Bioassay of Flavonoids Crataegus oxyacantha with Plant Bionanosensor

The Journal of Medical Entomology and Parasitology is one of the series issued quarterly by the Egyptian Academic Journal of Biological Sciences. It is an important specialist journal covering the latest advances in that subject. It publishes original research and review papers on all aspects of basic and applied medical entomology, parasitology and host-parasite relationships, including the latest discoveries in parasite biochemistry, molecular biology, genetics, ecology and epidemiology in the content of the biological, medical entomology and veterinary sciences. In addition to that, the journal promotes research on the impact of living organisms on their environment with emphasis on subjects such a resource, depletion, pollution, biodiversity, ecosystem.....etc.


INTRODUCTION
Hawthorn extract has a significant amount of flavonoid and is of high importance due to the antioxidant property.Flavonoids have two groups of anthocyanins and antoxantine.Anthocyanin has pigments and antoxantine is colorless (Kin and Young, 1999).Faramarz Moradi began his research of plant biosensors with the patent No. 26299 in Iran with the discovery of rotation organ (Moradi, 1999).Then, in 2000, he was chosen in Qwarizmi's Festival thanks to the invention named intelligent organ and could gain the third place (discovery of smart organ, 2000).He recorded the plant sensor No. 332/421 in Iran's scientific research and industrial organization in 2004.Foundation of the memory material was reported in Iran in 2005 (Moradi, 2005) and a type of bioreactor sensor was built (Moradi et al., 2003).From the intelligent organ, a plant engine generating bioenergy was created under Biotechnology Master's thesis at Tehran University with the participation of Sharif University and was patented in Iran (Moradi, 2005).Moreover, plant anchor cells as storage energy (Moradi, 2005) moisture signal sensor (Moradi, 2005) damp-proof Nano sensor of drugs and materials (Moradi, 2006) Mobile safety sensor (Recipient, grant and License, 2010) industry and materials micro-sensor (Moradi, 2005) were patented in Iran and one file named nanomotor was recorded in the US (Moradi, 2006).Foundation of plant sensors using discovering intelligent organ was accepted at the University of Arkansas of US (Moradi, 2009) and foundation of cellulosic rotating Nano composites, including NanoBiotech were accepted at Switzerland (Moradi, 2009) international nanotechnology conferences.Mobile safety sensor obtained the bronze medal in the Olympics (Recipient Bronze Medal from novel Technology, 2011) and was verified by the three levels of Iran's National Elite Foundation (Recipient, grant and License, 2011).
Biosensors are based on identifying the target material with high specific detection (Mello and Kubota, 2002) as follows: 1. Bio-recognition elements: in which enzymes, antibodies, a part of DNA, peptides, and even a tissue of microorganism are used (Gooding, 2006).2. Amperometric biosensors that act based on oxidation and electrode reduction coated with enzymes (Vastarella, 2001).Absorption biosensors bind to a molecule that is to be determined.The electrode with the coated enzyme is one of them.3. Carbon paste electrodes are widely used initially by Adams (Mailley et al., 2004).Carbon paste electrodes are simple and cheap (Ghobadi et al., 1996;Bolado et al., 2007).4. Electrochemical analysis of polyphenolic compounds: quality and quantity of polyphenols are done by spectrometry or HPLC that are expensive and time-consuming.Graphite powder compound with non-electrolytic paste is inexpensive and easy to use.For more polyphenolic compounds, antioxidants containing phenolic groups are commonly used, which are based on electrochemical biosensors activity.They are a.polyphenol in the oil extraction (Capannesi et al., 2000), b. red polyphenol compounds (Gomes et al., 2004), c. polyphenols of various teas (Campanella et al., 2003) Laccase biosensor directly causes the oxidation of phenolic compounds (Freire et al., 2001;Freire et al., 2003a).Figure . 1 shows oxidation of flavonoids with Laccase (Pourcel et al., 2007).

