Reichert 1310400A User guide

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REICHERT TS METER Refractometer Model 1310400A
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Reichert TS-METER Total Solids Refractometer Model 1310400A
Instruction Manual
1. 0 Introduction.................................... 2
2.0 Operating Instructions ...................... 2
4.0 T emperature Compensation ............... 4
5.0 Dipping Refractometry ..................... 5
6. 0 Air Bubble ...................................... 5
7.0 Non-Aqueous Solutions ................... 5
8.0 Use of TS METER Refractometer
Conversion T ables............................ 5
9.0 Serum T ot al Solids and Water Measure-
ments: Serum and Urine ................... 6
10.0 Estimation of Protein by
Refractometry .................................. 6
11.0 Estimation of Specific Gravity and T otal
Solids of Urine by Refractometry....... 7
12.0 Estimation of Concentration of Other
Body Fluids and of Pure Solutions .... 7
13.0 Refraction Scale ............................... 7
14.0 Information Obtainable with the
REICHERT TS METER Refractometer .......
1.0 INTRODUCTION
Determinations are precise and rapid and require only a drop of fluid sample. The value on the appropriate scale seen through the eye­piece, is read where the sharp boundary between dark and light fields crosses the scale. The instruments are temperature compensated for temperatures between 60°F (16°C) and 100°F (38°C). The reading does not need to be adjusted for the sample or ambient temperature.
The accuracy of the determination of total solids–or water–in plasma and urine from measurement of refractive index, and for the measurement of specific gravity of urine from refractive index, has been well documented.
1,2,3, 20
The estimation of plasma or serum protein concentration by refractometry has also been advocated for many years. accuracy of this determination is satisfactory for clinical use although it is not as reliable as the measurement of total solids and of specific gravity.
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The
2.0 OPERA TING INSTRUCTIONS
Hold the instrument in a horizontal position. T o minimize evaporation place the cover plate over the measuring prism, then place the sample liquid on the exposed top or bottom of the measuring prism. The liquid will be
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drawn into the space between the prism and the cover plate by capillary action. T ake care to avoid lifting the cover plate before the reading is made. A dropper may be used to transfer the sample to the measuring prism. The dropper should be plastic to minimize the possibility of scratching the prism surface.
The REICHERT TS METER Model 1310400A T ot al Solids Refractometer has been designed for simple, rapid microanalysis in biomedical, chemistry and classroom laborato­ries. The determination of concentrations of solutions is one of the oldest uses of refractometry . The TS METER Refractometer offers unprecedented simplicity of application, lending itself to checks and controls of many laboratory reagents, preparations, pharmaceuti­cals and specimens.
Scales are calibrated for protein concentration of plasma or serum (grams/100mL) and specific gravity of urine and refractive index
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Alternately , the instrument may be loaded by swinging the cover plate over the body of the instrument to expose the prism and the cover plate surfaces. A small sample is then placed on the measuring prism. T o minimize evapo­ration, close the cover plate over the measuring prism immediately .
T o hold the instrument for reading, place your middle finger on nameplate and press the plastic cover gently, but firmly . This spreads the sample in a thin, even layer over the prism. Point the instrument toward a bright window or other source of illumina­tion, such as a lamp. It can also be placed on
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a specially designed illuminated refractometer table stand. T o obtain the optimum contrast between light and dark boundary , the instru­ment must be properly tilted toward the window or light source. Increased contrast and sharpness of the boundary may be obtained by using a vertical, gold color fluorescent lamp.
Focus the scale seen in the eyepiece by rotating the eyepiece. This setting does not need to be changed as long as the same individual continues to use the instrument.
Read the appropriate scale at the point where the dividing line between bright and dark fields cross. Use the conversion tables in this manual, (pages 9-14), if required.
Use a soft cloth or tissue moistened with water to wipe the prism and dry thoroughly. If the prism surface or cover plate is not cleaned before the next sample is loaded, an erroneous or fuzzy reading may result. Do not immerse the eyepiece or the black focusing ring in water and do not use hot water. Never use gritty cleaning compounds to clean the prism. WARNING: Never expose the instrument to temperatures above 150°F (60°C).
3.0 ZERO SETTING
The zero setting of the TS METER Refracto­meter rarely needs adjustment. In order to check adjustment make sure the temperature of the instrument is between 70°F (21°C) and 85°F (29°C) and take a reading on distilled water as explained previously. If a reading departs from zero by more than .05% (1/2 division) gently pry through the cement prism seal and turn the adjustment screw, with the appropriate tool, clockwise to increase the reading, counter-clockwise to decrease it. Make sure that final motion is clockwise. Seal the
hole with caulking compound after correct reading has been obtained.
