CA1226794A - Stabilized multiparameter control product - Google Patents

Abstract

Abstract Compositions and methods for stabilized serum based multiparameter standards and controls. Stabilization of the chemistry controls and standards requires the addition of a plexiform stabilizing means such that the final percent concentration is in the range of 2%-8%. The most preferred embodiment employs lactose as the plexiform stabilizing means and chloremphenicol as a microbial growth inhibitor.

Description

AL

STABILIZED MULTI PARAMETER CONTROL PRODUCT

Field of the Invention _ ._ S This invention relates to the field of multi parameter control reagents useful for calibrating clinical chemistry analyzers and for manual testing methods. In particular, it relates to clinical control formulations and methods for producing stabilized clinical control reagents suitable for lyophilization or spray drying.

Back round of the invention Chemistry analyzers occupy a preeminent position in the clinical environment as they provide significant data regarding the diagnosis and treatment of patient illnesses. accordingly, there has been a concerted effort by many investigators to develop automated and manual methods for the determination and qualification of constituents in body fluids such as acid phosphates, ala nine amino~ransferase, albumin, aldolase, alkaline phosphates, alpha-hydroxybutyrate dehydrogenase, aimless, aspartame aminotransferase, bicarbonate, bilir~bin direct, bilirubin total, blood urea nitrogen, calcium total, carbon dioxide, chloride, total cholesterol, cholinesterase, courteously, creative Cannes, creatinine, dioxin, gamma glutamyl-transferase, globulin, glucose, hdl-cholesterol, iron, lactate dehydrogenase-l, lactate dehydrogenase-p, lactic acid, Lopez, lithium, magnesium, osmolality, phenylalanine, phosphorus, potassium, salicylate, sodium, To, To uptake, To, total iron binding capacity TIC total lipids, total protein, in-glycerides, UIBC, and uric acid, to name a few.

Both the manual and automated methods available for each one of the above analyzes and constituents have in common the requirement for a control reagent whereby the OARED

procedures and other variable parameters may be checked to ensure accuracy of the testing method or instrument It is an object of the present invention to provide such a control reagent useful for automated and manual methods.

Although such multi parameter control reagents are known and include the Ortho~ Normal Control Serum-Assayed, available from Ortho Diagnostic Systems Inc., Raritan, New Jersey, all such reagents have suffered, albeit to different extents, from stability related deficiencies.
Specifically, these reagents have heretofore been characterized by their limited shelf lives, both in their typically dry storage state and in their liquid testing format. For instance, COY levels and enzyme constituents are particularly prone to decomposition and deactivation.
Accordingly, the clinical researcher has heretofore been generally forced to prepare fresh chemistry controls for each testing batch or work shift as appropriate. The unfortunate consequence of this requirement is the increase in potential errors as well as the concomitant costs incurred.

It is another object of the instant invention to provide clinical chemistry controls having greatly increased stability whereby many of the above related stabilization problems are obviated.

Conventional attempts to prolong stability of chemistry controls and related reagents include reduction of the controls to a dry format which is then typically refrigerated at approximately 4C. Often, the dry format is accomplished by lyophilization rather than spray drying as the former operates at far lower temperatures than the latter. It is known that heat disadvantageously aggravates degradation of analyzes and proteins in OARED

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general and accordingly, cold processes such as freeze concentration and freeze drying, are generally preferred.
Although stability in a lyophilized state is enhanced over that associated with the aqueous format, significant losses in constituent activity levels of known chemistry controls are still incurred It is a yet further object of the present invention to provide methods and reagents which provide the required stability levels for substantially all constituents and analyzes in a clinical chemistry control reagent and which is suitable for storage in a dry state as attained by either lyophilization or spray drying methods.

the stability problems associated with such reagents has not however gone unrecognized and numerous attempts have been made to reduce these problems. Typically, however, these conventional methods have been limited to a very limited number of constituent (parameter) containing controls. Further and is the case with hemoglobin controls, these single constituents have usually been inherently stable. See underman et at., "A Lyophilized Hemoglobin Control Prepared From Stroma-free Hemolysates", Olin. Chum. 26/2:305-308 (1980); US. Patent No.

2,433,299 to Sewers; and Proxy et at., "reparation of Lyophilized Abnormal hemoglobin Controls For A Cellulose Acetate Electrophoresisn, A.J.C.P~ 74/1:64-67 (1980).

