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      Blumea Balsamifera Leaf is the dried mature leaf of Blumea balsamifera (L.) DC [Conyza balsamifera L., Placus balsamifer (L.) Baill.] (Family Compositae), Herbarium Specimen Number: DMSC 5330, Crude Drug Number: DMSc 1263.

Constituents  Blumea Balsamifera Leaf contains flavonoids such as blumeatin and quercetin. It also contains a volatile oil consisting of l-borneol and camphor. Other compounds are tannins, phenolics, sterols, etc.

Description of the plant  (Fig. 1)  Shrub or subshrub, up to 4 m tall; stem erect, bark greyish brown, tomentose, branched. Leave simple, alternate or spirally arranged, lanceolate, oblong-lanceolate, elliptic, or ovate-elliptic, 5 to 30 cm long, 1.5 to 12 cm wide, apex acute to acuminate, base attenuate or obtuse, margin serrulate to serrate, usually with upcurved teeth, rarely dentate or lacerate, coriaceous, bright green, upper surface rugose, pubescent and glandular hairs, lower surface tomentose, rarely woolly; petiole 1 to 4 cm long, with 1 to 6 pairs of appendages, 0.6 to 1.5 cm long. Inflorescence spreading panicle, terminal and/or axillary, 7 to 30 cm long; peduncle 0.2 to 1 cm long; capitula 4 to 7 mm in diameter; involucre campanulate; phyllaries in 3 to 5 series, outer one lanceolate or oblong-lanceolate, 1 to 3 mm long, 0.4 to 0.5 mm wide, apex  acute, pilose, inner one linear, 5 to 6 mm long, scarious with ciliate margin, pilose; receptacle 2.5 to 3 mm in diameter, slightly convex, alveolate, glabrous; marginal floret yellow, filiform, 4.5 to 6 mm long, 2- to 4-lobed, glabrous; disc floret yellow,   4 to 7 mm long, glabrous, lobe oblong-lanceolate, minute, pubescent with glandular hairs; anther 0.8 to 2 mm long, apical appendage round, base with branched tails; ovary inferior, style arm about 1 mm long, base swollen. Fruit an achene, brown, oblong, 1 to 2 mm long, about 0.3 mm wide, 5- to 10-ribbed, setuliferous; pappus reddish, bristly, 4 to 6 mm long, persistent. Seed 1.

Description  Odour, borneol-like, aromatic; taste, slightly bitter.

                        Macroscopical  (Fig. 1)  Whole or broken leaves; whole leaf petiolate, brownish green to brownish, oblong-elliptic, elliptic to lanceolate, apex acute or acuminate, base attenuate or obtuse, margin serrulate to serrate, tomentose on both surfaces with prominent veins, brittle.

                        Microscopical  (Figs. 2a‒2d)  Transverse section of the leaf through the midrib shows upper epidermis, mesophyll, vascular tissue, and lower epidermis. Upper epidermis: thick cuticle layer, a layer of rectangular cells, multicellular uniseriate trichomes, some with collapsed cells, glandular trichomes, and stomata. Mesophyll: a layer of rectangular palisade cells and loosen round spongy cells, and some containing starch grains; several layers of lamella and a few angular collenchyma cells in midrib. Vascular tissue: phloem and xylem. Lower epidermis: thin cuticle layer, a layer of rectangular cells, multicellular uniseriate trichomes, some with collapsed cells, glandular trichomes, and stomata.

     In surface view, the lamina shows wavy-walled epidermal cells, multicellular uniseriate trichomes, some with collapsed cells, glandular trichomes, and paracytic and anomocytic stomata.

Transverse section of the petiole shows epidermis, cortex, and vascular tissue. Epidermis: a layer of epidermal cells some with rosette aggregate crystals, numerous multicellular uniseriate trichomes, some with collapsed cells, glandular trichomes, and stomata. Cortex: collenchyma, several layers of lamella cells underneath epidermal layer, parenchyma some with rosette aggregate crystals or starch grains, angular collenchyma near vascular tissue, and schizogenous cavities. Vascular tissue: phloem and xylem.

                        Blumea Balsamifera Leaf in powder possesses the diagnostic microscopical characters of the unground drug. Multicellular uniseriate trichomes, some with collapsed cells, and glandular trichomes are characteristic.

Packaging and storage  Blumea Balsamifera Leaf shall be kept in well-closed containers, preferably of metal or glass, protected from light, and stored in a dry place.

Identification

A. To 200 mg of the sample, in powder, add 10 mL of ethanol, shake, allow to stand for 10 minutes, and filter. To 2 mL of the filtrate, add 0.5 mL each of a 5 per cent w/v solution of aluminium chloride, a 5 per cent w/v solution of sodium nitrite, and a 20 per cent w/v solution of sodium hydroxide successively. Mix well: a red colour develops.

B. Carry out the test as described in the “Thin-Layer Chromatography” (Appendix 3.1), using a high-performance plate with silica gel GF254 as the coating substance and a mixture of 94 volumes of chloroform, 5 volumes of methanol, and 1 volume of formic acid as the mobile phase and allowing the solvent front to ascend 8 cm above the line of application. Apply separately to the plate as bands of 8 mm, 10 μL of solution (A) and 2 μL of solution (B). Prepare solution (A) by adding 5 mL of methanol to 200 mg of the sample, in fine powder, shaking, allowing to stand for 10 minutes, and filtering. For solution (B), dissolve 1 mg of blumeatin 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. The chromatogram obtained from solution (A) shows a quenching band (hRf value 39 to 41) corresponding to the blumeatin band obtained from solution (B); several other quenching bands are observed. Subsequently, spray the plate with a 1 per cent w/v solution of aluminium chloride in ethanol and examine under ultraviolet light (366 nm). The band due to blumeatin shows green fluorescent. One yellow, three green, and three red fluorescent bands are also observed (Fig. 3).

Water  Not more than 9.0 per cent v/w (Azeotropic Distillation Method, Appendix 4.12).

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

Acid-insoluble ash  Not more than 6.0 per cent w/w (Appendix 7.6).

Total ash  Not more than 17.0 per cent w/w (Appendix 7.7).

Ethanol-soluble extractive  Not less than 6.0 per cent w/w (Appendix 7.12).

Water-soluble extractive  Not less than 13.0 per cent w/w (Appendix 7.12).

Volatile oil  Not less than 0.5 per cent v/w, calculated on the anhydrous basis (Appendix 7.3H). Use 20 g, in fine powder, freshly prepared and accurately weighed. Use 250 mL of water as the distillation liquid and a 500-mL round-bottomed flask. Distil at a rate of 2 to 3 mL per minute for 5 hours. Use 2.0 mL of xylene in the graduated tube.