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         Wild Spikenard is the dried aerial part of Hyptis suaveolens (L.) Poit. (Family Labiatae), Herbarium Specimen Number:  BKF 194900, Crude Drug Number:  DMSc 0436.

Constituents  Wild Spikenard contains terpenoids (e.g., betulinic acid, suaveolol, ursolic acid), phenolic compounds (e.g., caffeic acid, rosmarinic acid).  It also contains flavonoids, sterols, small amount of volatile oil, etc.

Description of the plant  (Figs. 1a, 1b)  Annual herb 0.6 to 1.6 m tall, strongly aromatic; stem erect, robust, quadrangular, branched, with long spreading white hairs.  Leaves simple, opposite decussate, ovate to broadly ovate, 2 to 5 cm long, 2 to 6 cm wide, apex subacute  to obtuse, base rounded to shallow cordate, oblique, margin serrulate, subcoriaceous,  gland-dotted, both surfaces hairy; petiole slender, 0.5 to 6 cm long.  Inflorescence axillary or terminal cyme, 1- or 2- to 5-flowered, in raceme or panicle; bract small.  Flower zygomorphic; calyx campanulate, 0.8 to 1(–1.2) cm long in fruit, 3 to 5 mm wide, villous, yellowish glandular, throat tufted villous, prominent 10-nerved, 5-toothed, spine-like, 1.5 to 2 mm long, with simple and glandular hairs; corolla blue, 6 to 8 mm long, puberulent except near base, tube about 2 mm wide at throat, upper lip 2-lobed, reflexed, lower lip 3-lobed, middle lobe shorter, lateral lobes similar to upper lip; stamens 4, filament flattened, minutely hairy, exserted from the corolla tube; ovary superior, 2-carpellate, each carpel deeply 2-lobed, style 1, bifid, stigma minute.  Fruit 2 nutlets, broadly obovoid, 3 to 4 mm long, 2 to 3 mm wide, apex truncate and emarginate, mucronate, laterally compressed, dark brown, dotted, with  2 basal white scars, within enlarged calyx tube. 

Description  Odour, aromatic; taste, slightly bitter. 

     Macroscopical  (Fig. 1a)  Stem quadrangular, greenish brown to brown; leaves opposite-decussately arranged, the complete ones ovate to broadly ovate, greenish, venation prominent; remnant inflorescence, persistent calyx, brown, villose. 

        Microscopical  (Figs. 2a, 2b, 2c, 2d)  Transverse sections of the leaf through the midrib  and the lamina show upper epidermis, mesophyll, vascular bundles, and lower epidermis.  Upper epidermis, a layer of wavy epidermal cells; stomata, mostly diacytic with some anomocytic; trichomes, numerous, multicellular uniseriate and glandular.  Mesophyll,  a layer of columnar palisade cells and several layers of spongy parenchyma, some containing rosette aggregate crystals.  Vascular bundle, composed of phloem and xylem; vessels, spiral and scalariform.  Collenchyma occurring in the midrib, beneath upper and lower epidermises, round and thick-walled cells.   

        Transverse section of the stem shows epidermis, cortex and vascular tissue.  Epidermis,  a layer of rectangular cells; trichomes, multicellular uniseriate, glandular and glandular with stalk. Cortex, layers of angular and lamella collenchyma cells, parenchyma and groups of fibres in the mature stem. Vascular tissue, composed of phloem and xylem; vessels, spiral and scalariform.  Pith, numerous round parenchyma. 

      Wild Spikenard in powder possesses the diagnostic microscopical characters of the unground drug.  Wavy epidermal cells, diacytic with some anomocytic stomata, numerous multicellular uniseriate trichomes with some collapsed cells, glandular trichomes, and collenchyma are distinguished features.  Sclereids of calyx, mucilage mass from seed coat and hexacolpate pollen grains are also unique. 

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

Identification 

      A. Reflux 500 mg of the sample, in powder, with 10 mL of methanol on a water-bath for 15 minutes and filter.  To the filtrate, add 300 mg of decolorizing charcoal, swirl for a few minutes and filter.  Evaporate the filtrate to dryness.  Dissolve the residue in 1 mL of acetic anhydride and slowly add 1 mL of sulfuric acid:  a reddish pink colour ring develops.

      B. Heat 2 g of the sample, in powder, with 20 mL of water on a water-bath for  30 minutes, and filter.  To 2 mL of the filtrate, add a few drops of a freshly prepared 1 per cent w/v solution of iron(III) chloride:  a greenish blue colour is produced.

      C. Carry out the test as described in the “Thin-Layer Chromatography” (Appendix 3.1), using silica gel G as the coating substance and a mixture of 70 volumes of dichloromethane, 30 volumes of ethyl acetate and 1 volume of water as the mobile phase and allowing the solvent front to ascend 12 cm above the line of application.  Apply separately to the plate, 2 μL each of solution (A) and solution (B).  Prepare solution (A) by refluxing 1 g of the sample, in powder, with 20 mL of methanol for 30 minutes, filtering while hot and evaporating the filtrate to dryness under reduced pressure at 50°.  Dissolve the residue in 10 mL of methanol.  For solution (B), dissolve 2 mg of ursolic acid in 1 mL of methanol.  After removal of the plate, allow it to dry in air.  Spray the plate with a 20 per cent v/v solution of sulfuric acid in ethanol and heat at 105° for 5 minutes.  The chromatogram obtained from solution (A) shows a red spot (hR_f value 73 to 74) corresponding to the ursolic acid spot from solution (B) and other several spots of different colours are also observed (Table 1); see also Fig. 3.

Table 1 hR_f Values of Components in Methanolic Extract of the Aerial Parts of Hyptis suaveolens (L.) Poit.

       *ursolic acid

Water  Not more than 10.0 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 2.0 per cent w/w (Appendix 7.6).

Total ash  Not more than 10.0 per cent w/w (Appendix 7.7).

Ethanol-soluble extractive  Not less than 4.0 per cent w/w (Appendix 7.12).

Water-soluble extractive  Not less than 7.0 per cent w/w (Appendix 7.12).