Abstract:
The invention relates to a method for determining biochemical parameters of a body fluid, wherein a sample of said body fluid in the form of a droplet is transported through a channel of a microfluidic system using a carrier liquid, mixed with a reagent thus initiating a chemical reaction between the sample and the reagent, and the result of the chemical reaction is measured, preferably with a spectrophotometer, whereby the said biochemical parameters of the body fluid are determined, characterised in that the material used for fabrication of the microfluidic system and the said carrier liquid is pair of Teflon and Fluorinert HFE-7100.

Description:
[0001]    The invention relates to a method for determining a set of biochemical parameters in body fluids such as blood, blood serum or blood plasma. The invention comprises also the use of specially selected pairs of materials and liquids for determining biochemical parameters of a body fluid. 
         [0002]    The state of the art knows many diagnostic methods in this study referred to as biochemical methods aiming at quantitative measurement (determination) of biochemical parameters of blood. At present, known groups of such assays of biochemical parameters include: assays of substrates, enzymes, electrolytes, specific proteins, monitoring of concentrations of drugs and intoxicants, concentrations of hormones, cancer markers, cytokines and other types of proteins, as well as all other parameters that can be determined using photometric methods. These assays can be performed in various materials of human or animal (veterinary) origin, including: whole blood, serum, plasma, cerebrospinal fluid, urine or other fluids from body cavities. These tests are usually performed in analytical laboratories by laboratory diagnosticians or medical testing technicians (material taken from humans) or in veterinary practices by trained personnel (material from animals). More and more often, however, such testing is performed as a part of a point-of-care or individual diagnostics outside the analytical laboratory, in emergency stations, intensive care units, specialised ambulances, directly by physicians, paramedics, nurses or other trained personnel. 
         [0003]    For obvious reasons, the equipment used for individual or point-of-care assays should be easily portable, capable of providing test results within single minutes, and first of all such equipment must be reliable. In this context reliability shall mean the metrological reliability, i.e., the level of accuracy and reproducibility of the results obtained with a given instrument. 
         [0004]    The development of microfluidics has enabled fabrication of a series of small, portable constructions that might be applied in medical diagnostics, especially in the area of individual or point-of-care diagnostics [e.g., A. Arora, G. Simone, G. B. Salieb-Beugelaar, J. T. Kim, A. Manz,  Analytical Chemistry  82, 4830 (2010)]. In the underlying assumptions, these constructions are capable of making use of droplet flows [e.g., A. B. Theberge, F. Courtois, Y. Schaerli, M. Fischlechner, C. Abell, F. Hollfelder, W. T. S. Huck,  Angewandte Chemie International Edition,  49, 5846 (2010)], i.e., such ones where two immiscible phases, such as water and oil, are introduced into a microfluidic system. In analogy to emulsification processes, the phase used to form droplets is referred to as the dispersed phase, and the phase in which the droplets are suspended is referred to as the continuous (dispersing) phase. The droplets (or bubbles), surrounded by another fluid spontaneously assume a spherical shape; when squeezed by channel walls they assume a shape of flattened ellipsoids or discs. It has been shown already that such systems are suitable for determining both single biochemical parameters and sets of multiple biochemical parameters using a single device. 
         [0005]    It is a necessary prerequisite for formation of and control over a droplet flow that the dispersed phase does not wet the walls of the system (channels), as opposed to the continuous phase that must superbly wet the walls. Since biochemical assays use a very broad variety of reagents, a very important technical issue is to select the polymer used to fabricate the microfluidic system and the continuous liquid so, that no reagent, or at most possibly a few reagents only wet the channel walls in the presence of the continuous liquid. Likely, the assayed material (e.g., human or animal blood serum) must not wet the channel walls in the presence of the continuous liquid. 
         [0006]    The Authors of the present invention have tested a very broad set of reagents for biochemical blood testing and a broad range of polymers and continuous liquids, and found unexpectedly preferred combination of polymer and continuous liquid that allow performing biochemical assays inside droplets formed and residing in the microfluidic systems, i.e., in microchannels inside the microfluidic cartridges. 
         [0007]    In particular, the Authors of the present invention have confirmed that particularly preferred combinations were composed of polymer materials commonly used in industry: polypropylene, polyethylene and cyclic olefin copolymer (COC). These materials performed very well in a combination with oil—hexadecane. The droplets formed on the surfaces of these materials in the presence of hexadecane had a large contact angle, and the parameter is of key importance in forming droplets in a system. Unexpectedly, satisfactory results were obtained for the combination of Teflon and Fluorinert. 
         [0008]    According to the invention, the method for determining biochemical parameters of a body fluid, wherein a sample of said body fluid in the form of a droplet is transported through a channel of a microfluidic system using a carrier liquid, mixed with a reagent thus initiating a chemical reaction between the sample and the reagent, and the result of the chemical reaction is measured, preferably with a spectrophotometer, whereby the said biochemical parameters of the body fluid are determined, is characterised in that the material used for fabrication of the microfluidic system and the said carrier liquid is pair of Teflon and Fluorinert. 
         [0009]    Preferably, the said reagent is selected from the group comprising: acp (acid phosphatase), alat (alanine aminotransferase), albumin, alp (alkaline phosphatase), alpha-fetoprotein, alpha-1-microglobulin, amylase, asat (aspartate transaminase), aso (anti-streptolysin O), bil direct (direct bilirubin), bil total (total bilirubin), calcium, ceruloplasmin, cholesterol, cholinesterase, ck (creatine kinase), ck MB (creatine kinase MB), complement C3, complement C4, crp (C-reactive protein), cystatin C, D-dimer D, ethanol, phenobarbital, ferrum, ferritin, fibrinogen, ggt (gamma-glutamyltransferase), glucose, haptoglobin, hbdh (α-hydroxybutyrate dehydrogenase), hdl cholesterol, HbA1C (haemoglobin), immunoglobulin A, immunoglobulin E, immunoglobulin M, carbamazepine, creatinine, alpha-1-acid glycoprotein, ldh (lactate dehydrogenase), Idl cholesterol, lipase, lipoprotein, Mg (magnesium), copper, myoglobin, lactates, paracetamol, phosphorus, potassium, rf (rheumatoid factor), salicylates, sodium, theophylline, tg (triglycerides), total protein, ua (uric acid), uibc (unsaturated iron binding capacity), urea, urine protein. 
         [0010]    The invention comprises also the use of a pair of the material and the liquid—Teflon and Fluorinert—for determining biochemical parameters of a body fluid. 
     
    
     
       DETAILED DESCRIPTION OF THE INVENTION 
         [0011]      FIG. 1  shows a picture (a) and a schematic drawing (b) of a droplet observed in static position (after placing directly on the plate). The digits in part (b) of  FIG. 1  have the following meaning: 1-polymer substrate; 2-tangent line to the interface of the dispersed phase; 3-static contact angle; 4-continuous fluid surrounding the droplet; 5-reagent droplet. 
           [0012]      FIG. 2  shows a picture (a) and a schematic drawing (b) of a droplet observed in dynamic situation (observation of the contact angle while changing the inclination of the plate). The digits in part (b) of  FIG. 2  have the following meaning: 1-polymer substrate; 2-tangent line to the interface of the dispersed phase; 4-continuous fluid surrounding the droplet; 5-reagent droplet; 6-parallel line to the polymer substrate in the initial position; 7-angle of inclination of the polymer plate (minimum angle required for a droplet to flow); 8-dynamic contact angle. 
       
    
    
       [0013]    In a non-limiting embodiment, the static  3  and the dynamic  8  contact angles are determined, said angles formed by the serum  5  and the biochemical reagents  5  with the surface  1  of the polymer plate in the atmosphere of the selected continuous fluid  4 . In a non-limiting example, the following continuous fluids  4  were tested: hexadecane, silicon oil with a viscosity of 20 cSt, paraffin oil, mineral oil, Fluorinert FC 3283, Fluorinert FC 40, Fluorinert HFE 7100. The following polymer substrates  1  were used in tests: Dyneon, Teflon, polydimethylsiloxane (PDMS), polystyrene, polyethylene, polypropylene (two types, in the following referred to as PP and PPR), styrene—prop-2-enonitrile copolymer (SAN), polystyrene GPPS and polycarbonate, cyclic olefin copolymer (two types, in the following referred to as COC 5013 and COC 6015). 
         [0014]    The wetting of substrates  1  by reference normal (HN) and pathological (HP) serum  5  was tested. The HN/HP serum is produced on the basis of the human serum. It is used as a measurement control of concentrations of organic and inorganic components, and of the activity of enzymes. Most parameters tested in the HN serum are within the range of normal values for adults, whereas the parameters obtained for HP mostly differ from the values considered as normal. The dynamic contact angle was also studied for serums with various dilutions. The dilution was performed using physiological saline (0.9% sodium chloride). 
         [0015]    Table 1 shows a list of biochemical assays and the volume ratio of reagents and serum used in the reaction (markings: S-serum, R1-reagent 1, R2-reagent 2). Depending on the parameter being determined, single-reagent (R1) or dual-reagent (R1 and R2) reagents were used. For most items, the Table shows English reagent abbreviations with full names given in parentheses. 
         [0000]    
       
         
               
               
               
               
               
             
               
               
               
               
               
             
           
               
                 TABLE 1 
               
               
                   
               
               
                 Tested parameter 
                 Method type 
                 S [μl] 
                 R1 [μl] 
                 R2 [μl] 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                 acp (acid phosphatase) 
                 colorimetric 
                 25 
                 250 
                   
               
               
                 alat (alanine aminotransferase) 
                 kinetic 
                 100 
                 1000 
                 250 
               
               
                 albumin 
                 colorimetric 
                 10 
                 2000 
               
               
                 alp (alkaline phosphatase) 
                 colorimetric 
                 20 
                 1000 
                 250 
               
               
                 alpha-fetoprotein 
                 immunoturbidimetric 
                 18 
                 150 
                 75 
               
               
                 amylase 
                 kinetic 
                 5 
                 250 
               
               
                 asat (aspartate transaminase) 
                 kinetic 
                 100 
                 1000 
                 250 
               
               
                 aso (anti-streptolysin O) 
                 immunoturbidimetric 
                 6 
                 200 
                 140 
               
               
                 bil direct (direct bilirubin) 
                 colorimetric 
                 50 
                 800 
                 100 
               
               
                 bil total (total bilirubin) 
                 colorimetric 
                 10 
                 280 
                 70 
               
               
                 calcium 
                 colorimetric 
                 3 
                 300 
               
               
                 ceruloplasmin 
                 immunoturbidimetric 
                 5 
                 250 
                 50 
               
               
                 chol (cholesterol) 
                 colorimetric, enzymatic 
                 3 
                 250 
               
               
                 cholinesterase 
                 kinetic 
                 4 
                 200 
                 50 
               
               
                 ck (creatine kinase) 
                 kinetic 
                 40 
                 1000 
                 200 
               
               
                 ck MB (creatine kinase MB) 
                 kinetic, immunoinhibition 
                 20 
                 200 
                 50 
               
               
                 complement C3 
                 immunoturbidimetric 
                 4 
                 250 
                 50 
               
               
                 complement C4 
                 immunoturbidimetric 
                 7 
                 250 
                 50 
               
               
                 crp (C-reactive protein) 
                 immunoturbidimetric 
                 4 
                 200 
                 200 
               
               
                 cystatin C 
                 immunoturbidimetric 
                 3 
                 180 
                 30 
               
               
                 D-dimer D 
                 immunoturbidimetric 
                 5 
                 150 
                 50 
               
               
                 ethanol 
                 enzymatic 
                 3 
                 300 
               
               
                 phenobarbital 
                 immunoturbidimetric 
                 2 
                 280 
                 75 
               
               
                 ferrum 
                 colorimetric 
                 20 
                 200 
                 50 
               
               
                 ferritin 
                 immunoturbidimetric 
                 10 
                 100 
                 50 
               
               
                 ggt (gamma-glutamyltransferase) 
                 colorimetric 
                 100 
                 1000 
                 250 
               
               
                 glucose 
                 colorimetric, enzymatic 
                 10 
                 1000 
               
               
                 haptoglobin 
                 immunoturbidimetric 
                 2 
                 250 
                 50 
               
               
                 hbdh (α-hydroxybutyrate 
                 kinetic 
                 20 
                 1000 
                 250 
               
               
                 dehydrogenase) 
               
               
                 hdl cholesterol 
                 enzymatic 
                 10 
                 100 
                 50 
               
               
                 HbA 1C  (haemoglobin) 
                 immunoturbidimetric 
                 5 
                 210 
                 70 
               
               
                 IgA (immunoglobulin A) 
                 immunoturbidimetric 
                 3 
                 250 
                 50 
               
