# Construction of Nanomaterials and the Role of Rutin in the Treatment of Cerebral Hemorrhage

Authors: Xin Li; Zhenzhong Gao
Journal: Advances in Materials Science and Engineering
 (2022)

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## Abstract

Cerebral hemorrhage generally refers to nontraumatic cerebral parenchymal hemorrhage. In China, about 20%–30% of all stroke in a month after the mortality rate can be as high as 40%. And most of the survivors have obvious neurological dysfunction, to the patient’s family and society from all walks of life caused great psychological pressure, which will seriously affect the human physical and mental health. This article aims at studying the construction of nanomaterials and the role of rutin in the treatment of ICH. It first introduces the characteristics of nanomaterials; nanomaterials have some special effects based on their special internal structural properties and surface states. These effects derive many physical and chemical properties that general substances do not possess, such as optical, electrical, magnetic, catalytic, and photocatalytic properties. A simple analysis of cerebral hemorrhage then gives the nanomaterials the first-principles algorithm. Practical application of nanomaterials is a class of nanomaterials, which have many strange chemical characteristics such as special light, electric, magnetic, thermal, mechanical, mechanical physical, and chemical characteristics. Then, the nanocomposite of rutin electrochemical detection is analyzed, and finally rutin and analog brain drug dynamics is discussed. The experimental results showed that the RSD of samples at room temperature, freeze-thaw cycles, and long-term stability of the mice were less than 5.0%, and the stability of mouse brain tissue homogenate samples completely meets the requirements of troxerutin, rutin, and troxerutin aglycone.


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## Body

## 1. Introduction

Patients with cerebral hemorrhage often have a sudden onset due to emotional agitation and exertion, and the early mortality is very high. Most of the survivors have sequelae of varying degrees of motor impairment, cognitive impairment, and speech and swallowing disorders. Cerebral hemorrhage is one of the most serious refractory diseases that endanger people’s lives. Its high case fatality rate and great mortality rate have brought a heavy burden to Chinese residents. In recent years, investigations have shown that stroke has surpassed cardiovascular disease and malignant cancer as the first fatal factor in Chinese adults, and cerebral hemorrhage is the second largest type of stroke, accounting for 10%–15% of all stroke. And the proportion of stroke in China is as high as 20%–30% and some even more than 50%.This article aims at studying the construction of nanomaterials and the role of rutin in the treatment of ICH. Cerebral hemorrhage is one of the most serious refractory diseases that endanger people’s lives. Its high case fatality rate and great mortality rate have brought a heavy burden to Chinese residents. Investigation in recent years has shown that stroke has surpassed cardiovascular disease and malignant cancer. As the first fatal factor of Chinese adults, it may be able to greatly shorten the treatment time window after stroke and get symptomatic treatment in an early stage, which has an open prospect of clinical transformation. This article aims at studying the construction of nanomaterials and the role of rutin in the treatment of ICH, in order to make a certain contribution in the treatment of ICH.The innovations of this article are as follows. (1) The characteristics of nanomaterials are analyzed. (2) The first-principles algorithm for nanomaterials is proposed. (3) The intracerebral pharmacokinetics of Troxerutin and its analogs are investigated experimentally.


## 2. Related Work

According to the progress of foreign research, different researchers have a corresponding cooperative research on the construction of nanomaterials and cerebral hemorrhage: Jiao et al. summarized the research work based on the impact of bile salt on the self-assembly of micro- and nanomaterials and related research worldwide. Dye molecules are considered ideal substrates for the construction of functional nanomaterials; as biosurfactants, bile salts are commonly used to assist in the synthesis of micronanomaterials. To gain a more comprehensive and deeper understanding of the preparation of micronanomaterials in the presence of bile salts, the research provides a solid foundation for exploring future applications [1]. Rincón-Morantes et al. aim at gathering different experiences using nanomaterials to stabilize or improve the mechanical and geotechnical properties of soils used in road construction, especially those related to manufacturing nanofibers for soils, finding that its applications in the road field will be studied and developed by different types of solutions [2]. Seevakan and Sheeba study and document applicable nanotechnology products that can enhance the overall competitiveness of the construction industry. The application of nanotechnology in architecture will focus on: (1) lighter and stronger structural composites, (2) low maintenance coatings, (3) better adhesive performance, (4) reduced heat transfer and insulation of flame retardants, (5) structurally related nanosensor [3]. The Lim et al. study aimed to investigate the possibility of using these carbon-based nanomaterials as building materials. The structural and electrical properties of cemented composites were studied based on carbon-based materials such as multiple, single, carbon nanotubes, graphene nanosheets, and conductive graphite powder. Microstructural analysis was also performed by noncovalent functionalization of carbon-based nanomaterials to check for dispersion [4]. The purpose of the Michael study is to evaluate resource utilization following the implementation of a treatment regimen for mild traumatic brain injury (TBI). Patients with isolated mild TBI both before and after the implementation of a mild TBI treatment regimen were retrospectively reviewed. Efficient care is critical in modern medicine, and this study shows that mild TBI treatment options significantly reduce resource utilization without compromising patient safety [5]. Target tissue dissection is an ultrasound-based treatment modality that relies on the generation of targeted cavitation bubble clouds that mechanically separate tissue. The purpose of the Sukovich et al. study is to investigate the in vivo feasibility of using tissue analysis techniques to produce localized destructive damage in the porcine brain, including dose requirements and safety. Lesions confined to the gyrus cannot cause the significant bleeding or edema response [6] at the treatment site. However, these scholars did not study the construction of nanomaterials and the role of rutin in the treatment of cerebral hemorrhage, but only unilaterally discussed its significance.


## 3. Construction of Nanomaterials and the Role of Rutin in the Treatment of Cerebral Hemorrhage

### 3.1. Characterization of the Nanomaterials

Small-scale effect is also known as small volume effect. When the scale of nanoparticles and transfer electron wavelength or superconducting coherent wavelength are under the same physical scale, even more periodic boundary conditions were broken, the temperature, magnetism, optical absorption, boundary thermal resistance, chemical reactivity, catalytic change, and the effect of nanomaterials have also opened up a wide range of new application fields [7]. Nanostructure is a new system constructed according to certain rules on the basis of nanoscale material units.Surface effect refers to the change in the ratio of the number of molecules to the total atomic order on the nanoparticle surface as the particle size decreases or increases rapidly. Because the application of nanomaterials is gradually smaller, the number of surface atoms increased rapidly, so the surface area and surface free energy of nanoparticles also increased rapidly, and the surface atoms field environment and binding freedom are different between the atoms. The surface molecules between there are no adjacent atoms, so many hanging bonds have unsaturated, which is easy to combine between other atoms and stabilize it, that show a strong catalytic activity [8].Quantum size effect is that the particle scales down to a value, around the fermi electron level from quasi-continuous electron-level into discrete electron-level special phenomenon. At the discrete quantization level, the electron shock of nanomaterials produced many specific properties, such as electron-specific catalytic and photocatalytic characteristics [9].The structure determines the properties. Because nanomaterials have a different internal structure and surface state from general substances, they will have special properties different from general substances, such as fluorescence properties, adsorption properties, and catalytic properties. Because nanomaterials based on their special internal structural properties and surface state will cause it to have some special effects, these effects derive many physical and chemical properties that general substances do not have, such as optical, electric, magnetic, catalytic, and photocatalytic properties [10]. The catalytic properties based on nanomaterials have been widely concerned and applied in various fields of production and life, such as the detection of heavy metal ions in water, degradation of sewage and organic dyes, sensitive detection of biomolecules, detection of toxic gases in the air (such as carbon monoxide and nitric oxide), and detection of additives in food. Sensors based on the properties of nanomaterials can achieve selective and highly sensitive detection [11]. Compared with conventional materials, nanomaterials have greatly improved toughness, strength, and hardness, so they are widely used in aviation, aerospace, navigation, oil drilling, and other fields. The nanomaterial assembly structure is shown in Figure 1.Figure 1
Nanomaterial assembly structure.


