Jahe (Zingiber officinale)

Zingiber officinale

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Zin giber giber off off icinale 

CONTENTS 1. Brief literature review on Zingiber . (Pg.1-4) Zingiber off icinale  a. Classification b. Vernacular names c. Part used d. Botanical description e. Geographical distribution f. Traditional use g. Anatomy of the Rhizome h. Pharmacology and Clinical studies i. Toxicity and Adverse reactions  j.  j. Phytochemistry k. Active Principles 2. Analytical specifications of Crude Drug (Pg.5-9) a. Macroscopic characterization characterization b. Identification of Crude drug by TLC c. TLC profile of Ginger oil d. GC profile of Ginger oil 3. Analytical specifications for Zin giber  extract.  extract. (Pg.10-14) iber offi cinale  cinale  a. Identification tests b. TLC profile c. Estimation of Gingerol in Zingiber in Zingiber officinale officinale extract  extract by HPLC 4. Analytical specifications for 6-Gingerol 6-Gingerol (Pg. 15-20) a. UV Spectrum  b. TLC and HPTLC c. FTIR and NMR Spectrum d. HPLC Chromatogram 5. Analytical specificati specifications ons for 8-Gingerol (Pg. 21-24) a. TLC and HPTLC  b. FTIR and NMR Spectrum c. HPLC Chromatogram 6. Analytical specifications for 6-Shogaol (Pg. 25-28) a. TLC and HPTLC  b. FTIR and NMR Spectrum c. HPLC Chromatogram 7. References (Pg. 29-31)

EMEDI ES – RESE ESE ARCH ARCH CENTRE NATURAL REMEDIES

Quality Control Department

 Zingiber officinale

Roscoe (a) Classification: Kingdom Division Class Order Family Genus Species

: Plantae : Angiosperma : Monocotyledoneae : Scitaminaea : Zingiberaceae : Zingiber : officinale

(b) Vernacular names: Sanskrit Hindi Kannada Marathi Gujarati English

(c) Part used

: Adrakam, Ardraka : Adrak,Sunthi,Sonth Adrak,Sunthi,Sonth : Sunthi : Nisam : Sunt : Ginger : Rhizome

(d) Botanical description: A herbaceous rhizomatous perennial, reaching up to 90 cm in height under cultivation. Rhizomes are aromatic, thick lobed, pale yellowish, bearing simple alternate distichous narrow oblong lanceolate leaves. The herb develops several lateral shoots in clumps, which begin to dry when the plant matures. Leaves are long and 2-3 cm broad with sheathing bases, the blade gradually tapering to a point. Inflorescence solitary, lateral radical pedunculate oblongcylindrical spikes. Flowers are rare, rather small, calyx superior, gamosepalous, gamosepalous, three toothed, open 1 splitting on one side, corolla of three subequal oblong to lanceolate connate greenish segments . (e) Geographical distribution: The distribution: The plant is widely cultivated all over India, Bangladesh, Taiwan, 1 Jamaica and Nigeria. This perennial grows in warm climates . (f) Traditional use: Ginger is carminative, pungent, stimulant, used widely for indigestion, stomachache, malaria and fevers. It is chiefly used to cure diseases due to morbidity of Kapha and Vata. Ginger with lime juice and rock salt increases appetite and stimulates the secretion of gastric  juices. It is said to be used for abdominal pain, anorexia, arthritis, atonic dyspepsia, bleeding, cancer, chest congestion, chicken pox, cholera, chronic bronchitis, cold extremities, colic, colitis, common cold, cough, cystic fibrosis, diarrhoea, difficulty in breathing, dropsy, fever, flatulent, indigestion, disorders of gallbladder, hyperacidity, hypercholesterolemia, hyperglycemia, indigestion, morning sickness, nausea, rheumatism, sore throat, throat ache, stomach ache and vomiting. Ginger forms an important constituent of many pharmacopoeial Ayurvedic 1,4 formulations . (g) Anatomy of the Rhizome: Rhizome: Scraped rhizome with buff external surface showing longitudinal striations and occasional loose fibers, outer surface dark brown and more or less covered with cork  which shows conspicuous, narrow, longitudinal and transverse ridges; the cork readily exfoliates from lateral surfaces but persists between between branches. Smoothed transversely transversely cut surface exhibiting a narrow cortex separated by an endodermis from a much wider stele, numerous widely scattered fibrovascular bundles, abundant scattered oleoresin cells with yellow contents. Starch abundant in the thin-walled ground tissue, as flattened, ovate to subrectangular, transversely straited, simple granules, each with the hilum in a projection towards one end. Pigments cells with dark reddish 1

 Zingiber officinale

brown contents occurring either singly in the ground tissue or in axial rows accompanying the vascular bundles. Vessels with spiral or reticulate thickening in the scattered vascular bundles are found. Irregularly shaped thin-walled fibers with delicate, transverse septa, yielding only slightly 1-2 the reaction characteristic of lignin. Sclereids and calcium oxalate crystals absent .

