Alkaloid Extraction using a Probe-Ultrasonicator

Alkaloids are bioactive compounds, which can be efficiently extracted from plants using ultrasonication. Alkaloids exhibit a wide array of biological effects and are thereby used as pharmaceuticals and therapeutics. Ultrasonic extraction is the preferred technique to produce high quality alkaloid extracts from plant material. Ultrasonic extraction results in very high alkaloid yields, whilst extraction time is short and the ultrasonic equipment is safe and easy to operate.

High-Efficient Alkaloid Extraction using Power-Ultrasound

How does ultrasonic extraction promote the isolation of alkaloids?
Ultrasonic extraction uses low-frequency ultrasound waves (e.g. 20-26kHz frequency) and ultrasound’s effect of acoustic cavitation. High-intensity ultrasound waves create cavitation bubbles that collapse, generating locally energy-dense pressure and temperature differentials as well as powerful shear forces, which perforate and break plant cell walls and plant tissue open. The liquid streaming generated by cavitation promotes mass transfer between the intracellular matrix and the surrounding solvent so that the alkaloids are efficiently released. For ultrasonic alkaloid extraction, the plant material (e.g. ground dried leaves) are placed in a solvent, which is typically a mixture of water and an organic solvent such as aqueous ethanol. Polar solvents such as methanol and ethanol have been found to be most efficient for high alkaloid yields. After a short ultrasonic treatment of the slurry – consisting in plant material and solvent – the alkaloids are released from the plant cells and are present in the solvent, from where they can be isolated and purified (e.g. using a rotor-evaporator).

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Ultrasonic batch extraction of alkaloids from plant material using the ultrasonic probe UP400St.

Ultrasonicator UP400St for alkaloid extraction from plant material.

Full Spectrum Extraction from Tobacco using Ultrasonics UP400StThe video shows the highly efficient and rapid ultrasonic extraction of alkaloids from tobacco. The ultrasonic processor used is a Hielscher UP400St ultrasonicator, which is ideal to prepare medium size batches of high-quality extracts. For large volumes, Hielscher Ultrasonics supplies industrial ultrasonic systems for continuous inline extraction.

Main Advantages of Ultrasonic Alkaloid Extraction

Ultrasonic extraction is considered as superior extraction technique for the isolation of alkaloids from plant materials as it offers several advantages, including:

  1. Increased extraction efficiency: Ultrasonic extraction can increase the yield of alkaloids by more than 200% compared to traditional extraction methods.
  2. Reduced extraction time: Ultrasonic extraction can significantly reduce the extraction time, allowing for faster and more efficient processing of large volumes of plant material.
  3. Lower solvent consumption: Ultrasonic extraction can reduce the amount of solvent required for extraction, resulting in reduced solvent consumption and decreased environmental impact.
  4. Selective extraction: Ultrasonic extraction can be used to selectively extract certain alkaloids by choosing the suitable solvent, process parameters and extraction temperature. As unwanted compounds are left behind, the purity of the final product is improved.
  5. Easy to scale-up: Ultrasonic extraction can be easily scaled up to handle larger volumes of plant material, making it suitable for commercial production. Ultrasonic processes can be scaled linearly, which facilitates the implementation on industrial scale.

Overall, ultrasonic extraction offers a faster, more efficient, and more environmentally friendly approach to isolating alkaloids from plant materials.