### 3.2. Cerebral Hemorrhage

Cerebral hemorrhage is a typical neurological disease. It is a high lethality, high mortality disease. But due to the developed social economy and further improvement of public quality of life, the age of hemorrhagic stroke is increasingly young. Scientific research confirmed that the average annual prevalence of hemorrhagic stroke is about 60-80-0/100000 people, accounting for 20%–30% of the ischemic stroke; the month after the average death rate of more than 40%, 3/4 of cerebral hemorrhage survivors lost life and work ability to varying degrees. Hypertension diseases are also the most common cause of cerebral hemorrhage. Head trauma, congenital cerebrovascular malformation, anticoagulation, or thrombolytic therapy are also the more common causes of cerebral hemorrhage. The primary principle of ICH treatment is to keep quiet, stabilize blood pressure, prevent continued bleeding, appropriately reduce intracranial pressure, prevent cerebral edema, and maintain the balance of water, electrolyte, blood sugar, and body temperature according to the situation. Age, genetic factors, smoking, alcohol abuse, eating habits, obesity, and diabetes are all risk factors for cerebral hemorrhage [12]. Hypertensive intracerebral hemorrhage often occurs in 50 to 70-year-old males, which is slightly more likely to occur in winter and spring; it usually occurs during activities and emotional agitation, and there is no warning before bleeding. Half of the patients had severe headache, vomiting was common, and blood pressure increased significantly after hemorrhage. As shown in Figure 2, cerebral hemorrhage will not only cause somatic motor function and swallowing function, and discourse dysfunction, but also lead to major obstacles to cognitive function, including attention, memory function, action function, and learning and other different dysfunctions, which severity affect the patient’s health process and survival ability [13]. Intracerebral hemorrhage refers to hemorrhage caused by rupture of blood vessels in the nontraumatic brain parenchyma. The most common causes are hypertension, cerebral arteriosclerosis, intracranial vascular malformations, etc., which are often induced by exertion, emotional agitation, and other factors. Therefore, most of the onset occurs suddenly during activities. Clinically, the onset of cerebral hemorrhage is very rapid, mainly manifested as disturbance of consciousness, limb hemiplegia, aphasia, and other nervous system damage.Figure 2
Causes of intracerebral hemorrhage.


### 3.3. First-Principles Calculation of Nanomaterials

Nanomaterials have many strange characteristics such as special light, electricity, magnetic, thermal, mechanical, and other physical and chemical characteristics, which makes nanotechnology quickly enter into all scientific research studies by a large number of physicists and chemists; nanomaterials gain home and abroad scholars’ general attention. It has become the global hot topic of scientific research. Physicists are interested in nanomaterials because of their special electromagnetic properties, and material chemists are interested in their physical and chemical activities and their potential use value, while material scholars are interested in its hardness, strength, and flexibility [14–16]. Undoubtedly, the new nanotechnology based on nanomaterials will have a huge and profound impact on the economic development and social progress in today’s world. Therefore, the scientific research of nanomaterials is of great importance. Among them, carbon nanomaterials are one of the most popular scientific research materials [17]. Figure 3 is the manufacturing process of nanometer.Figure 3
Nanofabrication process.When dealing with the related problems of the micro-multiparticle system, the basic starting point is to solve the following equation of the system:(1)Kψl,L=Wψl,L,where ψl,L as the wave function of the system and K is the Hamiltonian operator of the molecular system, which can be expressed as(2)Kl,L=Uwln+Rwwln+UPLN+RPPLN+RPwln,LN=−∑nk22qw∇ln2+∑nk22QN∇LN2+12∑N,MCNCMw2LN−LM−∑N,nCnw2ln−LN,12∑n,mw2ln−lmwhere qw is the mass of the electron, l is the electron position coordinate, QN is the nuclear mass, and L is the nuclear position coordinate. The meanings of the five terms of the Hamiltonian operator are the electron kinetic energy term, the Coulomb action between the electrons, the nuclear kinetic energy term, the action between the nucleus, and the action term between the electron and the nucleus, respectively [18].When using this equation to solve specific problems, there are many problems to be addressed. However, for many nanosystems, rigorously precisely solving the equations in many-particle systems is still a difficult career-long thing [19].The so-called first-principles method: starting from quantum mechanics, using numerical solution equations, all the physical and chemical properties of the system can be studied. The first-principles method can provide the electronic structure characteristics of the system, but also can describe the bond damage and reconstruction, as well as electronic rearrangement process (such as chemical reaction). Because the principle method usually gives the highest accuracy calculation method, in principle, it only requires the type and position between molecules as input, and can accurately estimate all the physical and chemical properties of the molecular system [20].


#### 3.3.1. The Born–Oppenheimer Approximation

The Born–Oppenheimer approximation is a commonly used approximation method for solving quantum mechanical equations of systems involving electrons and nuclei. In the Born–Oppenheimer approximation,(a)
Electronic equation of motion is given as(3)Kwψwl,L=Wwψwl,L,whereψw is the wave function for the electron, and Kw is the Hamiltonian operator for the electron, which can be expressed as(4)Kw=Uwln+Rwwln+RPwln,LN.(b)
Equation of nuclear motion is given as(5)KPψPl,L=WPψPl,L,whereψP is the wave function of the electron, and KP is the Hamiltonian operator of the nucleus, which can be expressed as(6)KP=UPLN+RPPLN+We.(c)
A brief discussion of the Born–Oppenheimer approximation is given as follows:The Born–Oppenheimer approximation is a very useful approximation, causing very small errors. Compared to other approximations that must be used to solve the multi-electron problem, the errors can usually be ignored. If the electron motion and nuclear motion interactions are strong (electron-vibration coupling), it must be considered, which generally use the perturbation theory [21].