               
                 IgE (immunoglobulin E) 
                 immunoturbidimetric 
                 5 
                 200 
                 100 
               
               
                 IgM (immunoglobulin M) 
                 immunoturbidimetric 
                 3 
                 250 
                 50 
               
               
                 carbamazepine 
                 immunoturbidimetric 
                 2 
                 220 
                 60 
               
               
                 creatinine 
                 colorimetric; Jaffe 
                 100 
                 1000 
                 250 
               
               
                 enzymatic creatinine 
                 enzymatic, colorimetric 
                 30 
                 900 
                 300 
               
               
                 alpha-1-acid glycoprotein 
                 immunoturbidimetric 
                 3 
                 250 
                 50 
               
               
                 ldh (lactate dehydrogenase) 
                 kinetic 
                 20 
                 1000 
                 250 
               
               
                 ldl cholesterol 
                 enzymatic 
                 10 
                 100 
                 50 
               
               
                 lipase 
                 colorimetric 
                 5 
                 100 
                 50 
               
               
                 lipoprotein 
                 immunoturbidimetric 
                 6 
                 180 
                 90 
               
               
                 Mg (magnesium) 
                 colorimetric 
                 3 
                 250 
               
               
                 myoglobin 
                 immunoturbidimetric 
                 5 
                 150 
                 50 
               
               
                 lactates 
                 colorimetric 
                 3 
                 300 
               
               
                 phosphorus 
                 colorimetric 
                 3 
                 300 
               
               
                 rf (rheumatoid factor) 
                 immunoturbidimetric 
                 8 
                 240 
                 80 
               
               
                 theophylline 
                 immunoturbidimetric 
                 3 
                 300 
                 50 
               
               
                 tg (triglycerides) 
                 enzymatic, colorimetric 
                 10 
                 1000 
               
               
                 total protein 
                 colorimetric 
                 3 
                 300 
               
               
                 ua (uric acid) 
                 enzymatic, colorimetric 
                 20 
                 1000 
                 250 
               
               
                 uibc (unsaturated iron binding 
                 colorimetric 
                 15 
                 200 
                 50 
               
               
                 capacity) 
               
               
                 urea 
                 enzymatic, kinetic 
                 10 
                 1000 
                 250 
               
               
                 urine protein 
                 colorimetric 
                 12.5 
                 250 
               
               
                   
               
             
          
         
       
     
         [0016]    In the following part of the description we refer to the measurements of the static angle  3 —the angle formed between the polymer substrate  1  and the plane  2  tangent to the interface of the dispersed phase  5  and the continuous phase  4  under static conditions, on a horizontally placed substrate ( FIG. 1 ), and to the measurements of the dynamic angle  8 , that is the largest angle between the interface  2  of the dispersed phase  5  and the continuous phase  4 , and the plane  1  of the polymer substrate, above which the droplet flew down from the substrate. The dynamic angle  8  was measured by slow inclining the substrate  1 . The dynamic angle  8  and the angle of inclination  7  of the substrate, at which the droplet starts to flow down the substrate, were measured. The measurements were carried out on substrates entirely immersed in a tub filled with continuous liquid  4 . 
         [0017]    In the case of some materials (polypropylene, polyethylene), two plate types were used in tests. In the first case, the surface of the plate was roughened, i.e., the plate had a number of unevennesses (notches) on the surface. In this case, reagent droplets were dispensed perpendicularly to the unevennesses (notches) to check the effect of surface unevennesses on the dynamic contact angle. The second type of the plate surface was the even surface. In a non-limiting embodiment, even plates are fabricated by casting the polymer on a polished metal matrix (e.g., aluminium or steel one). In a non-limiting embodiment, manipulations with droplets on the polymer substrates mentioned in the present patent application can be made at room temperature. 
         [0018]    To assure that the results are clearer, hereinafter we use the following markings coding the minimum inclination of the substrate 1, for which the droplet started to flow:
       −− the droplet did not flow down the plate even for the maximum inclination (90°),   − the droplet flew down for the plate inclination in the range 40-70°,   + the droplet flew down for the plate inclination in the range 10-40°,   ++ the droplet flew down for the plate inclination in the range 0-10°,   +++ the droplet flows immediately after placing.       
 
         [0024]    Where the dynamic angle  8  is not given in the tables below, then it means that for a given inclination  7  the droplet flew so fast that taking a picture was very difficult. 
         [0025]    In a non-limiting embodiment, the following, not much preferable angles were measured for reagents deposited on a substrate made of PDMS in the atmosphere of a silicon oil with a viscosity of 20 cSt (HP—pathological serum; HN—normal serum; e.g., x2—it means that the serum was diluted twice;) 
         [0000]    
       
         
               
               
               
               
               
             
           
               
                   
                   
               
               
                   
                   
                 Static 
                 Dynamic 
                 Substrate 
               
               
                   
                 Reagent 
                 angle 
                 angle 
                 inclination 
               
               
                   
                   
               
             
             
               
                   
                 Water 
                 150.6 
                 154.6 
                 − 
               
               
                   
                 Physiological 
                 152.3 
                 157.2 
                 − 
               
               
                   
                 saline 
               
               
                   
                 HPx4 
                 139.4 
                 145.6 
                 −− 
               
               
                   
                 HPx16 
                 142.6 
                 148.3 
                 − 
               
               
                   
                   
               
             
          
         
       
     
         [0026]    In a non-limiting embodiment, the following contact angles were measured for reagents deposited on a substrate made of PDMS and surface-modified with Aquapel (waterproof silane-siloxane sealer) in the atmosphere of Fluorinert 3283 fluorinated oil. 
         [0000]    
       
         
               
               
               
             
               
               
               
             
           
               
                   
                   
               
               
                   
                   
                 static 
               
               
                   
                 Reagent 
                 angle 
               
               
                   
                   
               
             
             
               
                   
               
             
          
           
               
                   
                 HN 
                 136.6 
               
               
                   
                 HNx2 
                 127.8 
               
               
                   
                 HNx8 
                 123.6 
               
               
                   
                 HNx16 
                 119.9 
               
               
                   
                 HP 
                 121.1 
               
               
                   
                 HPx2 
                 121.3 
               
               
                   
                 HPx4 
                 122.2 
               
               
                   
                 HPx8 
                 119.1 
               
               
                   
                 HPx16 
                 121.9 
               
               
                   
                   
               
             
          
         
       
     
         [0027]    In a non-limiting embodiment, the following contact angles were measured for reagents deposited on a substrate made of polypropylene (PPR) in the atmosphere hexadecane oil. The PPR substrate was roughened. 
         [0000]    
       
         
               
               
               
               
             
               
               
               
               
             
           
               
                   
                   
               
               
                   
                 Static 
                 Dynamic 
                 Substrate 
               
               
                   
                 angle 
                 angle 
                 inclination 
               
               
                   
                   
               
             
             
               
                   
               
             
          
           
               
                 Reagent 
                   
                   
                   
               
               
                 HN 
                   
                 162.6 
                 +++ 
               
               
                 HNx4 
                 152.6 
                   
                 +++ 
               
               
                 HNx16 
                 149 
                 152.4 
                 +++ 
               
               
                 HP 
                   
                 152.5 
                 +++ 
               
               
                 HPx4 
                 146 
                 154.1 
                 +++ 
               
               
                 HNx4 
                   
                   
                 +++ 
               
               
                 HPx16 
                 153.4 
                 156.1 
                 +++ 
               
               
                 Water 
                   
                 159.6 
                 +++ 
               
               
                 Physiological saline 
                 154.6 
                 157.2 
                 ++ 
               
               
                 ACP 
                 134.2 
                 145.2 
                 −− 
               
               
                 Mixture ACP + HN 
                 102.6 
                 107.6 
                 −− 
               
               
                 Alat R1 
                   
                 156.7 
                 +++ 
               
               
                 Alat R2 
                   
                   
                 +++ 
               
               
                 Mixture Alat + HN 
                   
                   
                 +++ 
               
               
                 Albumin 
                   
                   
                 +++ 
               
               
                 Mixture albumin + HN 
                   
                 153.5 
                 +++ 
               
               
                 ALP R2 
                 146.7 
                 158.6 
                 ++ 
               
               
                 Mixture ALP + HN 
                 149 
                 150.5 
                 ++ 
               
               
                 Alpha-1-acid glycoprotein R1 
                 155.2 
                 149.8 
                 + 
               
               
                 Alpha-1-acid glycoprotein R2 
                 150.2 
                 145.5 
                 + 
               
               
                 Mixture alpha-1-acid 
                   
                   
                 +++ 
               
               
                 glycoprotein + calibrator 
               
               
                 Alpha-fetoprotein R1 
                 132.4 
                 147.7 
                 + 
               
               
                 Alpha-fetoprotein R2 
                 142 
                 144.6 
                 + 
               
               
                 Mixture alpha-fetoprotein + 
                   
                   
                 +++ 
               
               
                 HN 
               
               
                 Amylase R1 
                   
                 158.2 
                 +++ 
               
               
                 Mixture amylase + HN 
                   
                   
                 +++ 
               
               
                 ASAT R1 
                   
                   
                 +++ 
               
               
                 ASAT R2 
                   
                 154.3 
                 +++ 
               
               
                 ASO R1 
                   
                 150.5 
                 +++ 
               
               
                 ASO R2 
                 151.9 
                 144.9 
                 ++ 
               
               
                 Mixture ASO + HN 
                 147.6 
                 119.6 
                 +++ 
               
               
                 Bil direct R1 
                   
                   
                 +++ 
               
               
                 Bil direct R2 
                   
                   
                 +++ 
               
               
                 Mixture bil direct + HN 
                   
                   
                 +++ 
               
               
                 Bil total R1 
                   
                   
                 +++ 
               
               
                 Bil total R2 
                 152.5 
                 148.3 
                 + 
               
               
                 Mixture bil total + HN 
                   
                   
                 +++ 
               
               
                 Calcium R1 
                 126.8 
                 104.5 
                 + 
               
               
                 Mixture calcium + HN 
                 131.8 
                 116.8 
                 ++ 
               
               
                 Ceruloplasmin R1 
                 148.7 
                 150.8 
                 ++ 
               
               
                 Ceruloplasmin R2 
                 136.2 
                 150.5 
                 ++ 
               
               
                 Mixture ceruloplasmin + HN 
                 151.4 
                   
                 ++ 
               
               
                 Cholesterol R1 
                 123.3 
                 127.5 
                 ++ 
               
               
                 Mixture cholesterol R1 
                 128.6 
                 139.9 
                 +++ 
               
               
                 Cholinesterase R1 
                 147.2 
                 151.8 
                 − 
               
               
                 Cholinesterase R2 
                 150.7 
                 149.9 
                 −− 
               
               
                 Mixture cholinesterase + HN 
                 147 
                 145.3 
                 − 
               
               
                 ck R1 
                   
                   
                 +++ 
               
               
                 ck R2 
                   
                   
                 +++ 
               
               
                 Mixture ck + HN 
                   
                   
                 +++ 
               
               
                 ck mb R1 
                   
                   
                 +++ 
               
               
                 ck mb R2 
                   
                   
                 +++ 
               
               
                 Mixture ck mb + HN 
                   
                   
                 +++ 
               
               
                 Complement C3 R1 
                 152.3 
                 149.3 
                 − 
               
               
                 Complement C3 R2 
                 149 
                 144.6 
                 −− 
               
               
                 Mixture complement C3 + HN 
                 149 
                 145.3 
                 − 
               
               
                 Complement C4 R1 
                 152.2 
                 153.5 
                 + 
               
               
                 Complement C4 R2 
                   
                 150.5 
                 +++ 
               
               
                 Mixture complement C4 + 
                 149.1 
                 142.2 
                 + 
               
               
                 HN 
               
               
                 CRP R1 
                 151.4 
                 141.9 
                 ++ 
               
               
                 CRP R2 
                   
                   
                 +++ 
               
               
                 Mixture CRP + HN 
                   
                   
                 +++ 
               
               
                 Cystatin C R1 
                 125.4 
                   
                 ++ 
               
               
                 Cystatin C R2 
                 122.2 
                   
                 + 
               
               
                 Mixture cystatin C + HN 
                   
                 140.5 
                 +++ 
               
               
                 D-dimer 
                   
                   
                 +++ 
               
               
                 D-dimer R1 
                   
                   
                 +++ 
               
               
                 D-dimer R2 
                   
                 155.9 
                 +++ 
               
               
                 Mixture D-dimer + HN 
                   
                   
                 +++ 
               
               
                 Ethanol 
                   
                   
                 +++ 
               
               
                 Mixture ethanol + HN 
                   
                   
                 +++ 
               
               
                 Ferritin R1 
                   
                   
                 +++ 
               
               
                 Ferritin R2 
                   
                   
                 +++ 
               
               
                 Mixture ferritin + HN 
                   
                   
                 +++ 
               
               
                 Ferrum R1 
                   
                   
                 +++ 
               
               
                 Ferrum R2 
                   
                 154 
                 +++ 
               
               
                 Mixture ferrum + HN 
                 133.8 
                 136.9 
                 +++ 
               
