Authors:V. Sarma, Pullela Srinivas, V. Anuradha, and J. Rao
The dried fruits of
are extensively used in Ayurvedic medicinal preparations. A simple and convenient HPTLC method has been developed for standardization of the plant material using the two major constituents, pellitorine and dihydropiperlonguminine, as markers. The stationary phase was silica gel 60 F
, hexane-ethyl acetate, 75 + 25 (
), was used as mobile phase, and detection was at
= 260 nm. The method is characterized by high sensitivity and linearity over a wide range of concentrations. The results obtained were evaluated statistically.
Authors:Kilambi Pundarikakshudu, Anilkumar Sharma, Chaitanya Bhatt, and Niranjan Kanaki
The fruit of Piper longum Linn. (family: Piperaceae), known as pippali in India, is a reputed drug of Ayurveda (the Indian system of medicine). It contains many amides of piperic acid among which piperine and piperlongumine are important from biological point of view. In the present work, we quantified these two marker compounds from the fruits of P. longum by high-performance thin-layer chromatography (HPTLC) with densitometry. The method was found to be precise. Relative standard deviation (RSD) values for intra-day analyses were in the range of 1.05 to1.07% (piperine) and 1.12 to 1.16% (piperlongumine) and, for inter-day analyses, the values were in the range of 1.11 to 1.21% (piperine) and 1.02 to1.18% (piperlongumine). Instrumental RSD values were 0.64 and 0.78% for piperine and piperlongumine, respectively. Accuracy of the method was evaluated by carrying a recovery study at three different levels for the two compounds. Average recoveries were found to be 99.23% for piperine and 99.26% for piperlongumine. P. longum fruits obtained from Bombay market showed 0.213 ± 0.009% w/w piperine and 0.364 ± 0.014% w/w piperlongumine when analyzed by the proposed method. Similarly, sample fruits obtained from Ahmedabad market had 0.85 ± 0.026% piperine and 2.70 ± 0.065% of piperlongumine. This TLC-densitometric method was found to be simple, precise, specific, sensitive, and accurate. So it can be used in routine quality control for the simultaneous analysis of piperine and piperlongumine from P. longum fruits and piperine alone from Piper nigrum fruits.
A simple, sensitive, and rapid high-performance thin layer chromatographic (HPTLC) method has been established for estimation of piperine in commercial Ayurvedic formulations and in the fruits of
Linn. Chromatography was performed on aluminum foil HPTLC plates coated with 0.2 mm layers of silica gel F
, with hexane-acetone 6.5: 3.5 (
) as mobile phase. The development distance was 76 mm, the temperature 25 ± 5°C, and the chamber was saturated for 5 min. Piperine was quantified at 340 nm, its wavelength of maximum absorbance. Under the conditions used the
of piperine was 0.33 and the limit of detection (LOD) was 4 ng per zone. The calibration plot was linear in the range of 10 to 60 ng per zone with a correlation coefficient of 0.9996. Recovery was in the range 98.76 to 100.70%. This HPTLC method was found to be reproducible, accurate, and precise and could be used to detect piperine at nanogram levels. The method is a very simple and cost-effective means of quantitative estimation of piperine in Ayurvedic formulations.
Authors:Anilkumar S. Sharma, Chaitanya J. Bhatt, and Kilambi Pundarikakshudu
Trikatu Churna is an important formulation in Ayurveda — the Traditional System of Indian Medicine. It consists of fine powders of fruits of Piper nigrum L., Piper longum L., and rhizomes of Zingiber officinale Roscoe in equal proportions. Piperine, present in both P. nigrum and P. longum, is considered to be responsible for the improvement of digestion and bioavailability enhancement of many medicaments. Gingerols and 6-shogaol are key chemical molecules in Z. officinale. Piperlongumine is present in P. longum fruits but absent in the fruits of P. nigrum. We report a validated high-performance thin-layer chromatography (HPTLC) method for the determination of piperine, piperlongumine, and 6-shogaol in these herbs and in Trikatu Churna. Piperine, piperlongumine, and 6-shogaol resolved well in n-hexane—ethyl acetate (8:2) on precoated silica gel 60 F254 plates. The absorption maxima for piperine, piperlongumine, and 6-shogaol were found to be 327, 272 and 235 nm, respectively. Linearity for the corresponding markers was observed between 0.1–0.5, 0.2–1.0, and 0.1–1.6 μg spot−1, respectively. The limit of detection (LOD) and limit of quantification (LOQ) were 28 and 100, 56 and 200, and 32 and 100 ng for piperine, piperlongumine, and 6-shogaol, respectively. Recovery experiments showed 99.6%, 99.5%, and 99.7% recoveries for piperine, piperlongumine, and 6-shogaol, respectively. P. nigrum fruits from Delhi and Ahmedabad had around 2.0% w/w piperine, while fruits of P. longum from these markets were analyzed for 0.8% and 0.6% w/w piperine. Piperlongumine was not found in both samples of P. nigrum, while the fruits of P. longum had 0.36% and 0.26% w/w piperlongumine. Z. officinale from Delhi had 0.19% w/w of 6-shogaol as against 0.16% w/w found in the sample from Ahmedabad. Plant materials procured from Delhi were employed for the preparation of Trikatu Churna which showed 96.5%, 95%, and 103% w/w of the expected values of piperine, piperlongumine, and 6-shogaol, respectively. The present method is simple, reproducible, and reliable which can be applied for the routine analysis of Trikatu Churna and its ingredients in polyherbal formulations.