#### 3.3.2. Hartree–Fock Approximation

Under the Born–Oppenheimer approximation, the Hamiltonian operator of the electrons in the system can be expressed as(7)Kl,L=−∑nk22qw∇ln2+12∑n,mw2ln−lm−∑n,mCNw2ln−LN.We know that the key point in solving these differential equations is to calculate the Coulomb term between electrons. Without examining the Pauli incompatibility principle between electrons and atoms, Hartree sees all electrons as operating in a mean potential field formed with other atoms, so that the situation of each atom can be explained only by the wavefunction of a single atom. Therefore, the multi-electronic problem of this treatment system can be simply approximated by treating the single-atom problem [22].Therefore, the wave function form of the system can also be written as a continuous product form of the single-electron wave function form:(8)ψl=∏n∅nln.Thus, the total energy of the system can be expressed as(9)W=ψKψ=∑n∅nlnKn∅nln.We know that Pauli’s incompatibility principle can also be examined in the Hartree equation. Therefore, Fock has modified the Hartree equation to change the wave function of the whole system from product type to Slater determinant, so that the system wave function satisfies the Pauli principle:(10)ψ≡1P!∅1a1∅2a1∅1a2∅2a2⋯∅Pa1⋯∅Pa2⋯⋯∅1aP∅2aP⋯⋯⋯∅PaPan=ln,δn.ln and δn are the coordinates and spin indicators of the electrons, respectively.Among them,(11)∅an=∅ln,δn=∅ln∙δn.Therefore, the total energy of the system can be expressed as(12)W=ψKψ=∑n∫dl∅n,δ∗an−12∇2+Dext∅n,δan+12∑n,m∫dlndlm∅n∗an∅m∗am∅man∅namln−lm12∑n,m∫dlndlm∅n∗an∅m∗am∅mam∅nanln−lm.Correspondingly, the Schrodinger equation for a single electron can be expressed as(13)−k22q∇2+Dextl∅nan+∑n≠m∫dlm∅mam2ln−lm∅nan−=∑mφnm∅man.∑n≠m∫dlm∅m∗am∅nanln−lm∅manIn conclusion, the Hartree–Fock equation is an adiabatic approximation for the multi-electron interaction system; that is, the electrons run in the average potential field of the nucleus. And then, the Hartree–Fock approximation considers the incompatibility principle of the electron and Pauli system and then transforms the multi-electron problem into a single electron in a certain effective potential field.


### 3.4. Structure and Properties of Graphene

With its unique electronic structure and properties, graphene is an ideal two-dimensional electronic system discovered by humans so far. Graphene is the thinnest and lightest, yet the strongest nanometer film material in the world. Figure4 shows the process of making graphene.Figure 4
Graphene production process.The surface of graphene is almost fully transparent, absorbing only 23% of the light, so its thermal conductivity is 5300 W/m ∗ K. At this temperature, its electron migration speed is 15000 cm2/V ∗ s, which is the nanomaterials with the least resistivity temperature coefficient known. Meanwhile, graphene also has a room-temperature quantum Hall effect and magnetic forces. These physicochemical properties of graphene are derived from its peculiar electronic structure. Graphene is a carbon molecule made at sp2. The honeycomb single-layer plane two-dimensional crystal is similar to the single-carbon source of graphite, as shown in Figure 5. Therefore, graphene should be considered a planar polycyclic aromatic hydrocarbon molecular lattice structure.Figure 5
Electronic band energy structure of graphene.As we can see from the lattice structure of graphene, it is composed of two sublatticesX and Y interspersed, and the two lattice targets can be expressed as(14)x1=x23,3x2=x23,−3,where x = 1.42°X is the C-C bond length in graphene. The inverted target of its inverted space can be expressed as(15)y1=2π3x1,3y2=2π3x1,−3.In addition, there are two very important high-symmetry pointsR and R′ in the Brillouin region of graphene, whose corresponding coordinates are(16)R=2π3x,2π33xR′=2π3x,−2π33xUsing a simple tight-binding approximation to investigate the small molecule interaction between the closest and subclose relatives, the HamiltonianT of graphene can be defined as(17)T=−s∑xδ,n+yδ,m+T.v.−s`∑xδ,n+xδ,m+yδ,n+yδ,m+T.v.,where xδ,mxδ,n+ and yδ,myδ,n+ represent the generation and annihilation operators of the sublattices X and Y. The band structure of graphene can be calculated from the tight-binding theory:(18)W±h=±s3+gh−s`ghgh=2cos3hbx+4cos32hbxcos32hbx.It is clear that graphene at two high-symmetry pointsH and H′ attachments, with a conical band structure, presents a linear dispersion relationship that can be approximately expressed as(19)W±u=±scGu+TuH2where cG≈106m/s are the Fermi velocities near H and H′, and these high-symmetry points H and H′ are called the Dirac points of graphene.


## 3.1. Characterization of the Nanomaterials

Small-scale effect is also known as small volume effect. When the scale of nanoparticles and transfer electron wavelength or superconducting coherent wavelength are under the same physical scale, even more periodic boundary conditions were broken, the temperature, magnetism, optical absorption, boundary thermal resistance, chemical reactivity, catalytic change, and the effect of nanomaterials have also opened up a wide range of new application fields [7]. Nanostructure is a new system constructed according to certain rules on the basis of nanoscale material units.Surface effect refers to the change in the ratio of the number of molecules to the total atomic order on the nanoparticle surface as the particle size decreases or increases rapidly. Because the application of nanomaterials is gradually smaller, the number of surface atoms increased rapidly, so the surface area and surface free energy of nanoparticles also increased rapidly, and the surface atoms field environment and binding freedom are different between the atoms. The surface molecules between there are no adjacent atoms, so many hanging bonds have unsaturated, which is easy to combine between other atoms and stabilize it, that show a strong catalytic activity [8].Quantum size effect is that the particle scales down to a value, around the fermi electron level from quasi-continuous electron-level into discrete electron-level special phenomenon. At the discrete quantization level, the electron shock of nanomaterials produced many specific properties, such as electron-specific catalytic and photocatalytic characteristics [9].The structure determines the properties. Because nanomaterials have a different internal structure and surface state from general substances, they will have special properties different from general substances, such as fluorescence properties, adsorption properties, and catalytic properties. Because nanomaterials based on their special internal structural properties and surface state will cause it to have some special effects, these effects derive many physical and chemical properties that general substances do not have, such as optical, electric, magnetic, catalytic, and photocatalytic properties [10]. The catalytic properties based on nanomaterials have been widely concerned and applied in various fields of production and life, such as the detection of heavy metal ions in water, degradation of sewage and organic dyes, sensitive detection of biomolecules, detection of toxic gases in the air (such as carbon monoxide and nitric oxide), and detection of additives in food. Sensors based on the properties of nanomaterials can achieve selective and highly sensitive detection [11]. Compared with conventional materials, nanomaterials have greatly improved toughness, strength, and hardness, so they are widely used in aviation, aerospace, navigation, oil drilling, and other fields. The nanomaterial assembly structure is shown in Figure 1.Figure 1
Nanomaterial assembly structure.