Fig. 1

Fig. 2

Fig. 3 Fig.1 : Transversely cut surface of the rhizome, vascular bundles represented by circles, secretion cells by dots(x3). Fig.2 : Transverse section in the region of the endodermis (x100) Fig.3 : Transverse section of the central part (x100) ct- cortex, end-endodermis, fr.b.vascular bundles, pg-pigment cell, S-stele, scl.f-sclerenchymatous fibres, secr.- secretion cell.

(h) Pharmacology and Clinical Studies Anti-emetic Activity: Early animal studies had demonstrated the anti-emetic property of fresh 1 ginger , but it was the clinical work of Mowrey and Clayson which generated a wider interest i n this 2 use of ginger . They compared the effects of 1.88g of dried powdered ginger, 100mg dimenhydrinate (Dramamine) and placebo on the symptoms of motion sickness in 36 healthy subjects who reported very high susceptibility to motion sickness. Motion sickness was induced by placing the blind folded subject in a tilted rotating chair. Ginger was found to be superior to dimenhydrinate and placebo in preventing the gastrointestinal symptoms of motion sickness and the authors postulated a local effort in the gastrointestinal tract for ginger. This was particularly likely since it was given as a powder only 25 minutes before the test. The gingerols and shogaols were 3 subsequently identified as the main anti-emetic compounds in ginger . Improvement of digestive function: Early Chinese and Japanese research found that oral and 1

intragastric application of fresh ginger decoction produced a stimulant action on gastric secretion .

German scientists found that chewing 9g of crystallised ginger had a profound effect on saliva production4. Amylase activity was also increased and the saliva was not more watery, although it contained slightly less mucroprotein. Intraduodenal doses of ginger extract increased bile secretion in rats5. Total secretion of bile solids was also increased, but not to the same extent as bile flow. 62

 Zingiber officinale

gingerol and 10-gingerol were identified as the active components 5. Fresh ginger also contains a 6 proteolytic enzyme . Ginger, in conjunction with other pungent Ayurvedic herbs, increased the bioavailability of a number of drugs by promoting their absorption and/or protecting them from being metabolized in 1 their first passage through the liver . 7

Oral doses of 6-shogaol accelerated intestinal transit in rats . Also an extract of ginger, and isolated 8 6-shogaol and gingerols, enhanced gastrointestinal motility in mice after oral doses . 9-10 Anti-ulcer Activity: Ginger and 6-gingerol inhibited experimental gastric ulcers in rats . Fresh 1 ginger decocted in water resulted in symptomatic improvement in 10 patients with peptic ulcers .

Antiplatelet Activity: Srivastava and co-workers found that aqueous extract of ginger inhibited 11 platelet aggregation induced by ADP, epinephrine, collagen and arachidonic acid in vitro . Ginger acted by inhibiting thromboxane synthesis 12-13. It also inhibited prostacyclin synthesis in rat aorta 11. The antiplatelet action of 6-gingerol was also mainly due to the inhibition of thromboxane 14 formation . Anti-inflammatory Activity: Ginger extract inhibited carrageenan-induced paw swelling and was as active as aspirin15. Essential oil of ginger inhibited chronic adjuvant arthritis in rats16. Ginger and its pungent components are dual inhibitors of arachiodonic acid metabolism. That is, they inhibit both cyclooxygenase (prostaglandin synthetase) and lipoxygenase enzymes of the 15,17-22 prostaglandin and leukotriene biosynthetic pathways . Antipyretic Activity: Ginger extract given orally reduced fever in rats by 38%, while the same 15 dose of aspirin was effective by 44% . The antipyretic activity of 6-shogaol and 6-gingerol has also been observed7. Cardiovascular Effects: Ginger exerted a powerful positive inotropic effect on isolated guinea pigs 23 23-24 left atria . Gingerols were identified as the active components . Antioxidant Activity: Extracts of ginger have pronounced antioxidant activity comparable to that 25 of synthetic antioxidant preservatives . 15

