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APOLLO 4000

NITRIC OXIDE DETECTION

WORLD PRECISION INSTRUMENTS

C-17

Place the distilled water vial in an ice-water bath. Reducing the temperature

increases the solubility of NO in solution. Thus when the solution is used at

room temperature you will be assured of a saturated NO solution.

Purge the system with argon (or nitrogen) gas for a period of 30 minutes at a

moderate flow rate such that the pressure is maintained at a safe level (1-2

psi). When purging it should be observed that gas is indeed bubbling through

the KOH solution as well as the distilled water. After 30 minutes turn off the

argon source, and switch the tee purge valve to the correct position for

purging with NO from the lecture bottle.

Purge the system with NO for 5-10 minutes if using a pure source (longer if the

NO source is not pure). Again make sure that gas is bubbling through both

solutions.

Warning: NO is now escaping from the pressure relief

needle in the stopper of the distilled water vial. It is imperative

that the fume hood be running at maximum capacity with the

front panel closed (in the down position).

After the time in step 7 has elapsed turn off the NO source.

Immediately remove the two needles from the distilled water vial.

10.

Set the tee purge valve for purging with argon (or nitrogen) gas, and turn on

the argon source. Purge the system for 5-10 minutes at a moderate flow rate.

Gas should be bubbling through the KOH and then escaping from the flask

into the atmosphere. Again be sure that the fume hood is ventilating well.

11.

Turn off the argon (or nitrogen) source, and allow the fume hood to continue to

ventilate for 10-15 minutes so as to ensure that all traces of NO gas are

removed from the atmosphere.

12.

The solution of distilled water should now be saturated with NO. The

concentration of NO produced by this saturation is dependent upon the

temperature. At 0

C, the concentration is approximately 3.3 mM, and at 20

the concentration is approximately 1.91 mM.

13.

Dilutions of known concentration can be prepared from this saturated solution

In preparing a dilution,

be careful not to unseal the vial,

for this

exposes the solution to atmospheric oxygen.

Once the dilutions are prepared, it is a simple matter to calibrate the instrument.

Содержание APOLLO 4000

Страница 1: ...the Internet Do not attach any peripheral device other than a USB printer Any change to the proprietary hard disk registry in this device whether by virus or by normally benign hardware or software in...

Страница 2: ......

Страница 3: ...CT NITRIC OXIDE C 1 Initial Set up C 1 Calibration of the NO Sensor C 1 The Calibration Kit C 1 Calibration by the chemical generation of NO C 2 Calibration of NO sensor by decomposition of SNAP C 7 C...

Страница 4: ......

Страница 5: ...lated multi channel electrode based free radical analyzer designed specifically for the detection of a variety of redox reactive species of biomedical importance The electrochemical amperometric detec...

Страница 6: ...based on a 24 bit A to D and 16 bit D to A conversion driven by a Digital Signal Processor DSP The User Interface is built on a standard PC platform with Windows 2000 operating system A standard seria...

Страница 7: ...erformance Faraday shield designed to minimize susceptibility to environmental noise The Apollo software can be programmed to display either redox current e g pA or concentration e g nM Output from th...

Страница 8: ...hannels Temperature LED red Sensor LED green Power Temperature Input Sensor Input DVD CD RW Drive Mains Power 110V 220V Switch mouse Power Supply for Monitor Keyboard Analog Outputs BNC USB Ethernet S...

Страница 9: ...nd Returns on the Warranty page of this manual Returns Do not return any goods to WPI without obtaining prior approval RMA required and instructions from our Returns Department Goods returned unautho...

Страница 10: ...APOLLO 4000 INTRODUCTION WORLD PRECISION INSTRUMENTS A 6...

Страница 11: ...ther end of the voltage converter to a matching 3 prong grounded wall receptacle and switch on 4 Connect the power supply cord to the back of the APOLLO 4000 and plug the other end into a matching 3 p...

Страница 12: ...by WPI vary in their fragility However at all times the user must exercise caution to avoid damaging the delicate polymeric membrane covering the end of each sensor This membrane prevents water and d...

