Research Articles

2018  |  Vol: 4(2)  |  Issue: 4(March-April)  |  https://doi.org/10.31024/ajpp.2018.4.2.23
Pharmacognostic Standardization of roots, stems, leaves and fruits of Fumaria parviflora Lam. (Fumariaceae)

Suresh Kumar1, 2*, Anjoo Kamboj3, Anil Kumar Sharma4

1Lord Shiva College of Pharmacy Sirsa, Haryana-125055 India

2Research Scholar, Department of Pharmacy, IK Gujral Punjab Technical University, Jalandhar, Punjab, India-144001

3Chandigarh College of Pharmacy, Landran, Mohali, Punjab-140110 India

4Former Director and Principal in CT Institute of Pharmaceutical Sciences, Jalandhar, Punjab-144020, India

*Corresponding author

Suresh Kumar

Lord Shiva College of Pharmacy, Sirsa, Haryana-125055, India


Abstract

Objective: Present studies deal the quality control parameters of a locally occurring Indian medicinal plant, Fumaria parviflora Lam. which is used as folk medicine in Haryana (India). Materials and methods: In this study the organoleptic, morphological, microscopical characteristics, and physicochemical evaluation (ash value, extractive value, foreign matters and moisture content), fluorescence analysis of the root, stem, leaf and fruit of Fumaria parviflora were investigated. Ethanolic extracts were prepared by extracting the ground powders of root, stem, leaf and fruit. Preliminary phytochemicals analysis of ethanolic extracts of different parts of Fumaria parviflora was carried out for qualitative analysis. Results and conclusion:  Preliminary phytochemicals analysis of ethanolic extracts of root, stem, leaf and fruit revealed the presence of carbohydrates, alkaloids, tannins and flavonoids. The pharmacognostic investigation of root, stem, leaf and fruit of Fumaria parviflora would be useful for maintaining the standards for the quality, purity and sample identification.

KeywordsFumaria parviflora, microscopy, pharmacognostic, flavonoids


Introduction                                    

Fumaria parviflora Lam. (Fumariaceae) commonly known as “fine leaf fumitory, earth smoke”, Indian fumitory, and wax doll in English (Rizvi et al., 2017). It is a loved medicinal herb in Indian system of medicine. It is well known annual weed growing throughout in India from Indus Ganga plain to down Nilgiris in South (Karuna modi et al., 2016). The major chemical constituents reported include alkaloids like Protopine, parfumine, d-bicuculline, hydrastine, N-methlhydrastine, N-methylhydrasteine, microcarpine, sanguinarine, adlumiceine, coptisine, fumaritine, sinactine, N-methylstylopine and sterols of plant are β-sitosterol, stigmasterol,and campesterol etc (Fafal et al., 2007; Paltinean et al., 2013). Entire herb has been widely used in Ayurvedic medicine system as bitter; cooling, expectorant, constipating, increases “vata” removes biliousness, fever, burning of the body, tired feeling, wandering of the mind, intoxication, urinary discharge, vomiting, thirst, enriches the blood, good in leprosy (API, 2004).

The plant shows many biological properties such as hepatoprotective (Khan et al., 2017), antipruritic, antifeedant, Antiprotozoal, antiparasitic, anthelmintic, antidiabetic, antieczema, antioxidant, Antinociceptive, antimicrobial, prokinetic, laxative and spasmodic effect (Kumar et al., 2017).

As plant was not explored in depth but it have lot of ethanopharmacological importance. Therefore, we carried out the present studies which deal with extremely important information on micro morphological characteristics of this medicinal plant which would help in identification and authentification as well as provide basic pharmacognostic parameters. The pharmacognostic and phytochemicals analysis is key steps to develops the herbal pharmacopoeia standards and helpful in identification and quality control of the medicinal plants.

Materials and methods

Plant materials

The fruits bearing plants of Fumaria parviflora Lam. were collected from district Sirsa, Haryana (India) in month of February-March 2015. The collected plant was authenticated from Raw Material Herbarium and Museum, Delhi (RHMD), CSIR-NISCAIR, New Delhi. A slide specimen/ sample have been deposited in the Raw Material Herbarium and Museum, Delhi (RHMD), CSIR-NISCAIR, Ref. No. NISCAIR/RHMD/Consult/2015/2811/04.