Other Effects: Ginger extract exhibited a prolonged hypoglycaemic activity in rabbits . Antihepatotoxic activities of gingerols and shogaols were observed using carbon tetrachloride and galactosamine induced cytotoxicity in cultured rat hepatocytes 26. Injection of 6-shogaol showed an 7 intense antitussive action in comparison with dihydrocodeine phosphate . Pharmacokinetics: After injection, 90% of 6-gingerol was bound to serum protein and elimination 27 was mainly via the liver . Oral or intraperitoneal dosage of zingerone resulted in the urinary excretion of metabolites within 24 hours, mainly as glucuronide and/or sulphate conjugates 28. 28 Appreciable biliary excretion (40% in 12 hours) also occurred . (i) Toxicity and Adverse Reactions: The mutagenic activity of ginger extracts has been observed 22,29 in several strains . As a result of component fractionation of ginger juice, it was found that 6gingerol was a potent mutagen 30. When mutagenicity of gingerol or shogaol was tested in the presence of zingerone, it was observed that zingerone suppressed the mutagenic activity of both compounds22,31. 3

 Zingiber officinale

Ginger extract caused no mortality at doses of up to 2.5g/kg in mice (equivalent to about 75g/kg of  15 fresh rhizome) . This low acute toxicity was confirmed in a separate study, which also found that 32 ginger extract at 100mg/kg per day for three months caused no signs of chronic toxicity . 33 Topical application of ginger may cause contact dermatitis in sensitive patients . (j) Phytochemistry: Ginger has been reported to contain usually 1-3% of volatile oil, pungent principles viz., gingerols and shogaols and about 6-8 lipids and others. Ginger oil contains Zingiberene and bisaboline as major constituents along with other sesqui and monoterpenes. Ginger oleoresin contains mainly the pungent principles gingerols and shogaols as well as zingiberone. 1-4 Shogaols have recently been found to be twice as pungent as gingerols . (k) Active principles: Gingerols, Shogaols

O CH3O

H

O

OH

CH3O

( )n

( )n

HO

HO [6] Gingerol [8] Gingerol [10] Gingerol

Fig.4

n 4 6 8

C17H26O4  C19H30O4  C21H34O4 

294.3 322.7 350.2

[6] Shogaol [8] Shogaol [10] Shogaol

n 4 6 8

C17H24O   3 C19H28O3  C21H32O3 

276.3 304.4 332.1

Fig.5

4

 Zingiber officinale

ANALYTICAL SPECIFICATION OF THE CRUDE DRUG Organoleptic Characters: Colour & Appearance

:

Odour Taste

: :

Yellowish Brown or light brown. Scraped rhizome with buff external surface showing longitudinal striations and occasional loose fibers, outer surface dark brown and more or less covered with cork which shows conspicuous, narrow, longitudinal and transverse ridges. Aromatic Pungent

Fig.6

TESTS

LIMITS

PROTOCOLS Quality Control Methods for Medicinal Plant Materials -WHO

Foreign matter

< 1.0%

-do-

Sand & Silica

Absent

-do-

Insect infestation

Nil

-do-

Rodent contamination

Nil

-do-

Ash content

< 8.0%w/w

-do-

Acid insoluble ash

< 1.0%w/w

-do-

Moisture content

< 12.0%w/w

-do-

0.5 – 1.5%w/w

-do-

1.5 – 2.5 % w/w 3.0 - 5.0 % w/w 2.0 – 4.0 % w/w > 7.0%w/w

-do-

-do-

> 14.0%w/w

-do-

0.6 - 1.5 %w/w

By HPLC

Tests for extraneous material

Physico-chemical analysis

Volatile oil content

Successive extractive value Petroleum ether extractive value Chloroform extractive value Methanol extractive value

Alcohol soluble extractive value Water soluble extractive value Phytochemical analysis 6-Gingerol

5

 Zingiber officinale

IDENTIFICATION OF CRUDE DRUG BY TLC Sample detail

:

 Zingiber officinale crude drug (dried rhizome)

Adsorbent

:

Precoated silicagel (Al - Sheet)

Mobile Phase

:

Toluene : Ethyl Acetate 93 : 7

Sample preparation

:

2 gms of Zingiber officinale rhizome powder was extracted with petroleum ether. The extract was concentrated and diluted with chloroform. 10µl was applied on different TLC plates.