               
                 GGT R1 
                   
                 154 
                 +++ 
               
               
                 GGT R2 
                 150.8 
                 142.5 
                 ++ 
               
               
                 Mixture GGT + HN 
                 147.4 
                 138.4 
                 ++ 
               
               
                 Glucose R1 
                   
                   
                 +++ 
               
               
                 Mixture glucose + HN 
                   
                   
                 +++ 
               
               
                 Haptoglobin R1 
                 148.1 
                 146.7 
                 −− 
               
               
                 Haptoglobin R2 
                 140.1 
                 143 
                 ++ 
               
               
                 Mixture haptoglobin + HN 
                 150.1 
                 143.2 
                 − 
               
               
                 HBDH R1 
                 149.3 
                 154.5 
                 + 
               
               
                 HBDH R2 
                 136.8 
                   
                 + 
               
               
                 Mixture HBDH + HN 
                   
                   
                 +++ 
               
               
                 Haemoglobin R1 
                 148.8 
                 147.6 
                 −− 
               
               
                 Haemoglobin R2 
                 122.5 
                   
                 +++ 
               
               
                 Haemoglobin R2a 
                   
                   
                 +++ 
               
               
                 Haemoglobin R2b 
                   
                   
                 +++ 
               
               
                 Mixture haemoglobin + HN 
                   
                   
                 +++ 
               
               
                 HDL R1 
                   
                 146.8 
                 +++ 
               
               
                 HDL R2 
                   
                   
                 +++ 
               
               
                 Mixture HDL + HN 
                   
                   
                 +++ 
               
               
                 Lipoprotein R1 
                 144.4 
                 143.8 
                 + 
               
               
                 Lipoprotein R2 
                 152.6 
                   
                 ++ 
               
               
                 Mixture lipoprotein + HN 
                   
                   
                 +++ 
               
               
                 Lipase R1 
                   
                 136.6 
                 +++ 
               
               
                 Lipase R2 
                   
                 143.5 
                 +++ 
               
               
                 Mixture lipase + HN 
                   
                   
                 +++ 
               
               
                 LDH R1 
                 156.3 
                   
                 + 
               
               
                 LDH R2 
                 148.6 
                 158.6 
                 + 
               
               
                 Mixture LDH + HN 
                   
                   
                 +++ 
               
               
                 LDL R1 
                   
                   
                 +++ 
               
               
                 LDL R2 
                 131.3 
                 133 
                 + 
               
               
                 Mixture LDL + HN 
                   
                 142.7 
                 +++ 
               
               
                 Mg R1 
                 121.2 
                 112.6 
                 ++ 
               
               
                 Mixture Mg + HN 
                 125.1 
                 118.2 
                 + 
               
               
                 Myoglobin R1 
                   
                   
                 +++ 
               
               
                 Myoglobin R2 
                   
                   
                 +++ 
               
               
                 Mixture myoglobin + HN 
                   
                   
                 +++ 
               
               
                 Phosphorus R1 
                 123.1 
                 128.3 
                 ++ 
               
               
                 Mixture phosphorus + HN 
                 128.3 
                 107.5 
                 ++ 
               
               
                 RF R1 
                   
                   
                 +++ 
               
               
                 RF R2 
                   
                 152.7 
                 +++ 
               
               
                 Mixture RF + HN 
                   
                   
                 +++ 
               
               
                 TG R1 
                 126.1 
                 130.5 
                 ++ 
               
               
                 TG R2 
                 134.8 
                 129.5 
                 + 
               
               
                 Mixture TG + HN 
                 126.1 
                 135.8 
                 ++ 
               
               
                 Total protein 
                 145.8 
                 152.2 
                 ++ 
               
               
                 Mixture total protein + HN 
                 152.5 
                   
                 ++ 
               
               
                 UA R1 
                 123.6 
                 125.8 
                 +++ 
               
               
                 UA R2 
                 126 
                 128.1 
                 ++ 
               
               
                 Mixture UA + HN 
                 124 
                 127.7 
                 +++ 
               
               
                 UIBC R1 
                 112.5 
                 126.5 
                 ++ 
               
               
                 UIBC R2 
                 148.6 
                 152.5 
                 ++ 
               
               
                 Mixture UIBC + HN 
                 127.2 
                 128.7 
                 ++ 
               
               
                 Urea R1 
                   
                   
                 +++ 
               
               
                 Urea R2 
                 158.3 
                 151.2 
                 + 
               
               
                 Mixture urea + HN 
                   
                   
                 +++ 
               
               
                 Urine protein 
                   
                   
                 +++ 
               
               
                 Mixture urine protein + 
                   
                   
                 +++ 
               
               
                 control 
               
               
                 Alpha-1-microglobulin R1 
                 136.2 
                 144.4 
                 + 
               
               
                 Alpha-1-microglobulin R2 
                 143.8 
                   
                 ++ 
               
               
                 Phenobarbital R1 
                   
                 159.0 
                 +++ 
               
               
                 Phenobarbital R2 
                 162.4 
                 152.5 
                 −− 
               
               
                 Mixture phenobarbital + HN 
                 152.6 
                 149.4 
                 ++ 
               
               
                 Fibrinogen R1 
                 115.3 
                 120.8 
                 ++ 
               
               
                 Fibrinogen R2 
                 148.6 
                 158.3 
                 ++ 
               
               
                 IgA R1 
                 147.6 
                 148.6 
                 ++ 
               
               
                 IgA R2 
                   
                   
                 +++ 
               
               
                 Mixture IgA + control 
                 153.6 
                 150.4 
                 − 
               
               
                 IgE R1 
                 155.3 
                 153.1 
                 − 
               
               
                 IgE R2 
                 154.1 
                 152.8 
                 + 
               
               
                 Mixture IgE + control 
                 149.2 
                 151.2 
                 ++ 
               
               
                 IgM R1 
                 151.5 
                 149.4 
                 − 
               
               
                 IgM R2 
                 154.1 
                 151.9 
                 −− 
               
               
                 Mixture IgM + control 
                   
                   
                 +++ 
               
               
                 Carbamazepine R1 
                 153.1 
                 149.0 
                 − 
               
               
                 Carbamazepine R2 
                 145.0 
                   
                 + 
               
               
                 Mixture carbamazepine + 
                 153.9 
                   
                 − 
               
               
                 control 
               
               
                 Reagents without 
               
               
                 surfactant 
               
               
                 Alat R1 
                 132.8 
                 149.1 
                 + 
               
               
                 Alat R2 
                 149.2 
                 142.3 
                 − 
               
               
                 Mixture Alat + HN 
                   
                   
                 +++ 
               
               
                 Albumin R1 
                 145 
                 147.8 
                 − 
               
               
                 Mixture albumin + HN 
                   
                   
                 +++ 
               
               
                 Ferrum R1 
                 155.8 
                 153 
                 − 
               
               
                 Ferrum R2 
                 152.5 
                 148.5 
                 −− 
               
               
                 Mixture ferrum + HN 
                   
                   
                 +++ 
               
               
                   
               
             
          
         
       
     
         [0028]    In a non-limiting embodiment, the following contact angles were measured for reagents deposited on an even substrate made of PPR in the atmosphere of hexadecane oil. 
         [0000]    
       
         
               
               
               
               
             
               
               
               
               
             
           
               
                   
               
               
                   
                 Static 
                 Dynamic 
                 Substrate 
               
               
                 Reagent 
                 angle 
                 angle 
                 inclination 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                 ACP 
                   
                   
                 +++ 
               
               
                 Mixture ACP + HN 
                   
                   
                 +++ 
               
               
                 ALP R2 
                   
                   
                 +++ 
               
               
                 Alpha-1-acid glycoprotein R1 
                 153.8 
                 148.1 
                 ++ 
               
               
                 Alpha-fetoprotein R1 
                 151.4 
                 147.2 
                 +++ 
               
               
                 Bil total R2 
                 156.6 
                 154.1 
                 ++ 
               
               
                 Cholinesterase R1 
                   
                   
                 +++ 
               
               
                 Mixture cholinesterase + HN 
                   
                   
                 +++ 
               
               
                 Complement C3 R1 
                 150.6 
                 146.5 
                 +++ 
               
               
                 Complement C3 R2 
                   
                   
                 +++ 
               
               
                 Mixture complement C3 + HN 
                   
                   
                 +++ 
               
               
                 Haptoglobin R1 
                   
                 152 
                 +++ 
               
               
                 Mixture haptoglobin + HN 
                   
                   
                 +++ 
               
               
                 Haemoglobin R1 
                   
                   
                 +++ 
               
               
                 HBDH R1 
                   
                   
                 +++ 
               
               
                 HBDH R2 
                   
                   
                 +++ 
               
               
                 Creatinine R1 
                   
                   
                 +++ 
               
               
                 Creatinine R2 
                   
                 153.6 
                 + 
               
               
                 Mixture creatinine + HN 
                   
                   
                 +++ 
               
               
                 LDH R2 
                   
                   
                 +++ 
               
               
                 Urea R2 
                   
                   
                 +++ 
               
               
                 Alpha-1-microglobulin R1 
                   
                   
                 +++ 
               
               
                 Creatinine enzymatic R1 
                   
                 151.3 
                 +++ 
               
               
                 Creatinine enzymatic R2 
                   
                 152.7 
                 +++ 
               
               
                 Mixture creatinine 
                   
                 153.5 
                 +++ 
               
               
                 enzymatic + HN 
               
               
                 Phenobarbital R2 
                   
                 160 
                 +++ 
               
               
                 Mixture IgA + control 
                   
                   
                 +++ 
               
               
                 IgE R1 
                   
                   
                 +++ 
               
               
                 IgM R1 
                   
                 156.6 
                 +++ 
               
               
                 IgM R2 
                   
                 154.1 
                 +++ 
               
               
                 Carbamazepine R1 
                   
                 161 
                 +++ 
               
               
                 Mixture carbamazepine + 
                   
                   
                 +++ 
               
               
                 control 
               
               
                 Copper R1 
                   
                 154.2 
                 +++ 
               
               
                 Paracetamol R1 
                   
                 149.2 
                 +++ 
               
               
                 Paracetamol R2 
                   
                   
                 +++ 
               
               
                 Potassium R1 
                   
                 155.9 
                 +++ 
               
               
                 Potassium R2 
                   
                   
                 +++ 
               
               
                 Salicylates R1 
                   
                   
                 +++ 
               
               
                 Salicylates R2 
                   
                   
                 +++ 
               
               
                 Sodium R1 
                   
                 153.9 
                 +++ 
               
               
                 Sodium R2 
                   
                   
                 +++ 
               
               
                 Alat R2 
                   
                   
                 +++ 
               
               
                 Albumin R1 
                   
                   
                 +++ 
               
               
                 Ferrum R1 
                   
                 158.4 
                 +++ 
               
               
                 Ferrum R2 
                 154.6 
                 154.2 
                 ++ 
               
               
                   
               
             
          
         
       
     
         [0029]    In a non-limiting embodiment the following contact angles were measured for reagents deposited on a substrate made of PPR in the atmosphere of mineral oil. 
         [0000]    
       
         
               
               
               
               
             
               
               
               
               
             
           
               
                   
                   
               
               
                   
                   
                 Dynamic 
                 Substrate 
               
               
                   
                 Reagent 
                 angle 
                 inclination 
               
               
                   
                   
               
             
             
               
                   
               
             
          
           
               
                   
                 Water 
                   
                 +++ 
               
               
                   
                 Physiological saline 
                 159.1 
                 − 
               
               
                   
                 HN 
                   
                 +++ 
               
               
                   
                 HNx4 
                 144.55 
                 ++ 
               
               
                   
                 HNx16 
                 163 
                 +++ 
               
               
                   
                 HP 
                 158.05 
                 +++ 
               
               
                   
                 HPx4 
                 161.55 
                 +++ 
               
               
                   
                 HPx16 
                 161.9 
                 +++ 
               
               
                   
                 Albumin R1 + HP 
                 156.8 
                 +++ 
               
               
                   
                 Albumin R1 
                 160.9 
                 +++ 
               
               
                   
                 Cholesterol R1 
                 153.6 
                 +++ 
               
               
                   
                   
               
             
          
         
       
