Terminalia bellerica (fruit)

Nomenclature

Terminalia bellerica (Gaertn.) Roxb.   Combretaceae  
Standardized common name (English): belleric myrobalan  
Ayurvedic name(s): bibhitaki  
Pinyin name(s): pi li le; mao he zi (fruit)

Microscopic Characteristics

A group of thick walled sclereids. Source: Elan M. Sudberg, Alkemist Laboratories	Covering trichomes showing tannic masses at the base. Source: Elan M. Sudberg, Alkemist Laboratories

High Performance Thin Layer Chromatographic Identification

Belleric myrobalan fruit, mao he zi (fruit) HPTLC ID - UV 254 nm Belleric myrobalan fruit, mao he zi (Terminalia bellerica) Lane Assignments Lanes, from left to right (Track, Volume, Sample): 5 µL Chebulinic acid, ellagic acid, and gallic acid (increasing Rf) 1 µL Belleric myrobalan fruit 1 µL Chebulic myrobalan fruit 1 µL Amla fruit 1 µL Triphala powder  Reference Sample(s) Reference: Dissolve 1 mg of chebulinic acid and 2 mg gallic acid in 10 mL of methanol. Optional: dissolve 1 mg of ellagic acid in 10 mL of methanol.  Stationary Phase Stationary phase, i.e. Silica gel 60, F254  Mobile Phase Ethyl formate, toluene, formic acid, water 30:1.5:4:3 (v/v/v/v)  Sample Preparation Method Sample: Mix 1.0 g of powdered sample with 10 mL of methanol and sonicate for 10 minutes, then centrifuge or filter and use the supernatants/filtrates as test solutions. Derivatization reagent: 1.) NP reagent, Preparation: 1 g of natural products reagent in 200 mL of ethyl acetate; 2.) PEG reagent, Preparation: 10 g of polyethylene glycol 400 in 200 mL of dichloromethane, Use: Heat plate for 3 min at 100°C, dip (time 0, speed 5) in NP reagent, dry, dip (time 0, speed 5) in PEG reagent.  Detection Method Saturated chamber; developing distance 70 mm from lower edge; relative humidity 33%  Other Notes Images presented in this entry are examples and are not intended to be used as basis for setting specifications for quality control purposes. System suitability test: Chebulinic acid: a quenching zone at Rf ~ 0.26 (UV 245 nm). Gallic acid: a quenching zone at Rf ~ 0.71 (UV 245 nm). Identification: Compare result with reference images. The fingerprint of the test solution is similar to that of the corresponding botanical reference sample. Additional weak zones may be present. Under UV 254 nm the test solution shows several quenching zones between Rf ~ 0.18 and 0.28. A characteristic well defined quenching zone at Rf ~ 0.53 is present (red arrow). At the position of gallic acid a faint dark zone is visible. Under UV 366 nm there is a blue fluorescent zone at Rf ~ 0.21 and a characteristic fluorescent zone at Rf ~ 0.53 (green arrow). Test for other species: Under UV 366 nm there are no fluorescent zones between Rf ~ 0.40 and 0.52 and below Rf ~ 0.14 (Chebulic myrobalan fruit, Amla fruit, pink arrows). Belleric myrobalan fruit, mao he zi (fruit) HPTLC ID - UV 254 nm Belleric myrobalan fruit, mao he zi (fruit) HPTLC ID - UV 366 nm, after derivatization Source: HPTLC Association

Triphala powder (mixture) HPTLC ID - UV 254 nm Triphala powder (mixture of Phyllanthus emblica, Terminalia bellerica, Terminalia chebula) Lane Assignments Lanes, from left to right (Track, Volume, Sample): 5 µL Chebulinic acid, ellagic acid, and gallic acid (increasing Rf) 1 µL Belleric myrobalan fruit 1 µL Chebulic myrobalan fruit 1 µL Amla fruit 1 µL Triphala powder 1 1 µL Triphala powder 2 1 µL Triphala powder 3 1 µL Triphala powder 4 1 µL Triphala tablets 1 1 µL Triphala tablets 2 1 µL Triphala tablets 3 1 µL Triphala tablets 4 1 µL Triphala tablets 5 2 µL Triphala powder extract 2 µL Triphala tablets 6  Reference Sample(s) Reference: Dissolve 1 mg of chebulinic acid and 2 mg of gallic acid in 10 mL of methanol. Optional: dissolve 1 mg of ellagic acid in 10 mL of methanol.  Stationary Phase Stationary phase, i.e. Silica gel 60, F254  Mobile Phase Ethyl formate, toluene, formic acid, water 30:1.5:4:3 (v/v/v/v)  Sample Preparation Method Sample: Mix 1.0 g of powdered sample with 10 mL of methanol and sonicate for 10 minutes, then centrifuge or filter and use the supernatants/filtrates as test solutions. Derivatization reagent: 1.) NP reagent, Preparation: 1 g of NP reagent in 200 mL of ethyl acetate; 2.) PEG reagent, Preparation: 10 g of polyethylene glycol 400 in 200 mL of dichloromethane, Use: Heat plate 3min at 100°C, dip (time 0, speed 5) in NP reagent, dry and dip (time 0, speed 5) in PEG reagent.  Detection Method Saturated chamber; developing distance 70 mm from lower edge; relative humidity 33%  Other Notes Images presented in this entry are examples and are not intended to be used as basis for setting specifications for quality control purposes. System suitability test: Chebulinic acid: a quenching zone at Rf ~ 0.26 (UV 245 nm). Gallic acid: a quenching zone at Rf ~ 0.71 (UV 245 nm). Identification: Compare result with reference images. The fingerprint of the test solution is similar to that of the corresponding botanical reference sample. Additional weak zones may be present. Under UV 254 nm the test solution shows several diffuse quenching zones above the application position. The two main quenching zones are detected at Rf ~ 0.19 and Rf ~ 0.28 (red arrows). Above these zones there are additional diffuse quenching zones. There are quenching zones at the position of ellagic acid and of gallic acid. Under UV 366 nm two blue fluorescent zones are detected right above the application position (yellow arrow). Several intense blue fluorescent zones are present between Rf ~ 0.16 and 0.40. At the position of gallic acid there is a fluorescent zone. Triphala powder (mixture) HPTLC ID - UV 254 nm Triphala powder(mixture) HPTLC ID - UV 366 nm, after derivatization Source: HPTLC Association