Assessment of pharmacognostic specification of Cannabis sativa leaves in Thailand.

Lack of quality control can affect the safety, efficacy, and acceptability of herbal products that may lead to health problems. Cannabis sativa L. (Cannabaceae) has been widely used as an ethnomedicinal practice for its medicinal values. This study aims to establish pharmacognostic specifications of C. sativa as per standard procedures. Macroscopic-microscopic characteristics, physicochemical parameters, thin-layer chromatography (TLC) fingerprinting, and phytochemical screening of C. sativa leaves collected from various locations throughout Thailand were investigated. Leaves are palmate consists of seven leaflets with green color, margin is serrate with acuminate apex. Anomocytic stomata were found in the upper epidermis while unicellular and glandular trichomes with cystolith were found in the lower epidermis and the epidermis layer covered with cuticle. The physicochemical analysis revealed that the loss on drying (4.068 ± 0.084 %w/w) was within acceptable limits, total ash (14.360 ± 0.084%w/w), acid insoluble ash (2.726 ± 0.080%w/w), ethanol-soluble extractive (11.101 ± 0.223%w/w), water-soluble extractive (23.038 ± 0.306%w/w), and water content (7.523 ± 0.524%w/w). TLC fingerprint showed nine spots with Rf value 0.14, 019, 0.23, 0.29, 0.32, 0.45, 0.58, 0.70, and 0.76. Phytochemical screening of Cannabis leaves indicated the presence of phenolic compounds, flavonoids, alkaloids, diterpenes, triterpenes, and steroids. This study provided referential data for the accurate plant identity, and establishment of cannabis leaves monograph in Thailand.

Quality evaluation with reference to clitoriacetal and in vitro antioxidant activities of Clitoria macrophylla root.

Clitoria macrophylla Wall. (syn. Clitoria hanceana Hemsl.), is commonly known in Thai as Nontai-yak, An-chan-pa, or Ueang-chan-pa, which belongs to Leguminosae family. According to traditional Thai medicine, the root has been used for the treatment of skin diseased as well as for pest control in horticulture and animal husbandry. The aim of this study is to investigate for the pharmacognostic specification, the clitoriacetal content, and in vitro antioxidant capacities of C. macrophylla roots from 12 different sources throughout Thailand. Clitoriacetal content was quantitatively analyzed by thin-layer chromatography (TLC) - densitometry with winCATS software and TLC image analysis with ImageJ software. Antioxidant activities were evaluated by 2, 2-diphenyl-1-picrylhydrazyl radical scavenging, ferric reducing antioxidant power assay, ß-carotene bleaching assay, total phenolic, and total flavonoid contents. The pharmacognostic specification of C. macrophylla roots in Thailand was established. The loss on drying, total ash, acid-insoluble ash, and water contents should be not more than 6.40%, 12.29%, 8.89%, and 8.16% of dry weight, whereas ethanol and water-soluble extractive values should be not less than 4.95% and 14.72% of dry weight, respectively. Furthermore, the clitoriacetal content of C. macrophylla roots determined by TLC-densitometry and TLC image analysis was found to be 2.20 ± 1.31 and 2.22 ± 1.16 g/100 g of dry roots, respectively. The clitoriacetal contents of both methods were not significantly different using paired t-test. Moreover, the ethanolic extract of C. macrophylla roots showed its antioxidant potential compared to the standard butylated hydroxyl toluene and quercetin.

Pharmacognostic Specifications and Lawsone Content of Lawsonia inermis Leaves.

BACKGROUND: Lawsonia inermis L. has been used as a traditional or folk medicine for the treatment of a wide range of skin infectious diseases. OBJECTIVE: The objective of this study was to determine the pharmacognostic specifications and lawsone contents of L. inermis leaves. MATERIALS AND METHODS: The pharmacognostic specifications of L. inermis leaves from 12 sources were evaluated according to the WHO guideline of quality control method for medicinal plant materials. The lawsone contents were analyzed by thin-layer chromatography (TLC) coupled with densitometry and image analysis. RESULTS: Microscopic evaluation of L. inermis powders showed the fragment of mesophyll, fragment of parenchyma, epidermis layer with stomata, and the rosette crystal of calcium oxalate. Physicochemical parameters revealed that total ash, acid-insoluble ash, loss on drying, and water content should be not <6.98, 1.12, 8.08, and 9.86% of dried weight, respectively, whereas ethanol and water extractive values should be not < 19.67 and 23.06% of dried weight, respectively. The content of lawsone in L. inermis leaves by TLC-densitometry was found to be 0.76 ± 0.05 g/100 g of dried crude drug, whereas the lawsone content evaluation by TLC image analysis was found to be 0.87 ± 0.11 g/100 g of dried crude drug. The validation of the methods revealed that both TLC-densitometry and TLC image analysis showed a good sensitivity and accuracy for lawsone quantitation in L. inermis. CONCLUSION: The pharmacognostic specifications could be used as the standardization data of L. inermis leaves, and the development of TLC method could be applied to determine lawsone content in this plant material. SUMMARY: The pharmacognostic specification of Lawsonia inermis leaves could be used as the standardization data of L. inermis leaves in Thailand.Both TLC-densitometry and TLC image analysis showed a good sensitivity and accuracy for lawsone quantitation. Abbreviations Used: LOD: Limit of detection; LOQ: Limit of quantitation; RSD: Relative standard deviation; TLC: Thin layer chromatography; UV: Ultraviolet; Rf value: Relative to front value.

Pharmacognostic Specification, Chlorogenic Acid Content, and In vitro Antioxidant Activities of Lonicera japonica Flowering Bud.