## 3.2. Cerebral Hemorrhage

Cerebral hemorrhage is a typical neurological disease. It is a high lethality, high mortality disease. But due to the developed social economy and further improvement of public quality of life, the age of hemorrhagic stroke is increasingly young. Scientific research confirmed that the average annual prevalence of hemorrhagic stroke is about 60-80-0/100000 people, accounting for 20%–30% of the ischemic stroke; the month after the average death rate of more than 40%, 3/4 of cerebral hemorrhage survivors lost life and work ability to varying degrees. Hypertension diseases are also the most common cause of cerebral hemorrhage. Head trauma, congenital cerebrovascular malformation, anticoagulation, or thrombolytic therapy are also the more common causes of cerebral hemorrhage. The primary principle of ICH treatment is to keep quiet, stabilize blood pressure, prevent continued bleeding, appropriately reduce intracranial pressure, prevent cerebral edema, and maintain the balance of water, electrolyte, blood sugar, and body temperature according to the situation. Age, genetic factors, smoking, alcohol abuse, eating habits, obesity, and diabetes are all risk factors for cerebral hemorrhage [12]. Hypertensive intracerebral hemorrhage often occurs in 50 to 70-year-old males, which is slightly more likely to occur in winter and spring; it usually occurs during activities and emotional agitation, and there is no warning before bleeding. Half of the patients had severe headache, vomiting was common, and blood pressure increased significantly after hemorrhage. As shown in Figure 2, cerebral hemorrhage will not only cause somatic motor function and swallowing function, and discourse dysfunction, but also lead to major obstacles to cognitive function, including attention, memory function, action function, and learning and other different dysfunctions, which severity affect the patient’s health process and survival ability [13]. Intracerebral hemorrhage refers to hemorrhage caused by rupture of blood vessels in the nontraumatic brain parenchyma. The most common causes are hypertension, cerebral arteriosclerosis, intracranial vascular malformations, etc., which are often induced by exertion, emotional agitation, and other factors. Therefore, most of the onset occurs suddenly during activities. Clinically, the onset of cerebral hemorrhage is very rapid, mainly manifested as disturbance of consciousness, limb hemiplegia, aphasia, and other nervous system damage.Figure 2
Causes of intracerebral hemorrhage.


## 3.3. First-Principles Calculation of Nanomaterials

Nanomaterials have many strange characteristics such as special light, electricity, magnetic, thermal, mechanical, and other physical and chemical characteristics, which makes nanotechnology quickly enter into all scientific research studies by a large number of physicists and chemists; nanomaterials gain home and abroad scholars’ general attention. It has become the global hot topic of scientific research. Physicists are interested in nanomaterials because of their special electromagnetic properties, and material chemists are interested in their physical and chemical activities and their potential use value, while material scholars are interested in its hardness, strength, and flexibility [14–16]. Undoubtedly, the new nanotechnology based on nanomaterials will have a huge and profound impact on the economic development and social progress in today’s world. Therefore, the scientific research of nanomaterials is of great importance. Among them, carbon nanomaterials are one of the most popular scientific research materials [17]. Figure 3 is the manufacturing process of nanometer.Figure 3
Nanofabrication process.When dealing with the related problems of the micro-multiparticle system, the basic starting point is to solve the following equation of the system:(1)Kψl,L=Wψl,L,where ψl,L as the wave function of the system and K is the Hamiltonian operator of the molecular system, which can be expressed as(2)Kl,L=Uwln+Rwwln+UPLN+RPPLN+RPwln,LN=−∑nk22qw∇ln2+∑nk22QN∇LN2+12∑N,MCNCMw2LN−LM−∑N,nCnw2ln−LN,12∑n,mw2ln−lmwhere qw is the mass of the electron, l is the electron position coordinate, QN is the nuclear mass, and L is the nuclear position coordinate. The meanings of the five terms of the Hamiltonian operator are the electron kinetic energy term, the Coulomb action between the electrons, the nuclear kinetic energy term, the action between the nucleus, and the action term between the electron and the nucleus, respectively [18].When using this equation to solve specific problems, there are many problems to be addressed. However, for many nanosystems, rigorously precisely solving the equations in many-particle systems is still a difficult career-long thing [19].The so-called first-principles method: starting from quantum mechanics, using numerical solution equations, all the physical and chemical properties of the system can be studied. The first-principles method can provide the electronic structure characteristics of the system, but also can describe the bond damage and reconstruction, as well as electronic rearrangement process (such as chemical reaction). Because the principle method usually gives the highest accuracy calculation method, in principle, it only requires the type and position between molecules as input, and can accurately estimate all the physical and chemical properties of the molecular system [20].


### 3.3.1. The Born–Oppenheimer Approximation

The Born–Oppenheimer approximation is a commonly used approximation method for solving quantum mechanical equations of systems involving electrons and nuclei. In the Born–Oppenheimer approximation,(a)
Electronic equation of motion is given as(3)Kwψwl,L=Wwψwl,L,whereψw is the wave function for the electron, and Kw is the Hamiltonian operator for the electron, which can be expressed as(4)Kw=Uwln+Rwwln+RPwln,LN.(b)
Equation of nuclear motion is given as(5)KPψPl,L=WPψPl,L,whereψP is the wave function of the electron, and KP is the Hamiltonian operator of the nucleus, which can be expressed as(6)KP=UPLN+RPPLN+We.(c)
A brief discussion of the Born–Oppenheimer approximation is given as follows:The Born–Oppenheimer approximation is a very useful approximation, causing very small errors. Compared to other approximations that must be used to solve the multi-electron problem, the errors can usually be ignored. If the electron motion and nuclear motion interactions are strong (electron-vibration coupling), it must be considered, which generally use the perturbation theory [21].


### 3.3.2. Hartree–Fock Approximation

Under the Born–Oppenheimer approximation, the Hamiltonian operator of the electrons in the system can be expressed as(7)Kl,L=−∑nk22qw∇ln2+12∑n,mw2ln−lm−∑n,mCNw2ln−LN.We know that the key point in solving these differential equations is to calculate the Coulomb term between electrons. Without examining the Pauli incompatibility principle between electrons and atoms, Hartree sees all electrons as operating in a mean potential field formed with other atoms, so that the situation of each atom can be explained only by the wavefunction of a single atom. Therefore, the multi-electronic problem of this treatment system can be simply approximated by treating the single-atom problem [22].Therefore, the wave function form of the system can also be written as a continuous product form of the single-electron wave function form:(8)ψl=∏n∅nln.Thus, the total energy of the system can be expressed as(9)W=ψKψ=∑n∅nlnKn∅nln.We know that Pauli’s incompatibility principle can also be examined in the Hartree equation. Therefore, Fock has modified the Hartree equation to change the wave function of the whole system from product type to Slater determinant, so that the system wave function satisfies the Pauli principle:(10)ψ≡1P!∅1a1∅2a1∅1a2∅2a2⋯∅Pa1⋯∅Pa2⋯⋯∅1aP∅2aP⋯⋯⋯∅PaPan=ln,δn.ln and δn are the coordinates and spin indicators of the electrons, respectively.Among them,(11)∅an=∅ln,δn=∅ln∙δn.Therefore, the total energy of the system can be expressed as(12)W=ψKψ=∑n∫dl∅n,δ∗an−12∇2+Dext∅n,δan+12∑n,m∫dlndlm∅n∗an∅m∗am∅man∅namln−lm12∑n,m∫dlndlm∅n∗an∅m∗am∅mam∅nanln−lm.Correspondingly, the Schrodinger equation for a single electron can be expressed as(13)−k22q∇2+Dextl∅nan+∑n≠m∫dlm∅mam2ln−lm∅nan−=∑mφnm∅man.∑n≠m∫dlm∅m∗am∅nanln−lm∅manIn conclusion, the Hartree–Fock equation is an adiabatic approximation for the multi-electron interaction system; that is, the electrons run in the average potential field of the nucleus. And then, the Hartree–Fock approximation considers the incompatibility principle of the electron and Pauli system and then transforms the multi-electron problem into a single electron in a certain effective potential field.