Ultrasonic Alkaloid Extraction from the Leaves of the Soursop Tree

The ultrasonic extractor UP400ST with temperature sensor was used for the extraction of alkaloids from dried leaves..Nolasco-González et al. (2022) investigated the effectiveness and efficiency of ultrasonic extraction of bioactive compounds including phenolic compounds, alkaloids, acetogenins (all with with high antioxidant capacity) from the leaves of the plant species Annona muricata (soursop tree). For ultrasonic extraction, they used the probe-type ultrasonicator UP400S at following conditions: 80% amplitude, 0.7 s pulse-cycle, for 4.54 min. Total soluble phenols (TSP) were extracted using 0.5 g of dried leaves powder and 20 mL of acetone: water (80:20 v/v). The ultrasonically produced extract contained 178.48 mg/100 mL of soluble phenols, 20.18 mg/100 mL of total flavonoids, 27.81 mg/100 mL of hydrolyzable polyphenols, 167.07 mg/100 mL of condensed tannins, 30.44 mg/100 mL of total alkaloids and 14.62 mg/100 mL of total acetogenins. The extract isolated with the U400S probe-type ultrasonicator (see picture left) exhibited a high antioxidant capacity and a higher content of bioactive compounds from ~6 to ~927-fold than decoction or infusion, depending on the type of compound. Twenty-four different phenolic compounds were identified in the samples, and the ultrasonically isolated extract produced the highest concentration.
[cf. Nolasco-González et al., 2022]

Ultrasonic alkaloid extraction gives superior yields with a high antioxidant capacity and excels extraction techniques such as decoction and infusion.

Antioxidant capacity of extracts obtained by the optimal conditions of ultrasound-assisted extraction (UAE), decoction and infusion. Ultrasonic extracts consistently show significantly higher antioxidant capacity.
(Study and graphic: ©Nolasco-González et al., 2022)

Ultrasonic extraction with the UP400S gave a full spectrum extract containing a high yield of numerous phenolic compounds.

HPLC chromatogram of phenolic compounds from ultrasonically isolated extract of Annona muricata leaves using the ultrasonicator UP400S.
(1) shikimic acid, (2) gallic acid, (3) protocatechuic acid, (4) neochlorogenic acid,
(5) 3,4-dihydroxyphenylacetic, (6) 4-hydroxybenzoic acid, (7) chlorogenic acid, (8) 4-hydroxyphenyl acetic, (9) vanillic acid, (10) syringic acid, (11) 3-hydroxybenzoic acid, (12) caffeic acid, (13) 4-hydroxybenzaldehyde, (14) homovanillic acid, (15) 3-(4-hydroxyphenyl), (16) p-coumaric acid, (17) trans-ferulic acid, (18) trans-cinnamic acid, (19) gallocatechin, (20) epigallocatechin, (21) catechin,(22) epicatechin, (23) rutin, (24) ellagic acid.
(Study and graphic: ©Nolasco-González et al., 2022)

Lee et al. (2021) had similar confirmed results as they report 2.3-fold more alkaloids from A. muricata leaves extracted under ultrasonication (340 W, 56◦C, 30 min) compared to Soxhlet extraction (7 h, 80◦C).

Ultrasonic probe-type extractor for the isolation of alkaloids from plant materials such as leaves, bark, roots and stems.

Ultrasonic extractor UIP1000hdT with stirrer for alkaloid extraction in batch mode.

Ultrasonic Extraction of Specific Alkaloids

Indole Alkaloid Extraction from Mimosa Root Bark and Ayahuasca using an Ultrasonicator

Mimosa root bark as well as ayahuasca leaves contains the alkaloid N,N-dimethyltryptamine (DMT). Dimethyltryptamine (DMT) is currently researched in clinical trials for its hallucinogenic effects and therefore as a potential drug to treat depression and post-traumatic stress disorders (PTSD). Ultrasonication effectively promotes the release and isolation of the alkaloid from the Mimosa hostilis roots and Psychotria viridis leaves (ayahuasca). Ultrasonic DMT extraction produces high yields within very short time. Compatible with almost any solvent, Hielscher probe-type ultrasonicators are used for the production of pharma-grade bioactive compounds for use under research and medical settings as well as for sample prep before analytical quality and potency testing. Therefore, Hielscher ultrasonicators can facilitate medical DMT and botanical research in numerous ways.
The most common source of the indole akaloid N,N-dimethyltryptamine (DMT) in jurema tree, it comes from Mimosa hostilis roots (commonly known as jurema preta). Ayahuasca is the traditional name for another plant species, in which the leaves of the shrub Psychotria viridis are rich in DMT.
Both, mimosa root bark (also mimosa tenuiflora, Mimosa Hostilis Root Bark; abbrev. MHRB) and ayahuasca leaves contain various bioactive compounds, which are currently researched in clinical trials regarding their potential as therapeutic drug. Mimosa root bark contains tannins, saponins, tryptamines, alkaloids, lipids, phytoindoles, xylose, phytosterols, glucosides, rhamnose, arabinose, methoxychalcones, lupeol and kukulkanins. These bioactive compounds can be efficiently extracted from the root bark using probe-type ultrasonicators. Ultrasonic extraction is the superior technique to isolate target substances such as alkaloids from botanical material such as mimosa root bark and ayahuasca leaves. Sonication excels by giving high yields within a very short extraction time. Mild process conditions ensure highest extract quality preventing unwanted degradation of the active molecules.