Authors:Y.T. Kamal, Mhaveer Singh, Shahana Salam, and Sayeed Ahmad
The alternative system of medicines like Unani and Ayurveda is preferred worldwide nowadays due to its therapeutic efficacy, lower side effects, holistic approach, psychological dimensions, and qualitative action of weather and seasonal requirement. A simple procedure is described for the simultaneous extraction and estimation of piperlongumine and piperine in a well-known Unani polyherbal formulation using reversed-phase high-performance liquid chromatography (HPLC). The chromatography was carried out on reversed-phase C18 (250 × 4.6 mm) column with a mobile phase containing acetonitrile—water (50:50 v/v). Detection was accomplished with ultraviolet (UV) detection at λ = 325 nm. The flow rate was kept as 1.0 mL−1. The proposed method was validated according to International Conference on Harmonization (ICH) guidelines for accuracy (94.4–105.0%), precision (0.37–2.17% RSD), and robustness (0.14–2.11% RSD). The limit of detection (LOD) values were found as 30 and 10 ng mL−1, while limit of quantification (LOQ) was 100 and 30 ng mL−1 for piperlongumine and piperine, respectively, which proved the sensitivity of the method satisfactory enough for accurate analysis of the both piperlongumine and piperine.
Authors:Anagha Rajopadhye, Anuradha Upadhye, and Arvind Mujumdar
Piperine, an alkaloid with diverse biological activity commonly occurring in fruits of Piper sp., has high commercial, economical, and medicinal value. In this communication densitometric estimation of piperine from fruits of Piper nigrum L., processed Piper nigrum L. (white pepper), Piper longum L., Piper retrofractum Vahl, Piper cubeba Hunter, and Piper betle L. is reported. Extracts of these fruits and a standard solution of piperine were applied to silica gel F254 HPTLC plates and the plates were developed in a twin-trough chamber with toluene-ethyl acetate-diethyl ether 6:3:1 as mobile phase. The plates were scanned at 337 nm and quantification of piperine was based on a predetermined calibration plot obtained by chromatography of 15 to 75 ng standard. The quantity of piperine was highest in fruits of Piper nigrum L. and least in Piper betle L. The trend was Piper nigrum L. > Piper longum L. > processed Piper nigrum L. > Piper retrofractum Vahl > Piper cubeba Hunter > Piper betle L. The HPTLC method was found to enable simple, convenient, rapid screening and quantification of the active marker piperine in six fruits of Piper sp. commonly used in the Indian traditional system of medicine.
Authors:Bokka Ramesh, Vanka Uma Maheswara Sarma, Katragunta Kumar, Katragadda Suresh Babu, and Potturi Sita Devi
The isolation and characterization of bioactive compounds from medicinal plants is usually a significant challenge in phytochemical analysis because of the natural chemical complexity of plant extracts. However, there exists a need for analytical tools which can quantitatively separate and characterize the components from these biosources with greater chromatographic selectivity and lesser analytical run times that facilitate the evaluation with enhanced separation profiles. Hyphenation of thin-layer chromatography (TLC/HPTLC) with mass spectrometry (MS) is an alternative for screening herbal extracts because of its rapid analysis and ability to aid structural characterization with powerful analytical capacity. The aim of the present study was to develop a sophisticated analytical method which utilizes HPTLC–MS coupling for the chromatographic profiling and evaluation of the therapeutically important genus Piper (Piperaceae). In this study, six marker compounds, namely, trichostachine, piperine, 4,5-dihydropiperlonguminine, guineensine, pellitorine, and sesamin were analyzed and quantified in extracts of Piper nigrum L. and compared with those of Piper longum L. and Piper chaba Hunter. All the samples tested showed similar phytochemical profiles, but the contents of the active ingredients varied. Additionally, HPTLC–MS further allowed confirming the identification of the constituents in the analyzed samples with greater chromatographic selectivity where HPTLC facilitated a selective chromatographic resolution, while MS offered an efficient characterization of the target compounds in one analytical run. The study finds a potential utility in adopting HPTLC–MS as a rapid and high throughput method for the efficient quantification and identification of marker compounds from medicinal plants.
]. Herbal preparations Herbal treatment approaches have been assessed as promising options for therapy-refractory giardiasis as well [ 121 ]. In India, Pippali rasayana, prepared from Piperlongum and Butea monosperma , has been tested for giardicidal