     
         [0030]    In a non-limiting embodiment, the following contact angles were measured for reagents deposited on a substrate made of PPR in the atmosphere of paraffin oil. 
         [0000]    
       
         
               
               
               
               
             
               
               
               
               
             
           
               
                   
                   
               
               
                   
                   
                 Dynamic 
                 Substrate 
               
               
                   
                 Reagent 
                 angle 
                 inclination 
               
               
                   
                   
               
             
             
               
                   
               
             
          
           
               
                   
                 Water 
                   
                 +++ 
               
               
                   
                 Physiological saline 
                 149.4 
                 − 
               
               
                   
                 HN 
                 158.8 
                 ++ 
               
               
                   
                 HNx4 
                 156.7 
                 ++ 
               
               
                   
                 HNx16 
                 153 
                 ++ 
               
               
                   
                 HP 
                 156.7 
                 +++ 
               
               
                   
                 HPx4 
                 162 
                 +++ 
               
               
                   
                 HPx16 
                 163 
                 +++ 
               
               
                   
                 ALAT R1 
                 159.6 
                 +++ 
               
               
                   
                 Albumin R1 + HN 
                 158.2 
                 +++ 
               
               
                   
                 Albumin R1 
                 154.3 
                 ++ 
               
               
                   
                 Cholesterol R1 
                 136.3 
                 ++ 
               
               
                   
                   
               
             
          
         
       
     
         [0031]    In a non-limiting embodiment, the following contact angles were measured for reagents, for which the results of the tests on a PPR substrate in hexadecane oil were unfavourable, on a polypropylene (PP) substrate in the atmosphere of the same oil (hexadecane). 
         [0000]    
       
         
               
               
               
               
             
           
               
                   
                   
               
               
                   
                   
                 Dynamic 
                 Substrate 
               
               
                   
                 Reagent 
                 angle 
                 inclination 
               
               
                   
                   
               
             
             
               
                   
                 ACP 
                   
                 +++ 
               
               
                   
                 Mixture ACP + HN 
                   
                 +++ 
               
               
                   
                 ALP R2 
                   
                 +++ 
               
               
                   
                 LDH R2 
                   
                 +++ 
               
               
                   
                 Bil total R2 
                 152.8 
                 +++ 
               
               
                   
                 Cholinesterase R1 
                   
                 +++ 
               
               
                   
                 Mixture cholinesterases + HN 
                 145.4 
                 +++ 
               
               
                   
                 Complement C3 R1 
                   
                 +++ 
               
               
                   
                 Complement C3 R2 
                   
                 +++ 
               
               
                   
                 Mixture complement C3 + HN 
                   
                 +++ 
               
               
                   
                 Haptoglobin R1 
                   
                 +++ 
               
               
                   
                 Mixture haptoglobin + HN 
                   
                 +++ 
               
               
                   
                 HBDH R1 
                   
                 +++ 
               
               
                   
                 HBDH R2 
                   
                 +++ 
               
               
                   
                 Urea R2 
                   
                 ++ 
               
               
                   
                   
               
             
          
         
       
     
         [0032]    In a non-limiting embodiment, the following contact angles were measured for reagents deposited on a substrate made of PP in the atmosphere of paraffin oil. 
         [0000]    
       
         
               
               
               
               
             
           
               
                   
                   
               
               
                   
                   
                 Dynamic 
                 Substrate 
               
               
                   
                 Reagent 
                 angle 
                 inclination 
               
               
                   
                   
               
             
             
               
                   
                 Water 
                 151.5 
                 ++ 
               
               
                   
                 Physiological saline 
                 154.1 
                 + 
               
               
                   
                 HN 
                 163.7 
                 ++ 
               
               
                   
                 HNx4 
                 165.6 
                 +++ 
               
               
                   
                 HNx16 
                 160.7 
                 ++ 
               
               
                   
                 HP 
                 158.6 
                 ++ 
               
               
                   
                 HPx4 
                 158.3 
                 +++ 
               
               
                   
                 HPx16 
                 162.3 
                 +++ 
               
               
                   
                 ALAT R1 
                 164.7 
                 ++ 
               
               
                   
                 Cholesterol R1 
                 140.6 
                 ++ 
               
               
                   
                   
               
             
          
         
       
     
         [0033]    In a non-limiting embodiment, the following contact angles were measured for reagents deposited on a polyethylene substrate containing small unevennesses in the atmosphere of hexadecane oil. 
         [0000]    
       
         
               
               
               
               
             
           
               
                   
                   
               
               
                   
                   
                 Dynamic 
                 Substrate 
               
               
                   
                 Reagent 
                 angle 
                 inclination 
               
               
                   
                   
               
             
             
               
                   
                 Water 
                   
                 +++ 
               
               
                   
                 Physiological saline 
                   
                 ++ 
               
               
                   
                 HPx16 
                   
                 +++ 
               
               
                   
                 HPx4 
                   
                 +++ 
               
               
                   
                 HP 
                   
                 ++ 
               
               
                   
                 HNx16 
                   
                 +++ 
               
               
                   
                 HNx4 
                   
                 +++ 
               
               
                   
                 HN 
                   
                 ++ 
               
               
                   
                 Alat R1 
                   
                 ++ 
               
               
                   
                 Alat R2 
                   
                 ++ 
               
               
                   
                 Mixture Alat + HN 
                   
                 +++ 
               
               
                   
                 Mixture albumin + HN 
                 152.9 
                 ++ 
               
               
                   
                 ASO R1 
                   
                 +++ 
               
               
                   
                 ASO R2 
                   
                 +++ 
               
               
                   
                 Mixture ASO + HN 
                   
                 +++ 
               
               
                   
                 Cholesterol R1 
                 122.6 
                 ++ 
               
               
                   
                 CK R2 
                 142.7 
                 +++ 
               
               
                   
                 CRP R1 
                 131.4 
                 + 
               
               
                   
                 CRP R2 
                   
                 ++ 
               
               
                   
                 Mixture CRP + HN 
                 124.1 
                 ++ 
               
               
                   
                 Ethanol 
                   
                 ++ 
               
               
                   
                 Mixture ethanol + HN 
                   
                 +++ 
               
               
                   
                 Ferrum R1 
                   
                 +++ 
               
               
                   
                 Ferrum R2 
                   
                 ++ 
               
               
                   
                 Mixture ferrum + HN 
                   
                 +++ 
               
               
                   
                 TG R1 
                   
                 + 
               
               
                   
                 TG R2 
                   
                 + 
               
               
                   
                 Mixture TG + HN 
                   
                 + 
               
               
                   
                 Total protein 
                   
                 ++ 
               
               
                   
                 Mixture total protein + HN 
                   
                 ++ 
               
               
                   
                 UA R1 
                   
                 +++ 
               
               
                   
                 UA R2 
                   
                 +++ 
               
               
                   
                 Mixture UA + HN 
                   
                 ++ 
               
               
                   
                 UIBC R1 
                   
                 + 
               
               
                   
                 UIBC R2 
                   
                 + 
               
               
                   
                 Mixture UIBC + HN 
                   
                 + 
               
               
                   
                   
               
             
          
         
       
     
         [0034]    In a non-limiting embodiment, the following contact angles were measured for reagents deposited on roughened polyethylene substrate in the atmosphere of hexadecane oil. 
         [0000]    
       
         
               
               
               
               
             
               
               
               
               
             
           
               
                   
                   
               
               
                   
                   
                 Dynamic 
                 Substrate 
               
               
                   
                 Reagent 
                 angle 
                 inclination 
               
               
                   
                   
               
             
             
               
                   
               
             
          
           
               
                   
                 Alat R1 
                 160.8 
                 +++ 
               
               
                   
                 Alat R2 
                 160.5 
                 +++ 
               
               
                   
                 Mixture ALAT + HN 
                 157.4 
                 +++ 
               
               
                   
                 Albumin R1 
                 154.4 
                 +++ 
               
               
                   
                 Mixture albumin + HN 
                 159 
                 +++ 
               
               
                   
                 Ferrum R1 
                 149.2 
                 +++ 
               
               
                   
                 Ferrum R2 
                 156.7 
                 +++ 
               
               
                   
                 Mixture ferrum + HN 
                 158.7 
                 +++ 
               
               
                   
                 ACP 
                 151.3 
                 +++ 
               
               
                   
                 Mixture ACP + HN 
                 152.2 
                 +++ 
               
               
                   
                 Amylase 
                   
                 +++ 
               
               
                   
                 Mixture amylase + HN 
                   
                 +++ 
               
               
                   
                 Albumin R1 
                 149.6 
                 +++ 
               
               
                   
                 Mixture albumin + HN 
                   
                 +++ 
               
               
                   
                 ALP R1 
                   
                 +++ 
               
               
                   
                 ALP R2 
                 157.9 
                 ++ 
               
               
                   
                 Mixture ALP + HN 
                   
                 +++ 
               
               
                   
                 Alpha-1-acid glycoprotein R1 
                 157.1 
                 +++ 
               
               
                   
                 Alpha-1-acid glycoprotein R2 
                 150.8 
                 +++ 
               
               
                   
                 Mixture alpha-1-acid 
                 149.4 
                 +++ 
               
               
                   
                 glycoprotein + HN 
               
               
                   
                 Alpha-fetoprotein R1 
                 153.2 
                 +++ 
               
               
                   
                 Alpha-fetoprotein R2 
                 152 
                 +++ 
               
               
                   
                 Mixture alpha-fetoprotein + HN 
                 140 
                 +++ 
               
               
                   
                 ASAT R1 
                   
                 +++ 
               
               
                   
                 ASAT R2 
                 135.7 
                 +++ 
               
               
                   
                 Mixture ASAT + HN 
                 138 
                 +++ 
               
               
                   
                 Bil direct R1 
                 161 
                 +++ 
               
               
                   
                 Bil direct R2 
                 159.8 
                 +++ 
               
               
                   
                 Mixture bil direct + HN 
                 150.4 
                 +++ 
               
               
                   
                 Bil total R1 
                 142.4 
                 +++ 
               
               
                   
                 Bil total R2 
                 141.6 
                 +++ 
               
               
                   
                 Mixture bil total + HN 
                 137.6 
                 +++ 
               
               
                   
                 Calcium R1 
                 133.3 
                 ++ 
               
               
                   
                 Mixture calcium + HN 
                 127.2 
                 + 
               
               
                   
                 Ceruloplasmin R1 
                 158.8 
                 +++ 
               
               
                   
                 Ceruloplasmin R2 
                 156.1 
                 +++ 
               
               
                   
                 Mixture ceruloplasmin + HN 
                   
                 +++ 
               
               
                   
                 Cholesterol 
                   
                 +++ 
               
               
                   
                 Mixture cholesterol + HN 
                 130.3 
                 +++ 
               
               
                   
                 Cholinesterase R1 
                 145.95 
                 +++ 
               
               
                   
                 Cholinesterase R2 
                 157.8 
                 +++ 
               
               
                   
                 Mixture cholinesterase + HN 
                 156 
                 +++ 
               
               
                   
                 CK R1 
                 149.2 
                 +++ 
               
               
                   
                 CK R2 
                   
                 +++ 
               
               
                   
                 Mixture CK + HN 
                 144 
                 +++ 
               
               
                   
                 Complement C3 R1 
                 156.6 
                 +++ 
               
               
                   
                 Complement C3 R2 
                   
                 +++ 
               
               
                   
                 Mixture complement C3 + HN 
                   
                 +++ 
               
               
                   
                 Complement C4 R1 
                 156.1 
                 +++ 
               
               
                   
                 Complement C4 R2 
                 147.5 
                 +++ 
               
               
                   
                 Mixture complement C4 + HN 
                 159 
                 +++ 
               
               
                   
                 Cystatin C R1 
                 143.4 
                 +++ 
               
               
                   
                 Cystatin C R2 
                 140.4 
                 +++ 
               
               
                   
                 Mixture cystatin C + HN 
                 150.2 
                 +++ 
               
               
                   
                 D-dimer R1 
                 154 
                 +++ 
               
               
                   
                 D-dimer R2 
                   
                 +++ 
               
               
                   
                 Mixture D-dimer + HN 
                   
                 +++ 
               
               
                   
                 Ferritin R1 
                 127.7 
                 +++ 
               
               
                   
                 Ferritin R2 
                 155.5 
                 +++ 
               
               
                   
                 Mixture ferritin + HN 
                   
                 +++ 
               
               
                   
                 GGT R1 
                   
                 +++ 
               
               
                   
                 GGT R2 
                 138.6 
                 +++ 
               
               
                   
                 Mixture GGT + HN 
                 155.4 
                 +++ 
               
               
                   
