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                Mammea Siamensis Flower is the dried flower of Mammea siamensis (Miq.) T. Anderson [Calysaccion siamense Miq., Ochrocarpos siamensis (Miq.) T. Anderson] (Family Calophyllaceae), Herbarium Specimen Number: DMSC 5362, Crude Drug Number: DMSc 1253.

Constituents  Mammea Siamensis Flower contains coumarins (e.g., kayeassamins and mammeasins). It also contains terpenoids, xanthones, sterols, etc.

Description of the plant  (Fig. 1)  Evergreen tree, up to 20 m tall, dioecious, sometimes polygamo-dioecious; outer bark dark grey, roughly cracked and flaking, sometimes smooth or slightly fissured, inner bark red with pale yellow latex; youngest twigs slightly flat. Leaves simple, opposite, oblanceolate, obovate or oblong to oblong-obovate, 7.5 to 25 cm long, 1.5 to 8 cm wide, apex obtuse to slightly cuspidate, base cuneate, margin entire, coriaceous, veins conspicuous, young leaves purple, dark green above when mature; petiole 0.5 to 1.5 cm long. Inflorescence cymose, mostly in axils of fallen leaves. Flower unisexual or bisexual, mostly male and perfect flowers on separate trees, 1.2 to 2.5 cm in diameter; pedicel up to 2 cm long; sepals 2, white-green, elliptic, 2 to 7 mm long, 4 to 8 mm wide; petals 4, white or pale yellow, oblong, 0.6 to 1 cm long, 5 to 9 mm wide; stamens 60 to 90, filament white, free, anther yellow to orange-yellow, 1 to 2 mm long; ovary superior, 2-loculed, ovary and style light green and turning dark green, style 1, short, stigma 2-lobed. Fruit a drupe, oval to ellipsoid, or rhomboid, 2 to 2.5(–4) cm long, 0.8 to 2.5 cm wide, with short blunt tip, green to yellow-orange. Seed 1, with thin yellow to orange aril.

Description  Odour, aromatic, and characteristic; taste, slightly bitter, and astringent.

                Macroscopical  (Fig. 1)  Complete or broken flowers, fragment of sepals, petals, filaments, stamens, and pedicels are commonly found. Sepals, petals, and filaments, pale yellow to brown; pedicels, dark brown.

                Microscopical  (Figs. 2a−2g)  Transverse section of the sepal shows upper epidermis, mesophyll, vascular tissue, and lower epidermis. Upper epidermis: a layer of oval cells and stomata. Mesophyll: collenchyma, 1 to 3 layers underneath epidermises; slightly thick-walled parenchyma, some containing rosette aggregate crystals and/or microcrystals, and schizogenous secretory ducts. Vascular tissue: phloem and xylem. Lower epidermis: a layer of rectangular epidermal cells and stomata.

                Transverse section of the petal shows upper epidermis, mesophyll, vascular tissue, and lower epidermis. Upper epidermis: a cuticle layer and a layer of oval cells. Mesophyll: collenchyma, 1 to 3 layers of small cells; parenchyma, some containing rosette aggregate crystals and/or microcrystals or brown substances, and schizogenous secretory ducts. Vascular tissue: phloem and xylem. Lower epidermis: a cuticle layer, layer of small rectangular epidermal cells, and stomata.

                Transverse section of the filament shows a layer of epidermis with papillae covered with cuticle layer, loosen parenchyma cells, and one strand of vascular tissue.

                Transverse section of the anther shows 4 pollen sacs containing a layer of round epidermal cells with papillae, a layer of endothecium, parenchyma, pollen grains, and vascular tissue.

                Transverse section of the ovary shows epidermis, cortex, vascular tissue, and ovules. Epidermis: a layer of small epidermal cells. Cortex: numerous parenchyma cells, some containing rosette aggregate crystals, secretory ducts. Vascular tissue: phloem and xylem. Ovule: 4.

                Transverse section of the pedicel shows epidermis, cortex, vascular tissue, and pith. Epidermis: a layer of small epidermal cells. Cortex: numerous parenchyma cells, some containing rosette aggregate crystals, and secretory ducts. Vascular tissue: phloem and xylem. Pith: parenchyma.

                Mammea Siamensis Flower in powder possesses the diagnostic microscopical of the unground drug. Secretory duct, endothecium, slightly thick-walled parenchyma of sepal, fragment of filament, and triporate pollen grains are characteristic.

Packaging and storage  Mammea Siamensis Flower shall be kept in well-closed containers, protected from light, and stored in a dry place.

Identification

A. To 500 mg of the sample, in powder, add 5 mL of ethanol, shake, allow to stand for 5 minutes, and filter. Drop the filtrate on a filter paper moistened with 1 M sodium hydroxide and examine under ultraviolet light (366 nm): a green fluorescence is produced.

B. Heat 1 g of the sample, in powder, with 20 mL of water in a water-bath for 10 minutes, allow to cool, and filter. To 2 mL of the filtrate, add a few drops of iron(III) chloride TS and mix well: a dark green colour is produced.

C. Carry out the test as described in the “Thin-Layer Chromatography” (Appendix 3.1),using silica gel GF254 as the coating substance and a mixture of 80 volumes of n-hexane and 20 volumes of ethyl acetate as the mobile phase and allowing the solvent front to ascend 12 cm above the line of application. Apply to the plate as a band of 8 mm, 20 μL of the test solution prepared by sonicating 1 g of the sample, in coarse powder, with 10 mL of methanol for 20 minutes and filtering. Evaporate the filtrate to dry under reduced pressure at 50°. Dissolve the residue in 1 mL of methanol. After removal of the plate, allow it to dry in air and examine the plate under ultraviolet light (254 nm), marking the quenching bands. Examine the plate under ultraviolet light (366 nm) through the cut-off filter; two green, two red, two orange, and six blue fluorescent bands are observed. Subsequently spray the plate with anisaldehyde TS and heat at 105° for 10 minutes; four violet and four purple bands are observed.

                Repeat the same procedure on another plate but spray with a 3 per cent w/v solution of potassium hydroxide, allow it to dry in air, and then examine under ultraviolet light (366 nm); one green and eight blue fluorescent bands are observed (Fig. 3).

Loss on drying  Not more than 7.0 per cent w/w after drying at 105° to constant weight (Appendix 4.15).

Foreign matter  Not more than 2.0 per cent w/w (Appendix 7.2).

Acid-insoluble ash  Not more than 2.0 per cent w/w (Appendix 7.6).

Total ash  Not more than 9.0 per cent w/w (Appendix 7.7).

Ethanol-soluble extractive  Not less than 12.0 per cent w/w (Appendix 7.12).

Water-soluble extractive  Not less than 21.0 per cent w/w (Appendix 7.12).