Hielscher – Ultrasound Technology

Ultrasonic Extraction of Cellular Matter

Ultrasonic extraction or sono-extraction is a process intensifying technology, which works by the coupling of high power ultrasonics into a slurry of plant or cell tissue. Hielscher Ultrasonics supplies reliable ultrasonicators for cell disruption and extraction from small lab samples up to high volumes in industrial processing. The appeal of the ultrasonically assisted extraction lies in the non-thermal treatment of the material, its easy application and the scaleability from test to production scale.
Hielscher’s ultrasonic devices generate high intense ultrasound that can be exactely controlled to your process requirements.

The list below gives you various ultrasonic processing protocols, which demonstrate the broad application field of power ultrasonics in biology, food processing and pharmaceutical production.
Ultrasonic homogenizers are powerful tools for extraction. (Click to enlarge!)

Probe-type ultrasonicator UP50H

Ultrasonic Extraction of Chilli Flakes

Boldine from Boldo leaves (Peumus boldus Molina)

Ultrasonic application:
An important active compound in boldo is boldine ((S)-2,9-dihydroxy-1,10-dimethoxiaporphine), which is besides catechin ((2S,3R)-2-(3,4-dihydroxy-phenyl)-3,4-dihydro-1(2H)-benzopyran-3,5,7-triol) the main components of the alkaloid and flavonoid fraction in boldo leaves. Boldine is a strong antioxidant agent that undergoes peroxidative free-radical-mediated damage and acts as an efficient hydroxyl radical scavenger.
Extraction Procedure: For a typical extraction procedure, samples of boldo leaves were extracted with 1L of distillated water at atmospheric pressure using the ultrasonicator UIP1000hd in batch and flow-through mode. Time of extraction ranges between 10 and 40 min., with an Ultrasonic intensity of 10 to 23 W/cm2, and a temperature range of 10 to 70°C. Best results wer achieved under following conditions: ultrasonic intensity of 23 W/cm2 for 40 min. at temperature of 36°C
Results: The analysis results show that high-power sonication enhances the analyte release of vegetal matrix material of Boldo at significantly better rates compared to conventional method: equal yield was released by sonication in 30 min. whilst conventional extraction time was 2h.
Chemat (2013) and co-workers have shown that ultrasonically-assisted extraction improves the efficiency of the plant extraction whilst reducing extraction time at increased concentration of the extracts (same amount of solvent and plant material). Analysis revealed that the optimized conditions were: sonication power 23 W/cm2 with UIP1000hd for 40 min. and a temperature of 36°C. The optimized parameters of the ultrasound extraction provide a better extraction compared to a conventional maceration in terms of process time (30 min. instead of 120 min.), higher yield, higher energy efficiency, improved cleanliness, higher safety and better product quality.
Device Recommendation:
UIP1000hd with sonotrode BS2d34 and flow cell
Reference/ Research Paper:
Petigny, L.; Périno-Issartier, S.; Wajsman, J.; Chemat, F. (2013): Batch and Continuous Ultrasound Assisted Extraction of Boldo Leaves (Peumus boldus Mol.). International Journal of Molecular Science 14, 2013. 5750-5764.

Chlorogenic acid from Tobacco leaves

Ultrasonic application:
For the extraction experiments, tobacco leaves from Nicotiana tabacum were grinded to smaller particle sizes of about >4 x ≧2 mm. For the ultrasonic extraction trials, 20g samples of dried tobacco leaf materials were extracted with distilled water at temperatures ranging from 5°C to 30°C. Extraction experiments were conducted for extraction cycle time intervals varying from 5 minutes to 30 minutes per cycle. About 10 to 15 drops of ethanol were added to the calculated 256 ml optimum volume of distilled water needed to extract all the soluble chlorogenic acid from 20 g samples of dried tobacco leaves. The 256 ml optimum volume of distilled water was then divided into fractional volumes of the extraction solvent for use in each ultrasonic extraction cycle.
For the ultrasonic irradiation, an ultrasonic processor UP400S (400 watts, 24 kHz) with a sonotrode H7 made of titanium, that had a tip diameter of 7mm, was used. The sonotrode H7 was inserted at half height of the extraction mixture.
Protocol: As optimal conditions for the ultrasonic extraction of chlorogenic acid from tobacco leaves were found: 20g dried tobacco leaves in distilled water (optimum liquid to solid ratio: 12,8mL/g) at 20 degC processing temperature, ultrasonic device UP400S (400W, 24kHz) for 3 sonication cycles of each 15min (total sonication duration: 45 min).
Device Recommendation:
UP400S with sonotrode H7
Reference/ Research Paper:
Mazvimba, Martin Tongai; Yu, Ying; Zhang, Ying (2011): Optimization And Orthogonal Design Of An Ultrasonic Assisted Aqueous Extraction Process For Extracting Chlorogenic Acid From Dry Tobacco Leaves. Chinese Journal of Natural Medicines 2011.