Страница 13: ...g LED indications will inform the user of the status of the system LED CONDITION INDICATION Temp LED red Steady RED light Electrode is performing normally No RED light Electrode is not connected or is...

Страница 14: ...re being turned off In the example shown the set up is for a four channel nitric oxide application with horizonal scale of 6 sec division and vertical scales 5000 pA div for channels 1 to 4 Data for e...

Страница 15: ...Open opens an existing data file Save saves the current experiment with the default name Save as allows the user to chose the name and the path of the saving data file Print prints the screen to the s...

Страница 16: ...le structure and it is therefore necessary to close the previous file Fig B4 This can interrupt the current experiment which may be undesirable Therefore the user must carefully select the sampling ra...

Страница 17: ...nA range is selected then the noise floor will be approximately 10 nA divided by 1 000 000 i e 10 fA Conversely if 10 A range is selected then the noise floor will be approximately 10 A divided by 1 0...

Страница 18: ...y configures the poise voltage i e 700 mV on the selected channel to measure oxygen Hydrogen Peroxide Automatically configures the poise voltage i e 400 mV on the selected channel to measure hydrogen...

Страница 19: ...submenu in the Setup control The Amplitude Unit Conversion screen appears Fig B13 Example The user defines a new unit called Custom with the conversion ratio 1 unit 186 pA Fig B14 If Add Unit button...

Страница 20: ...on pushed There are a total of two cursors The second cursor appears after the first one when the pointer device left button is released Calculate Menu includes five different self explanatory paramet...

Страница 21: ...set each channel individually all together or unzero all of previously zeroed channels The subtracting value is acquired at the moment of selecting the appropriate action Zero operation can be applied...

Страница 22: ...nt the Note to be added see Fig B22 This will create a mark at that place Double click the created mark A Note window will open where a message of up to 64 characters can be typed in see Fig B23 After...

Страница 23: ...a real signal application An example file is loaded in the Apollo 4000 Fig B24 Note the temperature sensor is also connected to channel 1 i e 25 C line However the sensor signal is not seen If we appl...

Страница 24: ...user must double click inside the appropriate channel plot The second window has a distinctive yellow background color Only one second window can be opened at the same time The size of the second wind...

Страница 25: ...sed on the decomposition of the S nitrosothiol NO donor SNAP using either Cu I as described in Method I or Cu II as outlined in Method II as a catalyst The NO liberated from SNAP is used to calibrate...

Страница 26: ...ate Standard Nitrite is available from WPI catalog 7357 Alternatively crystalline reagent KNO2 may be used but the user should note that KNO2 is extremely hygroscopic and degrades once exposed to atmo...

Страница 27: ...torage 2 Immerse the ISO NOP sensor tip in a strong saline solution 1 M and after waiting a few minutes for the current to stabilize record its value If the current is offscale or unstable after sever...

Страница 28: ...has undergone a large temperature change 8 If you feel it is necessary to degas solution 1 prior to calibration this can be done by inserting one of the long stainless steel needles included with the...

Страница 29: ...by simple dilution factors as follows 50 L of 50 M KNO2 solution 2 into 10 mL solution 1 1 201 dilution Hence amount of NO produced 50 M 201 0 2487 M 249 nM Likewise Addition 2 100 L of solution 2 add...

Страница 30: ...O3 to NO2 Researchers who use the Greiss method for NO2 determination will also be very interested to learn that using the ISO NOP and the titration method for calibration described above a detection...

Страница 31: ...rameters involved in SNAP decomposition neither is it intended to propose a model by which SNAP is decomposed Two methods are described here for the calibration of NO sensors based on decomposition of...

Страница 32: ...copper II sulfate CuSO4 or cupric II chloride CuCl2 in which Cu II is the active catalyst for the conversion of SNAP to NO Getting Started Prepare the following solutions 1 Saturated solution of cupr...

Страница 33: ...commend the use of high grade SNAP with minimal purity of 98 or better SNAP can be purchased from WPI in various amounts Calibration Procedure Within a nitrogen or argon environment place 10 0 mL of s...