NOTE: Caulking compound is supplied with the instrument.
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4.0 TEMPERA TURE COMPENSATION
The actual physical quantity measured by the TS METER is refractive index. The TS METER’s scales show this value as specific gravity, protein concentration and refraction. The relationship between refractive index and the quantity used is derived from an analysis of published and unpublished experimental data. Since refractive index changes appreciably with temperature, the conversion formulas are valid at a standard temperature only .
The TS METER Refractometer is temperature compensated to give correct readings directly on aqueous solutions at all temperatures ranging from 60°F (16°C) to 100°F (38°C). The maximum error at the extremes of the instrument and temperature ranges is 0.1% but the actual error over the most useful
portion is much less, as shown in Figure 3. The largest error occurs at the lowest scale readings, and particularly with pure water at temperatures of 65°F (18°C) and below. If water is used for calibration, the temperature should be 70°F (21°C) or above.
The TS METER Refractometer, provides temper­ature compensation and reads solid standards correctly only at temperatures between 65°F (18°C) and 70°F (21°C).
Figure 3 Temperature response of Model 1310400A TS METER
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5.0 DIPPING REFRACTOMETRY
7.0 NON-AQUEOUS SOLUTIONS
T o obtain a sharp shadow line and best possible compensation, the refractometer prism must be in temperature equilibrium with the sample to be measured or a sharp shadow line will not be produced. No special precautions are required if the sample is small because it will equilibriate to the instrument’s temperature immediately . However, if the refractometer is used as a dipping instrument, three to five minutes should be allowed if the temperature of the solution differs substantially from that of the instrument.
WARNING: It is extremely important that the hole of the zero setting screw be well caulked so that fluids do not enter and damage the instrument when immersed or when being washed. Use caulking compound supplied with the instrument.
6.0 AIR BUBBLE
T emperature compensation is produced with optical action of a liquid filled cavity arranged in the optical path. This cavity is hermetically sealed and cannot leak.
Thermal expansion of the liquid is accommodated with an air bubble which is kept out of the optical path by a bubble trap placed at the end of the cavity. In transit or under severe vibration, the bubble may escape the trap and appear in the visible portion of the refrac­tometer prism. If this occurs, the instrument should be held vertically, eyepiece down, and shaken lightly. This will allow the bubble to pass into the trap where it will be held during all normal operations.
The TS METER is temperature compensated for aqueous solutions. The refractive index temperature coefficient refractive index of oils, hydrocarbons or other liquid products is generally larger than that of water. Precautions should thus be taken if materials other than aqueous solutions are measured.
At temperatures between 65°F (18°C) and 70°F (21°C), the reading will generally be correct for all products. Above and below this range there will be an error in the same direction as that produced with the conven­tional, uncompensated type of refractometers, but of about half the magnitude. The tempera­ture correction necessary for specific non­aqueous solutions will be furnished upon request.
8.0 USE OF TS METER REFRACTOMETER CONVERSION TABLES
If using the direct reading scales in the instru­ment and the conversion tables in this manual, (pages 9-14), remember that these measurements are specific for two types: plasma/serum and urine. The scales are NOT a valid measure of other samples without conversion.
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9.0 SERUM TOTAL SOLIDS AND WA TER MEASUREMENTS: SERUM AND URINE SOLUTE CONCENTRATIONS BASED UPON WATER
The convenience of estimating serum total solids with the TS METER Refractometer makes it useful not merely as an absolute and comparative measure of solids, but also as a measure of serum water. The concentration of water in serum (g/100 mL) and percent water in serum (g/100 mL) are given in the conversion table. Individual solute concentrations are readily converted from a serum to a serum water base; e.g., serum sodium, 146mEq/L of serum; TS METER reading, 9.0%; water concentration (from conversion table), 93.3 gm/ 100 mL or .933 kg/mL; and 146/.933 = 156 mEq of sodium/kg of serum water. If desired, the concentration of sodium in extra-cellular fluid, corrected for Donnan ratio, can be found from 156 x 0.95 = 148 mEq of sodium/kg of water of extracellular fluid.
Similarly, the conversion t ables shown at the end of this manual can help you determine urinary water as a percent or as concentration; e.g., urine reading on the specific gravity scale,
1.035; actual urine solids, 8.5%; urine water,
91.5 gm/100 gm or 94.5 gm/100 mL.
