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Download Full Article As Article Alkaloids Detection in Commonly Found Medicinal Plants with Marquis Reagent Daniel Alejandro Ocampo-Bustos1 and María Elena Cano-Ruiz1 1 Tecnológico de Monterrey High School, Cuernavaca, Mexico. SUMMARY identity of social groups. Many of the medicinal plants have Alkaloids are a class of nitrogenous organic their healing properties known by empirical use through time, compounds of plant origin that may have important but these medicinal plants may contain active ingredients physiological actions on humans. They include many with tested pharmacological properties. One possibility is that drugs and poisons, but some alkaloids in low doses some of the active ingredients in medicinal plants belong to the have health benefits as well. Traditional medicinal group of alkaloids, which can be determined by a colorimetric plants may contain alkaloids as active ingredients, chemical reaction with the Marquis reagent. The reagent is but this is not well-understood. The Marquis reagent dripped onto the substance being tested, and if an alkaloid exists as a simple qualitative colorimetric method is present, a color change appears (5). The Marquis reagent to determine the presence of alkaloids in medicinal is traditionally composed of a mixture of formaldehyde and plants. The Marquis reagent test was assayed in concentrated sulfuric acid. medicinal plants by first optimizing the formulation Originally, the Marquis reagent was used for testing of the reagent using poppy seeds and lavender as many different alkaloids, and the results from those studies the positive and negative controls. Then using the were the base for developing the color scales that are optimized formulation of Marquis reagent in the extracts of 11 medicinal plants with known claims of used as a reference to determine the specific alkaloid health benefits. Four medicinal plants tested positive that is present in a solution (5, 6). The Marquis reagent for alkaloids, including a relaxing herbal mix (Tilia can give different color changes to hundreds of drugs cordata, Valeriana officinalis, Passiflora incarnata, (7). Purple-dark or blue-dark colors are observed with and Melissa officinalis), Turnera diffusa, Brickellia methylenedioxylphenyl groups in alkaloids, which is present cavanillesii, and Verbascum thapsus. These findings in methylenedioxymethamphetamine (i.e., MDMA, ecstasy); demonstrate the value of the Marquis reagent test to orange is observed with amphetamines; blue-black to yellow provide a rapid and simple method for screening for colors are observed with methylenedioxy-substituted analogs; potentially medicinal alkaloids of natural origin. and red-violet is observed with methylenedioxyphenyl amphetamine-type compounds (8, 9). INTRODUCTION In this study, a possible explanation for the medicinal Alkaloids are a class of nitrogenous organic compounds, properties of some plants was elicited by determining the usually with aromatic rings, that are widely known for their presence of alkaloids in 11 common medicinal plants that are effects on organisms. These effects are put to use in many found in local markets of Mexico. We hypothesized that plants kinds of medications, such as analgesics, relaxants, and with relaxing effects or claims of better sleep would contain even antiarrhythmics. They are also naturally present in some alkaloids, as many medications available in pharmacies for plants like Papaver somniferum, or poppy, from which several these purposes (e.g., clonazepam) are alkaloids. Using the opium alkaloids are extracted; for example, morphine is the Marquis reagent qualitative colorimetric test, we tested the 11 most abundant alkaloid in the poppy plant (1, 2). common medicinal plants for the presence of alkaloids. Four Alkaloids, in their vast majority, can cause addiction medicinal plants tested positive for alkaloids indicating the problems or toxicity when consumed by humans, which value of the Marquis reagent test as a rapid and economical is why most alkaloids are either prohibited, restricted, or, method for screening for potentially medicinal alkaloids of preferably, controlled by governments globally for availability natural origin. only with a medical prescription (3). On the other hand, in very low doses, alkaloids seem to have therapeutic effects RESULTS on certain conditions, such as different kinds of pain, stress, Marquis reagent is a colorimetric test commonly used for hyperactivity, or nervous system-related illnesses (4). the detection of alkaloids. To obtain the best preparation for the Phytotherapy is the use of medicinal plants or extracts of Marquis reagent, we tested several formulations in triplicate natural origin used to prevent and treat different diseases, varying the concentrations and amount of formaldehyde, which is also called folk medicine. In Mexico, this represents acetic acid, and sulfuric acid (10). a system rooted in the deepest part of the cosmovision and Comparing the different color tones that the mixture took Journal of Emerging Investigators • www.emerginginvestigators.org 21 DEC 2020 | VOL 3 | 1 Figure 1: Optimization of Marquis reagent preparation using poppy seed macerate as a positive control. A) (left). Poppy seed macerate was tested after 45 s with the different preparations and Figure 3: Representative images of tea preparations. From left concentrations of the Marquis reagent. First line, no acetic acid to right, tea preparations of Tilia cordata, Turnera diffusa, and the added; second line, no samples; third line, acetic acid added; bottom- relaxing herb mix, using 2 g of dried plant in 100 mL of water at 90°C. left sample, control sample with sulfuric acid only. First and third lines from left to right: 10%, 20%, 30%, and 38% of formaldehyde. tests because sulfuric acid in the reagent will continue reacting All preparations include 1 mL concentrated sulfuric acid and 0.1 mL with organic compounds, producing a dark color; therefore, formaldehyde. B) (right). Poppy seed macerate was tested after 45 s with the different preparations and concentrations of the Marquis the test must be completed in less than 60 seconds. The reagent (double formaldehyde). First line, no acetic acid added; most sensitive formulation of the Marquis reagent for color second line, no samples; third line, acetic acid added; bottom-left development contained 10% formaldehyde in 99% acetic acid sample, control sample with sulfuric acid only. First and third lines and 98% sulfuric acid (Figure 1A). We selected and used this from left to right: 10%, 20%, 30%, and 38% of formaldehyde. All preparations include concentrated 1 mL sulfuric acid and 0.2 mL formulation for determining the presence of alkaloids in the 11 formaldehyde. common Mexican medicinal plants. Dry lavender was used as a negative control in this with the poppy seed macerate when different formulations of experiment because it does not contain alkaloids (11). The the Marquis reagent were added, we determined that there was Marquis reagent did not present a change of color with an alkaloid present (morphine) in the poppy seed macerate lavender with any of the triplicate tests, where the color of the when the color changed from white to different tones of light macerate extract was dark-greenish-brown and remained the orange-brown. All poppy seed samples originated from the same color in all Marquis formulation tests (Figure 2A & B). same macerate, so the different tones observed are resultant We prepared teas from 11 common medicinal plants by of the different reagent formulations rather than variances in three different procedures: a standard preparation using 2 g the samples (Figure 1A & B). dried herbs in 100 mL of hot water, a concentrated preparation Reaction time must be measured during the colorimetric using 2 g dried herbs in 50 mL of hot water, and a macerated preparation with the 2 g of dried herbs blended in 100 mL of hot water (Figure 3). We added equal volumes of each tea preparation to porcelain trays (Figures 4A, 5A, and 6A). A sulfuric acid only control was used to control for any color changes due to the sulfuric acid and not the Marquis reagent with the macerated preparations (Figure 7A). We used the macerated preparation for the sulfuric acid control because, Figure 2: Optimization of Marquis reagent preparation using lavender macerate as a negative control. A) (left). Dry lavender macerate after 45 s with the different preparations and concentrations of the Marquis reagent. First line: no acetic acid added, second line, no samples; third line: acetic acid added; bottom left sample, control sample with sulfuric acid only. First and third lines from left to right: 10%, 20%, 30%, and 38% of formaldehyde. All preparations include 1 mL concentrated sulfuric acid and 0.1 mL formaldehyde. B) (right). Figure 4: Marquis reagent test for medicinal plants using a standard Dry lavender macerate after 45 s with the different preparations and tea preparation. A) (left). Tea samples of each of the 11 medicinal concentrations of the Marquis reagent (double formaldehyde). First plants before the Marquis reagent test. Standard preparation, with line: no acetic acid added; second line, no samples; third line: acetic 2 g of tea in 100 mL of water at 95°C and letting it rest for 5 min. acid added; bottom left sample, control sample with sulfuric acid B) (right). Tea samples of each of the 11 medicinal plants after the only. First and third lines from left to right: 10%, 20%, 30%, and 38% Marquis reagent test. Positive change in color was observed for the of formaldehyde. All preparations include 1 mL concentrated sulfuric relaxing herbs mix (first line, position one), Turnera diffusa (first line, acid and 0.2 mL formaldehyde. position three), and Verbascum thapsus (first line, position four). Journal of Emerging Investigators • www.emerginginvestigators.org 21 DEC 2020 | VOL 3 | 2 Figure 5: Marquis reagent test for medicinal plants using a concentrated tea preparation. A) (left). Tea samples of each of the Figure 7: Control test for medicinal plants using a macerated tea 11 medicinal plants before the Marquis reagent test. Concentrated preparation and sulfuric acid. A) (left).
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