Страница 34: ...ng Cu II as a catalyst This method of calibration relies on the use of Cu II for the partial but quantifiable conversion of SNAP to NO This procedure can be used as an alternative to the previous meth...

Страница 35: ...ains relatively stable for at least 5 hours if kept in refrigerator Note The purity of standard reagent SNAP is very important for the reported data Use high grade SNAP with minimal purity of 95 or be...

Страница 36: ...ing the signal output e g in pA vs the concentration of SNAP added at that time Note that every addition of SNAP solution corresponds to a particular NO concentration This will be discussed below Afte...

Страница 37: ...f detectable NO will be below the theoretical value because the copper sulfate solution was at equilibrium with ambient air and consequently a portion of the generated NO would have been immediately o...

Страница 38: ...0 6 X SNAP Output Current 232 4 nM 139 4 nM 230 pA 462 0 nM 277 2 nM 488 pA 916 8 nM 550 1 nM 1001 pA Y 32 038 1 88X The data from calibration curve indicates that this procedure allows an excellent...

Страница 39: ...standards prepared with NO Gas The following method can be used with all NO sensors Preparing an NO Standard This method has the advantage of allowing the user to calibrate NO sensors in the same env...

Страница 40: ...ion is used to remove other nitrogen oxides from the NO gas 4 Place 20 mL of distilled preferably deionized water in a small glass vial Seal the vial with a stopper and insert through the stopper a lo...

Страница 41: ...nning at maximum capacity with the front panel closed in the down position 8 After the time in step 7 has elapsed turn off the NO source 9 Immediately remove the two needles from the distilled water v...

Страница 42: ...should be placed as close as possible to the site of release of NO Durability and Handling The user must excercise caution when handling any NO sensor to avoid actions which could damage the sensor t...

Страница 43: ...rces may couple to the system and produce large extraneous signals in the output record The magnitude of this external noise depends on the environment of the laboratory If the interference introduced...

Страница 44: ...ng the end of the stainless sleeve The other end of the sleeve is flanged The locking cap is used to attach the sleeve to the probe handle When the sensor is fully assembled i e with locking cap and s...

Страница 45: ...or base solution at times both may be necessary for 10 seconds A good indication of a dirty membrane sleeve is a sluggish response or an unusually low sensitivity If these problems are not rectified b...

Страница 46: ...p into the handle The electrode should be observed to press gently against the membrane 9 The current displayed on the meter at this time will be high or offscale 10 Suspend the tip of the new assembl...

Страница 47: ...ilar device that permits accurate positioning The electrode should be cleaned periodically in distilled water and dried using soft tissue paper Organic contamination can be removed using a mild enzyma...

Страница 48: ...APOLLO 4000 NITRIC OXIDE DETECTION WORLD PRECISION INSTRUMENTS C 24...

Страница 49: ...f atmospheric pressure parts per million ppm or as a redox current in nanoamperes nA The ISO OXY 2 houses a platinum working electrode and a silver counter reference electrode inside a stainless steel...

Страница 50: ...is switched back on again Zero Oxygen Point Calibration After polarization of the ISO OXY 2 as described above a calibration for zero percent oxygen may be carried out in pure nitrogen gas or in wate...

Страница 51: ...vely air can be used as the calibration standard but since water vapor does affect the probe reading it is best to use dry air unless the ambient humidity is accurately known Dry air can be obtained b...

Страница 52: ...ed using the software At point 3 and point 4 the sensor is exposed to air 21 oxygen and 100 oxygen correspondingly By using the current at 0 0 nM and 100 236 nA a two point calibration curve is built...

Страница 53: ...e tip into the water via the top hole Aerate for a few minutes by bubbling air through the side arm of the bottle at a low pressure using a simple aquarium aeration pump The scale reading should be al...

Страница 54: ...oxygen concentration when the probe is in water is 8 4 ppm You do not need to correct for the water vapor effect for a ppm calibration since the values in Table 1a are obtained in water saturated air...

