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CL-10 Plus – User Manual 

26 / 75 

ECPUS vers 0.2 eng 

 
 

 

3.2.4 KINETIC 

measurement 

(KIN). 

. 

A practical example: Acetyl-cholinesterase determination in whole blood 

 
Sample:   Whole 

Blood 

Buffer Solution: 

Phosphate/Pyro-phosphate   (

Β

 

= 6.1 mM/L pH) 

Stopper:   volume 

1225 

µ

L 

Temperature: 37°C 

Micropipette:  

Gilson Microman M25 

Sample Volume: 

15 

µ

l of whole blood 

Starter: 

 

Acetylcholine (92 mM) 

 
Analysis of the measurement steps: 

 

                

electr. 1

electr. 2

starter 1

stirrer

Sample

E -E

2

1

D 0

 

 

 
PHASE ZERO

: the mixing chamber and the two electrodes are filled with the same buffer. 

 

PHASE 1

: Using the micropipette 15 

µ

L of blood are injected in the mixing chamber; the 

stirrer rotates and mixes  blood (which contains Acetyl-cholinesterase) for some 
seconds. Pumps 1 and 2 rotate and both the electrodes are filled with blood. 
 
The electrodes contain exactly the same solution, the 

∆

pH should be zero. 

 

Содержание CL-10 Plus

Страница 1: ...CL 10 Plus User Manual...

Страница 2: ..._______________________________14 2 8 Changing of application within the same working session _________________________________15 3 3 T Th he e I In ns st tr ru um me en nt t _ __ __ __ __ __ __ __ __...

Страница 3: ..._ __ __ __ __ __ __ __ __ __ __ __ __ __ __ __ __ __ __ __ __ __ __ __ __ __ __ __ __ __ __ __ __ __ __ __ __ __ __ __ __ __ __ __ __ __ __ __ __ __ __ _6 63 3 6 1 Electrical Section _________________...

Страница 4: ...t manual easier These products will be often mentioned when required for routine procedures maintenance and repair Product Code Use Polif Solution GEN718 wash solution to be used with all Milk and Win...

Страница 5: ...st of content 1 Instrument CL10 Plus 2 power 230 VAC 110 VAC 12 VDC 3 power cable 4 serial cable CL10 PC 5 Plastic bottles Kartel type for the buffer and or standby solution and for the waste 2x250 ml...

Страница 6: ...PC to a 500VA stabilizer In any case it is recommended to keep the CL10 Plus instrument under UPS Uninterruptible Power Supply for protection against accidental power loss 2 3 2 Mains voltage setting...

Страница 7: ...CL10Plus vs 4 3 CD ROM or superior version 4 Open the cl10plus vs 4 3 or sup folder and select the Setup exe icon 5 Automatically the program guided installation will start 6 Follow the instructions...

Страница 8: ...ess OK Methods setting 9 Check that the CL10Plus CD ROM is in the CD unit 10 Open the following folder D CD ROM S_Plus_Matrix Name_XXX CL10Plus vs 4 3 11 Open the folder containing the name of the app...

Страница 9: ...the correct tubing s assessment mark the thick ends of each tube before hooking it on the same side along the fusion line and check that once hooked it remains marked on the same side 2 Hook the 4 per...

Страница 10: ...set 8x SPP 311 2 Peristaltic Pump Tubing s Set 4x SPP 312 3 Enzyme Starter tubing only mono starter 1x SPP 313 4 Additional Tubing sample starter Wine Combi 1 1 SPP 314 5 Additional Tubing sample spec...

Страница 11: ...starter tube inside the vial Run Prime enzyme cycle to fill in the starter tube Take both ends of the starter tubing completely filled in with water from the vial and lift in the air the opposite end...

Страница 12: ...solution again and run 3 Clean cycles 9 Empty the waste bottle 2 5 4 Standby mode Rest For the CL 10 Plus instrument there are two available different standby mode REST mode 1 From the computer REST...

Страница 13: ...nts have mpH 0 5 mpH and the D1 of the last two measurements do not differ more than 1 mpH from each other 2 6 1 Instrument preparation Milk application The following procedure taken as an example app...

