A high-performance thin layer chromatography (HPTLC) method was used to standardize the 70% ethanol extract of A. javanica as described elsewhere (39 (link)). The chromatography was carried out on 10×10 cm precoated silica gel F254 RP-HPTLC plate, using rutin as the reference standard. Several mobile phases were tried to get good resolution and separation of different compounds present in the A. javanica ethanol extract. Based on our observations, we selected acetonitrile and water in the ratio of 4:6 as suitable mobile phase to carry out the standardization of the extract. The standard along with samples was applied on the HPTLC plate by CAMAG Automatic TLC Sampler-4. The plate was developed under controlled condition in CAMAG Automated Developing Chamber-2 and scanned by CAMAG TLC Scanner-3 (λ=360 nm).
Automated developing chamber 2
Manufactured by CAMAG
The Automated Developing Chamber-2 is a versatile piece of laboratory equipment designed to facilitate the development of thin-layer chromatography (TLC) plates. It automates the process of developing TLC plates, ensuring consistent and reproducible results. The device precisely controls the parameters of the developing process, such as solvent volume, developing time, and chamber saturation, to provide reliable and standardized TLC analyses.
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3 protocols using automated developing chamber 2
1
HPTLC Standardization of A. javanica Extract
2
HPTLC Standardization of 70% SSEE
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The high-performance thin layer chromatography (HPTLC) method was developed and validated to standardize the 70% SSEE as described elsewhere [28 (link)]. Briefly, HPTLC (10 × 10 cm pre-coated silica gel F254 plate) was carried out using β-sitosterol as reference biomarker. Of the several mobile phases tried to get good resolution and separation of different compounds present in SSEE, hexane and ethyl acetate (4:6; v/v) was selected as the best combination. The standard along with samples was applied on the HPTLC plate (CAMAG Automatic TLC Sampler-4), developed (CAMAG Automated Developing Chamber-2), and scanned (λ = 360 nm; CAMAG TLC Scanner-3) for densitometric analysis.
3
Standardization of S. irio Extracts by HPTLC
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A validated high-performance thin layer chromatography (HPTLC) method was used to standardize the total ethanolic extract, n-hexane, dichloromethane, ethyl acetate, and n-butanol fractions of S. irio. The chromatography was performed on 20 × 10 cm precoated silica gel F254 HPTLC plate, using β-sitosterol glucoside as the marker compound. Numerous combination of mobile phases was tried to get good separation and resolution of various compounds present in S. irio. Based on our observations, chloroform and methanol in the ratio of 16:4 were selected as mobile phase to accomplish the standardization. The marker along with different samples was applied by CAMAG automatic TLC sampler-4 on the HPTLC plate. CAMAG Automated Developing Chamber-2 was used to develop the HPTLC plate under controlled condition. The developed plate was dried and derivatized by spraying with p-anisaldehyde reagent and scanned by CAMAG TLC Scanner-3 (λ = 600 nm).
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