10.0 ESTIMATION OF PROTEIN BY REFRACTOMETRY
Although it is practical for many purposes to es­timate total serum protein by refractometric meth­ods, certain points should be kept in mind.
Measurement of serum protein depends on the high correlation between refraction and total solids. In the computation of the table, the two non-protein solids fractions are considered: ultrafiltrates proportional to water concentra­tion, and lipids (and other high molecular weight compounds proportional to the protein concentration). T otal non-protein solids is the sum of these two fractions. While the accuracy of estimate of total solids is approximately +.1 gm%, that of protein may be somewhat less This follows from the variability of concentra­tion of such serum solutes as glucose, urea, salts and lipids, especially in certain disease
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states. The TS METER Refractometer reticle and
conversion tables (pages 9—14) are scaled primarily for the measurement of serum or plasma protein and must not be applied indiscriminately to other protein-containing solutions. Pure water set and read at scale zero is sufficient to check the TS METER since the instrument is itself standardized optically . If further testing is desired, it is advisable to use an aqueous solution whose refractive index is well known from concentrative conversion
15,16
tables.
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The numerical relationship between refraction and total serum protein depends upon the factor used to convert human serum protein nitrogen to protein. The recommended factor, 6.54, provides values from protein concentration essentially consistent with serum total solids and nonprotein solids.
14,17
T otal protein in urine and certain other fluids can be measured refractometrically from the determination of total solids in the fluid before and after protein has been removed by heat.
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11.0 ESTIMATION OF SPECIFIC GRAVITY AND TOTAL SOLIDS OF URINE BY REFRACTOMETRY
12.0 ESTIMATION OF CONCENTRTION OF OTHER BODY FLUIDS AND OF PURE SOLUTIONS
In part because the TS METER Refractometer measures total solids in urine to an accuracy of
+
.lgm %, and since it requires a change of ca. .25gm % in total solids to change specific gravity by ca. .001 units, optical urinometry is excellent for clinical measurement of specific gravity. The reticle of the TS METER Refracto­meter, and conversion tables (pages 9–14) are scaled for human urine. Few normal human urines exceed 1.035; much higher values suggest the presence of unusual solutes in the specimen which have high specific gravity increments per osmol (e.g., glucose, sucrose, protein, radiopaque iodine compounds, sodium sulfate, etc.). Because refraction correlates relatively poorly with specific gravity, in contrast to total solids in solutions of variable composition, especially at high concentrations, refractometric readings will not provide reliable extrapolation of urine specific gravity to values in excess of 1.035. Such extrapolation is particularly not advised in animals which may produce exceedingly hypergravic and hypersteric urines. Urinary total solids or the refractive index difference as a measure of absolute and relative concentration is a preferred method.
Fluids such as pancreatic juice, saliva and prostatic fluid may also be analyzed refracto­metrically. However , interpretation of such refractions should not be made without reference to suitable standardization; it may be inappropriate to use the TS METER scale of serum directly for this purpose.
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13.0 REFRACTION SCALE
The refraction or refractive index difference scale (0-320) allows a check of a single solute concentration to be made in seconds.
Extensive data is given in the Manual “T ables of Properties of Aqueous Solutions Related to In­dex of Refraction,”16 (Cat. No. 10403), that tabu­lates the refraction of particular aqueous solu­tions against osmotic pressure, specific gravity, molarity , solute concentration and water concentration.
In addition, graphs for other solutions can be prepared by plotting points for several known concentrations against refraction.
If human urine being analyzed is hypogravic (spe­cific gravity less than 1.017) under concentration test, estimation of specific gravity by refracto­metric means is exceptionally accurate, regard­less of variation in relative composition (e.g., salt, urea). As a clinical measure of such renal dys­function, therefore, optical urinometry is an ex­cellent diagnostic tool. With progressive renal failure, diverse measures of urinary concentra­tion such as osmolality, tot al solids, specific grav­ity, and refraction converge to narrow ranges which are similarly pathognomonic.19 The fact that these measures diverge in the maximally con­centrated urine of normal individuals has no bear­ing on pathologic or diagnostic significance.
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14.0 INFORMATION OBTAINABLE
WITH THE REICHERT TS METER
PLASMA OR SERUM, 68°F (20°C).
1. T otal Solids % Composition by W eight, (TS%). Read instrument scale of protein and use Conversion T able.
2. Water % Composition by Weight, (Water %). Subtract total solids % composition above from 100%.
3. Tot al Solids Concentration, grams 100mL., 68°F (20°C), (CTS,g/100mL). Read instrument scale and use Conversion T able.
