The scope of our research is centered on conducting a resource survey of ethnic medicine. Specifically, this study aims to develop a rapid and convenient methods for the identification of Rhodiola crenulata. The protocol we offer presents a cost-effective, straightforward, and rapid method for identifying various medicinal herbal materials.
Our laboratory will concentrate its future research endeavors on investigating two primary areas, the distribution of research associated with Rhodiola crenulata and the pharmacological impacts of the species. To begin, observe the appearance traits of Rhodiola crenulata with the unaided eye and record the observations. Inhale near the plant to identify the fragrance smell.
Next place a small piece of the root in the mouth. Then sip and chew for taste identification. Using a brush, remove the soil on the surface of the plant.
Place the plant in an oven at 40 degrees Celsius and turn it every 24 hours. Then powder the dried medicinal materials using a powder machine and filter it through a medicated number three sieve. Spread the powder evenly on a clean slide using a dissecting needle covering one third of the slide within a two millimeter area.
Using a glass dropper, add a drop of deionized water to the powder and quickly place a cover glass using a tweezer. Next, place the slide onto the microscope platform. Adjust the light source and the coarse focus spiral to view the powder.
Switch to a 40X objective lens and observe the starch granules. Again, spread the powder evenly on a slide using a dissecting needle. Then add a drop of chloral hydrate to the powder.
Hold the slide with a tweezer and heat on the alcohol lamp thrice for one second each. Add a drop of glycerin to the powder and quickly place a cover glass onto the drop. After adjusting the microscope settings as demonstrated previously, select a 40X objective lens to visualize the catheter, cork cells, and fibers, along with the wood parenchyma cells and pigment block respectively.
Microscopic examination of our crenulata revealed key features with cork cells appearing brownish, yellow, or colorless with polygonal shapes. Starch grains were observed as single or multiples with distinctive umbilical points and closely arranged spiral vessels. Additionally, xylem parenchyma cells were visualized as aromatic and oval containing calcium oxalate sand crystals, and irregularly shaped brown red pigment blocks were identified.
To begin, weigh three grams of Rhodiola crenulata powder and transfer it into a 100 milliliter conical bottle. Using a large bowl pipette, add 25 milliliters of methanol into the bottle. Place the bottle in the ultrasonic instrument.
Set the power to 250 watts, frequency to 40 kilohertz. Time to 30 minutes, and start the ultrasonication. Next, rinse the outside of the bottle under running water until it reaches room temperature.
Filter the solution through a 0.22 micron microporous membrane. Using a one milliliter syringe aspirate 800 microliters of liquid from the bottle and collect 400 microliters of the midstream solution into a chromatographic sample bottle. Measure and add one milligram of Salidroside, 0.5 milligrams of tyrosyl, and 0.5 milligrams of gallic acid into three separate one milliliter volumetric flasks.
Then fill each flask to the scale mark with methanol. Sonicate the samples in the ultrasonic instrument using the conditions described previously. Following ultrasonication, aspirate 800 microliters of liquid from the flask.
Filter the solution through a 0.22 micrometer microporous membrane and collect 400 microliters of filtrate into the bottle. Add tri chloro methane, ethyl acetate, methanol, and formic acid on one side of the double tank chromatography cylinder. Shake the cylinder to mix the contents evenly and cover the upper cylinder head.
Arrange the gallic acid Salidroside, aerosol standard solutions, and our crenulata solution on the sample rack in positions A1 to A4.Place a five by 10 centimeter silicone sheet on the sampling table. Turn on the automatic sampling machine and open the air control valve. Press the continue button for automatic sampling.
After sampling, turn off the machine and air valve. Remove the silicone sheet from the machine. Put the sheet into the other side of the double tank chromatography cylinder.
Cover the upper cylinder head and pre-saturate for 20 minutes. Next, use a tweezer to hold the upper end of the thin layer plate and quickly put the silicone sheet into the development agent. Once the organic solvent evaporates, spray the silicone sheet with Chromogenic solution.
The thin layer chromatographic analysis showed that the crenulata sample appeared in spots of the same color corresponding to the gallic acid, Salidroside, and tyrosyl standards.