Solvent front run upto

:

9 cms

Application

:

CAMAG Linomat IV

Detection

: :

Anisaldehyde sulphuric acid (Fig. 7) Vanillin sulphuric acid (Fig. 8)

S

T Fig. 7

S

T Fig. 8

S- Standard T- Sample

6

 Zingiber officinale

IDENTIFICATION OF CRUDE DRUG BY TLC

Sample detail

:

 Zingiber officinale crude drug

Adsorbent

:

Precoated silicagel (Al - Sheet)

Mobile Phase

:

n-Hexane : Ether 40

: 60

Sample preparation

:

The mark from petroleum ether fraction of Zingiber officinale rhizome powder extract was successively extracted with chloroform and then by methanol. The individual extracts were concentrated and separately applied on different TLC plates.

Solvent front run upto

:

9 cms

Application

:

CAMAG Linomat IV

Detection

:

Vanillin sulphuric acid (Fig. 9&10)

Scanninig

:

254 nm (Fig. 10A)

Chloroform Fraction

Methanol fraction

Fig.9

Fig.10

Rf.

Fig. 10A - HPTLC of Methanolic fraction

0.35 – Gingerols (blue spot)

7

 Zingiber officinale

TLC PROFILE OF GINGER OIL Sample detail

:

 Zingiber officinale oil

Adsorbent

:

Precoated silicagel (Al - Sheet)

Mobile Phase

:

Toluene : Ethyl Acetate 93

:

7

Sample preparation

:

1 gm of  Zingiber officinale oil was dissolved in chloroform and 10µl applied on the TLC plate.

Solvent front run upto

:

9 cms

Detection

:

Anisaldihyde Sulphuric acid reagent (Fig.11) Vanillin sulphuric acid reagent (Fig.12)

Fig. 11 Rf.

Fig. 12

0.9 – Zingiberene (red spot)

8

 Zingiber officinale

GC PROFILE OF GINGER OIL TESTS

LIMITS

Colour and appearance Odour

Light yellow to greenish yellow liquid Sharp lemony top note with persistent dry spicy note

O

Relative density at 27  C

0.866 – 0.880

Chromatographic conditions 1. Column 2. Temperature Capillary injector port FID Detector Column oven Program > column oven Hold time Total time 3. Control mode Split ratio 4. Column pressure 5. Column flow 6. Detector 7. Carrier gas

: DB wax 0.25 mm (diameter) 30 mtr (length) : : : : : : : : : : : :

2600C 2750C 400 C initial for 2 min. 40C /min up to 2250C 5 min 53 min. Split 1 : 50 134 kPa 2.2 ml / min. F.I.D. Nitrogen

Fig.13

FIG. 13A - MASS SPECTRUM OF ZINGIBERENE

Fig. 13B - MASS SPECTRUM OF ar-CURCUMENE 9

 Zingiber officinale

ANALYTICAL SPECIFICATIONS FOR THE  Z INGIBER OFFICINALE - EXTRACT Item

:  Zingiber officinale extract (≥20% Gingerols)

Description

:  Brown to dark brown liquid with characteristic odour and taste.

Identification

:  1) Comparison with the standard TLC profile. 2) Positive for Gingerol

TESTS

LIMITS

PROTOCOL

Moisture content

< 4% w/w

As per I.P / B.P

Ash content

3.5 % w/w

As per I.P / B.P

Acid insoluble ash

< 1% w/w

As per I.P / B.P

Lead

< 10ppm

By A.A.S.

Cadmium

< 2ppm

By A.A.S.

Arsenic

< 2ppm

As per U.S.P

Physico-chemical analysis

Heavy metal analysis

Microbiological tests 4

-1

As per I.P / B.P

2

-1

As per I.P / B.P

2

-1

As per B.P.

Total Viable Aerobic Count

< 10  cfu g

Total Fungal count

< 10  cfu g

Total Enterobacteriaceae

< 10  cfu g

 E. Coli

Absent

As per I.P / B.P

Salmonella typhi

Absent

As per I.P / B.P

S. aureus

Absent

As per I.P / B.P

< 200 ppm

By GC

< 5 ppb

As per A.O.A.C

Solvent residues Organic solvents

Mycotoxin analysis Aflatoxins (Total B1,B2,G1,G2)

Phytochemical analysis

Gingerols

≥ 20.0%w/w

HPLC(High Performance Liquid Chromatography)

10

 Zingiber officinale

TLC PROFILE Sample detail

:

 Zingiber officinale extract

Adsorbant

:

Precoated silicagel (Al - Sheet)

Solvent system

:

n Hexane : Ether 40 : 60

Sample preparation

:

1 gm of Zingiber officinale extract was dissolved in 25 ml of methanol. 10 µl of this solution was applied on the TLC plate.