Other attempts at obtaining increased stablest have relied on surfactants such as Briton X-100, Proxy Pi at., Jan Optically Clear Hypercholesterolemic Hypertriglyceridemic Quality Control Material Prepared from Animal Lipid Sources", Clinical Chemistry, Vol.
27/3:468-471 (1981) or complex mixtures of mucilaginous gums, polymers containing hydroxy groups, hydroxy alkylamine buffers, ethylene Damon tetraacidic acid, bovine serum albumin, polyvalent anion salts, or * trade mark.
~RD-50 Jo Jo sulfhydryl compounds as taught by Douche in US. Patent Nos. 3,413,198i 3,527,331 and 3,539,450.

It is an object of the present invention to obtain superior stabilizing results by employing less complex stabilizing means.

Must in US. Patent No 3,133,001 discloses the stabilization of a select group of enzymes: ayatollahs, aldolase, and 17-hydroxy dehydrogenase, by the addition of sucrose, lactose or maltose at or near their saturation levels. Bonder man et at. claim stabilization of glucose-6-phosphate dehydrogenase by the addition of sucrose and lactose in an article in Clinical Chemistry Vol. 25/5:
815-816 (1977) entitled "Lyophilized Control for Determination of Glucose-6-phosphate Dehydrogenase Activity in Hemolysates".

It is still another object of the present invention to avoid adding stabilizing agents in concentrations which provide substantial, deleterious effects on other parameters normally found within a multi parameter clinical chemistry control reagent.

Multi et at. in US. Patent No. 4,127,502 attempt to improve optical clarifies of serum derived compositions by the addition of various sugars including minutely and lactose. Table II of that patent indicates, however, that minutely and lactose in fact failed to measurably reduce the optical density of the compositions. Hamburger et alto in US. Patent No. 4,056,484 disclosed the stabilization of a coagulation control by the addition of a protective gas containing at least 5% carbon dioxide in addition to various combinations of lactose succors or lactose.

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. 5_ It is a yet further object of the present invention to provide the correct methods for employing certain of the aforementioned sugars as stabilizing agents in multi-parameter control reagents to obtain substantially increased stability and optical Clara It is a still yet further object of the present invention to provide methods for stabilizing other types of control reagents such as coagulation control reagents, plasma resin control reagents, isoenzyme control reagents and the like.

Summary of the Invention In accordance with the principles and objects of the present invention, there are provided multi parameter control reagents having their inherent levels of analyzes and other constituents ("control" as used herein) or having specified amounts of analyzes added to a base material (a "standard" as used herein) in addition to plexiform stabilizing means selected from the group of reducing monosaccharide and reducing disaccharide sugars, present in the final volume concentration in the range of about 2~-8~. The preferred plexiform stabilizing means I and the reducing sugars are maltose, minutely syllables and lactose, the latter most being the most preferred on the basis of efficacy and economy. If cost is an insubstantial factor, however, then the penultimate sugar of those listed above is the most preferred.
The thusly stabilized reagent, whose source is a body fluid, may be advantageously adjusted by the addition or deletion of constituents so that the reagent contains those constituents, analyzes, and protein levels desired in accordance with the product's purpose and end use.

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-pa-Thus the present invention provides a serum based, multi-parameter control reagent comprising: (a) substantially normal blood having all normally present proteins, analyzes and enzymes but with substantial all red blood cells, white blood cells and platelets removed; and (b) plexiform stabilizing means selected from the group consisting of reducing monosaccharide sugars and reducing disaccharide sugars present in the range of about 2%-8% final volume percent whereby activity of substantially all proteins, analyzes and enzymes in solution is maintained for at least 10 days.
In another aspect the invention provides a stabilized multi-parameter control reagent comprising an aqueous solution of protein and nonprotein analyzes and plexiform stabilizing means selected from the group consisting of maltose, minutely, syllabus and lactose, said plexiform stabilizing means present in a concentration sufficient to obtain substantially increased stability over that without such plexiform stab-living means and wherein said concentration is further optic mixed to preserve maximum enzyme activity.
In still a further embodiment the invention provides serum based, multi parameter standard comprising: (a) sub-staunchly normal blood having substantially all red blood cells, white blood cells and platelets removed and having proteins, analyzes and enzymes present in known amounts;
and (b) plexiform stabilizing means selected from the group consisting of reducing monosaccharide sugar sand reducing disaccharide sugars present in the range of about 2~-8%
final volume percent whereby activity of substantially all proteins, analyzes and enzymes and solution is maintained for at least 10 days.
in another embodiment the invention provides a stabilized multi parameter standard comprising an aqueous solution of -5b~

known levels of protein and nonprotein analyzes and plucks-form stabilizing means selected from the group consisting of maltose, minutely, syllabus and lactose, said plucks-form stabilizing means present in a concentration sufficient to obtain substantially increased stability as compared to a standard without such plexiform stabilizing means.
In another embodiment the invention provides a method for stabilizing a multi parameter serum based control reagent obtained from substantially normal blood having all normally present proteins, analyzes and enzymes but with sub Stan-tidally all red blood cells, white blood cells and platelets removed comprising adding plexiform stabilizing means selected from the group consisting of reducing monosaccharide sugars and reducing disaccharide sugars in an amount sufficient so that the final volume percent of the plexiform stabilizing means is in the range of about I