Ultrasonic extraction and purification of phytochemicals from plants: The picture shows the probe-type ultrasonicator UP400ST, a Büchi vacuum filter and rotor-evaporator for the extraction of curcumin.

Extraction setup for botanical isolation: Probe-type ultrasonicator UP400ST, Büchi vacuum filter and rotor-evaporator for the extraction of phytochemicals such as alkaloids and phenols.

Ultrasonic extractors from Hielscher allow for the reliable and efficient production of large quantities of alkaloid extracts. Ultrasonic extraction gives high alkaloid yields of superior quality within short processing time.

Ultrasonic extractor UIP2000hdT with pharma-grade stainless steel batch for industrial production of alkaloid extracts.

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Hielscher Ultrasonicators for Efficient Alkaloid Extraction

Hielscher Ultrasonics designs, manufactures and distributes high-performance ultrasonic extractors, which are used for scientific research as well as the production of pharma-grade extracts.

Hielscher Ultrasonicators for Superior Extraction Efficiency

Ultrasound-assisted extraction is a scientifically proven technique with an extraordinary high efficiency regarding the extraction of bioactive compounds from plants. Whilst traditional methods often take a long time (several hours up to days), ultrasonic botanical extraction takes only a few minutes.

High Extraction Yield and Superior Extract Quality with Hielscher Ultrasonicators

Ultrasound-driven extraction allows for the complete release of bioactive compounds from the plant material. Ultrasonic cavitation breaks the cellular matrix and releases the bioactive molecules efficiently into the surrounding solvent. Comparative studies demonstrated that sonication gives higher extract yields than other extraction techniques (e.g. Soxhlet, maceration, decoction, infusion). Ultrasonic extraction is compatible with any solvent and allows thereby also for the use of very mild solvent such as water or aqueous ethanol. Regarding to specific extraction requirements, polar and non-polar solvents can be chosen and switching between solvents is no problem. Precise control over process parameters (e.g. amplitude), use of mild solvents and exact temperature control prevent the degradation and contamination of bioactive compounds (e.g. alkaloids, polyphenols, flavonoids, cannabinoids etc.) during ultrasonic extraction.

Versatile Application of Hielscher Ultrasonicators

Whilst alternative extraction techniques (e.g., supercritical CO2, solvent extraction, percolation etc.) are often only applicable to the extraction of specific molecules depending on molecular weight, solubility and heat-sensitivity, the extraction with Hielscher high-performance ultrasonicators allow for the versatile extraction of any botanical compound. Ultrasound-assisted extraction can be reliably applied for the extraction of broad-spectrum extracts containing bioactive compounds such as polyphenols, alkaloids, anthraquinones, flavonoids, glycosides, lipids, pectin, and polysaccharides.

Excellent Energy Efficiency and Environmental-Friendliness

The energy consumption of ultrasound-assisted extraction is less by far than the traditional methods in the extraction of medicinal compounds from plants.