Chemicals

All chemicals were analytical grade used in this study purchased from SRL, CDH, SD fine and HIMEDIA. Formalin, absolute alcohol, safranin, haematoxyllin, fast green, glacial acetic acid, clove oil, canada balsam, chloral hydrate, phloroglucinol, H2SO4, NaOH, NH3, lead acetate, FeCl3, potassium hydroxide, ethyl acetate and chloroform etc.

Macroscopic and microscopic examination

Morphological characters were done by using simple microscope. The color, size, shape, odour and taste of roots, stems, leaves and fruits were determined. Microscopic characters were done by preparing thin section of root, stem, leaf and fruit of Fumaria parviflora Lam. (Tekin et al., 2017).

Powdered microscopy

Air dried different parts of plant (leaves, stems, fruits and roots) were finely powdered (# 60) and observed under microscope. Small quantity of different plant parts powder were placed separately on slides and each slide was placed 2-3 drops of chloral hydrate solution and each slide was covered with cover slip then observed under microscope. Different cell contents i.e. epidermis, cork, collenchyma, schlerenchyma, parenchyma, reticulate vessels and stomatal cells were observed and snapshot was done by using digital camera (Sonibare et al., 2014).

Physiochemical examination

Physicochemical parameters of different plant parts of powdered as well as extract of crude drug such as foreign matter, total ash, water soluble ash, acid insoluble ash, alcohol and water soluble extractive values were determined. The moisture content of different plant parts of powdered and extracts were determined by using loss on drying method (Dhingra et al., 2014).

Fluorescence examination

Fluorescence examination of the different parts of plant powder was done by using standard method. The examination was done by treating the plant powder with different solvents including both acidic/basic and organic/inorganic. After treatment they were examined under visible light, short ultra-violet light and long ultra- violet light.

Fluorescence examination is an imperative mechanism for the screening of those constituents which have the assets of showing different colors under UV light. Some components are not fluorescent themselves but when they are reacted with solvents are converted into fluorescent derivatives. This phenomenon may be due to an exacting fluorescent substance or fluorescent derivative formed after treatment with reagents. Still many natural drugs are assessed qualitatively by using this parameter. Powdered roots, stems, leaves and fruits materials were observed under visible light, short ultra-violet light and long ultra-violet light simultaneously after treatment with different organic and inorganic reagents like KOH, NaOH, H2SO4, HCl, FeCl3, iodine solution and HNO3 (Akbar et al., 2014).

Microchemistry examinations of roots, stems, leaves and fruits powder

Microchemistry evaluations of different parts of plant with different chemical reagents were observed. Screening showed the presence of different phytoconstituents along with colour changes under ordinary day light by standard procedure.

Preparation of extract and preliminary phytochemicals examination

The different parts of the plant were air dried in shadow followed by grinding. Extraction of each part was performed with ethanol as solvent on soxhlet apparatus. The each extract was evaporated in rotary evaporator apparatus and air dried at room temperature for 2-3 days. These extracts were stored in refrigerator for further study. The preliminary phytochemicals examination of different plant parts extract of Fumaria parviflora was done by using the standard procedure.

Results

Macroscopic examination of roots, stems, leaves and fruits

The organoleptic and morphological study of Fumaria parviflora leaves as well as powder showed green colour. Leaves are compound, pinnatifid, 5 to 7 cm long, apex acute; petiole is very thin, 2.5 to 4.0 cm long and bitter in taste. The root was branched, cylindrical, about 8-10 cm long, 3 mm thick, cream coloured, and bitter in taste. The stem was pentagonal, pale green, smooth, hollow, about 2-4 mm thick, bitter and slightly acrid in taste. Fruits Capsule, are single seeded, 2 mm long and obovate, subtruncate, obscurely, apiculate, rugose and bitter in taste (Figure 1).

Figure 1. Fumaria parviflora

 

 

Microscopic examination

Roots microscopy

Transverse section of root shows a single layered epidermis. The cortex (ct) consisting of thin walled, rectangular, parenchymatous cells, outer 1 or 2 layers irregular and brown in colour. Endodermis is not distinct, secondary phloem very narrow and central part shows a wide zone of xylem and consists of common elements. Xylem vessels (xyv) mostly single having reticulate and spiral thickening, medullary rays (mr) are less developed and mostly uniseriate (Figure 2).

Figure 2. T.S. of Fumaria parviflora roots

 

 

Stems microscopy

Transverse section of Stem shows a single layered thin walled epidermis (e) of rectangular cells, covered with thin cuticle (cl), cortex (ct) narrow brown pitted parenchymatous (bpp). Pentagonal outline, of stem having well-known collenchymatous hypodermis (hyp); vascular bundles (vb) collateral, 5 or 6 arranged in a ring; each vascular bundle capped with pericyclic fibres (per). It has centrally hollow wide pith (pi) (Figure 3).

