23.

februar 2012 15:20

Bitter-fennel fruit oil Foeniculi amari fructus aetheroleum

DEFINITION Essential oil obtained by steam distillation from the ripe fruits of Foeniculum vulgare Miller, ssp. vulgare var. vulgare . Content: fenchone: 12.0 per cent to 25.0 per cent, trans-anethole: 55.0 per cent to 75.0 per cent. CHARACTERS Appearance: clear, colourless or pale yellow liquid. Characteristic odour. IDENTIFICATION First identification: B. Second identification: A. A. Thin-layer chromatography (2.2.27). Test solution. Dissolve 0.1 mL of the oil to be examined in 5 mL of toluene R. Reference solution. Dissolve 10 L of fenchone R and 80 L of anethole R in 5 mL of toluene R. Plate: TLC silica gel plate R. Mobile phase: ethyl acetate R, toluene R (5:95 V/V). Application: 10 L as bands. Development: over a path of 15 cm. Drying: in air. Detection: spray with a freshly prepared 200 g/L solution of phosphomolybdic acid R in ethanol (96 per cent) R and heat at 150 C for 15 min; examine in daylight. Results: see below the sequence of the zones present in the chromatograms obtained with the reference solution and the test solution. Furthermore, other zones may be present in the chromatogram obtained with the test solution.
Top of the plate Anethole: a dark blue to dark violet zone A dark blue to dark violet zone (anethole) _______ Fenchone: a blue or bluish-grey zone _______ Reference solution Test solution _______ A blue or bluish-grey zone (fenchone) _______

B. Examine the chromatograms obtained in the test for chromatographic profile. Results: the characteristic peaks in the chromatogram obtained with the test solution are similar in retention time to those in the chromatogram obtained with the reference solution. TESTS Relative density (2.2.5): 0.961 to 0.975. Refractive index (2.2.6): 1.528 to 1.539. Optical rotation (2.2.7): + 10.0 to + 24.0. Chromatographic profile. Gas chromatography (2.2.28): use the normalisation procedure.

1. -pinene 3. fenchone 5. cis-anethole

7. anisaldehyde

2. limonene 4. estragole 6. trans-anethole

Figure 1826.-1. Chromatogram for the test for chromatographic profile of bitter-fennel fruit oil Test solution. Dissolve 0.20 mL of the oil to be examined in heptane R and dilute to 10.0 mL with the same solvent. Reference solution. Dissolve 20 L of -pinene R, 20 L of limonene R, 50 L of fenchone R, 20 L of estragole R, 100 L of anethole R and 20 L of anisaldehyde R in heptane R and dilute to 10.0 mL with the same solvent. Column: material: fused silica, size: l = 60 m, = 0.25 mm, stationary phase: macrogol 20 000 R (film thickness 0.25 m). Carrier gas: helium for chromatography R. Flow rate: 1 mL/min. Split ratio: 1:200. Temperature:
Time (min) Column 04 4 26 26 41 Temperature (C) 60 60 170 170

Injection port Detector

220 270

Detection: flame ionisation. Injection: 1.0 L. Elution order: order indicated in the composition of the reference solution. Record the retention times of these substances. System suitability: reference solution: resolution: minimum 5.0 between the peaks due to estragole and trans-anethole. Using the retention times determined from the chromatogram obtained with the reference solution, locate the components of the reference solution on the chromatogram obtained with the test solution and locate cis-anethole using Figure 1826.-1. (Disregard the peak due to heptane). Determine the percentage content of each of these components. The percentages are within the following ranges: -pinene: 1.0 per cent to 10.0 per cent, limonene: 0.9 per cent to 5.0 per cent, fenchone: 12.0 per cent to 25.0 per cent, estragole: maximum 6.0 per cent, cis-anethole: maximum 0.5 per cent, trans-anethole: 55.0 per cent to 75.0 per cent,

Foeniculi fructus side 1

 trans-anethole: 55.0 per cent to 75.0 per cent, anisaldehyde: maximum 2.0 per cent. The ratio of -pinene content to limonene content is greater than 1.0. STORAGE At a temperature not exceeding 25 C.
Pasted from <http://online6.edqm.eu/ep703/NetisUtils/srvrutil_getdoc.aspx/2L3CsDZWvCbmnE38sPIveT6q0/1826e.htm?ts= 00HifCyG6lC9EqitsBq~DmX~n94R7-i3UzJxOp-zu1llGC8kl8Tj5mH2k&00HifCyG6lC9EqitsBq~DmX~n94R7-i3UzJxOp-zu1llGC8kl8Tj5mH2k>