## 3.3.1. The Born–Oppenheimer Approximation

The Born–Oppenheimer approximation is a commonly used approximation method for solving quantum mechanical equations of systems involving electrons and nuclei. In the Born–Oppenheimer approximation,(a)
Electronic equation of motion is given as(3)Kwψwl,L=Wwψwl,L,whereψw is the wave function for the electron, and Kw is the Hamiltonian operator for the electron, which can be expressed as(4)Kw=Uwln+Rwwln+RPwln,LN.(b)
Equation of nuclear motion is given as(5)KPψPl,L=WPψPl,L,whereψP is the wave function of the electron, and KP is the Hamiltonian operator of the nucleus, which can be expressed as(6)KP=UPLN+RPPLN+We.(c)
A brief discussion of the Born–Oppenheimer approximation is given as follows:The Born–Oppenheimer approximation is a very useful approximation, causing very small errors. Compared to other approximations that must be used to solve the multi-electron problem, the errors can usually be ignored. If the electron motion and nuclear motion interactions are strong (electron-vibration coupling), it must be considered, which generally use the perturbation theory [21].


## 3.3.2. Hartree–Fock Approximation

Under the Born–Oppenheimer approximation, the Hamiltonian operator of the electrons in the system can be expressed as(7)Kl,L=−∑nk22qw∇ln2+12∑n,mw2ln−lm−∑n,mCNw2ln−LN.We know that the key point in solving these differential equations is to calculate the Coulomb term between electrons. Without examining the Pauli incompatibility principle between electrons and atoms, Hartree sees all electrons as operating in a mean potential field formed with other atoms, so that the situation of each atom can be explained only by the wavefunction of a single atom. Therefore, the multi-electronic problem of this treatment system can be simply approximated by treating the single-atom problem [22].Therefore, the wave function form of the system can also be written as a continuous product form of the single-electron wave function form:(8)ψl=∏n∅nln.Thus, the total energy of the system can be expressed as(9)W=ψKψ=∑n∅nlnKn∅nln.We know that Pauli’s incompatibility principle can also be examined in the Hartree equation. Therefore, Fock has modified the Hartree equation to change the wave function of the whole system from product type to Slater determinant, so that the system wave function satisfies the Pauli principle:(10)ψ≡1P!∅1a1∅2a1∅1a2∅2a2⋯∅Pa1⋯∅Pa2⋯⋯∅1aP∅2aP⋯⋯⋯∅PaPan=ln,δn.ln and δn are the coordinates and spin indicators of the electrons, respectively.Among them,(11)∅an=∅ln,δn=∅ln∙δn.Therefore, the total energy of the system can be expressed as(12)W=ψKψ=∑n∫dl∅n,δ∗an−12∇2+Dext∅n,δan+12∑n,m∫dlndlm∅n∗an∅m∗am∅man∅namln−lm12∑n,m∫dlndlm∅n∗an∅m∗am∅mam∅nanln−lm.Correspondingly, the Schrodinger equation for a single electron can be expressed as(13)−k22q∇2+Dextl∅nan+∑n≠m∫dlm∅mam2ln−lm∅nan−=∑mφnm∅man.∑n≠m∫dlm∅m∗am∅nanln−lm∅manIn conclusion, the Hartree–Fock equation is an adiabatic approximation for the multi-electron interaction system; that is, the electrons run in the average potential field of the nucleus. And then, the Hartree–Fock approximation considers the incompatibility principle of the electron and Pauli system and then transforms the multi-electron problem into a single electron in a certain effective potential field.


## 3.4. Structure and Properties of Graphene

With its unique electronic structure and properties, graphene is an ideal two-dimensional electronic system discovered by humans so far. Graphene is the thinnest and lightest, yet the strongest nanometer film material in the world. Figure4 shows the process of making graphene.Figure 4
Graphene production process.The surface of graphene is almost fully transparent, absorbing only 23% of the light, so its thermal conductivity is 5300 W/m ∗ K. At this temperature, its electron migration speed is 15000 cm2/V ∗ s, which is the nanomaterials with the least resistivity temperature coefficient known. Meanwhile, graphene also has a room-temperature quantum Hall effect and magnetic forces. These physicochemical properties of graphene are derived from its peculiar electronic structure. Graphene is a carbon molecule made at sp2. The honeycomb single-layer plane two-dimensional crystal is similar to the single-carbon source of graphite, as shown in Figure 5. Therefore, graphene should be considered a planar polycyclic aromatic hydrocarbon molecular lattice structure.Figure 5
Electronic band energy structure of graphene.As we can see from the lattice structure of graphene, it is composed of two sublatticesX and Y interspersed, and the two lattice targets can be expressed as(14)x1=x23,3x2=x23,−3,where x = 1.42°X is the C-C bond length in graphene. The inverted target of its inverted space can be expressed as(15)y1=2π3x1,3y2=2π3x1,−3.In addition, there are two very important high-symmetry pointsR and R′ in the Brillouin region of graphene, whose corresponding coordinates are(16)R=2π3x,2π33xR′=2π3x,−2π33xUsing a simple tight-binding approximation to investigate the small molecule interaction between the closest and subclose relatives, the HamiltonianT of graphene can be defined as(17)T=−s∑xδ,n+yδ,m+T.v.−s`∑xδ,n+xδ,m+yδ,n+yδ,m+T.v.,where xδ,mxδ,n+ and yδ,myδ,n+ represent the generation and annihilation operators of the sublattices X and Y. The band structure of graphene can be calculated from the tight-binding theory:(18)W±h=±s3+gh−s`ghgh=2cos3hbx+4cos32hbxcos32hbx.It is clear that graphene at two high-symmetry pointsH and H′ attachments, with a conical band structure, presents a linear dispersion relationship that can be approximately expressed as(19)W±u=±scGu+TuH2where cG≈106m/s are the Fermi velocities near H and H′, and these high-symmetry points H and H′ are called the Dirac points of graphene.