Страница 55: ...factor to nA or ppm to nA sensitivity of the probe is determined from the slope of the calibration curve and must be typed into the corresponding field see Fig B15 Only after selecting ppm or unit wi...

Страница 56: ...r depending on the unit of pressure chosen 1 for atm pt and pH2O are the total pressure of gas and the partial pressures of water respectively pO2 is the partial pressure of oxygen in blood obtained f...

Страница 57: ...let water get into the handle The current on the Apollo 4000 should go offscale when the electrode is being rinsed 4 Gently dry the electrode with a soft tissue Kimwipes Be sure to dry thoroughly the...

Страница 58: ...Y 2 is not being used it should be stored with the tip suspended in distilled water Clean the Membrane The membrane sleeve itself requires very little maintenance The primary concern is to avoid to da...

Страница 59: ...it s recommended that the electrode be disconnected from the instrument and stored with the tip immersed in distilled water This practice will reduce the possibility of a gradual reduction of electro...

Страница 60: ...10 0 6 11 8 11 1 10 5 9 8 7 11 5 10 9 10 2 9 6 8 11 2 10 6 10 0 9 4 9 11 0 10 4 9 8 9 2 10 10 7 10 1 9 6 9 0 11 10 5 9 9 9 4 8 8 12 10 3 9 7 9 2 8 6 13 10 1 9 5 9 0 8 5 14 9 9 9 3 8 8 8 3 15 9 7 9 1...

Страница 61: ...500 668 0 88 4000 656 0 86 4500 644 0 85 5000 632 0 83 5500 621 0 82 6000 609 0 80 Table 3 Water vapor partial pressure in mm Hg Temp Pv Temp Pv C mm Hg C mm Hg 0 5 20 18 2 5 22 20 4 6 24 22 6 7 26 25...

Страница 62: ...Hb g 100 mL C Plasma 5 g 10 g 15 g 20 g 15 0 0302 0 0310 0 0312 0 0316 0 0323 20 0 0277 0 0282 0 0284 0 0287 0 0293 25 0 0257 0 0261 0 0263 0 0265 0 0271 28 0 0246 0 0249 0 0251 0 0253 0 0259 30 0 02...

Страница 63: ...00012 which must contain fresh refillable electrolyte WPI 100042 and is separated from the external environment by membrane covering the end of the stainless sleeve The other end of the sleeve is flan...

Страница 64: ...chambers WPI NOCHM or NOCHM 4 can be used as an alternative calibration kit specifically for use at different temperature condition Calibration temperatures from 4 40 C can be controlled using an ext...

Страница 65: ...he calibration solution 1 After the current is stable zero the baseline 2 Add a known volume of the H2 O2 standard solution to the PBS buffer solution For example add 0 5 10 20 40 80mL H2 O2 standard...

Страница 66: ...ate use Clean the Membrane The membrane sleeve itself requires very little maintenance The primary concern is to avoid to damage the membrane and to keep it as clean as possible After each use the mem...

Страница 67: ...lue e g 0 20 pA If this occurs then the electrode is in good working order 5 If the electrode is not clean repeat steps 3 and 4 If necessary the ISO NOP Rejuvenator WPI JUV can be used to restore sens...

Страница 68: ...h time It may take several hours for the sensor current to reach a low stable value at which time it will be ready for use 12 If the current observed after a few minutes in the saline solution increas...

Страница 69: ...ion procedure are correct b The membrane may be heavily contaminated Clean the membrane as described in the section Cleaning the Membrane c The electrolyte inside the sleeve may no longer be functioni...

Страница 70: ...APOLLO 4000 TROUBLESHOOTING WORLD PRECISION INSTRUMENTS F 2...

Страница 71: ...the probe is immersed into the effluent of a perfusion system or placed directly into the vein of the heart of an animal In this case grounding the external bathing fluid vein or tissue with a stainle...

Страница 72: ......

Страница 73: ...quested All claims for shortage or damage must be made within 10 days after receipt of shipment Claims for lost shipments must be made within 30 days of invoice or other notification of shipment Pleas...

Страница 74: ......

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