Brief Desert lion of the-Drawings and Tables P

Further understanding of the principles of the present invention may be had by reference to the Figures and Tables wherein FIGURE 1 shows a comparison of LDH units per liter of a 14 day liquid stability study in controls having various lactose concentrations;

FIGURE 2 graphically depicts the differences in absorbency of reagents containing different lactose concentration solutions over time;

15 FIGURE 3 shows creatinine levels over time in reagents having various lactose concentrations;

Table 1 shows lyophilization losses for various analyzes in normal and abnormal levels for controls and for the quality control stabilized product of the present invention;

Table 2 shows wet and dry interference studies in a control product with normal analyze levels;
Table 3 presents the same data as Table 2 but for products with abnormal levels;

Table 4 presents wet accelerated stability data for various constituents;

Table 5 presents data for wet accelerated stability studies for abnormal controls listing various constituents;

~RD-50 Jo Table 6 presents reconstitution stability recovery data for normal controls; and Table 7 presents reconstitution stability recovery data for products with abnormal analyze levels.

Detailed Description of the Drawings, Tables and Best Mode ---- - ----- -The reagent products of the present invention are useful for manual methods and automated methods in chemistry analysis and in particular may be used with multi channel chemistry analyzers such as the Adam from Du Pont or the SMACK from Technic on. The control reagent material is a patient-like sample characterized by a range of values for each constituent, enzyme and analyze found therein. The closer the control material simulates a patient's sample in providing all unknowns at their proper revels and in appearance (i.e., optical clarity and the like), the more useful it is. Similarly, the longer it presents such characteristics, i.e. the greater the time period it can hold the activity level for each constituent in a stable fashion, the more valuable the control material.

As has been previously intimated, there has heretofore been great difficulty in stabilizing such a multi parameter product and in particular, stabilizing various analyzes such as CPK, LDH, CO,, bilirubin, among others without deleteriously affecting other constituents or the tests therefore With the addition of the plexiform stabilizing means of the present invention, these analyzes or constituents have now been stabilized and may be maintained in solution for at least ten days. In dry form, i.e. substantially all water has been removed, this stability period is greatly increased.

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For instance, and with reference to Figure 1, lactate dehydrogenase ~LDH), typically measured in units per liter, has heretofore generally been present in the initial solutions at approximately 190 units per liter.
As may be apparent by reference to the control material (no plexiform stabilizing means added), typically a loss of approximately 20 units per liter was sustained upon lyophilization and reconstitution. Previously, such losses could not be avoided and it was hoped that they recurred on a consistent basis. Figure 1 shows the affects of the addition of lactose as a plexiform stabilizing means in various concentrations. As may be readily apparent, there is a significant increase in LDH
stability over time.
Additional surprising affects were encountered with the addition of lactose and include the reduced absorbency with increasing lactose concentrations See Figure 2. In fact, it has been discovered that the reconstituted materials typically exhibit two or more times clearer optical density readings than without the addition of such plexiform stabilizing means.

However, and as may be seen by reviewing Figure 2, there is little absorbency gain with an increase of the concentration from 6% to 8% and indeed, as the percentages increase to 8% and beyond, other disadvantages become more apparent. For example, the high percentage levels become quite significant with respect to the limits of volubility and thus problems with solubilizing the sugar. Further, and as may be expected with the addition of sugar to any media, an increase in bacterial growth may be expected and this is only exacerbated with higher concentration levels.
Accordingly, it may be desirable to add an antimicrobial substance in sufficient concentration to effectively OARED