Figure. 1. Oxidation of flavonoids with Laccase
There are various methods for measuring antioxidant biological compounds.Qualitative and quantitative techniques of polyphenols measurement include chromatography such as HPLC and GC-MS.These techniques are expensive and time-consuming so the development of biosensors can overcome these limitations.In order to prevent damage by free radicals, the body has a defense system of antioxidants (Antolovich et al., 2002).Some plants have antioxidant properties, especially biological flavonoids are considered as neutralizing free radicals due to the antioxidant, natural origin and the effective ability of activity (Katalinic et al., 2006).The flavonoids (Urquiaga and Leighton. 2000), which can be seen in Fig. 2 are used to eliminate free radicals (Simone, 1992) which can be seen in Fig. 3. Plant bionanosensor is a certain type and new approach of biosensors that has been used by the Faramarz Moradi as inventor of this paper.The study was used to determine two types of concentrations of flavonoids in hawthorn extract through the rotation.So far, no report was made about such a plant biosensor at international level from other researchers.

MATERIALS AND METHODS
Plant Nano-structure used in plant bionanosensor is the rotating Nano-structures as storage (i.e., it can turn to its previous memory such as memory metal) that has been fabricated by the Faramarz Moradi.
According to the following figure, plant bionanosensor has a graded plane divided by 310 sections, in which the amount of rotation could be read by a hand connected to a plant nano-structure (Figure 4and Figure 6).In the bottom of the bionanosensor, test site of the extract flavonoids can be seen (Figure . 5).Using insulin syringe, a drop of treatment inserted at the test site and the test site was placed on the paper to read the degree of each treatment.2-2.5 mg, 1-1.25 mg, 0.625-0.5 mg and 0 mg of flavonoid on hyperosoide/1 ml of hawthorn (Crataegus oxyacantha) in ethanol 70% was purchased by a pharmaceutical company of Iran Daru.
With regard to injection of 1.100 cc, the treatments were divided into 100 and the effect of treatments was analyzed by 02-0.025mg, 0.0125-0.01mg, 0.0625-0.005mg and 0 mg flavonoid in 1.100 cc as a completely randomized experiment with three iterations by software sas9.1.Comparison of average rotation was performed with LSD test at 1% level.3 shows that the A treatment has the greatest effect on rotational memory that is related to control and showed the greatest rotation by the greatest uptake by bionanosensor and the D treatment has the lowest effect on the rotational memory because it has the highest concentration.Therefore, it has low absorption and low rotation.

RESULTS AND DISCUSSION
According to above tables, plant bionano sensor is capable to detect hawthorn extract with a probability of 99% (70% ethanol as a control), 0. 025-0.02mg, 0.0125-0.01mg, 0.00625-0.005mg and 0 mg flavonoid on hyperosoide.The higher the flavonoid concentration, the lower the degree of rotation sensors will be.To examine the cause of rotation through an electron microscope manufactured by Zeiss German Factory, model 60A at Tehran University by SEM of plant sensor used in plant biosensor, a scan was run.The following picture is one of the images obtained.The picture shows that the lower part of the picture magnified 1,500 times, in which nanoparticles less than the mean diameter of a nanometer or nano-holes can be seen that contribute to its rotation.Plant sensor longitudinal section shows that nanoholes and nanoparticles with absorption treatments convert the absorption treatment energy into the rotation with the help of specific structural features.
Plant bio-nanosensor advantage includes: 1.It is inexpensive compared to chromatography, spectroscopy techniques and other biosensors.2. It has been the result of years of research since 1999.3. It is simple and easy transportation makes its use in the desert.4. It does not use any electrical energy source unlike other methods use it as flavonoid extract acts as an energy source of the biosensor.5.If it is recorded at the international level, it can be commercialized at the international level.6.If it is optimized, plant secondary metabolites can be directly studied by intact leaves and other physiological properties.7. It is inexpensive and easy to use. 8. Any material is used in its performance.9.The use of other methods requires purification of flavonoids.

Table 1 .
Table1shows the results of the rotation bioassay.The Degree of plant bio nanosensor rotation

Table 2 .
Analysis of variance of the treatments rotation of flavonoids extract