Solvent front run upto

:

9 cms

Detection

:

Vanillin sulphuric acid reagent. (Fig.14)

Scanning

:

Densitometer 254 nm (Fig. 15)

Application

:

CAMAG Linomat IV

S Fig. 14 S = Standard

T Fig. 15

T = Sample Rf.

0.9 – Zingiberene (dark purple spot) Fig.14

Rf.

0.3 – Gingerols (violet spot) Fig.14 11

 Zingiber officinale

ESTIMATION OF GINGEROL IN ZINGIBER OFFICINALE EXTRACT BY HPLC Summary:  Gingerol is separated from its related compounds like Shagaol, by reverse phase chromatography. The separated compounds are identified with the retention times in comparison with the pure compound and quantified with the corresponding peak area. The results were found to be accurate and reproducible. Analysis Chromatographic system: High Performance Liquid Chromatographic system equipped with LC8A pump, SPD-M 10A vp Photo Array Detector in combination with Class LC 10A software. Mobile phase:

Acetonitrile 55

Column

: C18 - ODS (Octadecylsilane) (Lichrocart 250-4, Lichrospher RP-18e-5µ (Merck) Art.No: 1.50216

Detector

: SPD-M 10Avp Photo diode array detector

Flow rate

: 1.3 ml/min

Wave length

: 280 nm

Injection volume

: 10 µL

: :

Water 45

Standard preparation: Weigh accurately 100mg of working standard (contains 40% w/w of  Gingerol) to a 25ml volumetric flask. Dissolve and make upto 25ml with HPLC grade methanol.

Sample preparation: Weigh accurately a sample quantity equivalent to 40 mg of gingerol to a 25 ml volumetric flask. Dissolve and make upto 25ml with HPLC grade methanol. Procedure: Set the instrument as per the chromatographic condition prescribed above. By means of suitable syringe inject 10 µl of standard solution. Record the chromatogram repeat the injections for another 4 times and calculate the RSD of the area. It should not be more than 2%. Inject 10 µL of sample preparation and record the chromatogram. Calculate the % of gingerol from the peak areas.

12

 Zingiber officinale

HPLC PROFILE WORKING STANDARD

Fig.16 Chromatogram

Fig.17 PDA Spectrum of 6-Gingerol

Fig.18

3-D View

13

 Zingiber officinale

HPLC PROFILE SAMPLE ( Zingiber officinale extract 20% Gingerols)

Fig.19 Chromatogram

Fig.20 PDA Spectrum of 6-Gingerol  

Fig.21

3-D View PKNO R.TIME 1 4.75

k' 4.7

2

3.47

10.275

NAME 6-Gingerol

Plate # Tailing 7357 1.02

11281 0.91

14

 Zingiber officinale

ANALYTICAL SPECIFICATION OF 6-GINGEROL (S)-5-Hydroxy-1-(4-hydroxy-3-methoxyphenyl)-3-decanone 6-Gingerol is the major phenol and most important pungent and active principle isolated from the 1 rhizome of Zingiber officinale Roscoe. Normally obtained as yellow oil . 

  

   

Mol. Formula: C17H26O4

Mol. Wt. 294.39

TESTS

RESULTS Yellow oily liquid

Description 1

Sparingly soluble in petroleum ether, soluble in alcohol, chloroform, ether and almost insoluble in water.

Solubility

Identification (by Spectroscopy & Chromatography) Light absorption 1 (UV absorption) 2 TLC & HPTLC

Exhibits maxima at 282nm Gives a single spot

2

Characteristic of 6-Gingerol

NMR

2

Characteristic of 6-Gingerol

HPLC

UV spectra of the principal peak matches with the 6-Gingerol

FTIR

(using PDA detector)

MASS SPECTRUM

Characteristic of 6-Gingerol

Moisture content

< 0.5%w/w

Purity

> 97%w/w

References: th

1. Merck Index. 12  edn., 1996 2. D.W. Connell et al., Aust. J. Chem., 1969; 22 : 1033-1043

15

 Zingiber officinale

UV SPECTRUM

Fig. 22 UV absorption of 0.01788 % w/v solution in alcohol ε reported at

: 282nm 2560

ε calculated at : 282nm 2620

Created

:

09:06 09/07/99

Data

:

Original

Measuring mode

:

Abs.