## 4. Nanomaterials and Their Role in the Treatment of Cerebral Hemorrhage

### 4.1. Electrochemical Detection of Rutin by Nanocomposites

Lutin, also known as vitamin P, belongs to flavonoids and is widely found in the leaves of some plants such as Sophora japonica rice and buckwheat. Lutin is usually considered a kind of active therapeutic drugs to prevent and treat diseases such as hypertension and cerebral hemorrhage. Meanwhile, and as a natural drug, rutin has obvious antibacterial, antiallergic, antioxidant, and other physical and chemical properties. However, the overdose of rutin drugs can produce serious side effects and pose a threat to human life safety. Therefore, the development of simple, economical, and sensitive rutin detection methods is of great significance for human health. So far, the laboratory detection methods used in rutin include HPLC, reverse-phase HPLC, UV visible spectrophotometry, and chemical luminescence method, but some of these methods are complicated predetection and require a long detection time; on the contrary, the electrochemical method is convenient, and has characteristics such as quick response, low detection limit, and good stability; therefore, compared with these common methods, electrochemical detection method is a good choice for Lutin detection [23].We experimentally explored the effect of solution pH on the electrochemical reaction during rutin detection. As can be seen in Figure6, a clear linear relationship between the oxidative voltage and pH with solution pH increases, and the number of electron transport and proton transfer is the same during the redox process of rutin. In addition, the relationship curve between the solution pH value and the oxidation peak current value can also be seen. When the pH value of the solution is less than 6.0, rutin is hydrolyzed into rhamnose, glucose, and cutin under acidic conditions, so the oxidation peak current is lower to detect than the other oxidation ones. When the pH value of the solution is greater than 6.0, the oxidation peak current also appears decreasing, possibly because the flavonoids’ parent core is damaged under alkaline conditions, which affects the electrochemical catalysis of rutin. Therefore, we chose pH = 6.0 as the best pH value for the detection solution.Figure 6
The linear relationship between oxidation voltage, current, and pH value and the linear relationship between redox peak current and scan rate.
(a)(b)(c)(d)As can be seen from Figure6, the redox current of rutin is correlated with the sweep speed, while the peak potential of the redox peak is slightly changed, and a good linear relationship between the oxidation peak current and the pH value is shown. The linear relationship between them proves that the catalytic reaction of rutin on a platinum/graphene-modified glass carbon electrode is an adsorption-controlled process.Under the appropriate experimental conditions, we used the differential pulse voltammetry of the prepared electrode. In the range of 0.057–102.59 uM, the oxidation peak current value of rutin and the concentration of rutin solution were positively correlated and showed a two-stage linear relationship. In the concentration range of 0.57–5.47 uM, the oxidation peak current increases rapidly; with the concentration of rutin increasing from 5.47 uM to 102.59 uM, the oxidation peak current slows down, and the minimum detection limit of the electrode reaches 0.02 uM (S/N = 3), which is much lower than the other reported detection results. As shown in Table 1, it proves that our platinum/graphene-modified glass carbon electrode shows good detection performance for rutin.Table 1
Comparison of rutin determination results with different electrodes.
ElectrodeLine range (uM)LOD (uM)Au/en/MWNTsa/GCEb0.047–0.0960.031GR/CILEc0.06–10.023IL/GCE0.2–10.08PtNPs/RGO/GCE0.056–102.610.02In the repetitive test, five prepared PtNPs/RGO/GCE electrodes were tested for cyclic voltammetry in 10 uM rutin solution, with the relative standard deviation of the oxidation peak current of 1.5%, showing good repeatability of the electrodes. In addition, the prepared electrode was placed at room temperature for one week and tested every other day after the results of DPV, whose oxidation peak current value remained at 91.08% of the initial value, showing good stability of the prepared electrode. To study the anti-interference properties of PtNPs/RGO/GCE, we added 10 times the concentration of KCI, NagSO4, and NH4 to 10uM rutin solution; PO4, DA, AA, UA, glucose, and no obvious interference peak, as shown in Figure7, indicating that the prepared electrode has good anti-interference performance. The recovery rate on PtNPs/RGO/GCE is 98.60%, 106.25%, and 98.90%, respectively, which shows that the prepared electrode can be effectively applied to the detection of the rutin content in commercial rutin tablets.Figure 7
Response results of PtNPs/RGO/GCE in rutin solution to interfering ions.
(a)(b)The composite graphene nanochip material is modified and applied to rutin detection. During the electrochemical detection of rutin, platinum nanoparticles/graphene composites effectively enhance the electrochemical catalytic response and improved detection performance compared with pure platinum and graphene, probably due to its large flexural area, strong conductivity, and catalytic activity. In the preparation of the electrode, the measurement range of rutin is 0.057–102.59 uM, and the minimum measurement limit is 0.02 uM. Meanwhile, the electrode also showed good repetition, stability, and interference resistance. This indicates that the described method for rutin detection and the prepared electrodes have great potential applications in the actual rutin detection.


### 4.2. Intracerebral Pharmacokinetics of Trekrutin and Analogs

#### 4.2.1. Experimental Materials

Clean-grade healthy mice, male and generally weighing between 18 g and 22 g, were purchased at a laboratory animal management center at a medical institution. Mice were housed in a plastic feeding cage, bedding was replaced once the other day, and water was consumed freely, after 1 week of temperature 22∼25°C and humidity around 40%∼60%.


#### 4.2.2. Experimental Method

Internal standard material: quercetin: chromatographic analysis column: ® (50 mm × 2.1 mm, 1.7 m); flow phase: acetamide: 0.1% formic acid water; gradient elution, as shown in Table2. Flow rate: 0. 3 mL·min−1; column temperature: 30°C; mass of inflow sample: 1 L.Table 2
Mobile phase ratios.
Mobile phase ratioTime (min)Methanol0.1% formic acid water034642.534644.549496.5346412.5 mg of troxerutin, rutin, and troxerutin aglycone were taken as the reference product, weighed accurately, and put into a 50-mL brown volumetric flask. It was hydrolyzed with an acetylpropylamine ultrasonic tube and fixed to volume to prepare a stock solution with a content of about 250μg ∗ mL−1, which was frozen and stored in a refrigerator at 4°C for later use.200 L of blank brain tissue was taken and blow-dried with nitrogen, and a series of 200 L of troxerutin, rutin, and troxerutin aglycone were added to prepare low, medium, and high concentrated water solvents of 10,50,200 ng·mL−1 and 72 ng ∗ mL−1. The aqueous solution (n = 6) is the difference between the peak building area measured and the peak area measured with the same content in the blank EP tube.


