ, Joshi BA, Gajjar MN, Vaidya RA, et al.
Conceptual and methodologic challenges of assessing the short-term efficacy of guggulu in obesity: Data emergent from a naturalistic clinical trial. J Postgrad Med 1995;41:5-7.
] [Full text]
Kumar S, Shankar V. Medicinal plants of Indian desert: Commiphora wightii
(Arnott) Bhand. J Arid Environ 1982;5:1-11.
El Ashry ES, Rashed N, Salama OM, Saleh A. Components, therapeutic value and uses of myrrh. Pharmazie 2003;58:163-8.
Abbas FA, Al-Massarany SM, Khan S, Al-Howiriny TA, Mossa JS, Abourashed EA. Phytochemical and biological studies on Saudi Commiphora opobalsamum
L. Nat Prod Res 2007;21:383-91.
Massoud AM, El Ebiary FH, Abd El Salam NF. Effect of myrrh extract on the liver of normal and bilharzially infected mice. An ultrastructural study. J Egypt Soc Parasitol 2004;34:1-21.
Shen T, Wan W, Yuan H, Kong F, Guo H, Fan P, et al.
Secondary metabolites from Commiphora opobalsamum
and their antiproliferative effect on human prostate cancer cells. Phytochemistry 2007;68:1331-7.
Satyavati GV. Use of plant drugs in Indian traditional systems of medicine and their relevance to primary health care. In: Wagner H, Farnsworth NR, editors. Economic and Medicinal Plant Research. London: Academic Press; 1990. p. 39-56.
Goyal C, Ahuja M, Sharma SK. Preparation and evaluation of anti-inflammatory activity of gugulipid-loaded proniosomal gel. Acta Pol Pharm 2011;68:147-50.
Szapary PO, Wolfe ML, Bloedon LT, Cucchiara AJ, DerMarderosian AH, Cirigliano MD, et al.
Guggulipid for the treatment of hypercholesterolemia: A randomized controlled trial. JAMA 2003;290:765-72.
Zhu N, Rafi MM, DiPaola RS, Xin J, Chin CK, Badmaev V, et al.
Bioactive constituents from gum guggul (Commiphora wightii
). Phytochemistry 2001;56:723-7.
Mouli KC, Rao SD. Phytoresources as potential therapeutic agents for cancer treatment and prevention. J Glob Pharm Tech 2009;1:4-18.
Hasan MK, Kabir AK, Mistr S. Chemical and biological investigation of leaves of Polygonum plebejum.
S J Pharm Sci 2009;2:66-71.
Vaidya AD, Vaidya RA, Nagral SI. Ayurveda and a different kind of evidence: From Lord Macaulay to Lord Walton (1835 to 2001 AD). J Assoc Physicians India 2001;49:534-7.
Tripathi YB, Malhotra OP, Tripathi SN. Thyroid stimulating action of Z-guggulsterone obtained from Commiphora mukul
. Planta Med 1984;50:78-80.
Tripathi YB, Tripathi P, Malhotra OP, Tripathi SN. Thyroid stimulatory action of (Z)-guggulsterone: Mechanism of action. Planta Med 1988;54:271-7.
Chander R, Khanna AK, Kapoor NK. Lipid lowering activity of guggulsterone from Commiphora mukul
in hyperlipaemic Rats. Phytother Res 1996;10:508-11.
Beg M, Singhal KC, Afzaal S. A study of effect of guggulsterone on hyperlipidemia of secondary glomerulopathy. Indian J Physiol Pharmacol 1996;40:237-40.
Jariwala JK, Saluja AK, Anajwala CC, Dakhara SL. Simultaneous estimation of guggulsterone E & Z and tinosporaside in jivitprada vati by HPTLC method. Int J Green Pharm 2011;5:113-7. [Full text]
Shelke NN, Kumar VV, Bhope SG, Jadhav RB. Simultaneous HPTLC analysis of E-guggulsterone, Z-guggulsterone, 11-keto-β-boswellic acid, and 3-acetyl-11-keto-β-boswellic acid in an anti-arthritic formulation. Journal of Planar Chromatography - Modern TLC 2011;24:242-7.
Dhalwal K, Biradar YS, Shinde VM, Mahadik KR, Rajani M. Phytochemical evaluation and validation of a polyherbal formulation using HPTLC. Pharmacogn Mag 2008;4:89-95.
Roy SK. TLC separation and quantitative determination of guggulsterones. Indian J Pharm Sci 1989;11:251-3.
Agrawal H, Kaul N, Paradkar AR, Mahadik KR. HPTLC method for guggulsterone. I. Quantitative determination of E- and Z-guggulsterone in herbal extract and pharmaceutical dosage form. J Pharm Biomed Anal 2004;36:33-41.
