Patent ID: 11921123
Assignee: WUHAN UNIVERSITY OF TECHNOLOGY
Field: Measurement (Instruments)
Classification: CPC G | IPC G

Claim 0:
1. A quantitative detection method of rare earth doped calcium phosphate fluorescent nanoparticles (RE-nCaP) in at least one biological tissue of one of blood, heart, liver, spleen, lung, kidney, pancreas, brain, lymph, skin, and nerve of an animal, comprising steps of:
(a) establishing a fluorescent intensity-concentration standard curve of rare earth (RE) ions in a fluorescent enhancement liquid, wherein the fluorescent intensity-concentration standard curve satisfies a relationship of Y=B+N·X, wherein Y and X are respectively a fluorescent intensity and an RE concentration of the fluorescent enhancement liquid, and B and N are constants and respectively an intercept and a slope of the fluorescent intensity-concentration standard curve;
(b) separately treating at least one test biological tissue sample of a group to be tested, and at least one control biological tissue sample of a blank control group with an acid solution to obtain homogenates of the group to be tested and the blank control group, wherein the at least one test biological tissue sample of the group to be tested is obtained from said at least one biological tissue of the animal, and the at least one control biological tissue sample of the blank control group is obtained from at least one control biological tissue of another animal containing no RE-nCaP, wherein said at least one control biological tissue is a tissue of one of blood, heart, liver, spleen, lung, kidney, pancreas, brain, lymph, skin, and nerve of said another animal that is corresponding to said one of said blood, heart, liver, spleen, lung, kidney, pancreas, brain, lymph, skin, and nerve of the animal;
performing centrifugal separation on the homogenates of the at least one test biological tissue sample of the group to be tested and the at least one control biological tissue sample of the blank control group separately to obtain supernatants of the at least one test biological tissue sample of the group to be tested and the at least one control biological tissue sample of the blank control group;
diluting the supernatants of the at least one test biological tissue sample of the group to be tested and the at least one control biological tissue sample of the blank control group with the fluorescent enhancement liquid;
detecting fluorescent intensities of the diluted supernatants of the at least one test biological tissue sample of the group to be tested and the at least one control biological tissue sample of the blank control group;
calculating values, T and T0, of a fluorescent intensity per unit mass or volume of the at least one test biological tissue sample of the group to be tested and the at least one control biological tissue sample of the blank control group according to formula (I):, T
      =
      
       
        y
        -
        B
       
       W
      
     
     ;
     
      
       and
       ⁢
          
       
        T
        0
       
      
      =
      
       
        
         y
         0
        
        -
        B
       
       
        W
        0
       
      
     
    
   
   
    
     (
     I
     )
    
   
  
 

wherein y and y0 are respectively the detected fluorescent intensities of the diluted supernatants of the at least one test biological tissue sample of the group to be tested and the at least one control biological tissue sample of the blank control group, W is a weight or volume of the at least one test biological tissue sample of the group to be tested, and W0 is a weight or volume of the at least one control biological tissue sample of the blank control group;
performing significant difference analysis on the values T and T0 of the fluorescent intensity per unit mass or volume between the at least one test biological tissue sample of the group to be tested and the at least one control biological tissue sample of the blank control group, so as to determine if the at least one test biological tissue sample in the group to be tested contain the RE-nCaP; and
continuing to perform a step (c) and a step (d) to further determine an amount of the RE-nCaP when the at least one test biological tissue sample in the group to be tested are determined to contain the RE-nCaP;

(c) separately adding the RE-nCaP to the homogenate of the at least one control biological tissue sample of the blank control group and to the acid solution having a same volume as that of the homogenate of the at least one control biological tissue sample of the blank control group;
performing centrifugal separation on the RE-nCaP added homogenate of the at least one control biological tissue sample of the blank control group and the RE-nCaP added acid solution separately to obtain supernatants of the RE-nCaP added homogenate of the at least one control biological tissue sample of the blank control group and the RE-nCaP added acid solution;
diluting the supernatants of the RE-nCaP added homogenate of the at least one control biological tissue sample of the blank control group and the RE-nCaP added acid solution with the fluorescent enhancement liquid;
detecting fluorescent intensities of the diluted supernatants of the RE-nCaP added homogenate of the at least one control biological tissue sample of the blank control group and the RE-nCaP added acid solution;
calculating a tissue retention rate, S, per unit mass or volume according to formula (II):, S
     =
     
      
       (
       
        
         A
         0
        
        -
        
         A
         1
        
       
       )
      
      
       
        A
        0
       
       ⁢
       
        W
        0
       
      
     
    
   
   
    
     (
     II
     )
    
   
  
 

wherein A1 is the detected fluorescent intensity of the RE-nCaP added homogenate of the at least one control biological tissue sample of the blank control group and A0 is the detected fluorescent intensity of the RE-nCaP added acid solution, and W0 is the weight or volume of the at least one control biological tissue sample of the blank control group; and
calculating a tissue extraction rate, R, of the group to be tested according to a weight or volume of the at least one test biological tissue sample in the group to be tested according to formula (III):

R=(1−S·W)×100%  (III), wherein W is the weight or volume of the at least one test biological tissue sample in the group to be tested; and
(d) calculating an amount, M, of the RE-nCaP in the at least one test biological tissue sample of the group to be tested according to formula (IV):, M
     =
     
      
       (
       
        
         T
         1
        
        -
        
         
          T
          0
         
         _
        
       
       )
      
      ·
      
       
        k
        ⁢
        V
       
       RN
      
     
    
   
   
    
     (
     IV
     ), wherein T1 is the value of the fluorescent intensity per unit mass or volume of the at least one test biological tissue sample of the group to be tested, T0 is an average value of the fluorescent intensity per unit mass or volume of the at least one control biological tissue sample of the blank control group, k is a ratio of a dilution ratio of the homogenate of the at least one test biological tissue sample to the diluted supernatants of the at least one test biological tissue sample to a mole amount of rare earth elements in calcium phosphate, V is a homogenate volume of the at least one test biological tissue sample in the group to be tested, and R is the tissue extraction rate R of the group to be tested.