                 Glucose R1 
                 144.4 
                 +++ 
               
               
                   
                 Mixture glucose + HN 
                   
                 +++ 
               
               
                   
                 Haptoglobin R1 
                 63.6 
                 +++ 
               
               
                   
                 Haptoglobin R2 
                 155 
                 +++ 
               
               
                   
                 Mixture haptoglobin + HN 
                   
                 +++ 
               
               
                   
                 Haemoglobin R1 
                 158 
                 +++ 
               
               
                   
                 Haemoglobin R2 
                 143.6 
                 +++ 
               
               
                   
                 Haemoglobin R2a 
                 144.6 
                 +++ 
               
               
                   
                 Haemoglobin R2b 
                 154.2 
                 +++ 
               
               
                   
                 Mixture haemoglobin + HN 
                 142.6 
                 +++ 
               
               
                   
                 HBDH R1 
                 157 
                 +++ 
               
               
                   
                 HBDH R2 
                 154 
                 +++ 
               
               
                   
                 Mixture HBDH + HN 
                 138.7 
                 +++ 
               
               
                   
                 HDL R1 
                 143.9 
                 +++ 
               
               
                   
                 HDL R2 
                 139.4 
                 +++ 
               
               
                   
                 Mixture HDL + HN 
                 149.6 
                 +++ 
               
               
                   
                 Creatinine R1 
                 156.6 
                 +++ 
               
               
                   
                 Creatinine R2 
                 158.6 
                 +++ 
               
               
                   
                 Mixture creatinine + HN 
                 156.4 
                 +++ 
               
               
                   
                 LDH R1 
                 158.8 
                 +++ 
               
               
                   
                 LDH R2 
                 150.4 
                 +++ 
               
               
                   
                 Mixture LDH + HN 
                 150.9 
                 +++ 
               
               
                   
                 LDL R1 
                 146.4 
                 +++ 
               
               
                   
                 LDL R2 
                 143 
                 +++ 
               
               
                   
                 Mixture LDL + HN 
                   
                 +++ 
               
               
                   
                 Lipase R1 
                 134.1 
                 +++ 
               
               
                   
                 Lipase R2 
                 141.6 
                 +++ 
               
               
                   
                 Mixture lipase + HN 
                 147.6 
                 +++ 
               
               
                   
                 Lipoprotein R1 
                 141.4 
                 +++ 
               
               
                   
                 Lipoprotein R2 
                 159 
                 +++ 
               
               
                   
                 Mixture lipoprotein + HN 
                   
                 +++ 
               
               
                   
                 CK MB R1 
                 157.9 
                 +++ 
               
               
                   
                 CK MB R2 
                 154 
                 +++ 
               
               
                   
                 Mixture CK-MB + HN 
                 151.4 
                 +++ 
               
               
                   
                 MG R1 
                 133.65 
                 +++ 
               
               
                   
                 Mixture Mg + HN 
                 135.1 
                 +++ 
               
               
                   
                 Myoglobin R1 
                 153.6 
                 +++ 
               
               
                   
                 Myoglobin R2 
                 156.9 
                 +++ 
               
               
                   
                 Mixture myoglobin + HN 
                 153.4 
                 +++ 
               
               
                   
                 Phosphorus 
                   
                 ++ 
               
               
                   
                 Mixture phosphorus + HN 
                 146 
                 +++ 
               
               
                   
                 RF R1 
                 152.6 
                 +++ 
               
               
                   
                 RF R2 
                 159 
                 +++ 
               
               
                   
                 Mixture RF + HN 
                 155.4 
                 +++ 
               
               
                   
                 Urine proteins 
                   
                 +++ 
               
               
                   
                 Mixture urine proteins + HN 
                 145.4 
                 +++ 
               
               
                   
                 UA R1 
                 139.85 
                 +++ 
               
               
                   
                 UA R2 
                 144.3 
                 +++ 
               
               
                   
                 Mixture UA + HN 
                   
                 +++ 
               
               
                   
                 Urea R1 
                 147.8 
                 +++ 
               
               
                   
                 Urea R2 
                   
                 +++ 
               
               
                   
                 Mixture urea + HN 
                 151.7 
                 +++ 
               
               
                   
                 Alpha-1-microglobulin R1 
                 165.9 
                 +++ 
               
               
                   
                 Alpha-1-microglobulin R2 
                 168.3 
                 +++ 
               
               
                   
                 Phenobarbital R1 
                   
                 ++ 
               
               
                   
                 Phenobarbital R2 
                   
                 ++ 
               
               
                   
                 Mixture phenobarbital + control 
                 159.7 
                 +++ 
               
               
                   
                 Fibrinogen R1 
                 152.3 
                 ++ 
               
               
                   
                 Fibrinogen R2 
                 161 
                 +++ 
               
               
                   
                 IgA R1 
                 163.3 
                 +++ 
               
               
                   
                 IgA R2 
                 160.8 
                 +++ 
               
               
                   
                 Mixture IgA + control 
                 157.9 
                 +++ 
               
               
                   
                 Ig E R1 
                   
                 +++ 
               
               
                   
                 Ig E R2 
                 161.7 
                 +++ 
               
               
                   
                 Mixture IgE + control 
                 162.5 
                 +++ 
               
               
                   
                 Ig M R1 
                   
                 +++ 
               
               
                   
                 IgM R2 
                 150.9 
                 ++ 
               
               
                   
                 Mixture IgM + control 
                 165.5 
                 +++ 
               
               
                   
                 Carbamazepine R1 
                 160.2 
                 +++ 
               
               
                   
                 Carbamazepine R2 
                 153.7 
                 +++ 
               
               
                   
                 Mixture carbamazapine + control 
                 161 
                 +++ 
               
               
                   
                 Creatinine enzymatic R1 
                 156 
                 +++ 
               
               
                   
                 Creatinine enzymatic R2 
                 160.8 
                 +++ 
               
               
                   
                 Mixture creatinine enzymatic + HN 
                 156.8 
                 +++ 
               
               
                   
                 Lactates 
                   
                 +++ 
               
               
                   
                 Mixture Lactates + HN 
                   
                 ++ 
               
               
                   
                 Theophylline R1 
                 166 
                 +++ 
               
               
                   
                 Theophylline R2 
                   
                 +++ 
               
               
                   
                 Mixture theophylline + control 
                 157.1 
                 +++ 
               
               
                   
                 UA without surfactant R1 
                 160.2 
                 +++ 
               
               
                   
                 UA without surfactant R2 
                   
                 ++ 
               
               
                   
                 Mixture UA without surfactant + HN 
                 159.4 
                 +++ 
               
               
                   
                   
               
             
          
         
       
     
         [0035]    In a non-limiting embodiment, the following contact angles were measured for reagents deposited on a substrate made of polyethylene in the atmosphere of mineral oil. 
         [0000]    
       
         
               
               
               
             
           
               
                   
                   
               
               
                   
                   
                 Substrate 
               
               
                   
                 Reagent 
                 inclination 
               
               
                   
                   
               
             
             
               
                   
                 Water 
                 +++ 
               
               
                   
                 Physiological saline 
                 ++ 
               
               
                   
                 HNx16 
                 +++ 
               
               
                   
                 HNx4 
                 ++ 
               
               
                   
                 HN 
                 +++ 
               
               
                   
                 HPx16 
                 ++ 
               
               
                   
                 HPx4 
                 ++ 
               
               
                   
                 HP 
                 ++ 
               
               
                   
                 HNx4 
                 ++ 
               
               
                   
                 HNx16 
                 ++ 
               
               
                   
                 Mixture albumin + HN 
                 ++ 
               
               
                   
                 Albumin 
                 ++ 
               
               
                   
                 Cholesterol 
                 ++ 
               
               
                   
                 Alat R1 
                 ++ 
               
               
                   
                   
               
             
          
         
       
     
         [0036]    In a non-limiting embodiment, the following contact angles were measured for reagents deposited on a substrate made of GPPS in the atmosphere of hexadecane oil. 
         [0000]    
       
         
               
               
               
               
               
             
               
               
               
               
               
             
           
               
                   
                   
               
               
                   
                   
                 Static 
                 Dynamic 
                 Substrate 
               
               
                   
                 Reagent 
                 angle 
                 angle 
                 inclination 
               
               
                   
                   
               
             
             
               
                   
               
             
          
           
               
                   
                 Water 
                 141.4 
                   
                 ++ 
               
               
                   
                 Physiological saline 
                 136.7 
                   
                 −− 
               
               
                   
                 HN 
                 121.2 
                 133.8 
                 − 
               
               
                   
                 HNx4 
                 138.6 
                 137.6 
                 + 
               
               
                   
                 HNx16 
                 149.5 
                 139.7 
                 − 
               
               
                   
                 HPx4 
                 86.2 
                 134.0 
                 − 
               
               
                   
                 HPx16 
                 113.4 
                 111.3 
                 + 
               
               
                   
                   
               
             
          
         
       
     
         [0037]    In a non-limiting embodiment, the following contact angles were measured for reagents deposited on a substrate made of GPPS polystyrene in the atmosphere of mineral oil. 
         [0000]    
       
         
               
               
               
               
               
             
               
               
               
               
               
             
           
               
                   
                   
               
               
                   
                   
                 Static 
                 Dynamic 
                 Substrate 
               
               
                   
                 Reagent 
                 angle 
                 angle 
                 inclination 
               
               
                   
                   
               
             
             
               
                   
               
             
          
           
               
                   
                 Water 
                 68.8 
                 104.2 
                 −− 
               
               
                   
                 Physiological saline 
                 106.2 
                 107.2 
                 − 
               
               
                   
                 HN 
                 52.6 
                 106.5 
                 −− 
               
               
                   
                 HNx4 
                 97.8 
                 100.5 
                 −− 
               
               
                   
                 HNx16 
                 121.4 
                 120.6 
                 + 
               
               
                   
                   
               
             
          
         
       
     
         [0038]    In a non-limiting embodiment, the following contact angles were measured for serum (HN—normal control serum, and HP—pathological control serum) and serum dilutions deposited on substrate made of dedecylamine-modified polycarbonate in the atmosphere of hexadecane oil. 
         [0000]    
       
         
               
               
               
             
               
               
               
             
           
               
                   
                   
               
               
                   
                   
                 Static 
               
               
                   
                 Reagent 
                 angle 
               
               
                   
                   
               
             
             
               
                   
               
             
          
           
               
                   
                 HN 
                 135.0 
               
               
                   
                 HNx2 
                 134.4 
               
               
                   
                 HNx4 
                 141.8 
               
               
                   
                 HNx8 
                 140.4 
               
               
                   
                 HNx16 
                 142.5 
               
               
                   
                 HP 
                 144.6 
               
               
                   
                 HPx2 
                 146.5 
               
               
                   
                 HPx4 
                 142.3 
               
               
                   
                 HPx8 
                 147.7 
               
               
                   
                 HPx16 
                 146.9 
               
               
                   
                   
               
             
          
         
       
     
         [0039]    In a non-limiting embodiment, the following contact angles were measured for reagents deposited on a substrate made of PS polystyrene in the atmosphere of hexadecane oil. 
         [0000]    
       
         
               
               
               
               
             
           
               
                   
               
               
                   
                 Static 
                 Dynamic 
                 Substrate 
               
               
                 Reagent 
                 angle 
                 angle 
                 inclination 
               
               
                   
               
             
             
               
                 Water 
                 131.6 
                 137.2 
                 −− 
               
               
                 Physiological saline 
                 148.4 
                 155.6 
                 + 
               
               
                 HN 
                   
                   
                 +++ 
               
               
                 HNx4 
                 142.2 
                 130.4 
                 ++ 
               
               
                 HNx16 
                 146.8 
                 139.8 
                 ++ 
               
               
                 HP 
                   
                   
                 +++ 
               
               
                 HPx4 
                   
                   
                 +++ 
               
               
                 HPx16 
                   
                   
                 +++ 
               
               
                 Mixture albumin + HN 
                   
                   
                 +++ 
               
               
                 Albumin (without surfactant) 
                   
                   
                 +++ 
               
               
                 Haptoglobin R1 
                 147.6 
                 142.7 
                 ++ 
               
               
                 Haemoglobin R2 
                 104.2 
                 105.4 
                 − 
               
               
                   
               
             
          
         
       
     
         [0040]    In a non-limiting embodiment, the following contact angles were measured for reagents deposited on a substrate made of SAN polymer in the atmosphere of mineral oil. 
         [0000]    
       
         
               
               
               
               
               
             
               
               
               
               
               
             
           
               
                   
                   
               
               
                   
                   
                 Static 
                 Dynamic 
                 Substrate 
               
               
                   
                 Reagent 
                 angle 
                 angle 
                 inclination 
               