#### 4.2.3. Results

The brain tissue matrix has less influence on troxerutin, rutin, and troxerutin aglycone and internal target; the mean matrix effect is between 90% and 100%; and the RSD is less than 2%, which can be considered to have basically no matrix effect. The specific results are shown in Table3.Table 3
Investigation of brain tissue matrix effect.
SamplesConcentration (ng ∗ ml−1)Matrix effect (%)RSD(%)Troxerutin1093.11.35090.620091.5Rutin1091.51.65092.420091.4Troxerutin aglycone1092.81.15091.120090.7The extraction recovery rate meets the requirements of pharmacokinetics. Intraday and daytime precision suggested that the RSD of tretin and rutin at high, medium, and low concentrations was <6.0%. The specific results are shown in Figures8 and 9 and Table 4.Figure 8
Extraction precision of troxerutin in mouse brain tissue.
(a)(b)Figure 9
The extraction precision of rutin in mouse brain tissue.
(a)(b)Table 4
Extraction precision of troxerutin aglycone in mouse brain tissue.
Concentration (ng/mL)Within the day (ng ∗ mL−1)RSD (%)Interday (ng/mL)RSD (%)109.511.959.372.875049.172.5148.9698.2200196.081.24194.1598.7Room temperature, freeze-thaw cycles, and long-term stability were investigated, and the results showed that the RSD of rutin and low concentrations was less than 5.0%, and the stability of troxerutin, rutin, and troxerutin aglycone met the requirements.


## 4.1. Electrochemical Detection of Rutin by Nanocomposites

Lutin, also known as vitamin P, belongs to flavonoids and is widely found in the leaves of some plants such as Sophora japonica rice and buckwheat. Lutin is usually considered a kind of active therapeutic drugs to prevent and treat diseases such as hypertension and cerebral hemorrhage. Meanwhile, and as a natural drug, rutin has obvious antibacterial, antiallergic, antioxidant, and other physical and chemical properties. However, the overdose of rutin drugs can produce serious side effects and pose a threat to human life safety. Therefore, the development of simple, economical, and sensitive rutin detection methods is of great significance for human health. So far, the laboratory detection methods used in rutin include HPLC, reverse-phase HPLC, UV visible spectrophotometry, and chemical luminescence method, but some of these methods are complicated predetection and require a long detection time; on the contrary, the electrochemical method is convenient, and has characteristics such as quick response, low detection limit, and good stability; therefore, compared with these common methods, electrochemical detection method is a good choice for Lutin detection [23].We experimentally explored the effect of solution pH on the electrochemical reaction during rutin detection. As can be seen in Figure6, a clear linear relationship between the oxidative voltage and pH with solution pH increases, and the number of electron transport and proton transfer is the same during the redox process of rutin. In addition, the relationship curve between the solution pH value and the oxidation peak current value can also be seen. When the pH value of the solution is less than 6.0, rutin is hydrolyzed into rhamnose, glucose, and cutin under acidic conditions, so the oxidation peak current is lower to detect than the other oxidation ones. When the pH value of the solution is greater than 6.0, the oxidation peak current also appears decreasing, possibly because the flavonoids’ parent core is damaged under alkaline conditions, which affects the electrochemical catalysis of rutin. Therefore, we chose pH = 6.0 as the best pH value for the detection solution.Figure 6
The linear relationship between oxidation voltage, current, and pH value and the linear relationship between redox peak current and scan rate.
(a)(b)(c)(d)As can be seen from Figure6, the redox current of rutin is correlated with the sweep speed, while the peak potential of the redox peak is slightly changed, and a good linear relationship between the oxidation peak current and the pH value is shown. The linear relationship between them proves that the catalytic reaction of rutin on a platinum/graphene-modified glass carbon electrode is an adsorption-controlled process.Under the appropriate experimental conditions, we used the differential pulse voltammetry of the prepared electrode. In the range of 0.057–102.59 uM, the oxidation peak current value of rutin and the concentration of rutin solution were positively correlated and showed a two-stage linear relationship. In the concentration range of 0.57–5.47 uM, the oxidation peak current increases rapidly; with the concentration of rutin increasing from 5.47 uM to 102.59 uM, the oxidation peak current slows down, and the minimum detection limit of the electrode reaches 0.02 uM (S/N = 3), which is much lower than the other reported detection results. As shown in Table 1, it proves that our platinum/graphene-modified glass carbon electrode shows good detection performance for rutin.Table 1
Comparison of rutin determination results with different electrodes.
ElectrodeLine range (uM)LOD (uM)Au/en/MWNTsa/GCEb0.047–0.0960.031GR/CILEc0.06–10.023IL/GCE0.2–10.08PtNPs/RGO/GCE0.056–102.610.02In the repetitive test, five prepared PtNPs/RGO/GCE electrodes were tested for cyclic voltammetry in 10 uM rutin solution, with the relative standard deviation of the oxidation peak current of 1.5%, showing good repeatability of the electrodes. In addition, the prepared electrode was placed at room temperature for one week and tested every other day after the results of DPV, whose oxidation peak current value remained at 91.08% of the initial value, showing good stability of the prepared electrode. To study the anti-interference properties of PtNPs/RGO/GCE, we added 10 times the concentration of KCI, NagSO4, and NH4 to 10uM rutin solution; PO4, DA, AA, UA, glucose, and no obvious interference peak, as shown in Figure7, indicating that the prepared electrode has good anti-interference performance. The recovery rate on PtNPs/RGO/GCE is 98.60%, 106.25%, and 98.90%, respectively, which shows that the prepared electrode can be effectively applied to the detection of the rutin content in commercial rutin tablets.Figure 7
Response results of PtNPs/RGO/GCE in rutin solution to interfering ions.
(a)(b)The composite graphene nanochip material is modified and applied to rutin detection. During the electrochemical detection of rutin, platinum nanoparticles/graphene composites effectively enhance the electrochemical catalytic response and improved detection performance compared with pure platinum and graphene, probably due to its large flexural area, strong conductivity, and catalytic activity. In the preparation of the electrode, the measurement range of rutin is 0.057–102.59 uM, and the minimum measurement limit is 0.02 uM. Meanwhile, the electrode also showed good repetition, stability, and interference resistance. This indicates that the described method for rutin detection and the prepared electrodes have great potential applications in the actual rutin detection.


## 4.2. Intracerebral Pharmacokinetics of Trekrutin and Analogs

### 4.2.1. Experimental Materials

Clean-grade healthy mice, male and generally weighing between 18 g and 22 g, were purchased at a laboratory animal management center at a medical institution. Mice were housed in a plastic feeding cage, bedding was replaced once the other day, and water was consumed freely, after 1 week of temperature 22∼25°C and humidity around 40%∼60%.


### 4.2.2. Experimental Method

Internal standard material: quercetin: chromatographic analysis column: ® (50 mm × 2.1 mm, 1.7 m); flow phase: acetamide: 0.1% formic acid water; gradient elution, as shown in Table2. Flow rate: 0. 3 mL·min−1; column temperature: 30°C; mass of inflow sample: 1 L.Table 2
Mobile phase ratios.
Mobile phase ratioTime (min)Methanol0.1% formic acid water034642.534644.549496.5346412.5 mg of troxerutin, rutin, and troxerutin aglycone were taken as the reference product, weighed accurately, and put into a 50-mL brown volumetric flask. It was hydrolyzed with an acetylpropylamine ultrasonic tube and fixed to volume to prepare a stock solution with a content of about 250μg ∗ mL−1, which was frozen and stored in a refrigerator at 4°C for later use.200 L of blank brain tissue was taken and blow-dried with nitrogen, and a series of 200 L of troxerutin, rutin, and troxerutin aglycone were added to prepare low, medium, and high concentrated water solvents of 10,50,200 ng·mL−1 and 72 ng ∗ mL−1. The aqueous solution (n = 6) is the difference between the peak building area measured and the peak area measured with the same content in the blank EP tube.