Figure 3. T.S. of Fumaria parviflora stems

 

 

Fruits microscopy

Cross section of fruit wall we have observed 3 layers. Outer layer of fruit showed epidermis (e) with thin cuticle cells and some hypodermal cells. Mesocarp (mc) with schlerenchymatous cells. The endocarp (ec) one to many layers of endosperm cells developed under the apical region and folded on inner side of the fruit (Figure 4).

Figure 4. T.S. of Fumaria parviflora fruits

 

 

Powder microscopy

The powder characteristics of the root, stem, leaf and fruit were study under microscope and shown in the (Figure 5 a-d).

Figure 5 (a-d). Powder microscopy of Fumaria parviflora (a) roots, (b) stems, (c) leaves, (d) fruits

 

Physicochemical examination

Present study deals the various physicochemical parameters such as foreign matter, loss on drying, total ash, water soluble ash, acid insoluble ash values, and extractive values were determined in duplicate as depicted in table 1.

Table 1. Physicochemical examination of roots, stems, leaves and fruits of Fumaria parviflora Lam.

S.N.

Parameters

Mean ± SD (%W/W)

Roots

Stems

Leaves

Fruits

  Ash value                  

1

Total ash

16.23±0.21

15.23±0.21

15.50±0.41

8.30±0.22

2

Water soluble ash

8.5±0.41

8.3±0.08

9.4±0.29

6.16±0.24

3

Acid insoluble ash

2.33±0.24

1.32±0.23

5.30±0.22

1.17±0.24

 Extractive value

1

Water soluble

12.28±0.21

26.19±0.05

29.73±0.25

18.03±0.21

2

Alcohol soluble

10.19±0.05

12.03±0.02

14.56±0.40

20.83±0.12

 Moisture content

1

Moisture content

0.59±0.01

0.45±0.02

0.82±0.02

0.16±0.01

 Foreign matter

1

Foreign matter

1.03±0.12

0.90±0.08

1.33±0.09

0.8±0.08

Fluorescence investigation

The fluorescence characteristics of the powder with different chemical reagents are depicted in table 2,3,4,5.

Table 2. Fluorescence examination of powdered roots

S.N.

Reagents

Colour observed in visible light

Colour observed under UV light

Short (254nm)           

Long (365nm)

1

Powder

Light brown

Light brown

Light brown

2

NaOH (1N)

Brown

Yellowish green

Pale yellow

3

KOH (1N)

Brown

Green

Yellow

4

Conc. H2SO­4

Pale yellow

Yellow

Yellow

5

Conc. HNO3

Reddish brown

Green

Green

6

Conc. HCl

Brown

Yellow

Yellow

7

50% H2SO­4

Reddish brown

Yellow

Yellow

8

Iodine solution

Dull Yellow

Black

Green

9

5% FeCl3

Brownish black

Black

Black

10

Picric acid

Dull yellow

Green

Brown

Table 3. Fluorescence examination of powdered stems

S.N.

Reagents

Colour observed in visible light

Colour observed under UV light

Short (254nm)          

Long (365nm)

1

Powder

Yellowish green

Green

Light green

2

NaOH (1N)

Sand

Lemon

Creamy

3

KOH (1N)

Sand

Lemon

Yellow

4

Conc. H2SO­4

Reddish brown

Green

Yellow

5

Conc. HNO3

Reddish

Yellow

Pale yellow

6

Conc. HCl

Pale yellow

Creamy

Creamy

7

50% H2SO­4

Reddish

Lemon

Yellow

8

Iodine solution

Brown

Black

Black

9

5% FeCl3

Black

Black

Black

10

Picric acid

Yellow

Walnut brown

Walnut brown

Table 4. Fluorescence examination of powdered leaves

S.N.

Reagents

Colour observed in visible light

Colour observed under UV light

Short (254nm)            

Long (365nm)

1

Powder

Green

Green

Dull green

2

NaOH (1N)

Pale yellow

Light green

Light green

3

KOH (1N)

Pale yellow

Light green

Light green

4

Conc. H2SO­4

Yellowish green

Green

Green

5

Conc. HNO3

Reddish brown

Green

Green

6

Conc. HCl

Light brown

Purple

Purple

7

50% H2SO­4

Yellow

Green

Green

8

Iodine solution

Blood red

Blood red

Blood red

9

5% FeCl3

Bluish black

Black

Black

10

Picric acid

Yellowish green

Green

Green

Table 5. Fluorescence examination of powdered fruits

S.N.