BACKGROUND: Lonicera japonica Thunb. or Japanese Honeysuckle has been widely used in traditional medicine for antipyretic. OBJECTIVE: To establish the pharmacognostic specification of L. japonica flowering bud in Thailand and to determine its chlorogenic acid content and in vitro antioxidant activities. MATERIALS AND METHODS: Dried L. japonica flowering bud from 15 various herbal drugstores throughout Thailand were investigated for pharmacognostic specification. Their chlorogenic acid contents were quantitatively analyzed by thin layer chromatography (TLC) densitometry with winCATS software. The mobile phase for TLC development consisted of ethyl acetate: formic acid: acetic acid: water (10:1.1:1.1:2.6). Antioxidant activities were investigated by 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging, ferric ion reducing antioxidant power assay, nitric oxide scavenging assay, and ß-carotene bleaching assays. RESULTS: Qualified L. japonica flowering bud in Thailand was presented that the contents of loss on drying, total ash, acid-insoluble ash, and water should not be >10.11%, 6.59%, 1.14%, and 10.82% by weight, respectively. The ethanol and water soluble extractive values should not be < 16.46% and 28.88% by weight, respectively. Chlorogenic acid content in L. japonica flowering bud was found to be 2.24 ± 0.50 g/100 g of crude drug. L. japonica flowering bud showed DPPH and nitric oxide scavenging activities as well as reducing power property. CONCLUSION: This pharmacognostic specification with special reference to the chlorogenic acid content can be used for quality control of L. japonica flowering bud in Thailand. The potential antioxidant of this crude drug was demonstrated in vitro. SUMMARY: Pharmacognostic specification of Lonicera japonica flowering bud in Thailand has been establishedThe chlorogenic acid content has been quantified by thin layer chromatography-densitometryThe ethanolic extract of L. japonica flowering bud showed antioxidation potential, especially on reducing power property. Abbreviations Used: TLC: Thin layer chromatography, DPPH: 2,2-diphenyl-1-picrylhydrazyl, FRAP: Ferric ion Reducing Antioxidant Power, WHO: World Health Organization, ICH: International Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use; LOD: Limit of detection; LOQ: Limit of quantitation; BHT: Butylated hydroxytoluene; FeSO4: Iron(II) sulfate; DMSO: Dimethyl sulfoxide; TPTZ: 2,4,6-tripyridyl-s-triazine.

Pharmacognostic specification of Zanthoxylum limonella (Dennst.) Alston: Fruits and seeds in Thailand.

Zanthoxylum limonella (Dennst.) Alston. (Rutaceae) or Ma-khwaen is one of the medicinal plants in Thai traditional medicine. To investigate the pharmacognostic specifications and chemical constituents of Z. limonella fruits and seeds. Fruits and seeds of Z. limonella were collected from 15 sources throughout Thailand; then examined the pharmacognostic specification following WHO guideline of quality control method for medicinal plant materials. Microscopic determination of Z. limonella powders demonstrated fragment of mesocarp, fragment of brown vitta, oil glands, fragment of endocarp, and endosperm containing oil globule, trichome and pale brown stone cells. Stomatal index and pellucid dots in mm2 were 19.87 and 4.2 respectively. Physico-chemical parameters unveiled that loss on drying, water content, total ash, and acid-insoluble ash should be not >17.90%, 9.18%, 4.50%, and 0.60% of dried weight respectively; while ethanol, water, and hexane extractive values and volatile oil content should be not <2.24%, 2.27%, 1.57% and 9.63% of dried weight respectively. Rf values of thin-layer chromatographic fingerprint of Z. limonella fruits and seeds ethanolic extract were 0.38, 0.45, 0.90, and 0.97 detected ultraviolet (UV) light 254 nm, 0.30, 0.44, 0.67, and 0.77 detected UV light 366 nm, and 0.24, 0.73, 0.78, and 0.93 detected 10% sulfuric acid. There are three main chemical compounds in Z. limonella oil including limonene (43.63%), (+)-sabinene (16.72%), and terpinen-4-ol (10.95%). The result gained from pharmacognostic specifications and chemical fingerprints could be used as standardization data of Z. limonella fruits and seeds to apply or provide for guarantee of quality.

Pharmacognostic specifications and quantification of (+)-catechin and (-)-epicatechin in Pentace burmanica stem bark.

BACKGROUND: According to Thai traditional medicine, Pentace burmanica Kurz. stem bark has been used as crude drug for treating diarrhea. However, the crude drug is also found susceptible to adulteration. OBJECTIVES: To develop specific standardization parameters of P. burmanica stem bark in Thailand and to determine the (+)-catechin and (-)-epicatechin contents of P. burmanica stem bark by HPLC analysis. MATERIALS AND METHODS: P. burmanica stem barks from various sources throughout Thailand were investigated according to WHO guideline of the pharmacognostic specification. High performance liquid chromatography (HPLC) was performed for (+)-catechin and (-)-epicatechin quantification. RESULTS: Macroscopic evaluation was demonstrated as whole plant drawing. Microscopic evaluation of stem bark powdered drug showed fragment of fibers, resin masses, tannin masses, starch grain, calcium oxalate, and fragment of parenchyma. Physico-chemical parameters revealed that total ash, acid insoluble ash, loss on drying, and water content should be not more than 3.58, 0.50, 8.40, and 9.70% of dry weight respectively; while ethanol and water soluble extractive values should not be less than 21.90 and 19.06% of dry weight respectively. Both (+)-catechin and (-)-epicatechin were existed in P. burmanica ethanolic extract. Owing to the small amount of (+)-catechin, quantitation of its content was omitted. However, (-)-epicatechin contents was found as 59.74 ± 1.69µg/mg of crude extract. CONCLUSION: The pharmacognostic investigations can be used to set the standard parameters of P. burmanica stem bark in Thailand. HPLC method can be applied to determine (+)-catechin and (-)-epicatechin content in plant materials.