GC-MS and HPLC Analysis of Chlorophytum
Borivilianum (Safed Musli) A Plant from
Ayurveda- Herbal Viagra
Ruchi Vyas, Garima Sharma, Adya Chaturvedi, Devki and Rashmi Sisodia*
Department of Zoology, University of Rajasthan, Jaipur, India
Submission: June 03, 2020; Published:June 09, 2020
*Corresponding author:Rashmi Sisodia, Department of Zoology, University of Rajasthan, Jaipur 302004, India
How to cite this article:Ruchi V, Garima S, Adya C, Devki S, Rashmi S. GC-MS and Hplc Analysis of Chlorophytum Borivilianum (Safed Musli) A Plant
from Ayurveda- Herbal Viagra.Glob J Reprod Med. 2020; 7 (5): 5556723. DOI: 10.19080/GJORM.2020.07.555723.
The objective of this study was to determine the photochemical content of Chlorophytum borivilianum by GC-MS analysis and HPLC. Chlorophytum borivilianum is an endangered medicinal plant having several phyto constituents, which makes it suitable for many therapeutic applications. GC-MS analysis was carried out on a Shimadzu GCMSQP2010 Ultra system. Methanol is used as a solvent. The injector temperature was 280°C. The samples were injected in the split mode with split ratio 1 to 60. Injection volume was 1 μl. A capillary column Rtx-5MS (5% Diphenyl-95% Dimethyl Polysiloxane), 30 m x 0.25mm x 0.25 μm, was used. The carrier gas was helium with a constant flow of 1.00 ml/min.GC-MS analysis of drug led to the identification of 9 compounds viz., 5-methylhex-2-yl pentadecyl ester (23.69%), 9,10-Anthracenedione (19.02%), Phthalic acid (18.63%) etc. High-performance liquid chromatography was conducted to enumerate active ingredient in the plant extracts. These analyses help us to understand the importance of various phyto chemicals and also open up new ways to exploit a particular plant for various therapeutic and restorative purposes.
Keywords: Chlorophytum borivilianum; Gas chromatography; Phthalic acid; HPLC
Chlorophytum borivilianum belongs to family Liliaceae Which is an herb found in the tropical wet forests in peninsular India. It is commonly called as Musli or Safed Musli in Hindi language. In the Ayurvedic literature, Safed Musli is celebrated as a Divya Aushad with unparalleled medicinal properties and therapeutic applications. It is considered an elixir to increase the general body immunity and has been reported to possess several biological activities including antimicrobial, anti-inflammatory, antipyretic, hepato protective, antioxidant, hypolipidemic and anti-diabetic  thus also known as ‘white gold’  Its aphrodisiac properties have proved very much useful for the people suffering from Erectile Dysfunction and to increase male potency, it is extensively used as herbal Viagra due to its peculiar and It has sperm atogenic property helps cure impotency as they are rich in glycosides  Safed Musli Which is also gaining increasing acceptance as a vitalizer and health-giving tonic, acurative for pre-natal and post-natal problems, a restorative for immunity-improvement. The recent discovery of Safed Musli as a natural and safe aphrodisiac agent has also opened up new channels of application and new demand for it. Safed Musli is a herb and is also an ideal aphrodisiac with no negative side-effects associated with chemical-based aphrodisiac.
Roots are claimed to be useful to treat oligospermia, pre- and post-natal symptoms, arthritis, diabetes and dysuria . It is a threatened species and named ‘Chlorophytum’ due to the white milky texture of its tubers after peeling. Since ages, tribal communities in India used Safed Musli for enhancing their virility and thus due to its overexploitation it has reached the rare status in nature. It is used for rheumatism and joint pains, increases lactation in feeding mothers, diarrhoea, dysentery, gonorrhoea, leucorrhea etc. Its root contains steroidal and triterpenoid saponins, sapogenins and fructans which act as therapeutic agents and play a vital role in many therapeutic applications. It is a rich source of over 25 alkaloids, vitamins, proteins, carbohydrates, steroids, saponins, potassium, calcium, magnesium, phenol, resins, mucilage, and polysaccharides and also contains a high quantity of simple sugars, mainly sucrose, glucose, fructose, galactose, mannose and xylose . Steroidal saponins, polysaccharides, fructans and fructo oligosaccharides, Beta-sitosterol, stigmasterol, hecogenin and mucilage are major constituents of C. borivilianum that act as therapeutic agents.The in vitro propagated roots were tested for the production of pharmaceutically important secondary metabolites stigmasterol and hecogenin. A maximum of 46.4 mg/gDCW of stigmasterol and 685 mg/gDCW of hecogenin was
evident in these cultures . Gas chromatography has a very wide
field of applications. It is used is in the separation and analysis
of multi-component mixtures such as essential oils, hydrocarbons
and solvents. Intrinsically, with the use of the flame ionization
detector and the electron capture detector (which have very high
sensitivities) gas chromatography can quantitatively determine
materials present at very low concentrations . HPLC is ideally
suited for the rapid processing of the multi component samples
on both an analytical and preparative scale. Chemical separations
can be accomplished using HPLC by utilizing the fact that certain
compounds have different migration rates given a particular
column and mobile phase .