Fennel, bitter Foeniculi amari fructus

DEFINITION Dry cremocarps and mericarps of Foeniculum vulgare Miller sp. vulgare var. vulgare . Content: essential oil: minimum 40 mL/kg (anhydrous drug); anethole: minimum 60.0 per cent in the essential oil; fenchone: minimum 15.0 per cent in the essential oil. CHARACTERS Bitter fennel is greenish-brown, brown or green. IDENTIFICATION A. The fruit of bitter fennel is a cremocarp, of almost cylindrical shape with a rounded base and a narrower summit crowned with a large stylopod. It is generally 3-12 mm long and 3-4 mm wide. The mericarps, usually free, are glabrous. Each bears 5 prominent slightly carenated ridges. When cut transversely, 4 vittae on the dorsal surface and 2 on the commissural surface may be seen with a lens. B. Reduce to a powder (355) (2.9.12). The powder is greyish-brown or greyish-yellow. Examine under a microscope using chloral hydrate solution R. The powder shows the following diagnostic characters: yellow fragments of wide secretory canals, often made up of yellowish-brown-walled polygonal secretory cells, frequently associated with a layer of thin-walled transversely elongated cells 2-9 m wide, having a parquetry arrangement; reticulate parenchyma of the mesocarp; numerous fibre bundles from the ridges, often accompanied by narrow spiral vessels; very numerous endosperm fragments containing aleurone grains and very small calcium oxalate microrosette crystals, as well as some fibre bundles from the carpophore. C. Thin-layer chromatography (2.2.27). Test solution. Shake 0.3 g of the freshly powdered drug (1400) (2.9.12) with 5.0 mL of methylene chloride R for 15 min. Filter and carefully evaporate the filtrate to dryness on a water-bath at 60 C. Dissolve the residue in 0.5 mL of toluene R. Reference solution. Dissolve 50 L of anethole R and 10 L of fenchone R in 5.0 mL of hexane R. Plate: TLC silica gel GF254 plate R. Mobile phase: hexane R, toluene R (20:80 V/V). Application: 10 L, as bands of 20 mm by 3 mm. Development: over a path of 10 cm. Drying: in air. Detection A: examine in ultraviolet light at 254 nm. Results A: the chromatograms show in the central part a quenching zone due to anethole. Detection B: spray with sulfuric acid R and heat at 140 C for 5-10 min until a yellow zone due to fenchone appears in the lower third of the chromatograms. Results B: anethole appears as a violet band in the central part. The chromatogram obtained with the test solution also shows a reddish-brown zone in its upper third (terpenes). TESTS Estragole. Gas chromatography (2.2.28): use the normalisation procedure. Test solution. Dilute the mixture of essential oil and xylene R obtained in the determination of essential oil to 5.0 mL with xylene R, by rinsing the apparatus. Reference solution. Dissolve 5 mg of estragole R in 0.5 mL of xylene R. Column: size: l = 30-60 m, = 0.3 mm; stationary phase: macrogol 20 000 R. Carrier gas: nitrogen for chromatography R. Flow rate: 0.40 mL/min. Split ratio: 1:200.
Time (min)
Column 0-4 4 - 26 26 - 41 Injection port Detector

Temperature (C)
60 60 170 170 220 270

Fennel, sweet Foeniculi dulcis fructus

DEFINITION Dry cremocarps and mericarps of Foeniculum vulgare Mill. subsp. vulgare var. dulce (Mill.) Batt. & Trab. Content: essential oil: minimum 20 mL/kg (anhydrous drug); anethole: minimum 80.0 per cent in the essential oil. CHARACTERS Sweet fennel is pale green or pale yellowish-brown. IDENTIFICATION A. The fruit of sweet fennel is a cremocarp of almost cylindrical shape with a rounded base and a narrowed summit crowned with a large stylopod. It is generally 3-12 mm long and 3-4 mm wide. The mericarps, usually free, are glabrous. Each bears 5 prominent, slightly carenated ridges. When cut transversely, 4 vittae on the dorsal surface and 2 on the commissural surface may be seen with a lens. B. Microscopic examination (2.8.23). The powder is greyish-brown or greyish-yellow. Examine under a microscope using chloral hydrate solution R. The powder shows the following diagnostic characters (Figure 0825.-1.): yellow fragments of wide secretory canals, often made up of yellowish-brown-walled polygonal secretory cells [D, H]; reticulate parenchyma of the mesocarp [B]; numerous fibre bundles [G] from the ridges [Ga], often accompanied by narrow spiral vessels [Gb]; very numerous endosperm fragments [F] containing aleurone grains [Fb] and very small calcium oxalate cluster crystals [Fa]; some fibre bundles from the carpophore [E]; fragments of the endocarp, in surface view [K, A], consisting of thin-walled, transversely elongated cells 2-9 m wide, having a parquetry arrangement, sometimes accompanied by the inner layer of the mesocarp [Aa]; fragments of the epicarp with stomata accompanied by oil droplets [C]; very numerous oil droplets [J].