               
                   
                   
               
             
             
               
                   
               
             
          
           
               
                   
                 Water 
                 135.4 
                 132.6 
                 − 
               
               
                   
                 Physiological saline 
                 136.7 
                 135.5 
                 − 
               
               
                   
                 HN 
                 154 
                 156.5 
                 ++ 
               
               
                   
                 HNx4 
                   
                   
                 +++ 
               
               
                   
                 HNx4 
                 95.8 
                 154 
                 − 
               
               
                   
                 HNx16 
                 70.5 
                   
                 − 
               
               
                   
                 HP 
                 113.2 
                 140.7 
                 − 
               
               
                   
                 HPx4 
                 142.3 
                 154.5 
                 ++ 
               
               
                   
                 HPx16 
                 112.6 
                 116.7 
                 ++ 
               
               
                   
                   
               
             
          
         
       
     
         [0041]    In a non-limiting embodiment, the following contact angles were measured for reagents deposited on a substrate made of SAN polymer in the atmosphere of paraffin oil. 
         [0000]    
       
         
               
               
               
               
               
             
           
               
                   
                   
               
               
                   
                   
                 Static 
                 Dynamic 
                 Substrate 
               
               
                   
                 Reagent 
                 angle 
                 angle 
                 inclination 
               
               
                   
                   
               
             
             
               
                   
                 Water 
                 128.8 
                 125.4 
                 −− 
               
               
                   
                 Physiological saline 
                 112.8 
                 121.8 
                 −− 
               
               
                   
                 HNx16 
                 155.1 
                 157.5 
                 − 
               
               
                   
                   
               
             
          
         
       
     
         [0042]    In a non-limiting embodiment, the following contact angles were measured for reagents deposited on a substrate made of Dyneon polymer in the atmosphere of Fluorinert FC-40 fluorinated oil. 
         [0000]    
       
         
               
               
               
               
               
             
               
               
               
               
               
             
           
               
                   
                   
               
               
                   
                   
                 Static 
                 Dynamic 
                 Substrate 
               
               
                   
                 Reagent 
                 angle 
                 angle 
                 inclination 
               
               
                   
                   
               
             
             
               
                   
               
             
          
           
               
                   
                 Water 
                 98.8 
                 110.4 
                 −− 
               
               
                   
                 Physiological saline 
                 102.6 
                 110.9 
                 − 
               
               
                   
                 HN 
                 122.4 
                 128.4 
                 − 
               
               
                   
                 HNx4 
                 138.1 
                 136.4 
                 − 
               
               
                   
                 HNx16 
                 132.6 
                 124.3 
                 − 
               
               
                   
                 HPx4 
                 128.2 
                 123.8 
                 − 
               
               
                   
                 HPx16 
                 110 
                 130.3 
                 − 
               
               
                   
                   
               
             
          
         
       
     
         [0043]    In a non-limiting embodiment, the following contact angles were measured for reagents deposited on a substrate made of Dyneon polymer in the atmosphere of Fluorinert FC 3283 fluorinated oil: 
         [0000]    
       
         
               
               
               
               
               
             
               
               
               
               
               
             
           
               
                   
                   
               
               
                   
                   
                 Static 
                 Dynamic 
                 Substrate 
               
               
                   
                 Reagent 
                 angle 
                 angle 
                 inclination 
               
               
                   
                   
               
             
             
               
                   
               
             
          
           
               
                   
                 Water 
                 143 
                 135.3 
                 + 
               
               
                   
                 Physiological saline 
                 139.7 
                 131.4 
                 − 
               
               
                   
                 HN 
                 147 
                 120 
                 −− 
               
               
                   
                 HNx4 
                 137.5 
                 133.5 
                 −− 
               
               
                   
                 HNx16 
                 143.3 
                 123.1 
                 − 
               
               
                   
                 HP 
                 110 
                 114 
                 −− 
               
               
                   
                 HPx2 
                 125 
                 133.4 
                 −− 
               
               
                   
                 HPx4 
                 136.5 
                 133.6 
                 − 
               
               
                   
                 HPx8 
                 113.7 
                 133.8 
                 −− 
               
               
                   
                 HNx16 
                 112.2 
                   
                 − 
               
               
                   
                   
               
             
          
         
       
     
         [0044]    In a non-limiting embodiment, the following contact angles were measured for reagents deposited on a substrate made of Dyneon polymer in the atmosphere Fluorinert FC-7100 fluorinated oil. 
         [0000]    
       
         
               
               
               
               
               
             
               
               
               
               
               
             
           
               
                   
                   
               
               
                   
                   
                 Static 
                 Dynamic 
                 Substrate 
               
               
                   
                 Reagent 
                 angle 
                 angle 
                 inclination 
               
               
                   
                   
               
             
             
               
                   
               
             
          
           
               
                   
                 Water 
                 75.1 
                 107.7 
                 −− 
               
               
                   
                 Physiological saline 
                 152.9 
                 148.4 
                 + 
               
               
                   
                 HN 
                 70.6 
                   
                 − 
               
               
                   
                 HNx4 
                 79.2 
                 80.7 
                 − 
               
               
                   
                 HNx16 
                 126.7 
                 112 
                 − 
               
               
                   
                   
               
             
          
         
       
     
         [0045]    In a non-limiting embodiment, the following contact angles were measured for reagents deposited on a substrate made of Teflon polymer in the atmosphere of Fluorinert THF 7100 fluorinated oil. 
         [0000]    
       
         
               
               
               
               
             
               
               
               
               
             
           
               
                   
               
             
             
               
                   
                 Static 
                 Dynamic 
                 Substrate 
               
               
                 Reagent 
                 angle 
                 angle 
                 inclination 
               
               
                   
               
               
                 Water 
                   
                   
                 +++ 
               
               
                 Physiological saline 
                 142 
                   
                 +++ 
               
               
                 HN 
                 151.7 
                 117.6 
                 +++ 
               
               
                 HNx4 
                 139.9 
                 133 
                 +++ 
               
               
                 HNx16 
                   
                   
                 +++ 
               
               
                 HP 
                   
                 151.7 
                 +++ 
               
               
                 HPx4 
                   
                   
                 +++ 
               
               
                 HPx16 
                   
                   
                 +++ 
               
               
                 ACP R1 
                   
                 154.9 
                 +++ 
               
               
                 Mixture ACP + HN 
                   
                   
                 +++ 
               
               
                 Alat R1 
                   
                 158.8 
                 +++ 
               
               
                 Alat R2 
                   
                   
                 +++ 
               
               
                 Mixture Alat + HN 
                   
                   
                 +++ 
               
               
                 Albumin R1 
                   
                 160.4 
                 +++ 
               
               
                 Mixture albumin + HN 
                   
                   
                 +++ 
               
               
                 ALP R1 
                   
                 152.2 
                 +++ 
               
               
                 ALP R2 
                   
                 159.1 
                 +++ 
               
               
                 Mixture ALP + HN 
                   
                 147.7 
                 +++ 
               
               
                 Alpha-1-acid glycoprotein R1 
                   
                   
                 +++ 
               
               
                 Alpha-1-acid glycoprotein R2 
                   
                   
                 +++ 
               
               
                 Mixture alpha-1-acid 
                   
                   
                 +++ 
               
               
                 glycoprotein + HN 
               
               
                 Alpha-fetoprotein R1 
                   
                   
                 +++ 
               
               
                 Alpha-fetoprotein R2 
                   
                   
                 +++ 
               
               
                 Mixture alpha-fetoprotein + 
                   
                   
                 +++ 
               
               
                 HN 
               
               
                 Amylase R1 
                   
                   
                 ++ 
               
               
                 Mixture amylase + HN 
                   
                   
                 +++ 
               
               
                 Asat R1 
                 151.5 
                 153.7 
                 +++ 
               
               
                 Asat R2 
                 157.3 
                 154.4 
                 + 
               
               
                 Mixture Asat + HN 
                 149 
                 139.9 
                 ++ 
               
               
                 ASO R1 
                   
                   
                 +++ 
               
               
                 ASO R2 
                   
                 160.6 
                 +++ 
               
               
                 Mixture ASO + HN 
                   
                   
                 +++ 
               
               
                 Bil total R1 
                   
                   
                 +++ 
               
               
                 Bil total R2 
                   
                 157.3 
                 +++ 
               
               
                 Mixture bil total + HN 
                   
                   
                 +++ 
               
               
                 Calcium R1 
                 135.6 
                 141.5 
                 ++ 
               
               
                 Mixture calcium + HN 
                   
                 144.9 
                 +++ 
               
               
                 Ceruloplasmin R1 
                   
                   
                 +++ 
               
               
                 Ceruloplasmin R2 
                   
                 160.5 
                 +++ 
               
               
                 Mixture ceruloplasmin + HN 
                   
                   
                 +++ 
               
               
                 Cholesterol R1 
                 142.9 
                 144.2 
                 ++ 
               
               
                 Mixture cholesterol + HN 
                 140.8 
                 144.6 
                 ++ 
               