Mangiferin acylation

Ultrasonic application:
Mangiferin (1,3,6,7-Tetrahydroxy-2-[3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]xanthen-9-one; formula: C19H18O11) is a polyphenol of C-glycosylxanthone structure that can be found in many plant species. Mangiferin shows various pharmacological activities. The regioselective acylation of mangiferin can be very efficiently catalyzed by lipase under ultrasonication. Compared with the conventional methods, the ultrasonically-assisted catalysis excels by the advantages of shorter reaction time and higher yields. The optimum conditions for the ultrasonic mangiferin acylation were found as following:
lipase: PCL, acyl donor: vinyl acetate; reaction solvent: DMSO, reaction temperature: 45 degC, ultrasonic power: 200W; substrate ratio: acyl donor/mangiferin 6/1, enzyme loading: 6 mg/ml
The regioselective acylation yield was up to 84%.
Device Recommendation:
UP200St or UP200Ht
Reference/ Research Paper:
cp.: Wang, Z.; Wang, R.; Tian, J.; Zhao, B; Wei, X.F.; Su, Y.L.; Li, C.Y.; Cao, S.G.; Wang, L. (2010): The effect of ultrasound on lipase-catalyzed regioselective acylation of mangiferin in non-aqueous solvents. J. Asian Nat Prod. Res. 12/1, 2010. 56-63.

Capsaicinoids

Ultrasonic application:
Extraction of capsaicinoids (capsaicin, nordihydrocapsaicin) from chili peppers: Capsaicinoids from Capsicum frutescens peppers was obtained via ultrasonic extraction under following conditions: solvent: 95% (v/v) ethanol, solvent/ mass ratio of 10 ml/g, 40 min. sonication extraction time, 45°C extraction temperature. Extractant yield: 85% of the capsaicinoids
Device Recommendation:
UP400S

Silymarin from Milk Thistle Seeds

Ultrasonic application:
Ultrasound-assisted extraction was performed by using an ultrasonicator UP400S. In this study a horn-type ultrasonic probe was used with a diameter of 1.5 cm. 10 g of nonfat milk thistle powder was weighed accurately and dissolved in 100 mL of methanol. The beaker was kept in a water bath and maintained at a temperature of 25°C. The milk thistle powder dissolved in solvent was sonicated for different time intervals (30, 60, 90, 120 and 150 min.) After each time interval, the solution was filtered through a Whatman filter paper and each filtrate (about 80mL) was evaporated using rotary evaporator until reached 30 mL solution and the absorbance of phenolic compounds was measured with UV-Visible Spectrophotometer at wavelength of 517 nm according to DPPH method. As expected increasing extraction times resulted in increasing silymarin contents.
The authors of the study found that ultrasonic extraction in order to obtain the alcoholic extracts is one of the best techniques to replace the conventional techniques. The ultrasonic extraction was proven to obtain a better extract quality. Further benefits are time-saving being more efficient etc.
Device Recommendation:
UP400S
Reference/ Research Paper:
Çağdaş, E.; Kumcuoğlu, S.; Güventürk, S.; Tavman, S. (2011): Ultrasound-Assisted Extraction of Silymarin Components from Milk Thistle Seeds (Silybum Marianum L.). GIDA 36/6, 2011. 311-318.

Stevioside

Ultrasonic application:
Samples of 10g dry and ground stevia leaves were extracted in 100mL water under continuous stirring (with a magnetic stirrer). The pH value was controlled with 0.01 M pH 7 sodium phosphate. The sample was put in a 150 mL glass beaker and sonicated with an probe-type ultrasonicator (UIP500hd, 20kHz, 500W). The tip of the sonotrode was immersed about 1.5 cm into the slurry of stevia leaves. The ultrasonic device was set for a power output of 350W. A mild sonication treatment of 350 W for 5-10 min. at a constant process temperature of 30°C gave a rebaudioside A yield of 30-34g per 100g sample. After sonication, the extract solution was centrifuged and filtered off through 0.45 μm microporous membrane; the filtrate was taken for total rebaudioside A content analysis. The extraction yield of total rebaudioside A content was analyzed by HPLC.
By the solvent-free ultrasonically-assisted extraction, a high yield of rebaudioside A was obtained in comparison to traditional extraction methods such as heat extraction or maceration.
Device Recommendation:
UIP500hd