The pure dried root powder of C. borivilianum was purchased
commercially from Naturemed, Hyderabad, AP. Then plant root
extract (CBE) was prepared by mixing 1-5% of plant root powder
with deionized water in a 250 ml of (Borosil, India) conical flask.
The solution was boiled in a water bath and then incubated at 50-
70°C for an hour followed by filtering from What man paper .
GC-MS analysis was carried out on a Shimadzu GCMSQP2010
Ultra system. Methanol was used as a solvent. The injector
temperature was 280°C. The samples were injected in the split
mode with split ratio 1 to 60. Injection volume was 1 μl. A capillary
column Rtx-5MS (5% Diphenyl-95% Dimethyl Polysiloxane),
30 m x 0.25mm x 0.25 μm, was used (Figure 1). The carrier
gas was helium with a constant flow of 1.00 ml/min.The oven
temperature was as follows: initial temperature of 60°C, held for
2 min, increased to 10°C/min up to 260°C and held for 10 min.
The MS ionization potential was 70 eV, and the temperatures
were as follows: interface 260°C, Ion source 280°C. Mass scan
range 40-550. GC-MS analysis of C. Bori vilianum root extract led
to the identification of the following compounds (Table 1). Highperformance
liquid chromatography (HPLC) is a versatile, robust,
and widely used technique for the isolation of natural products
 Currently, this technique is gaining popularity among various
analytical techniques as the main choice for fingerprinting study
for the quality control of herbal plants . Natural products
are frequently isolated following the evaluation of a relatively
crude extract in a biological assay to fully characterize the
active entity. The method followed for High-performance liquid
chromatography was according to Kardani et al.  as published.
HPLC was carried out for detection of active ingredients
in aqueous extracts of C. borivilianum. Gallic acid was used as a
standard marker in this experiment. A peak was observed at 3.08
min retention time (RT), with an area of 2.713 mAU, for standard
Gallic acid. For the test sample, a 20-μl solution injected and ran
for 10 min (Figure 2a-2j). Two prominent peaks were observed,
among which one matched the RT of gallic acid (3.08 min), whereas,
another peak of greater concentration was observed at retention
time 2.5 min. Besides these two peaks, 6 small peaks were also
observed at RT 1.8 min, 3.12 min, 4.7 min, 6 min, 6.8 min and 7.2
min. Considering injected sample without any contamination a
total of 8 active compounds were present in the aqueous solution
of C. borivilianum (Figure 3).
The results of GC-MS analysis led to the identification of
several compounds from the GC fractions of the extract of C.
borivilianum. These compounds were identified through mass
spectrometry attached with GC. The results of the GC-MS was
interpreted by using the database of the National Institute of
Standards and Technology (NIST) library having more than
2,00,000 patterns. The GC-MS spectrum of the unknown
constituent was then compared with the known components
stored in the NIST-14 library. The results of the present study
are tabulated in Table 1. The identified constituents from the
root extract of C. borivilianum like a saponin glycoside 4H-Pyran-
4-one, 2,3-dihydro-3,5-dihydroxy-6-methyl possess antifungal
activity [13,14] 9,10-Anthracenedione possess anticancer activity
, butyl isopropyl ester as an antimicrobial agent  Diethyl
phthalate (DEP) is known as a colourless liquid which has a
slight aromatic odour. It is used in the various applications such
as sprays of insecticides, as an ingredient in coatings of aspirin,
cosmetics, food and pharmaceutical packaging, preparation
of skincare products [17,18] 2-fluorenecarboxaldehyde have
antioxidant activity  5-methylhex-2-yl pentadecyl ester shows
antimicrobial activity .
The result of GC-MS and HPLC indicated the presence of
vital bioactive phyto constituents belonging to various groups
of alkaloids, glycosides, steroids, etc. Based on the results it can
be concluded that C. borivilianum is the plant with potential
therapeutic and nutritional importance thuscould be used as a
natural source of antioxidants and its regular consumption in diet
could provide health benefits to humans by the protection against
The authors are thankful to the Department of Zoology, Centre
for Advanced Studies, University of Rajasthan, Jaipur for providing
necessary infrastructure & facility. We are also thankful DSTInspire
fellowship program for continuous funding.
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