Reagents

Colour observed in visible light

Colour observed under UV light

Short (254nm)            

Long (365nm)

1

Powder

Dark brown

Black

Black

2

NaOH (1N)

Pale yellow

Parrot colour

Green

3

KOH (1N)

Reddish brown

Light green

Light green

4

Conc. H2SO­4

Reddish

Purple

Purple

5

Conc. HNO3

Light brown

Green

Green

6

Conc. HCl

Dark brown

Green

Green

7

50% H2SO­4

Red

Reddish brown

Reddish brown

8

Iodine solution

Red

Parrot colour

Green

9

5% FeCl3

Black

Dark Green

Green

10

Picric acid

Pale yellow

Green

Green

Table 6. Microchemistry investigation of powdered roots, stems, leaves and fruits

S.N.

Reagents

Roots

Stems

Leaves

Fruits

Colour/ppt

Constituents

Colour/ppt

Constituents

Colour/ppt 

Constituents

Colour/ppt

Constituents

1

Iodine solution

Pale yellow

Cellulose (+)

Pale yellow colour

Cellulose (+)

Pale yellow

Cellulose (+)

Pale yellow

Cellulose (-)

2

Iodine solution+66% H2SO4

Bright blue

Cellulose (+)

Bright blue colour

Cellulose (+)

Bright blue

Cellulose (+)

Bright blue

Cellulose (-)

3

Phloroglucinol + HCl

Red

Lignin (+)

Red colour

Lignin (+)

Red

Lignin (+)

Red

Lignin (+)

4

Water

No change

Saponin(-)

No change

Saponin(-)

No change

Saponin(-)

No change

Saponin(-)

5

Molisch’S reagent

 Violet colour

Carbohydrate(+)

 Violet colour

Carbohydrate(+)

 Violet colour

Carbohydrate(+)

 Violet colour

Carbohydrate(+)

6

Aqueous FeCl3

Black

Tannin(+)

Black

Tannin(+)

Black

Tannin(+)

Black colour

Tannin(+)

7

Mg-HCl

No change

Flavonoid(-)

Orange colour

Flavonoid(+)

Red to purple colour

Flavonoid(+)

Red colour

Flavonoid(+)

8

Picric acid

Yellow ppt

Alkaloid(+)

Yellow ppt

Alkaloid(+)

Yellow ppt

Alkaloid(+)

Yellow ppt

Alkaloid(+)

Microchemistry investigation of powdered roots, stems, leaves, and fruits

The different parts of the plant powder was treated with different chemical reagent showed the presence of carbohydrate, lignin, tannin, alkaloid, saponin and flavonoids were shown in table 6.

Extractive values and preliminary phytochemicals examination

The extractive values of ethanolic extract of root, stem, leaf and fruit of Fumaria parviflora were calculated (table 7). Preliminary phytochemicals examination of ethanolic extracts of different parts of Fumaria parviflora revealed the presence of carbohydrates, proteins, steroids, alkaloids, glycosides, flavonoids, and tannins (table 8).

Table 7. Extractive values of different parts of Fumaria parviflora extract

Extracts

Yield (%W/W)

Colour of extract

Consistency

Root ethanolic extract

6.25

Light brown

Solid

Stem ethanolic extract

9.75

Yellowish green

Viscous

Leaf ethanolic extract

17

Blackish brown

Gummy

Fruit ethanolic extract

14.65

Dark brown

Viscous

Table 8. Preliminary phytochemicals examination of ethanolic extracts of different parts of Fumaria parviflora

