Phyllanthus emblica (fruit)

Nomenclature

Phyllanthus emblica L.   Euphorbiaceae  
Syn. Emblica officinalis Gaertn.  
Standardized common name (English): amla  
Ayurvedic name(s): amalaki  
Pinyin name(s): yu gan zi (fruit)

Botanical Voucher Specimen

Source: MOBOT, Tropicos.org

Microscopic Characteristics

Extremely large rosette/shards of calcium oxalate (some as small as a few 1μm in the form of sand) observed at 400x with Acidified Chloral Hydrate Glycerol Solution. Source: Elan M. Sudberg, Alkemist Laboratories	Large parenchyma with wrinkled cells walls and crystal sand observed at 400x with Acidified Chloral Hydrate Glycerol Solution. Source: Elan M. Sudberg, Alkemist Laboratories

High Performance Thin Layer Chromatographic Identification

Phyllanthus emblica HPTLC ID - 10% Methanolic Sulfuric Acid UV 365 nm Amla (fruit) (Phyllanthus emblica) Lane Assignments Lanes, from left to right (Track, Volume, Sample): 2 μL Gallic Acid ~0.1% in CH3OH 2 μL Phyllanthus emblica-1 2 μL Phyllanthus emblica-2 1 μL Phyllanthus emblica-3 1 μL Phyllanthus emblica-3 3 μL Phyllanthus emblica-4 3 μL Phyllanthus emblica-4 2 μL Gallic Acid ~0.1% in CH3OH Reference materials used here have been authenticated by macroscopic, microscopic &/or TLC studies according to the reference source cited below held at Alkemists Laboratories, Costa Mesa, CA.  Stationary Phase Silica gel 60, F254, 10 x 10 cm HPTLC plates  Mobile Phase toluene: ethyl formate: formic acid [5/4/1]  Sample Preparation Method 0.3g+3mL 70% grain EtOH sonicate/heat @ 50° C ~ 1/2 hr  Detection Method 10% Methanolic H2SO4 -> 115° C 15 min -> UV 365 nm  Reference see Indian Herbal Pharmacopoeia Vol 2 1999 Source: Elan M. Sudberg, Alkemist Laboratories

Phyllanthus emblica (fruit) HPTLC ID - UV 254 nm Amla (fruit) (Phyllanthus emblica) Lane Assignments Lanes, from left to right (Track, Volume, Sample): 5 µL Chebulinic acid, ellagic acid, and gallic acid (with 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 two characteristic quenching zones, one at Rf ~ 0.68 (red arrow) below the zone corresponding to gallic acid and the other at Rf ~ 0.14 (yellow arrow). After derivatization under UV 366 nm the test solution shows two characteristic and very intense fluorescent zones at Rf ~ 0.08 and 0.20 (pink arrows). Test for other species: No quenching zone is seen under UV 254 nm at Rf ~ 0.39 (Chebulic myrobalan fruit; black arrow). No blue fluorescent zone is seen under UV 366 nm at Rf ~ 0.52 (Belleric myrobalan fruit, green arrow). Phyllanthus emblica (fruit) HPTLC ID - UV 254 nm Phyllanthus emblica (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