Страница 14: ...he AUX tubing according to the package insert and run Calibrate cycle F7 once If the SLOPE value obtained is OUT OF RANGE the result will appear in red see package insert and 4 4 3 of this manual in t...

Страница 15: ...ed by the next method Leave the enzyme vial pump 6 of the method in use in its place 2 From File menu Select the ENZY_REC_R3 method 3 Run Sample once using the GO icon or the F5 function key Press Sta...

Страница 16: ...e CL10 Plus is a compact and easy to operate instrument It is made of a differential amplifier and a microprocessor which calculates the analyte concentration or enzyme activity in the sample and cont...

Страница 17: ...e point of neutralisation A plot of pH of the solution vs the amount of NaOH added to this point is called titration curve Let us take the titration curve of acetic acid a typical weak acid At the beg...

Страница 18: ...pH is the pH increment induced by the addition of an amount of base whereas H is the relative change of free H in solution A large Buffer Capacity of the medium corresponds to a large amount of base...

Страница 19: ...ethod and wait until it is visualised on the screen Inject sample 10 L of HCl 100 mM in the mixing chamber Run Sample command or press the GO icon or the F5 function key Press Start Measure then Accep...

Страница 20: ...ration 0 81 mM L 4 38 mM L pH pH pH variation 0 185 pH 3 2 3 END POINT measurement A practical example urea determination in milk The heart of the differential pH apparatus are the mixing chamber and...

Страница 21: ...k E E 2 1 D1 1 2 At this stage the instrument reads for 5 seconds the pH difference between EL 2 EL 1 mpH called D1 The electrodes contain exactly the same solution the pH difference should be zero in...

Страница 22: ...his stage the instrument reads for 30 seconds the mpH difference between EL 2 and EL 1 but while EL 1 does not modify its pH value because there is no reaction inside in the EL 2 the urease catalyses...

Страница 23: ...calibration The previous example was useful to present the enzymatic reaction that occurs in the differential pH apparatus A number of simplifications have been made first of all the D1 value obtained...

Страница 24: ...enzyme into the mixer and only the electrode 2 takes the solution buffer urease while the electrode 1 contains only buffer the urease may have a little difference of pH compared to the buffer due to...

Страница 25: ...rd urease enzyme which promotes the reaction DL1 D2 D1 30 20 10 mpH But also in this case not all the 10 mpH are caused by the urea hydrolysis we have to subtract 1 mpH the acidity of urease STD 100 m...

Страница 26: ...d Starter Acetylcholine 92 mM Analysis of the measurement steps electr 1 electr 2 starter 1 stirrer Sample E E 2 1 D 0 PHASE ZERO the mixing chamber and the two electrodes are filled with the same buf...

Страница 27: ...tarter whilst EL 1 stays in the solution buffer blood E E 2 1 D2 1 2 PHASE 3 the instrument reads for 70 seconds the mpH between EL 2 and EL 1 The calculation of the reaction rate slope of pH curve is...

Страница 28: ...ty is the number of enzyme units per milligram of protein The specific activity is a measurement of enzyme purity it increases during purification of an enzyme and becomes maximal and constant when th...

Страница 29: ...determination with Ist order i e la analyte concentration Km of the enzyme Analysing the steps of the measurement cycle electr 1 electr 2 starter 1 stirrer Sample E E 2 1 D 0 PHASE ZERO the mixing cha...

Страница 30: ...the EL 1 still remains buffer sample E E 2 1 D2 1 2 PHASE 3 the instrument reads the mpH between EL 2 and EL 1 for some seconds While the EL 1 works as reference electrode in the EL 2 the starter cat...

Страница 31: ...he curve b will be obtained with mpH b is lower than mpH PHASE 4 the instrument is automatically washed and the mixing chamber refilled with the new buffer Lag Phase t s Lag Phase mpH Starter t0 t1 t2...

Страница 32: ...n order to enter the menu ex ALT f to access File submenu press instead the combination CTRL the underlined letter to open the chosen submenu example to Select method press CTRL s B Toolbar Contains a...

Страница 33: ...and maximum is not visualised because under the File window During alkaline reactions OH which increases the pH value the baseline on the screen goes to higher values During reactions which release hy...