Phytochemical constituents

Chemical tests

Roots ethanolic extract

Stems ethanolic extract

Leaves ethanolic extract

Fruits ethanolic extract

Alkaloids

Dragendorff’s reagent

+

+

+

+

Mayer’s reagent

+

+

+

+

Wagner’s reagent

+

+

+

+

Hager’s reagent

+

+

+

+

Cabohydrates

Molisch’s reagent

+

+

+

+

Proteins

Biuret reagent

  -

  -

  -

  -

Lipids

Paper Staining test

  -

  -

  -

+

Saponins

Foam test

  -

  -

  -

  -

Glycosides

Borntrager’s test

  -

  -

  -

  -

Keller- Killiani test

  -

  -

  -

  -

Tannins

Ferric chloride test

+

+

+

+

Lead acetate test

+

+

+

+

Flavonoids

Shinoda test

+

+

+

+

66% H2SO4

+

+

+

+

Steroids

Salkowski test

+

-

-

+

Conc. H2SO4

+

-

-

+

Discussion

As quality control of the allopathic medicine is essential, likewise the pharmacognostic standardization of herbal drug is also necessary for the quality control because substitute or adulterated plant drugs are mainly present in the market. This type of investigation will help to make sure the identity, quality, purity and safety of herbal drug for the medicinal use. The different parameters studied are organoleptic characters, macroscopic analysis, microscopic analysis and fluorescence analysis. Macroscopic and microscopic analysis is one of the easiest and cheapest methods to correctly identify the authentic herbal drug and the surety of raw material. Morphological and microscopical studies of root stem, leaf and fruit will be helpful in the identification of these parts of Fumaria parviflora Lam. Physicochemical analysis of root, stem, leaf and fruit are helpful to establish quality standards of the plant. Therefore, various parameters used for identification of different plant parts are important for drug evaluation. The results of all types of analysis are helpful in establishing quality control standards and purity assurance of drugs. Phytochemicals analysis is also the important part of herbal drug quality parameters. These simple, economical but consistent principles can be useful even for an inexpert person whenever using the drug as folk medicine. This investigation will also be helpful for producer for assessing the quality and purity of raw material. Concisely, the different parameters were described here can be considered as distinguishing to identify and authenticate this herbal drug.

References

Akbar S, Hanif U, Ali J, Ishtiaq S. 2014. Pharmacognostic studies of stem, roots and leaves of Malva parviflora L. Asian Pacific Journal of Tropical Biomedicine, 4(5):410-415.

Ayurvedic Pharmacopoeia of India. 2004. Part-I, Volume-IV, 1st ed., pp. 84-86, Government of India, Ministry of Health and Family Welfare, Department of Ayush, Delhi, The Controller of Publications, Civil Lines.

Babu K, Shankar SG, Rai S. 2010. Comparative pharmacognostic studies on the barks of four Ficus species. Turkish Journal of Botany, 34(3):215-224.

Dhingra V, Dhingra S, Singla A. 2013. Forensic and pharmacognostic studies of the Terminalia Arjuna bark. Egyptian Journal of Forensic Sciences, 3:15–19.

Fafal T, Ali M. 2007. Determination of protopine in Fumaria densiflora dc. by TLC- densitometric and spectrophotometric method. Journal of Faculty of Pharmacy, Ankara, (4):223 – 235.

Khan HM, IQBAL S. 2017. Comparative hepatoprotective activity of Fumaria parviflora Lam. leaf extract and silymarin on isoniazid and rifampicin-induced hepatotoxic rats. Indian Journal of Pharmaceutical Sciences, 79(1):124-130.

Kumar S, Kamboj A, Sharma AK. 2017. Fumaria parviflora Lam. (Fumitory): A traditional herbal medicine with modern evidence. Asian Journal of Pharmacy and Pharmacology, 3(6):200-207.

Modi K, Amin A, Shah M. 2016. A pharmacognostical study on Fumaria parviflora Lam. Journal of Natural Remedies, 16: 1–6.

Paltinean R, Toiu A, Wauters JN, Frederich M, Tits M, Angenot L, Tamas M, and Crisan G. 2013. Identification and determination of alkaloids in fumaria species from Romania. Digest Journal of Nanomaterials and Biostructures, 8(2): 817-824.

Rizvi W, Fayazuddin M, Singh O, Naeem SS, Moin S, Akhtar K, Kumar A. 2017. Anti-inflammatory effect of Fumaria parviflora leaves based on TNF-α, IL-1, IL-6 and antioxidant potential. Avicenna Journal of Phytomedicine, 7(1): 37-45.

Sharma V, Janmeda P. 2014. Extraction, isolation and identification of flavonoids from Euphorbia neriifolia leaves. Arabian Journal of Chemistry, 1-6.

Sonibare MA, Oke TA, Soladoye MO. 2014. A pharmacobotanical study of two medicinal species of Fabaceae. Asian Pacific Journal of Tropical Biomedicine, 4(2): 131-136.

Tekin M. 2017. Pharmacobotanical study of Hypericum thymopsis. Revista Brasileira de Farmacognosia, 27: 143–152.

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