### 4.2.3. Results

The brain tissue matrix has less influence on troxerutin, rutin, and troxerutin aglycone and internal target; the mean matrix effect is between 90% and 100%; and the RSD is less than 2%, which can be considered to have basically no matrix effect. The specific results are shown in Table3.Table 3
Investigation of brain tissue matrix effect.
SamplesConcentration (ng ∗ ml−1)Matrix effect (%)RSD(%)Troxerutin1093.11.35090.620091.5Rutin1091.51.65092.420091.4Troxerutin aglycone1092.81.15091.120090.7The extraction recovery rate meets the requirements of pharmacokinetics. Intraday and daytime precision suggested that the RSD of tretin and rutin at high, medium, and low concentrations was <6.0%. The specific results are shown in Figures8 and 9 and Table 4.Figure 8
Extraction precision of troxerutin in mouse brain tissue.
(a)(b)Figure 9
The extraction precision of rutin in mouse brain tissue.
(a)(b)Table 4
Extraction precision of troxerutin aglycone in mouse brain tissue.
Concentration (ng/mL)Within the day (ng ∗ mL−1)RSD (%)Interday (ng/mL)RSD (%)109.511.959.372.875049.172.5148.9698.2200196.081.24194.1598.7Room temperature, freeze-thaw cycles, and long-term stability were investigated, and the results showed that the RSD of rutin and low concentrations was less than 5.0%, and the stability of troxerutin, rutin, and troxerutin aglycone met the requirements.


## 4.2.1. Experimental Materials

Clean-grade healthy mice, male and generally weighing between 18 g and 22 g, were purchased at a laboratory animal management center at a medical institution. Mice were housed in a plastic feeding cage, bedding was replaced once the other day, and water was consumed freely, after 1 week of temperature 22∼25°C and humidity around 40%∼60%.


## 4.2.2. Experimental Method

Internal standard material: quercetin: chromatographic analysis column: ® (50 mm × 2.1 mm, 1.7 m); flow phase: acetamide: 0.1% formic acid water; gradient elution, as shown in Table2. Flow rate: 0. 3 mL·min−1; column temperature: 30°C; mass of inflow sample: 1 L.Table 2
Mobile phase ratios.
Mobile phase ratioTime (min)Methanol0.1% formic acid water034642.534644.549496.5346412.5 mg of troxerutin, rutin, and troxerutin aglycone were taken as the reference product, weighed accurately, and put into a 50-mL brown volumetric flask. It was hydrolyzed with an acetylpropylamine ultrasonic tube and fixed to volume to prepare a stock solution with a content of about 250μg ∗ mL−1, which was frozen and stored in a refrigerator at 4°C for later use.200 L of blank brain tissue was taken and blow-dried with nitrogen, and a series of 200 L of troxerutin, rutin, and troxerutin aglycone were added to prepare low, medium, and high concentrated water solvents of 10,50,200 ng·mL−1 and 72 ng ∗ mL−1. The aqueous solution (n = 6) is the difference between the peak building area measured and the peak area measured with the same content in the blank EP tube.


## 4.2.3. Results

The brain tissue matrix has less influence on troxerutin, rutin, and troxerutin aglycone and internal target; the mean matrix effect is between 90% and 100%; and the RSD is less than 2%, which can be considered to have basically no matrix effect. The specific results are shown in Table3.Table 3
Investigation of brain tissue matrix effect.
SamplesConcentration (ng ∗ ml−1)Matrix effect (%)RSD(%)Troxerutin1093.11.35090.620091.5Rutin1091.51.65092.420091.4Troxerutin aglycone1092.81.15091.120090.7The extraction recovery rate meets the requirements of pharmacokinetics. Intraday and daytime precision suggested that the RSD of tretin and rutin at high, medium, and low concentrations was <6.0%. The specific results are shown in Figures8 and 9 and Table 4.Figure 8
Extraction precision of troxerutin in mouse brain tissue.
(a)(b)Figure 9
The extraction precision of rutin in mouse brain tissue.
(a)(b)Table 4
Extraction precision of troxerutin aglycone in mouse brain tissue.
Concentration (ng/mL)Within the day (ng ∗ mL−1)RSD (%)Interday (ng/mL)RSD (%)109.511.959.372.875049.172.5148.9698.2200196.081.24194.1598.7Room temperature, freeze-thaw cycles, and long-term stability were investigated, and the results showed that the RSD of rutin and low concentrations was less than 5.0%, and the stability of troxerutin, rutin, and troxerutin aglycone met the requirements.


## 5. Discussion

Rutin has antioxidative effect against the decline of antioxidant capacity caused by the decline of endogenous estrogen. At present, the pharmacokinetics reports of troxerutin, rutin, and troxerutin aglycone are mostly detected in the blood concentration, and there is no literature on the concentration determination of trekrutin and analogs in brain tissue or brain pharmacokinetics, and the detection methods are mostly HPLC. The liquid mass combination combines the characteristics of chromatography and mass spectroscopy, which combines the high separation of chromatography and the high-resolution characteristics of mass spectrometry. It has recently become a tool with high sensitivity and high specificity, and is widely used for biological sample analysis and pharmacokinetic research. In general, the mass spectrometry detection method mostly selects the MRM (multireaction monitoring) method, and the MRM method has higher sensitivity, stronger specificity, and lower quantitative limit. In the first-level full scanning of trekrutin and analogs, it was not easy. When secondary fragment ion scanning, it was found that rutin is the same at the ion source, and too low voltage to test ionization will cause low response, and high voltage of source cracking will also cause low response; multiple groups of voltage were tried. Finally, we choose 3.49 kV capillary voltage, and used acetpropyamine and low strength formic acid as the flow phase peak tail and facilitate ionization, and then, a better peak shape and mass spectrum response were gotten.


## 6. Conclusions

Tricludin, rutin, and tricluditidine in the mouse brain tissue were extracted by methanol protein precipitation, and the UPLC-M S/MS analysis method of trecludin and analogs was first established and successfully used for the brain pharmacokinetic study of trecludin and analogs in the mouse brain tissue. The tail vein injection of troxerutin, rutin, and troxerutin aglycone peaked rapidly and eliminated more slowly in the brain. It is much higher than the other two drug groups, suggesting that the least polar logP crosses the BBB, providing a reference to study the structure of drugs passing through the BBB. However, due to the limitations of time and technology, we have not carried out research on the deeper application of rutin. We will carry out further experiments to explore this in the follow-up.



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*Source: 7044153-2022-08-18.xml*