Detection: flame ionisation. Injection: 1 L. Limits: estragole: maximum 5.0 per cent in the essential oil obtained in the assay. Foreign matter (2.8.2): maximum 1.5 per cent of peduncles and maximum 1.5 per cent of other foreign matter. Water (2.2.13): maximum 80 mL/kg, determined on 20.0 g of the powdered drug (710) (2.9.12). Total ash (2.4.16): maximum 10.0 per cent. ASSAY Essential oil. Carry out the determination of essential oils in herbal drugs (2.8.12). Use a 500 mL round-bottomed flask and 200 mL of water R as the distillation liquid. Reduce the drug to a coarse powder (1400) (2.9.12) and immediately use 5.0 g for the determination. Introduce 0.50 mL of xylene R in the graduated tube. Distil at a rate of 2-3 mL/min for 2 h. Anethole and fenchone. Gas chromatography (2.2.28) as described in the test for estragole with the following modifications. Reference solution. Dissolve 5 mg of fenchone R and 5 mg of anethole R in 0.5 mL of xylene R. Elution order: the order indicated in the composition of the reference solution. Record the retention times of these substances. STORAGE Protected from moisture.
Pasted from <http://online6.edqm.eu/ep703/NetisUtils/srvrutil_getdoc.aspx/2L3CsDZaqCrmmE38qPIveT6q0/0824e.htm?ts= 00HifCyG6lC9EqitsBadBnX~n94R7-i3UzJxOp-zu1llGC8kl8Tf8m12m&00HifCyG6lC9EqitsBadBnX~n94R7-i3UzJxOpzu1llGC8kl8Tf8m12m>

Foeniculi fructus side 2

zu1llGC8kl8Tf8m12m>

Figure 0825.-1. Illustration for identification test B of powdered herbal drug of sweet fennel C. Thin-layer chromatography (2.2.27). Test solution. Shake 0.3 g of the freshly powdered herbal drug (1400) (2.9.12) with 5.0 mL of methylene chloride R for 15 min. Filter and carefully evaporate the filtrate to dryness on a water-bath at 60 C. Dissolve the residue in 0.5 mL of toluene R. Reference solution. Dissolve 60 L of anethole R in 5.0 mL of hexane R. Plate: TLC silica gel GF254 plate R. Mobile phase: hexane R, toluene R (20:80 V/V). Application: 10 L as bands of 20 mm by 3 mm. Development: over a path of 10 cm. Drying: in air. Detection A: examine in ultraviolet light at 254 nm. Results A: the chromatograms show in the central part a quenching zone due to anethole. Detection B: spray with sulfuric acid R and heat at 140 C for 5 min; examine in daylight. Results B: the chromatograms show in the central part a violet band due to anethole; the chromatogram obtained with the test solution also shows a reddish-brown zone in the upper third (terpenes). TESTS Estragole and fenchone. Gas chromatography (2.2.28): use the normalisation procedure. Test solution. Dilute the mixture of essential oil and xylene R obtained in the assay of essential oil to 5.0 mL with xylene R, by rinsing the apparatus. Reference solution. Dissolve 5 mg of estragole R and 5 mg of fenchone R in 0.5 mL of xylene R. Column: size: l = 30-60 m, = 0.3 mm; stationary phase: macrogol 20 000 R. Carrier gas: nitrogen for chromatography R. Flow rate: 0.40 mL/min. Split ratio: 1:200.
Time (min)
Column 0-4

Temperature (C)
60

4 - 26 26 - 41
Injection port Detector

60 170 170
220 270

Detection: flame ionisation. Injection: 1 L. Limits: estragole: maximum 10.0 per cent in the essential oil; fenchone: maximum 7.5 per cent in the essential oil. Foreign matter (2.8.2): maximum 1.5 per cent of peduncles and maximum 1.5 per cent of other foreign matter. Water (2.2.13): maximum 80 mL/kg, determined on 20.0 g of the powdered herbal drug (710) (2.9.12). Total ash (2.4.16): maximum 10.0 per cent. ASSAY Essential oil (2.8.12). Use 10.0 g of the herbal drug reduced to a coarse powder (1400) (2.9.12) immediately before the assay, a 500 mL round-bottomed flask, 200 mL of water R as the distillation liquid, and 0.50 mL of xylene R in the graduated tube. Distil at a rate of 2-3 mL/min for 2 h. Anethole. Gas chromatography (2.2.28) as described in the test for estragole and fenchone with the following modification. Reference solution. Dissolve 5 mg of anethole R in 0.5 mL of xylene R. STORAGE Protected from moisture.
Pasted from <http://online6.edqm.eu/ep703/NetisUtils/srvrutil_getdoc.aspx/2L3CsDZaqD5mmE38rPIveT6q0/0825e.htm?ts= 00HifCyG6lC9EqitsBad8n1~n94R7-i3UzJxOp-zu1llGC8kl8Tf8nn2-&00HifCyG6lC9EqitsBad8n1~n94R7-i3UzJxOpzu1llGC8kl8Tf8nn2->

Foeniculi fructus side 3