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1

Stability-indicating TLC—image analysis method for determination of andrographolide in bulk drug and Andrographis paniculata formulations

100%

Phattanawasin P. , Burana-Osot J. , Sotanaphun U. , Kumsum A.

Acta Chromatographica

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2016

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tom Vol. 28, no. 4

525--540

EN

A simple and rapid thin-layer chromatographic (TLC)–image analysis method was developed for stability-indicating studies and determination of andrographolide in bulk drug and in Andrographis paniculata formulations. Good chromatographic separation of andrographolide and the degradation products under various stress conditions was achieved on a silica gel G60 F254 TLC plate with the use of two mobile phases, dichloromethane—toluene—ethanol (6:3:1, v/v/v) and toluene—ethyl acetate—formic acid (5:3.5:1.5, v/v/v) and p-anisaldehyde as visualization reagent. Image analysis of the scanned TLC plate was performed by Sorbfil TLC Videodensitometer, UN-SCAN-IT, JustTLC, and ImageJ software to measure the quantity of the dark bluecolored band of andrographolide on a TLC plate. The TLC—image analysis method was validated in terms of specificity, linearity, sensitivity, precision, accuracy, and robustness and was also applied for determination of the amount of andrographolide in A. paniculata formulations and the content of andrographolide remained in the samples under forced degradations. The analytical results determined by the TLC—image analysis method, TLC—densitometry, and high-performance liquid chromatography (HPLC) methods were compared and found to be not significantly different.

2

Simultaneous estimation of anti-cancer terpenoids in pharmaceutical nanoformulation by RP-HPLC and HPTLC

100%

Parveen R. , Ahmad F. J. , Iqbal Z. , Singh M. , Kamal Y. , Ahmad S.

Acta Chromatographica

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2014

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tom Vol. 26, no. 2

391--400

EN

Andrographolide and betulinic acid are the terpenoids having potential anti-cancer activity. The cytotoxicity activity of both the drugs was carried out separately and in combination on liver cancer HepG2 cell lines. High-performance liquid chromatography (HPLC) and high-performance thin-layer chromatography (HPTLC) methods were developed and validated for simultaneous estimation of these two terpenoids as per the International Conference on Harmonization (ICH) guidelines, which was applied for quantification in nanoformulation. The retention time by HPLC and retardation factor by HPTLC for andrographolide and betulinic acid were found to be 2.2 and 6.6 min, and 0.24 ± 0.01 and 0.66 ± 0.01, respectively. Both the methods were validated for accuracy, precision, repeatability, robustness, limit of detection (LOD), and limit of quantitation (LOQ). The content of andrographolide and betulinic acid in nanoformulation was found to be 96.0% and 98.0% by HPLC and 96.59% and 98.33% by HPTLC, respectively, of labelled claim.

3

Ultra-rapid determination of andrographolide and dehydroandrographolide in Andrographis Herba by solid phase extraction and liquid chromatography with mass spektrometry

88%

Qian Z. , Chen J. , Lei Q. , Li D. , Tan G. , Xie J. , Li W.

Acta Chromatographica

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2024

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tom Vol. 36, no. 3

279-–285

EN

An ultra-rapid analytical method for determination of andrographolide and dehydroandrographolide in Andrographis Herba (AH) was developed by liquid chromatography with mass spectrometry (LC-MS). The sample was ultrasonically extracted with 10 mL 40% (v/v) methanol, and then purified with a C18 solid phase extraction column. The LC separation was performed on a Poroshell 120 EC-C18 column (3032.1 mm, 2.7 μm) and eluted with 0.5 mmol L1 ammonium acetate aqueous solution and acetonitrile (65:35) at a flow rate of 0.7 mL min1, and detected by mass spectrometry (MS). The LC-MS analytical time was less than 1 min. The new developed method presented a good linearity (r > 0.9900), precision and repeatability (RSD < 2.0%). The recoveries for andrographolide and dehydroandrographolide were 93.5% (RSD 5 2.2%) and 97.7% (RSD 5 2.4%), respectively. The developed method was successfully applied in determination of andrographolide and dehydroandrographolide in seven batches of AH samples, and the contents of analytes in all samples were complied with the relative acceptance criteria in Chinese Pharmacopeia (>0.8%). This new developed LC-MS method is an ultrarapid assay method for AH, which will help to improve the efficiency and reduce the cost of AH sample test.

4

Development and validation of RP-HPLC method for simultaneous analysis of andrographolide, phyllanthin, and hypophyllanthin from herbal hepatoprotective formulation

88%

Bhope S. G. , Kuber V. V. , Nagore D. H. , Gaikwad P. S. , Patil M. J.