Scan speed

:

Fast

Slit width

:

1.0

Sampling interval

:

0.2

No.

Wavelength (nm)

Abs.

1

281.60

1.596

2

225.20

3.109

16

 Zingiber officinale

TLC PROFILE Sample detail

:

6-Gingerol 

Adsorbant

:

Precoated silicagel (Al - Sheet)

Solvent system

:

n Hexane : Ether 40 : 60

Sample preparation

:

10 mg of 6-Gingerol was dissolved in 1 ml of   methanol. 10µl of this solution was applied on the TLC plate.

Solvent front run upto

:

9 cms

Detection

:

Vanillin sulphuric acid reagent. (Fig.22)

Scanning

:

Densitometer 280 nm (Fig. 23)

Application

:

CAMAG Linomat IV

Fig. 23

Fig. 24

17

 Zingiber officinale

FTIR SPECTRUM

Fig. 25 - FTIR SPECTRUM OF LIQUID FILM IN KBr DISK FTIR SPECTRUM MATCHES WITH THE PEAK POSITION PROVIDED IN D.W. CONNELL, AUST. J. CHEM., 1969; 22 : 1040-1041 3400s(br), 2930s, 1710s, 1603, 1515s, 1465, 1453, 1430, 1370s, 1275s, 1240s, 1210s, 1151s, 1123s, 1090, -1 1033s, 935w, 850w, 813, 793, 723 cm

NMR SPECTRUM

Fig. 26- NMR SPECTRUM IN CDCl 3 NMR SPECTRUM MATCHES WITH THE DETAILS PROVIDED IN D.W. CONNELL, AUST. J. CHEM., 1969; 22 : 1041 6.35 to 6.75 (3H, multiplet), 3.7(3H, s), 2.6(4H, s), 2.4(2H, d, j6 c/s), 1.25 (8-9H, 0.87 p.p.m(3H).

18

 Zingiber officinale

HPLC PROFILE USING PDA-DETECTOR

CHROMATOGRAM

PDA SPECTRUM

  Fig. 27

Fig. 28

CHROMATOGRAM IN 3D VIEW

Fig. 29

PEAK TABLE

19

 Zingiber officinale

HPLC PROFILE USING UV-DETECTOR

CHROMATOGRAM

Fig. 30

Fig. 31

PEAK TABLE

MASS SPECTRUM

Fig. 32 20

 Zingiber officinale

ANALYTICAL SPECIFICATION OF 8-GINGEROL (S)-5-Hydroxy-1-(4-hydroxy-3-methoxyphenyl)-3-hexanone 8-Gingerol is another important pungent and active principle isolated from the rhizome of  Zingiber  officinale Roscoe. Normally obtained as yellow oil.





     

Mol. Formula: C19H30O4

TESTS

Mol. Wt. 322.7

RESULTS

Description

Yellow oily liquid

Solubility

Sparingly soluble in petroleum ether, soluble in alcohol, chloroform, ether and almost insoluble in water.

Identification (by Spectroscopy & Chromatography) TLC & HPTLC

Gives a single spot

FTIR

Characteristic of 8-Gingerol

NMR

Characteristic of 8-Gingerol

HPLC

UV spectra of the principal peak matches with the 8-Gingerol

(using PDA detector)

MASS SPECTRUM

Characteristic of 8-Gingerol

Moisture content

< 0.5%w/w

Purity

> 96%w/w

21

 Zingiber officinale

TLC PROFILE Sample detail

:

8-Gingerol 

Adsorbant

:

Precoated silicagel (Al - Sheet)

Solvent system

:

n Hexane : Ether 40 : 60

Sample preparation

:

10 mg of 8-Gingerol was dissolved in 1 ml of   methanol. 10µl of this solution was applied on the TLC plate.

Solvent front run upto

:

9 cms

Detection

:

Vanillin sulphuric acid reagent. (Fig.33)

Scanning

:

Densitometer 280 nm (Fig. 34)

Application

:

CAMAG Linomat IV

Fig. 33

Fig. 34

22

 Zingiber officinale

FTIR SPECTRUM

FTIR SPECTRUM OF LIQUID FILM IN KBr DISK

Fig. 35

NMR SPECTRUM

NMR SPECTRUM IN CDCl 3

Fig. 36

MASS SPECTRUM

Fig. 37 23

 Zingiber officinale

HPLC PROFILE USING PDA-DETECTOR

CHROMATOGRAM

PDA SPECTRUM

Fig. 38

Fig. 39

CHROMATOGRAM IN 3D VIEW

Fig. 40 ** Peak Report ** GINGE28.K02 99/09/28 19:38:24 PKNO ChNO

TIME

AREA

HEIGHT

CONC

NAME

1

10.463

2308591

100744

100.0000

8-Gingerol

TOTAL

2308591

100744

100.0000

 

1

24

 Zingiber officinale

ANALYTICAL SPECIFICATION OF 6-SHOGAOL

6-Shogaol is another important pungent and active principle isolated from the rhizome of  Zingiber  officinale Roscoe. Normally obtained as yellow oil.