Reliability due to Process Standardization with Hielscher Ultrasonics

Hielscher ultrasonicators can be remotely controlled via browser control. Sonication parameters can be monitored and adjusted precisely to the process requirements.Hielscher ultrasonicators allow for the precise control over all ultrasonic process parameters such as amplitude, intensity, duration, and temperature. The process parameters can be easily controlled and are continuously monitored. Additionally, all process parameters are automatically protocolled as CSV file on an built-in SD-card. This enables the operator to revise sonication runs, to optimize the extraction process and to ensure consistent extract quality. Hielscher ultrasonicators enable manufacturers to produce botanical extracts of superior quality and facilitate the fulfilment of Good Manufacturing Practice (GMP).

The table below gives you an indication of the approximate processing capacity of our ultrasonicators:

Batch Volume Flow Rate Recommended Devices
1 to 500mL 10 to 200mL/min UP100H
10 to 2000mL 20 to 400mL/min UP200Ht, UP400St
0.1 to 20L 0.2 to 4L/min UIP2000hdT
10 to 100L 2 to 10L/min UIP4000hdT
15 to 150L 3 to 15L/min UIP6000hdT
n.a. 10 to 100L/min UIP16000
n.a. larger cluster of UIP16000

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Ultrasonic Extraction of Botanicals - 30 Liter per 8 Gallon BatchUltrasonic botanical extraction gives higher yields. The Hielscher UIP2000hdT, 2000 watts homogenizer is powerful enough to extract batches from 10 liters to 120 liters easily.
Ultrasonic high-shear homogenizers are used in lab, bench-top, pilot and industrial processing.

Hielscher Ultrasonics manufactures high-performance ultrasonic homogenizers for mixing applications, dispersion, emulsification and extraction on lab, pilot and industrial scale.

Literature / References

Facts Worth Knowing

What are Alkaloids?

Alkaloids form a class of naturally occurring nitrogenous compounds of low molecular weight that are frequently found as secondary metabolites in the plant kingdom. An alkaloid contains at least one nitrogen atom. Alkaloids are biochemically synthesized from one of the following three common amino acids – lysine, tyrosine and tryptophan.
In plants, alkaloids generally exist as salts of organic acids like acetic, malic, lactic, citric, oxalic, tartaric, tannic and other acids. Some weak basic alkaloids (such as nicotine) occur freely in nature. A few alkaloids also occur as glycosides of sugar such as glucose, rhamnose and galactose, for example, alkaloids of the solanum group (solanine), as amides (piperine) and as esters (atropine, cocaine) of organic acids (cf. Ramawat et al., 2009).
Many alkaloids are bioactive compounds which exhibit medicinal or therapeutic effects. For instance, specific types of alkaloids are used to treat various diseases including malaria, diabetics, specific cancer types, cardiac dysfunction, depression etc.
Since many alkaloids show strong bioactive properties, their use as pharmaceuticals, narcotics, stimulants and poisons has been either adopted or is under scientific research. Morphine, a well-known alkaloid from Papaver somniferum poppy, is used as pharmaceutical drug in today’s medicine. Quinine and codeine are two other alkaloid types used for medical purposes. Under medical supervision and at medically prescribed doses, alkaloids are pharmacologically useful. However, when used wrongly some alkaloids can be toxic at high doses. The alkaloid strychnine is for instance used as a highly effective toxin used as pesticide against small vertebrae.

Examples of some alkaloids in plants

Compounds Plant Source Effects and Applications
Morphine Papaver somniferum Analgesic
Camptothecin Camptotheca acuminata Anticancer
Atropine Hyoscyamus niger Treatment of intestinal spasms, antidote to other poisons
Vinblastine Catharanthus roseus Anticancer
Codeine Papaver somniferum Analgesic, antitusive
Caffeine Coffea arabica Stimulant, natural pesticide
Nicotine Nicotiana tabacum Stimulant, tranquillizer
Cocaine Erythroxylon coca Stimulant of the central nervous system, local anesthetic
N,N-dimethyltryptamine Mimosa hostilis, Psychotria viridis Hallucinogenic, entheogenic
Mitragynine Mitragyna speciosa Analgesic, psychoactive

High performance ultrasonics! Hielscher's product range covers the full spectrum from the compact lab ultrasonicator over bench-top units to full-industrial ultrasonic systems.

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