Acta Chromatographica

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2013

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tom Vol. 25, no. 1

159--169

EN

A simple, sensitive, and accurate liquid chromatographic method with photodiode array detector was developed for the determination of andrographolide, phyllanthin, and hypophyllanthin. The separation was carried out on a reverse-phase 250 mm × 4.6 mm, 5μ symmetry C8 column (Waters). The gradient was prepared from 0.1% orthophosphoric acid (solvent A) and (1:1) acetonitrile:methanol (solvent B) as mobile phase delivered at a flow rate of 1 mL min -1. A linear behavior was observed between observed peak area response, and concentration of analytes was investigated, with good correlation coefficient. The method established was successfully applied to quantify andrographolide, phyllanthin, and hypophyllanthin from the herbal hepatoprotective formulation.

5

Isolation, characterization and evaluation of anti-proliferative properties of andrographolide isolated from Andrographis paniculata on cultured HaCaT cells

75%

Jindal S. , Awasthi R. , Singare D. , Kulkarni G. T.

Herba Polonica

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2021

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tom 67

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nr 1

6

In vitro α-glucosidase and α-amylase enzyme inhibitory effects of Andrographis paniculata extract and andrographolide

75%

Subramanian R. , Asmawi M. , Sadikun A.

Acta Biochimica Polonica

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2008

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tom 55

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nr 2

391-398

EN

There has been an enormous interest in the development of alternative medicines for type 2 diabetes, specifically screening for phytochemicals with the ability to delay or prevent glucose absorption. The goal of the present study was to provide in vitro evidence for potential inhibition of α-glucosidase and α-amylase enzymes, followed by a confirmatory in vivo study on rats to generate a stronger biochemical rationale for further studies on the ethanolic extract of Andrographis paniculata and andrographolide. The extract showed appreciable α-glucosidase inhibitory effect in a concentration-dependent manner (IC50=17.2±0.15 mg/ml) and a weak α-amylase inhibitory activity (IC50=50.9±0.17 mg/ml). Andrographolide demonstrated a similar (IC50=11.0±0.28 mg/ml) α-glucosidase and α-amylase inhibitory activity (IC50=11.3±0.29 mg/ml). The positive in vitro enzyme inhibition tests paved way for confirmatory in vivo studies. The in vivo studies demonstrated that A. paniculata extract significantly (P<0.05) reduced peak blood glucose and area under curve in diabetic rats when challenged with oral administration of starch and sucrose. Further, andrographolide also caused a significant (P<0.05) reduction in peak blood glucose and area under the curve in diabetic rats. Hence α-glucosidase inhibition may possibly be one of the mechanisms for the A. paniculata extract to exert antidiabetic activity and indicates that AP extract can be considered as a potential candidate for the management of type 2 diabetes mellitus.

7

In vitro alpha-glucosidase and alpha-amylase enzyme inhibitory effects of Andrographis paniculata extract and andrographolide

63%

Subramanian R. , Asmawi M. Z. , Sadikun A.

Acta Biochimica Polonica

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2008

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tom 55

|

nr 2

391-398

EN

There has been an enormous interest in the development of alternative medicines for type 2 diabetes, specifically screening for phytochemicals with the ability to delay or prevent glucose absorption. The goal of the present study was to provide in vitroevidence for potential inhibition of α-glucosidase and α-amylase enzymes, followed by a confirmatory in vivostudy on rats to generate a stronger biochemical rationale for further studies on the ethanolic extract of Andrographis paniculata and andrographolide. The extract showed appreciable α-glucosidase inhibitory effect in a concentration-dependent manner (IC50=17.2±0.15 mg/ml) and a weak α-amylase inhibitory activity (IC50=50.9±0.17 mg/ml). Andrographolide demonstrated a similar (IC50=11.0±0.28 mg/ml) α-glucosidase and α-amylase inhibitory activity (IC50=11.3±0.29 mg/ml). The positive in vitroenzyme inhibition tests paved way for confirmatory in vivostudies. The in vivostudies demonstrated that A. Paniculata extract significantly (P<0.05) reduced peak blood glucose and area under curve in diabetic rats when challenged with oral administration of starch and sucrose. Further, andrographolide also caused a significant (P<0.05) reduction in peak blood glucose and area under the curve in diabetic rats. Hence α-glucosidase inhibition may possibly be one of the mechanisms for the A. paniculataextract to exert antidiabetic activity and indicates that AP extract can be considered as a potential candidate for the management of type 2 diabetes mellitus.