               
                 Cholinesterase R1 
                   
                 146.9 
                 +++ 
               
               
                 Cholinesterase R2 
                   
                 159.8 
                 +++ 
               
               
                 Mixture cholinesterase + HN 
                   
                 153.4 
                 +++ 
               
               
                 CK R1 
                   
                 157.2 
                 +++ 
               
               
                 CK R2 
                   
                 152.7 
                 +++ 
               
               
                 Mixture CK + HN 
                   
                   
                 +++ 
               
               
                 ck-mb R1 
                   
                   
                 +++ 
               
               
                 ck-mb R2 
                   
                   
                 +++ 
               
               
                 Mixture ck-mb + HN 
                   
                   
                 +++ 
               
               
                 Complement C3 R1 
                   
                 156.4 
                 +++ 
               
               
                 Complement C3 R2 
                   
                 152.7 
                 ++ 
               
               
                 Mixture complement C3 + 
                   
                   
                 +++ 
               
               
                 HN 
               
               
                 Complement C4 R1 
                   
                 148 
                 +++ 
               
               
                 Complement C4 R2 
                   
                   
                 +++ 
               
               
                 Mixture complement C4 + 
                   
                   
                 +++ 
               
               
                 HN 
               
               
                 CRP R1 
                   
                   
                 +++ 
               
               
                 CRP R2 
                   
                 161.4 
                 +++ 
               
               
                 Mixture CRP + HN 
                   
                   
                 +++ 
               
               
                 Cystatin C R1 
                   
                   
                 +++ 
               
               
                 Cystatin C R2 
                   
                   
                 +++ 
               
               
                 Mixture cystatin C + HN 
                   
                   
                 +++ 
               
               
                 D-dimer R1 
                   
                 154.6 
                 +++ 
               
               
                 D-dimer R2 
                   
                 150.1 
                 +++ 
               
               
                 Mixture D-dimer + HN 
                   
                 148.5 
                 +++ 
               
               
                 Ethanol R1 
                   
                 158.1 
                 +++ 
               
               
                 Mixture ethanol + HN 
                   
                 150.5 
                 +++ 
               
               
                 Ferritin R1 
                   
                   
                 +++ 
               
               
                 Ferritin R2 
                   
                 157.3 
                 +++ 
               
               
                 Mixture ferritin + HN 
                   
                   
                 +++ 
               
               
                 Ferrum R1 
                   
                 136.7 
                 +++ 
               
               
                 Ferrum R2 
                   
                 147.4 
                 +++ 
               
               
                 Mixture ferrum + HN 
                   
                 140.6 
                 +++ 
               
               
                 GGT R1 
                   
                   
                 +++ 
               
               
                 GGT R2 
                   
                   
                 +++ 
               
               
                 Mixture GGT + HN 
                   
                   
                 +++ 
               
               
                 Glucose R1 
                   
                 149.4 
                 +++ 
               
               
                 Mixture glucose + HN 
                   
                 148.4 
                 +++ 
               
               
                 Haptoglobin R1 
                   
                   
                 +++ 
               
               
                 Haptoglobin R2 
                   
                   
                 +++ 
               
               
                 Mixture haptoglobin + HN 
                   
                   
                 +++ 
               
               
                 Haemoglobin R1 
                   
                 157.4 
                 +++ 
               
               
                 Haemoglobin R2 
                   
                 155.7 
                 +++ 
               
               
                 Haemoglobin R2a 
                   
                   
                 +++ 
               
               
                 Haemoglobin R2b 
                   
                   
                 +++ 
               
               
                 Mixture haemoglobin + HN 
                   
                   
                 +++ 
               
               
                 HBDH R1 
                   
                 167.9 
                 +++ 
               
               
                 HBDH R2 
                   
                 161.3 
                 +++ 
               
               
                 Mixture HBDH + HN 
                   
                 145.7 
                 +++ 
               
               
                 HDL R1 
                   
                 155.6 
                 +++ 
               
               
                 HDL R2 
                   
                 160.1 
                 +++ 
               
               
                 Mixture HDL + HN 
                   
                 152.2 
                 +++ 
               
               
                 Creatinine R1 
                   
                   
                 +++ 
               
               
                 Creatinine R2 
                   
                   
                 +++ 
               
               
                 Mixture creatinine + HN 
                   
                 162.7 
                 +++ 
               
               
                 LDH R1 
                   
                   
                 +++ 
               
               
                 LDH R2 
                 145.2 
                 161.8 
                 ++ 
               
               
                 Mixture LDH + HN 
                   
                 157.6 
                 +++ 
               
               
                 LDL R1 
                   
                 153.9 
                 +++ 
               
               
                 LDL R2 
                   
                 139.2 
                 +++ 
               
               
                 Mixture LDL + HN 
                   
                 142.3 
                 +++ 
               
               
                 Lipase R1 
                   
                 156.1 
                 +++ 
               
               
                 Lipase R2 
                   
                 151.6 
                 +++ 
               
               
                 Mixture lipase + HN 
                   
                 148.7 
                 +++ 
               
               
                 Lipoprotein R1 
                   
                   
                 +++ 
               
               
                 Lipoprotein R2 
                   
                   
                 +++ 
               
               
                 Mixture lipoprotein + HN 
                   
                   
                 +++ 
               
               
                 MG R1 
                 143.2 
                 138.8 
                 +++ 
               
               
                 Mixture Mg + HN 
                   
                 144.1 
                 ++ 
               
               
                 Myoglobin R1 
                   
                   
                 +++ 
               
               
                 Myoglobin R2 
                   
                   
                 +++ 
               
               
                 Mixture myoglobin + HN 
                   
                   
                 +++ 
               
               
                 Phosphorus R1 
                   
                 143.1 
                 +++ 
               
               
                 Mixture phosphorus + HN 
                   
                 143 
                 +++ 
               
               
                 RF R1 
                   
                   
                 +++ 
               
               
                 RF R2 
                   
                   
                 +++ 
               
               
                 Mixture RF + HN 
                   
                   
                 +++ 
               
               
                 TG R1 
                 126.8 
                 138 
                 ++ 
               
               
                 TG R2 
                 141.5 
                 111.8 
                 ++ 
               
               
                 Mixture TG + HN 
                 130.8 
                 129.4 
                 ++ 
               
               
                 Total protein R1 
                   
                   
                 +++ 
               
               
                 Mixture total protein + HN 
                   
                   
                 +++ 
               
               
                 UA R1 
                 140.8 
                 142.6 
                 ++ 
               
               
                 UA R2 
                 142.9 
                 143.8 
                 ++ 
               
               
                 Mixture UA + HN 
                 139.3 
                 137.1 
                 ++ 
               
               
                 UIBC R1 
                 128.5 
                 138.4 
                 ++ 
               
               
                 UIBC R2 
                   
                 160.5 
                 +++ 
               
               
                 Mixture UIBC + HN 
                   
                 137.9 
                 +++ 
               
               
                 Urea R1 
                   
                 143.8 
                 +++ 
               
               
                 Urea R2 
                   
                 158.6 
                 +++ 
               
               
                 Mixture urea + HN 
                   
                 155.5 
                 +++ 
               
               
                 Urine proteins R1 
                 139.2 
                 141.9 
                 ++ 
               
               
                 Mixture urine proteins + HN 
                 145.3 
                 141.9 
                 +++ 
               
               
                   
               
             
          
           
               
                   
                 Reagents without 
                 Dynamic 
                 Substrate 
               
               
                   
                 surfactant 
                 angle 
                 inclination 
               
               
                   
                   
               
               
                   
                 Alat R1 
                 166.8 
                 +++ 
               
               
                   
                 Alat R2 
                   
                 +++ 
               
               
                   
                 Mixture Alat + HN 
                 160 
                 +++ 
               
               
                   
                 Albumin R1 
                   
                 +++ 
               
               
                   
                 Mixture albumin + HN 
                 157.9 
                 +++ 
               
               
                   
                 Ferrum R1 
                   
                 +++ 
               
               
                   
                 Ferrum R2 
                 148 
                 +++ 
               
               
                   
                 Mixture ferrum + HN 
                 146.4 
                 +++ 
               
               
                   
                   
               
             
          
         
       
     
         [0046]    In a non-limiting embodiment, the following contact angles were measured for reagents deposited on a substrate made of Teflon polymer in the atmosphere of Fluorinert FC-3283 fluorinated oil. 
         [0000]    
       
         
               
               
               
               
               
             
               
               
               
               
               
             
           
               
                   
                   
               
               
                   
                   
                 Static 
                 Dynamic 
                 Substrate 
               
               
                   
                 Reagents 
                 angle 
                 angle 
                 inclination 
               
               
                   
                   
               
             
             
               
                   
               
             
          
           
               
                   
                 Water 
                 136.9 
                 138.4 
                 ++ 
               
               
                   
                 Physiological saline 
                 134.5 
                 136.5 
                 ++ 
               
               
                   
                 HN 
                 146 
                 121.5 
                 + 
               
               
                   
                 HNx4 
                 143.4 
                 138.4 
                 − 
               
               
                   
                 HNx8 
                 130.6 
                 113.4 
                 + 
               
               
                   
                 HNx16 
                 134.5 
                 136.7 
                 − 
               
               
                   
                 HP 
                 134.5 
                 136 
                 + 
               
               
                   
                 HPx2 
                 126.6 
                 123.4 
                 + 
               
               
                   
                 HPx4 
                 140.7 
                 137.8 
                 + 
               
               
                   
                 HPx8 
                 146.9 
                 142.4 
                 + 
               
               
                   
                 HPx16 
                 142.5 
                 129.9 
                 + 
               
               
                   
                   
               
             
          
         
       
     
         [0047]    In a non-limiting embodiment, the following contact angles were measured for reagents deposited on a substrate made of cyclic olefin copolymer (COC) 5013 in the atmosphere of hexadecane oil. 
         [0000]    
       
         
               
               
               
               
               
             
               
               
               
               
               
             
           
               
                   
                   
               
               
                   
                   
                 Static 
                 Dynamic 
                 Substrate 
               
               
                   
                 Reagent 
                 angle 
                 angle 
                 inclination 
               
               
                   
                   
               
             
             
               
                   
               
             
          
           
               
                   
                 Alat R1 
                 163.3 
                 161.7 
                 +++ 
               
               
                   
                 Alat R2 
                 164.3 
                 164.3 
                 +++ 
               
               
                   
                 Albumina R1 
                 156.0 
                 154.2 
                 ++ 
               
               
                   
                 α-fetoprotein R1 
                 162.5 
                 160.7 
                 +++ 
               
               
                   
                 α-fetoprotein R2 
                 158.6 
                 141.0 
                 −− 
               
               
                   
                 Alp R1 
                 163.9 
                 163.9 
                 +++ 
               
               
                   
                 Alp R2 
                 163.7 
                 163.7 
                 +++ 
               
               
                   
                 Amylase R1 
                 162.7 
                 163.5 
                 +++ 
               
               
                   
                 Asat R1 
                 162.4 
                 162.2 
                 +++ 
               
               
                   
                 Asat R2 
                 164.3 
                 164.3 
                 +++ 
               
               
                   
                 Aso R1 
                 159.8 
                 159.8 
                 +++ 
               
               
                   
                 Aso R2 
                 162.1 
                 162.1 
                 +++ 
               
               
                   
                 Bil direct R1 
                 150.8 
                 150.8 
                 + 
               
               
                   
                 Bil direct R2 
                 153.5 
                 153.5 
                 +++ 
               
               
                   
                 Cholesterol R1 
                 129.4 
                 127.8 
                 +++ 
               
               
                   
                 CK R1 
                 152.1 
                 152.1 
                 +++ 
               
               
                   
                 CK R2 
                 143.7 
                 143.7 
                 +++ 
               
               
                   
                 CK MB R1 
                 147.9 
                   
                 +++ 
               
               
                   
                 CK MB R2 
                 146.1 
                 146.1 
                 +++ 
               
               
                   
                 CRP R1 
                 159.4 
                 143.6 
                 − 
               
               
                   
                 CRP R2 
                 153.0 
                 150.5 
                 ++ 
               
               
                   
                 Cystatin R1 
                 158.5 
                 158.5 
                 +++ 
               
               
                   
                 Cystatin R2 
                 143.4 
                 143.4 
                 +++ 
               
               
                   
                 D-dimer Diluent 
                 145.6 
                   
                 −− 
               
               
                   
                 D-dimer R1 
                 161.1 
                 162.3 
                 + 
               
               
                   
                 D-dimer R2 
                 162.4 
                   
                 −− 
               
               
                   
                 Ethanol R1 
                 161.5 
                 161.5 
                 +++ 
               
               
                   
                 Ferrum R1 
                 163.4 
                 163.4 
                 +++ 
               
               
                   
                 Ferrum R2 
                 160.2 
                 160.2 
                 +++ 
               
               
                   
                 Ferritin R1 
                 158.4 
                 157.2 
                 +++ 
               
               
                   
                 Ferritin R2 
                 158.6 
                 158.6 
                 +++ 
               
               
                   
                 GGT R1 
                 162.3 
                 162.3 
                 +++ 
               
               
                   
                 GGT R2 
                 160.3 
                 160.3 
                 +++ 
               
               
                   
                 Glucose R1 
                 152.9 
                 151.6 
                 + 
               
               
                   
                 HBDH R1 
                 163.5 
                 163.5 
                 +++ 
               
               
                   
                 HBDH R2 
                 162.4 
                 126.7 
                 + 
               
               
                   
                 HDL R1 
                 162.0 
                 162.2 
                 +++ 
               
               
                   
                 HDL R2 
                 157.3 
                 157.3 
                 +++ 
               
               
                   
                 HbA1c R1 
                 163.7 
                 163.7 
                 +++ 
               
               
                   
                 HbA1c R2a 
                 149.3 
                 149.3 
                 +++ 
               
               
                   
                 HbA1c R2b 
                 151.2 
                 156.5 
                 +++ 
               
               
                   
                 Creatynine R1 
                 147.5 
                 147.5 
                 +++ 
               
               
                   
                 Creatynine R2 
                 158.4 
                 158.4 
                 +++ 
               
               
                   
                 LDH R1 
                 162.7 
                 162.7 
                 +++ 
               
               
                   
                 LDH R2 
                 161.3 
                 161.3 
                 +++ 
               
               
                   
                 LDL R1 
                 156.8 
                 156.8 
                 +++ 
               
               
                   
                 LDL R2 
                 134.6 
                 131.7 
                 + 
               
               
                   
                 Lipase R1 
                 151.2 
                 151.2 
                 +++ 
               
               
                   
                 Lipase R2 
                 147.2 
                 147.2 
                 +++ 
               
               
                   
                 Lipoprotein R1 
                 137.0 
                 137.0 
                 + 
               
               
                   
                 Lipoprotein R2 
                 158.0 
                 142.1 
                 + 
               
               
                   
                 Mioglobin R1 
                 161.8 
                 161.8 
                 +++ 
               
               
                   
                 Mioglobin R2 
                 160.3 
                 160.3 
                 +++ 
               
               
                   
                 RF R1 
                 156.6 
                 156.6 
                 +++ 
               
               
                   
                 RF R2 
                 150.1 
                 150.1 
                 +++ 
               
               
                   
                 TG R1 
                 136.7 
                 136.7 
                 +++ 
               
               
                   
                 TG R2 
                 142.5 
                 142.5 
                 +++ 
               
               
                   
                 Total protein R1 
                 163.5 
                 156.8 
                 +++ 
               
               
                   
                 UA R1 
                 140.6 
                 138.9 
                 +++ 
               
               
                   
                 UA R2 
                 134.0 
                 134.0 
                 + 
               
               
                   
                 UIBC R1 
                 138.4 
                   
                 + 
               
               
                   
                 UIBC R2 
                 149.9 
                 151.1 
                 + 
               
               
                   
                 Urea R1 
                 155.7 
                 159.6 
                 +++ 
               
               
                   
                 Urea R2 
                 161.8 
                   
                 + 
               
               
                   
                 Urine protein R1 
                 151.8 
                 151.8 
                 +++ 
               
               
                   
                 ACP 
                 148.6 
                 148.6 
                 + 
               
               
                   
                 Calcium R1 
                 145.5 
                   
                 + 
               
               
                   
                 Ceruloplasmin R1 
                 157.5 
                 157.5 
                 +++ 
               