Terpenes

Ultrasonic application:
The extraction of volatile compounds, such as terpenes, from the inflorescences of a fiber type Cannabis sativa L. cultivar can be ultrasonically enhanced. Studies have shown that after a short sonication the targeted terpenes are already released from the plant material.
The results show that ultrasonic treatment not longer than 5 min allows to obtain an enhanced concentration of terpenes in comparison with maceration. Instead, an ultrasonic treatment longer than 5 min increased the concentration of δ-9-tetraidrocannabinol (THC).
Sonication Protocol:
The ultrasound-assisted extraction of terpenes was carried out using an 200W probe-type ultrasonicator. The ultrasonic device was set at 25% amplitude. Three aliquots of 50g of dried inflorescences were added each with 250mL of 70% ethanol v/v, which was used extracting solvent. Each beaker and its content were immersed into an ice-bath. Sonication was carried out for 5, 10 and 15 min. During the ultrasonic extraction procedure, a continuous rapid heat dissipation was ensured to maintain the process temperature below 30°C. After extraction, the mixtures were filtered under vacuum through Whatman No. 3 paper, and the solvent was removed by a rotary vacuum evaporation. Each extraction trial was performed in triplicate using three different samples.
Ultrasonication was found to be an interesting alternative to maceration for extracting volatile compounds from cannabis inflorescence, but shows also that the ultrasonic treatment must be not longer than 5 minutes to obtain an enhanced recovery of terpenes. Instead, an ultrasonic treatment longer than 5 minutes increased the concentration of δ-9-tetraidrocannabinol (THC). The extract of Cannabis sativa inflorescences obtained by ultrasonic extraction carried out for 5 min could be used as ingredient for perfumes or flavoring for beverages. It can be used as alternative for industrial hemp, where stems are used for the fibre industry and inflorescences extracts for cosmetic and food industry.
Device Recommendation:
UP200St or UP200Ht with sonotrode/ probe S26d14
Reference/ Research Paper:
Da Porto, C.; Decorti, D.; Natolino, A. (2014): “Ultrasound-assisted extraction of volatile compounds from industrial Cannabis sativa L. inflorescences”. IJARNP 2014, 7/ 1. 8-14.

Vanillin extraction from cured vanilla beans

Ultrasonic application:
Vanillin extraction was optimized under sonication. Therefore, the 100 watts ultrasonic device (20kHz) was operated in pulsed mode (cycles: 5 sec. on followed by 5 sec. off).
Best results were achieved for ultrasonic extraction in with 40% ethanol for 1h at 30°C. To compare the results with a conventional extraction methods, water bath and ultrasound bath extractions were also carried out. Results showed that the optimization of vanillin extraction by ultrasonic horn with 40% ethanol for 1h at 30°C was comparative to water bath extraction with 40% ethanol at 56°C for 15h.
Device Recommendation:
UP100H
Reference/ Research Paper:
Rasoamandrary, N.; Fernandes, A. M.; Bashari, M.; Masamba, K.; Xueming, X. (2013): Improved Extraction of Vanillin 4-Hydroxy-3-methoxybenzaldehyde from Cured Vanilla Beans Using Ultrasound-Assisted Extraction: A Comparison of Ultrasound-Assisted and Hot Water Bath Extraction. Akademik Gıda 11/1, 2013. 6-12.

Ziziphus Jujube - extraction of phenolic compounds of jujube

Ultrasonic application:
For the ultrasonically assisted extraction of phenolic compounds of jujube (Ziziphus Jujube), a high intensity probe-type ultrasonicator UP200H with 200W power and 24kHz frequency was used. The ultrasonic device was equipped with the micro-tip sonotrode S2 (tip diameter 2 mm), which was immersed in a water bath in which a precipitate glass with the sample was placed (internal dimensions: 280:195:135 mm). Amplitude of ultrasonic vibrations was 100% of nominal power (maximum amplitude of 260 μm) and acoustic power of 0.171402 W and intensity of 21.8346 W/cm2 .The ultrasonic intensity was determined calorimetrically by measuring the time–temperature increase of the suspension under adiabatic conditions. The ultrasonic extraction procedure was used for the extraction of total phenolics from samples according to the experimental design (for optimization).
Device Recommendation:
UP200H with sonotrode S2
Reference/ Research Paper:
Fooladi, H.; Mortazavi, S. A.; Rajaei, A.; Elhami Rad, A.H.; Salar Bashi, D.; Savabi Sani Kargar, S. (2013): Optimize the extraction of phenolic compounds of jujube (Ziziphus Jujube) using ultrasound-assisted extraction method. IECFP 2013.



Click here to find more sonication protocols for the ultrasonic treatment of biological matter!

Ultrasonic extraction can be carried out in batch operation and continuous flow-through mode. (Click to enlarge!)

Sonication setup with UIP1000hd for the extraction from boldo leaves in a batch. [Petigny et al. 2013]

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Facts Worth Knowing

Ultrasonic tissue homogenizers are often referred to as probe sonicator, sonic lyser, ultrasound disruptor, ultrasonic grinder, sono-ruptor, sonifier, sonic dismembrator, cell disrupter, ultrasonic disperser or dissolver. The different terms result from the various applications that can be fulfilled by sonication.