O CH3O

( )n

HO [6] Shogaol

n 4

TESTS

C17H24O   3

276.3

RESULTS

Description

Yellow oily liquid

Solubility

Sparingly soluble in petroleum ether, soluble in alcohol, chloroform, ether and almost insoluble in water.

Identification (by Spectroscopy & Chromatography) TLC & HPTLC

Gives a single spot

1

Characteristic of 6-Shogaol

NMR

1

Characteristic of 6-Shogaol

HPLC

UV spectra of the principal peak matches with the 6-Shogaol

FTIR

(using PDA detector)

MASS SPECTRUM

Characteristic of 6-Shogaol

Moisture content

< 0.5%w/w

Purity

> 96%w/w

References: 1. D.W. Connell et al., Aust. J. Chem., 1969; 22 : 1033-1043

25

 Zingiber officinale

TLC PROFILE Sample detail

:

6-Shogaol 

Adsorbant

:

Precoated silicagel (Al - Sheet)

Solvent system

:

n-Hexane : Ether 40 : 60

Sample preparation

:

10 mg of 6-Shogaol was dissolved in 1 ml of   methanol. 10µl of this solution was applied on the TLC plate.

Solvent front run upto

:

9 cms

Detection

:

Vanillin sulphuric acid reagent. (Fig.41)

Scanning

:

Densitometer 280 nm (Fig. 42)

Application

:

CAMAG Linomat IV

Fig. 41

Fig. 42

26

 Zingiber officinale

FTIR SPECTRUM

FTIR SPECTRUM OF LIQUID FILM IN KBr DISK

Fig. 43

NMR SPECTRUM

Fig. 44

MASS SPECTRUM

Fig. 45 27

 Zingiber officinale

HPLC PROFILE USING PDA-DETECTOR

CHROMATOGRAM

PDA SPECTRUM

Fig. 46

Fig. 47

CHROMATOGRAM IN 3D VIEW

Fig. 48

28

 Zingiber officinale

References: Botanical Description 1. Schauenberg P & Paris F, Guide to Medicinal Plants, Keats Publishing,New Canaan CT,1977

Geographical Distribution 1. Kokate C.K, et.al, Pharmacognosy, Nirali Prakashan, 4th edition, 1996 Traditional Use 1. Misra B, Bhavaprakasha Nighantu , 5th edition,1969, p.14 2. Sharma P.V. Dravyagunavigna, Part II, Chowkamba Publications, 1993, p. 331 3. Indian Medicinal Plants, A Compendium of 500 species, Part V, by Orient Longman Publications, 1997, p. 431. 4. Nadakarni, Indian Materia Medica, Vol. I, 1993, p. 1308.

Anatomy of the Rhizome 1.  British Herbal Pharmacopoeia, B.H.M.A., 1983, 239-240 2. Wallis T.E., Textbook of Pharmacognosy, C.B.S. Publishers, 5th edition, 1985

Pharmacology and Clinical Studies, Toxicity and Adverse Reactions 1. Chang, H.M et al, “Pharmacology and Applications of Chinese Materia Medica.” World Scientific, Singapore, 1987 2. Mowrey, D.B, et al, “ Motion Sickness, Ginger, and Psychophysics”  Lancet , 1982, 1(8273); 655-657 3. Kawai, T, et al, “Anti-emetic principles of Magnolia obovata Bark and Zingiber officinale Rhizome”, Planta Medica, 1994, 60; 17-20 4. Blumberger, W, et al, “The Physiology of Spices and Condiments. The Action of Ginger on the Amount and Condition of the Saliva” ,  Nutritio et Dieta (European Review of Nutrition and Dietetics), 1965, 7(3); 222-237 5. Yamahara, J, et al, “Cholagogic Effect of Ginger and its Active Constituents”,  Journal of   Ethnopharmacology, 1985, 13(2); 217-225 6. Thompson, E.H, et al, “Ginger Rhizome : A New Source of Proteolytic Enzyme”,  Journal of Food  Science, 1973, 38; 652-655 7. Suekawa, M, et al, “Pharmacological Action of Pungent Constituents, (6)-Gingerol and (6)-Shogaol”,  Journal of Pharmacobio-Dynamics”, 1984, 7(11), 836-848 8. Desai, H.G, et al, “Effect of Ginger and Garlic on DNA Content of Gastric Aspirate”,  Indian Journal of   Medical Research, 1990, 92; 139-141 29