               
                   
                 Ceruloplasmin R2 
                 154.4 
                 154.4 
                 +++ 
               
               
                   
                 Cholinesterase R1 
                 158.9 
                 158.9 
                 +++ 
               
               
                   
                 Cholinesterase R2 
                 157.7 
                 157.7 
                 +++ 
               
               
                   
                 Complement C3 R1 
                 158.0 
                 158.0 
                 +++ 
               
               
                   
                 Complement C3 R2 
                 157.5 
                 159.4 
                 +++ 
               
               
                   
                 Complement C4 R1 
                 156.2 
                 156.2 
                 +++ 
               
               
                   
                 Complement C4 R2 
                 154.6 
                 154.6 
                 +++ 
               
               
                   
                 HN 
                 157.6 
                 157.6 
                 +++ 
               
               
                   
                 Water 
                 155.2 
                 155.2 
                 +++ 
               
               
                   
                   
               
             
          
         
       
     
         [0048]    In a non-limiting embodiment, the following contact angles were measured for reagents deposited on a substrate made of cyclic olefin copolymer 6015 in the atmosphere of hexadecane oil. 
         [0000]    
       
         
               
               
               
               
               
             
               
               
               
               
               
             
           
               
                   
                   
               
               
                   
                   
                 Static 
                 Dynamic 
                 Substrate 
               
               
                   
                 Reagent 
                 angle 
                 angle 
                 inclination 
               
               
                   
                   
               
             
             
               
                   
               
             
          
           
               
                   
                 Alat R1 
                 162.7 
                 164.1 
                 + 
               
               
                   
                 Alat R2 
                 162.3 
                 147.0 
                 + 
               
               
                   
                 Albumin R1 
                 160.2 
                 134.6 
                 + 
               
               
                   
                 α-fetoprotein R1 
                 155.6 
                 149.8 
                 −− 
               
               
                   
                 α-fetoprotein R2 
                 159.2 
                 159.2 
                 +++ 
               
               
                   
                 Alp R1 
                 151.3 
                 148.4 
                 −− 
               
               
                   
                 Alp R2 
                 152.2 
                 152.2 
                 + 
               
               
                   
                 Asat R1 
                 160.8 
                   
                 −− 
               
               
                   
                 Asat R2 
                 159.9 
                 162.4 
                 − 
               
               
                   
                 Aso R1 
                 163.5 
                 131.7 
                 − 
               
               
                   
                 Aso R2 
                 157.9 
                 157.9 
                 + 
               
               
                   
                 Bil direct R1 
                 146.6 
                 146.6 
                 +++ 
               
               
                   
                 Bil direct R2 
                 152.1 
                 152.1 
                 +++ 
               
               
                   
                 Cholesterol R1 
                 126.8 
                 141.5 
                 +++ 
               
               
                   
                 CK R1 
                 140.7 
                 147.2 
                 + 
               
               
                   
                 CK R2 
                 143.6 
                 143.6 
                 +++ 
               
               
                   
                 CK MB R1 
                 139.9 
                 139.9 
                 +++ 
               
               
                   
                 CK MB R2 
                   
                 130.8 
                 +++ 
               
               
                   
                 D-dimer Diluent 
                 155.7 
                   
                 −− 
               
               
                   
                 D-dimer R1 
                 153.8 
                 148.3 
                 −− 
               
               
                   
                 D-dimer R2 
                 135.5 
                   
                 −− 
               
               
                   
                 Ethanol R1 
                 162.1 
                 162.1 
                 +++ 
               
               
                   
                 Ferrum R1 
                 134.7 
                 134.7 
                 +++ 
               
               
                   
                 Ferrum R2 
                 156.3 
                 157.4 
                 + 
               
               
                   
                 Ferritin R1 
                 158.8 
                 152.9 
                 + 
               
               
                   
                 Ferritin R2 
                 157.8 
                 157.8 
                 +++ 
               
               
                   
                 GGT R1 
                 164.0 
                 164.0 
                 −− 
               
               
                   
                 GGT R2 
                 160.1 
                 146.7 
                 +++ 
               
               
                   
                 Glucose R1 
                 162.6 
                 162.6 
                 +++ 
               
               
                   
                 HBDH R1 
                 146.4 
                 140.7 
                 −− 
               
               
                   
                 HBDH R2 
                 148.3 
                 137.1 
                 −− 
               
               
                   
                 HDL R1 
                 160.4 
                 160.4 
                 +++ 
               
               
                   
                 HDL R2 
                 152.6 
                 152.6 
                 +++ 
               
               
                   
                 HbA1c Hemolysing 
                 150.7 
                 139.7 
                 +++ 
               
               
                   
                 HbA1c R2a 
                 151.2 
                 138.8 
                 ++ 
               
               
                   
                 HbA1c R2b 
                 130.6 
                 132.0 
                 +++ 
               
               
                   
                 Creatynine R1 
                 158.9 
                 158.9 
                 +++ 
               
               
                   
                 Creatynine R2 
                 158.0 
                 158.0 
                 +++ 
               
               
                   
                 LDL R1 
                 128.2 
                 129.2 
                 + 
               
               
                   
                 LDL R2 
                 99.4 
                 99.4 
                 +++ 
               
               
                   
                 Lipase R1 
                 120.0 
                 120.0 
                 +++ 
               
               
                   
                 Lipase R2 
                 136.0 
                 131.6 
                 ++ 
               
               
                   
                 RF R1 
                 141.4 
                 134.7 
                 +++ 
               
               
                   
                 RF R2 
                 160.9 
                 160.9 
                 +++ 
               
               
                   
                 HN 
                 154.9 
                 157.0 
                 +++ 
               
               
                   
                 TG mono 
                 126.4 
                 128.7 
                 ++ 
               
               
                   
                 Water 
                 160.5 
                 160.5 
                 +++ 
               
               
                   
                   
               
             
          
         
       
     
         [0049]    Considering the conditions mentioned in the introduction, based on the measurements of contact angles, the Authors of the present invention have unexpectedly discovered that the most preferred combination of polymer and continuous liquid for performing biochemical assays in droplets manipulated inside microfluidic cartridges is Teflon and Fluorinert HFE-7100. 
         [0050]    The Authors of the present invention have unexpectedly discovered that the combination of Teflon and Fluorinert HFE-7100 enable controlled formation of droplets of biochemical reagents (Table 2) and manipulating these droplets inside microchannels or microchambers in the microfluidic cartridges. 
         [0000]    
       
         
               
               
               
             
           
               
                   
                 TABLE 2 
               
               
                   
                   
               
               
                   
                 Tested parameter 
                 Method type 
               
               
                   
                   
               
             
             
               
                   
                 acp (acid phosphatase) 
                 colorimetric 
               
               
                   
                 alat (alanine aminotransferase) 
                 kinetic 
               
               
                   
                 albumin 
                 colorimetric 
               
               
                   
                 alp (alkaline phosphatase) 
                 colorimetric 
               
               
                   
                 alpha-fetoprotein 
                 immunoturbidimetric 
               
               
                   
                 alpha-1-microglobulin 
                 immunoturbidimetric 
               
               
                   
                 amylase 
                 kinetic 
               
               
                   
                 asat (aspartate transaminase) 
                 kinetic 
               
               
                   
                 aso (anti-streptolysin O) 
                 immunoturbidimetric 
               
               
                   
                 bil direct (direct bilirubin) 
                 colorimetric 
               
               
                   
                 bil total (total bilirubin) 
                 colorimetric 
               
               
                   
                 calcium 
                 colorimetric 
               
               
                   
                 ceruloplasmin 
                 immunoturbidimetric 
               
               
                   
                 cholesterol 
                 colorimetric, enzymatic 
               
               
                   
                 cholinesterase 
                 kinetic 
               
               
                   
                 ck (creatine kinase) 
                 kinetic 
               
               
                   
                 ck MB (creatine kinase MB) 
                 kinetic, immunoinhibition 
               
               
                   
                 complement C3 
                 immunoturbidimetric 
               
               
                   
                 complement C4 
                 immunoturbidimetric 
               
               
                   
                 crp (C-reactive protein) 
                 immunoturbidimetric 
               
               
                   
                 cystatin C 
                 immunoturbidimetric 
               
               
                   
                 D-dimer D 
                 immunoturbidimetric 
               
               
                   
                 ethanol 
                 enzymatic 
               
               
                   
                 phenobarbital 
                 immunoturbidimetric 
               
               
                   
                 ferrum 
                 colorimetric 
               
               
                   
                 ferritin 
                 immunoturbidimetric 
               
               
                   
                 fibrinogen 
                 immunoturbidimetric 
               
               
                   
                 ggt (gamma-glutamyltransferase) 
                 colorimetric 
               
               
                   
                 glucose 
                 colorimetric, enzymatic 
               
               
                   
                 haptoglobin 
                 immunoturbidimetric 
               
               
                   
                 hbdh (α-hydroxybutyrate 
                 kinetic 
               
               
                   
                 dehydrogenase) 
               
               
                   
                 hdl cholesterol 
                 enzymatic 
               
               
                   
                 HbA 1C  (haemoglobin) 
                 immunoturbidimetric 
               
               
                   
                 immunoglobulin A 
                 immunoturbidimetric 
               
               
                   
                 immunoglobulin E 
                 immunoturbidimetric 
               
               
                   
                 immunoglobulin M 
                 immunoturbidimetric 
               
               
                   
                 carbamazepine 
                 immunoturbidimetric 
               
               
                   
                 creatinine 
                 colorimetric; Jaffe 
               
               
                   
                 creatinine 
                 enzymatic, colorimetric 
               
               
                   
                 alpha-1-acid glycoprotein 
                 immunoturbidimetric 
               
               
                   
                 ldh (lactate dehydrogenase) 
                 kinetic 
               
               
                   
                 ldl cholesterol 
                 enzymatic 
               
               
                   
                 lipase 
                 colorimetric 
               
               
                   
                 lipoprotein 
                 immunoturbidimetric 
               
               
                   
                 Mg (magnesium) 
                 colorimetric 
               
               
                   
                 copper 
                 colorimetric 
               
               
                   
                 myoglobin 
                 immunoturbidimetric 
               
               
                   
                 lactates 
                 colorimetric 
               
               
                   
                 paracetamol 
                 colorimetric 
               
               
                   
                 phosphorus 
                 colorimetric 
               
               
                   
                 potassium 
                 colorimetric 
               
               
                   
                 rf (rheumatoid factor) 
                 immunoturbidimetric 
               
               
                   
                 salicylates 
                 colorimetric 
               
               
                   
                 sodium 
                 colorimetric 
               
               
                   
                 theophylline 
                 immunoturbidimetric 
               
               
                   
                 tg (triglycerides) 
                 enzymatic, colorimetric 
               
               
                   
                 total protein 
                 colorimetric 
               
               
                   
                 ua (uric acid) 
                 enzymatic, colorimetric 
               
               
                   
                 uibc (unsaturated iron binding 
                 colorimetric 
               
               
                   
                 capacity) 
               
               
                   
                 urea 
                 enzymatic, kinetic 
               
               
                   
                 urine protein 
                 colorimetric 
               
               
                   
                   
               
             
          
         
       
     
       Preferred Embodiment 
       [0051]    A microfluidic system has been fabricated from polypropylene. The scheme of the system is shown in  FIG. 3 . The system contains channels with diameter of 400 and 800 μm, and comprises, among others, T-junctions connecting channels with each other. A sample in the form of a serum portion (marked “Serum” in  FIG. 3 ) was introduced into the system and a 100 nl droplet was produced in a T-junction. A portion of reagent (“Reagent R1” in  FIG. 3 ) for amylase assay was introduced into the second channel of the system and a 5 μl droplet was produced in a T-junction. Using hexadecane as a continuous liquid, the sample droplet and the reagent droplet (“Oil 3”, “Oil 2” in  FIG. 3 , respectively) were transported to the location, where the sample droplet and the reagent droplet merged. The mixing was effected by further pumping the merged droplet through a meandering channel between the outlet 1 and the outlet 2. As a result, chemical reaction took place, and the result of the reaction was measured with a spectrophotometer. Based on the spectrophotometric measurement, the amylase content in the sample was determined. 
         [0052]    The microfluidic systems and the method for transporting microdroplets using carrier liquids (continuous liquids) in these systems are known in the state of the art, e.g., from a patent application WO2011/090396. Likely, the method for determining concentrations of, for instance, albumin, bilirubin or creatinine, and many other biochemical parameters in a sample using spectrophotometric analysis is known in the state of the art, whereas the selection of the material for fabrication of the microfluidic system, the carrier liquid and the reagent constitute the element of the present invention.