4. Water Concentration, grams/100mL, 68°F (20°C), (Cw, g/100mL). Read instrument scale and use Conversion T able.
5. Specific Gravity , 68°F (20°C), (D Read instrument scale and use Conversion T able.
6. Protein Concentration, grams/100mL, 68°F (20°C), (CPR, g/100mL). Direct scale reading with TS Meter .
7. Concentration T otal Solids relative to water (CTS ÷ Cw, x 102 = grams/100g water).
REFRACTIVE INDEX, 20°C. (REFRACTIVE INDEX AND REFRACTION)
20/20
sp. g).
URINE, 68°F (20°C).
1. Specific Gravity, 68°F (20°C), (D
20/20 sp.
g). Direct scale reading, TS Meter.
2. Total Solids % Composition by Weight, (TS%). Read instrument scale and use Conversion T able.
3. Water % Composition by Weight, (Water %). Subtract total solids % composition above from 100%.
4. Total Solids Concentration, grams/ 100mL, 68°F (20°C), (CTS, g/100mL). Read in­strument scale and use Conversion Table.
5. Water Concentration, grams/100mL, 68°F (20°C), (Cw, g/100ml.). Read instrument scale and use Conversion T able.
6. Concentration Total Solids relative to water (CTS = Cw X 102 = grams/100g water).
1. Refractive Index, (n), of aqueous or other solutions, 68°F (20°C). With the TS Meter read the instrument scale and use the Conver­sion T able.
2. Refractive Index Difference x 104, between aqueous solutions and water, 68°F (20°C), also called Refraction, r = (n -
1.3330) 104. Direct scale reading with TS Meter.
3. Concentrative Properties of aqueous, such as % composition, Concentration, Specific Gravity, Freezing Point Depression, V iscosity and Electrical Conductivity may be obtained with special Concentrative, Conversion T ables Based upon refractive index and refraction. Direct scale reading of refraction with TS METER.
891011121314
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REFERENCES

1. Adolph, E. F., 1943, Physiological Regulations, Lancaster, p. 219.
2. Blohm, G. J., 1918, Upsala lakf. forf. 23,
283.
3. Rubini, M. E. and A. V. Wolf, 1957, J. Biol. Chem. 225 (2) 869.
4. Gettler, A. O. and W. Baker, 1916, J. Biol. Chem. 25, 11.
5. Sunderman, F. W ., 1944, J.Biol. Chem. 153 (1) 139.
6. Sunderman, F. W. and F. Boerner , 1950, Normal Values in Clinical Medicine, Saunders, Philadelphia.
7. Barry, K. G., A. W. McLaurin, and B. L. Parnell, 1960, J. Lab and Clin. Med. 55, 803.
8. Drickman, A., and F.A. McKeon, Jr., 1962. Am. J. Clin. Path. 38, 392.
9. Wolf, A. V., J. B. Fuller , E. J. Goldman, and T. D. Mahony, 1962. Clin. Chem. 8, 158.
10. Wolf A. V., 1955-1964. Personal Communications to American Optical Co.
11. Holmes, J. H., P. James, and H. Bivens, Use of the TS-METER Meter in the clinical laboratory with special reference to the serum protein determination. Unpublished report to American Optical Co.
12. Holmes, J. H., P. James, and H. Bivens, Use of Refractometric methods for determining urine specific gravity in the clinical laboratory. Unpublished report to American Optical Co.
13. Remp, D. G., and V. Schelling. 1960. Clin. Chem. 6, 400.
14. Chiaraviglio, E. C., A. V. Wolf, and P. G. Prentiss, 1963. Am. J. Clin. Path. 39, 42.
15. Handbook of Chemistry and Physics, 52nd ed., Chemical Rubber Publishing Co., 1971.
16. Tables of Properties of Aqueous Solutions Related to Index of Refraction, American Optical Co. 1964, 1974. Cat. No. 10403.
17. Sunderman, F. W. Jr ., F . W . Sunderman, E. A. Falvo, and C. J. Kallic, 1958. Am. J. Clin. Path. 30, 112.
18. Wolf, A. V ., 1962. T echnique Manual, Workshop on Urinalysis and Renal Function Studies, Council on Clin. Chem. of Am. Soc. Clin. Path., p. 39.
19. Wolf A. V., 1962. Am. J. Med. 32, 329.
20. Wolf, A. V., 1966. Aqueous Solutions and Body Fluids, Harper and Row, New York.
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Reichert Analytical Instruments
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©2003 Reichert, Inc. Depew NY, USA 10400A-101 Rev. D
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