 Zingiber officinale

9. Yamahara, J, et al, “The Anti-ulcer Effect in Rats of Ginger Constituents”,  Journal of   Ethnopharmacology, 1988, 23(2-3); 299-304 10. al-Yahya, M.A, et al, “Gastroprotective Activity of Ginger (Zingiber officinale Rose.) in Albino Rats”,  American Journal of Chinese Medicine, 1989, 17(1-2); 51-56 11. Srivastava, K.C, “Effects of Aqueous Extracts of Onion, Garlic and Ginger on Platelet Aggregation and Metabolism of Arachidonic Acid in the Blood Vascular System; In vitro Study”, Prostaglandins  Leukotrienes and Medicine, 1984, 13(2); 227-235 12. Srivastava, K.C, “Aqueous Extracts of Onion, Garlic and Ginger Inhibit Platelet Aggregation and Alter Arachidonic Acid Metabolism”, Biomedica, Biochimica Acta, 1984, 43(8-9); S335-346 13. Srivastava, K.C, “Isolation and Effects of Some Ginger Components of Platelet Aggregation and Eicosanoid Biosynthesis”, Prostaglandins Leukotrienes and Medicine, 1986, 25(2-3); 187-198 14. Guh, J.H, et al, “Antiplatelet Effect of Gingerol Isolated from Zingiber officinale”,  Journal of Pharmacy and Pharmacology”, 1995, 47(4); 329-332 15. Mascolo, N, et al, “Ethnopharmacologic Investigation of Ginger (Zingiber officinale)”,  Journal of   Ethnopharmacology, 1989, 27; 129-140 16. Sharma, J.N, et al, “Suppressive Effects of Eugenol and Ginger Oil on Arthritic Rats”, Pharmacology, 1984, 49(5), 314-318. 17. Flynn, D.L, et al, “ Inhibition of Neutrophil 5-Lipoxygenase Activity by Gingerdione, Shogaol, Capsaicin and Related Pungent Compounds”,  Prostaglandins Leukotrienes and Medicine, 1986, 24(226), 195-198 18. Kiuchi, F, et al, “Inhibitors of Prostaglandin Biosynthesis from Ginger”, Chemical and Pharmaceutical  Bulletin, 1982,(30(2); 754-757 19. Iwakmi, S, et al, “Inhibition of Arachidonate 5-Lipoxygenase by Phenolic Compounds”, Chemical and  Pharmaceutical Bulletin, 1986, 34(9); 3960-3963 20. Kiuchi, F, et al, “Inhibition of Prostaglandin and Leukotriene Biosynthesis by Gingerols and Diarylheptanoides”, Chemical and Pharmaceutical Bulletin, 1992, 40(2); 387-391 21. Suekawa, N, et al, “Pharmacological Studies on Ginger IV. Effect of (6)-Shogaol on the Arachidonic Cascade”,  Nippon Yakurigakuzasshi (Folia Pharmacologica Japonica), 1986, 88(4); 263-269 (In Japanese) 22. Farnsworth, N.R, et al, Bunyapraphatsara, N, eds Thai Medicinal Plants. Medicinal Plant Information Centre, Bangkok, 1992. 23. Shoji, N, et al, “Cardiotonic Principles of Ginger (Zingiber officinale Roscoe.)”,  Journal of  Pharmaceutical Sciences, 1982, 71(10), 1174-1175 2+

24. Kobayashi, M, et al, “Cardiotonic Action of (8)-Gingerol, An Activator of Ca   pumping Adenosine Triphosphatase of Sarcoplasmic Reticulum in Guinea Pig Atrial Muscle”,  Journal of Pharmacology and   Experimental Therapeutics”, 1988, 246(2), 667-673 25. Govindarajan, V.S, “Ginger- Chemistry, Technology and Quality evaluation; Parts 1 & 2”, CRC Critical  Reviews in Food, Science and Nutrition, 1982, 17(1); 1-96: 1982, 17(3), 189-258 30