Страница 34: ...d to select the desired method for the routine All default methods are supplied with the installation disk Attention select a method only after choosing the kit intended to be used for the routine in...

Страница 35: ...rameters of methods and possibly to insert new values for upgrades as described in the package insert of the kits Moreover to change the user unit upon needs Access the Edit Method command from file m...

Страница 36: ...ibed in 3 3 STD units calibrators concentrations expressed in standard unit SI according to the international standards mM M meq L User units used to obtain different measure units from standard units...

Страница 37: ...panel The Slope value is automatically set during the Calibrate measure The value must be within the range defined by Minimum slope and Maximum slope and it can be a positive or negative number depend...

Страница 38: ...e movements represent a measure cycle and a cleaning cycle for a specific method Each method is characterized by only one dedicated string or code vice versa the same string can characterize few metho...

Страница 39: ...However it is possible to express the erythrocyte enzymes concentration also in U g Hb directly from CL 10 Plus Software modifying the method file MD For this function it is crucial to know the Haemo...

Страница 40: ...t Name field G6PD Select Edit method and rename the new file G6PD as G6PD_HbR1 MD right hand button of the mouse placed on the document Rename command From File menu enter Select method and choose G6P...

Страница 41: ...unactivate Concentration given in U L 4 2 5 Clear graph This function is accessible trough the Clear Graph command from File menu or can also be selected by the icon 4 2 6 Print graph This function is...

Страница 42: ...u item Instead of opening the menu and choosing a command you simply press the key combination SHORTCUT KEYS LIST F2 Prime Enzyme F3 Clean F4 Run Start Up F5 Sample F6 Blank F7 Calibrate F8 Rest Mode...

Страница 43: ...nsecutive cycles Maximum Error mpH it allows to set the maximum error allowed i e the maximum mpH possible range between 0 2 and 10 0 mpH The last three consecutive GO results of the Start up must not...

Страница 44: ...CL 10 Plus User Manual 44 75 ECPUS vers 0 2 eng First case Maximum error 0 5 The startup procedure passed Second case Maximum error 0 5 The startup procedure failed...

Страница 45: ...mple analysis after the calibration procedure and control inject the sample then select icon or press F5 function key Samples can be identified After the GO command is given before starting the measur...

Страница 46: ...od Minimum offset e Maximum offset 4 2 3 in order to be automatically saved by the program The icon for this command is or F6 function key 4 4 3 Calibrate C icon or F7 This function necessarily associ...

Страница 47: ...d it is possible to produce the printout of data stored in the archive The paper layout of the printed results can be customised in the following way 1 Exit the CL 10 Plus program 2 Opne the CL 10 plu...

Страница 48: ...ot Entry key make the same for all the data to be plotted Once all the desired data have been plotted select Done key to close the window 4 5 3 Erase Worklist This command is used to delete completely...

Страница 49: ...ct find enter the destination folder 6 Change file type file type from txt to All Files 7 Give the name to the file from export to desired name followed by xls extension 8 Press OK follows the informa...

Страница 50: ...ation fields it is possible to select the columns width the separation symbol is 13 During the third and last step you can decide which column to transfer or delete which format to give After these st...

Страница 51: ...e This command activates the standby mode for the instrument halt period normally at least 8 10 hours for longer periods than 2 weeks see 5 5 With the rest program in active mode one measure cycle of...

Страница 52: ...edium volume low volume refer to previous CL 10 versions Sample Pump Timing This function is not active Misc parameters This function is not active Rest parameters This function is used to define rest...

Страница 53: ...on parameters This window is used if it is required to configure and select the serial port RS 232 used by CL10 Plus The preferential channel is COM1 the alternative one is COM2 in case COM 1 is engag...

Страница 54: ...irrer motor working performances it manages the stirrer bar rotation in the mixing chamber for obtaining homogeneous solutions 1 peristaltic pumps working performance hall sensors From Service window...

Страница 55: ...2 2 control of correct stirrer motor operation ATTENTION in case the sensors have passed peristaltic pumps working performances check it is required to exit the Terminal Emulation and CL 10 working pr...