Agrawal H, Kaul N, Paradkar AR, Mahadik KR. HPTLC method for guggulsterone. II. Stress degradation studies on guggulsterone. J Pharm Biomed Anal 2004;36:23-31.
Kulhari A, Sheorayan A, Saxena N, Mohan C, Mangal M, Chaudhury A, et al
. HPTLC analysis of guggulsterone isomers in Commiphora wightii
(Arn.) Bhandari: An endangered oleo-gum resin species heading towards extinction. Genet Resour Crop Evol 2013;60:1173-80.
Musharraf SG, Iqbal N, Gulzar U, Ali A, Choudhary MI, Atta-Ur-Rahman. Effective separation and analysis of E- and Z-guggulsterones in Commiphora mukul
resin, guggulipid and their pharmaceutical product by high performance thin-layer chromatography-densitometric method. J Pharm Biomed Anal 2011;56:240-5.
Indrayanto G. Analytical aspects of high performance thin layer chromatography. In: Srivastava MM, editor. High-Performance Thin-Layer Chromatography (HPTLC). Berlin Heidelberg: Springer-Verlag; 2011. p. 179-202.
Jain S, Kumar V, Aina, Singh R. Development and validation of HPLC-UV method for the determination of E and Z gugglesterone in rat plasma: Application to plasma protein binding study. World J Pharm Res 2014;3:1616-27.
Sane RT, Bhate VR, Malkar VB, Nayak VG, Kothurkar RM. Reversed-phase high performance liquid chromatography determination of guggulsterons from pharmaceutical preparation and from blood plasma. Indian Drugs 1990;28:86-9.
Singh SK, Verma N, Gupta RC. Sensitive high-performance liquid chromatographic assay method for the determination of guggulsterone in serum. J Chromatogr B Biomed Appl 1995;670:173-6.
Verma N, Singh SK, Gupta RC. Simultaneous determination of the stereoisomers of guggulsterone in serum by high-performance liquid chromatography. J Chromatogr B Biomed Sci Appl 1998;708:243-8.
Nagarajan M, Waszkuc TW, Sun J. Simultaneous determination of E- and Z-guggulsterones in dietary supplements containing Commiphora mukul
extract (guggulipid) by liquid chromatography. J AOAC Int 2001;84:24-8.
Gottumukkala AL, Ramakrishna T, Babu SK, Subbararaju GV. Determination of E & Z- guggulsterons in guggul formulations by high performance liquid chromatography. Asian J Chem 2005;17:1789-93.
Kamal YT, Mohammed Musthaba S, Singh M, Parveen R, Ahmad S, Baboota S, et al.
Development and validation of HPLC method for simultaneous estimation of piperine and guggulsterones in compound Unani formulation (tablets) and a nanoreservoir system. Biomed Chromatogr 2012;26:1183-90.
Akhade MS, Agrawal PA, Laddha KS. Development and validation of RP-HPLC method for simultaneous estimation of Picroside I, Plumbagin, and Z-guggulsterone in tablet formulation. Indian J Pharm Sci 2013;75:476-82.
] [Full text]
Agrawal P, Vegda R, Laddha K. Simultaneous estimation of withaferin A and Z-guggulsterone in marketed formulation by RP-HPLC. J Chromatogr Sci 2015;53:940-4.
Patil P, Nipanikar S, Bhusnar H, Nagore D. Simultaneous estimation of guggulsterone E, guggulsterone Z, KBA, AKBA, withaferin A and 6-gingerol by using HPLC-DAD method from ariflex tablet formulation. Int J Pharm Sci Drug Res 2015;7:89-95.
Mesrob B, Nesbitt C, Misra R, Pandey RC. High-performance liquid chromatographic method for fingerprinting and quantitative determination of E- and Z-guggulsterones in Commiphora mukul
resin and its products. J Chromatogr B Biomed Sci Appl 1998;720:189-96.
Musharraf SG, Iqbala N, Ahmeda MA, Mazhara S, Choudharya MI. Screening of E- and Z-guggulsterones in the gum-resin exudates of some common plants and method validation in raw, extracted, and pharmaceutical formulations of Commiphora Mukul
by HPLC. J Liq Chromatogr Relat Technol 2011b; 34:2103-17.
Mishra SK, Kumar A. Biosynthesis of guggulsterone in the callus culture of Commiphora wightii
. Arnott. Bhandari (Burseraceae
). Res J Pharm Biol Chem Sci 2010;1:35-41.
Mathur M, Jain AK, Dass S, Ramawat KG. Optimization of guggulsterone production in callus cultures of Commiphora wightii
(Arnott.) Bhandari. Indian J Biotechnol 2007;6:525-31.
[Figure 1], [Figure 2], [Figure 3], [Figure 4], [Figure 5], [Figure 6]
[Table 1], [Table 2]