Страница 56: ...tip and feeling the collisions of the bar with the tip 4 6 8 Electrodes Test 1 and 2 This command is used to check the electrodes performances Normally it is used by service personnel only unless dir...

Страница 57: ...utes more 6 Replace the REGENERATING SOLUTION with reconstituted wash solution 7 Run 1 Prime enzyme and 2 Clean 8 Leave the instrument in Rest mode see 4 6 2 5 2 Routine Maintenance Run the following...

Страница 58: ...bserve if during the wash cycle Clean or F3 the liquid comes up to the top of the mixing chamber If not tubing s are not in proper sealing conditions Replace all pump tubing s with a new set not only...

Страница 59: ...pf the mixing chamber 3 Fill in the chamber with 1 mL of HCL 0 1M and run a measurement GO or F5 4 Inject 1 mL milk any kind and run 3 Clean cycles F3 5 Reconnect the enzyme tube to the top of the mi...

Страница 60: ...CL 10 Plus User Manual 60 75 ECPUS vers 0 2 eng Figure 1 Figure 2...

Страница 61: ...water vials both from buffer and starter positions 4 Empty the fluidics repeating the sequence 1 and 2 and empty the waste bottle 5 Unhook all 6 peristaltic pumps tubing s from their seats around the...

Страница 62: ...8 Place a vial containing distilled water 2 mL minimum in starter position and insert the starter tube needle connected to pump 6 and in case also the AUX starter needle connected to pump 5 9 Run two...

Страница 63: ...ping inside the circuit damages the rotor circuit communication or a whole set of pumps is not responding Vice versa it may happen that one of the rotors once activated is never stopping regardless th...

Страница 64: ...or following box appears on the screen Check that the instrument is on Check that the CL 10TBL_BUILDER program is off if not turn it off Check the connections between the serial port RS 232 COM 1 in t...

Страница 65: ...t rotating at the beginning of the measurement for approx 4 seconds If you cannot hear it insert a pipette tip in the chamber you should feel the collision with the bar If not ask for technical servic...

Страница 66: ...e problem persists call for technical assistance 6 3 Electrodes 6 3 1 Control of electrodes performances The following procedure is used and recommended by in order to control electrodes performances...

Страница 67: ...in a vial containing distilled water the same where also the other end C is placed 3 Follow the instructions of package insert and fill in the form Report Electrodes performance CL 10 Plus available...

Страница 68: ...l discharge all the water and flush inside only air to be ready for the next steps 4 Remove the steel cover of the mixing chamber Fig 4 part A by unscrewing the black stopper on the top Fig 4 part B a...

Страница 69: ...FIGURE 3 Electrodes Installation A B C D E F G A B C D A MEASURING CHAMBER B SILICON TIPS C FLUIDIC S D GLASS ELECTRODES E ELECTRODES HOLDER SCREW F ELECTRODE CABLE G ELECTRODES HOLDER A STEEL COVER B...

Страница 70: ...s User Manual 70 75 ECPUS vers 0 2 eng FIGURE 4 MIXING CHAMBER C B A D E F FIGURE 5 PERISTALTIC PUMP GROUP A CHASSIS B LEFT PERUISTALTIC PUMP GROUP C RIGHT PERISTALTIC PUMP GROUP D EXHAGONAL SCREW E F...

Страница 71: ...operly With the starter tube full of water disconnect both ends and run F2 Prime enzyme once If the liquid inside is not flowing change the peristaltic pump of the tube if the problem persist change a...

Страница 72: ...ts Carry out initialise cycle Prime Enzyme or F2 Insert the needle in the vial and run Prime Enzyme or F2 cycle Replace the starter vial and run one Prime Enzyme or F2 cycle Inspect and if necessary c...

Страница 73: ...ume below the initial value shown on the volume come slowly to the new value making sure not to pass the required volume To set a volume above the initial value shown on the volume pass the required v...

Страница 74: ...the vessel and press the push on button slowly to the first stop Move the capillary away from the side wall keeping the push button pressed then withdraw the pipette from the vessel Release the push...

Страница 75: ...ette Aspirate slowly the sample taking care not to have air bubbles into the capillary Wipe with laboratory paper any fluid outside the capillary Insert the capillary into the CL10 measuring chamber u...

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