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reduce or eliminate the possibility of microbial growth and resultant contamination. Such an antimicrobial should, of course, not interfere with any of the other constituents or with the testing methods. One preferred antimicrobial is chloremphenicol present, in a final concentration in the range of about 10 Pam to 1000 Pam.
In this regard, it should be noted that the choice of plexiform stabilizing means will have a substantial affect on the types and amount of microbial growth to be 10 encountered. For instance, sucrose is susceptible to fermenting bacteria and yeasts which typically more difficult to control. Indeed, are virtually impossible to eliminate by filtering due to the presence of spores Rand other methods either destroy the product or are 15 prohibitively expensive Conversely, lactose fermenters are much more susceptible to antimicrobial and thus may be more easily controlled. Accordingly, lactose is greatly preferred over sucrose as a plexiform stabilizing means.
With the addition of the plexiform stabilizing means of the present invention, it has been noted that there is a positive bias affect on creatinine as shown in Figure 3.
Indeed, with the addition of lactose in concentration 25 ranges of approximately 4% to 8%, an increase in creatinine levels of approximately lug per deciliter results. Although this type of an interference is preferably to be avoided, it is not significantly deleterious as the positive bias merely puts the 30 creatinine levels at the top of the normal range.
Similarly, as may be expected whenever material is added, the osmolality increases as does total protein levels when tested by refractometers. Accordingly, and in order to limit these undesirable affects, the high concentration 35 ranges and in particular those significantly in excess of I are to be avoided. It is also noted that if sucrose is chosen as the plexiform stabilizing means, then any of oared the glucose oxidize procedures will in effect actually be 79f~

measuring the amount of sucrose, an invert sugar, in addition to the levels previously present. Thus, glucose is a sugar to be preferably avoided.

The few, limited disadvantages incurred by addition of the plexiform stabilizing means of the present invention are, however, far outweighed by the numerous and significant advantages gained thereby. For instance, and in addition to the aforementioned stability increases, many constituents previously unstable or requiring augmentation, may now be measured over significant periods of time in their normal ranges without the addition of extraneous materials. For instance, many of the enzymes, such as CPK, incurred 50% or greater losses in the lyophilization step alone. These enzymes are now present in stable configuration and within normal levels even following lyophilization. Similarly, COY is maintained in solution for significantly greater periods of time thus also limiting the otherwise concomitant rise in phi In fact, CO has been such a difficult constituent to measure heretofore, that many of the instruments presently available are incapable of detecting COY with any great accuracy. It should be noted that bacterial growth and the like can result in the disadvantageous alteration of COY levels.

Another unexpected surprise concerns the constituent bilirubin. This component is normally stable only a high pus and accordingly, one would not expect stability at physiological pi (the pi of the control reagent; in Volvo it is constantly being removed by the liver), however, with the addition of lactose, bilirubin becomes stabilized.

Still other unexpected results were discovered. As expected, with the addition of the plexiform stabilizing OARED

--if--means, the actual freezing point is depressed.
Depression is generally associated with slower freezing rates however, faster freezing of the product of the present invention has been observed. The inventor attributes this in part to the loss of the eutectic point plateau. In conjunction with this, it has been noted that without the addition ox lactose, previous lyophilization methods on otherwise similar products resulted in a 2-1 ratio of powder versus crystalline cake configurations.
The explanation of this phenomenon is as yet, totally unknown. With the addition of lactose, however, and the resultant faster freezing, it has been noted that all of the cakes are crystalline in nature and upon reconsti-tuition, the absorbency measured on a vial to vial basis is far more uniform.

Although the inventor does not wish to be held to a theory, he has postulated that the plexiform stabilizing means of the present invention acts to provide a three dimensional structure which assists in holding the analyzes in juxtaposition vis-a-vis one another while water is sublimed. Thus, water may be more quickly removed during lyophilization and more quickly reinserted into the structure during reconstitution. Thus, the plexiform stability means results in significant cold stability. Lyophilization with or without freeze concentration has been predominantly used to remove water as it operates at low temperatures, typically associated with decreased problem liability, disassociation, degradation and the like, with the plexiform stab living means of the present invention, however, like stability advantages are also associated with increased heat. Thus, the water may now also be removed with "hot" means (less than boiling) such as by spray drying techniques, and the like, commonly known ~RD-50 Another important characteristic of the plexiform stabilizing means of the present invention is the associated reducing nature In particular, the monosaccharide and disaccharide reducing sugars are preferred and of those, maltose, minutely, syllabus and lactose are employed in the preferred embodiments. If cost is not a factor regarding the constitution of the controls, then syllabus is the most preferred reducing sugar as it subjectively produces the most advantageous results with limited or insignificant disadvantages or affects. Syllabus is, however, an expensive commodity as compared to other preferred reducing sugars and in view of modern economic trends, lactose is employed in the most preferred embodiment as it closely parallels syllabus in lo all categories except price. Indeed, the selection of the proper sugar for the plexiform stabilizing means is critical in order to obtain the advantages of a stabilized multi parameter control reagent. For instance, isomaltose provides some increased stabilization as well as and many of the other advantages previously discussed, however, with this sugar, all saccarogenic aimless tests are eliminated. Typically, non reducing sugars do not work well with respect to stability and accordingly, those sugars are to be preferably avoided.
Other surprising results include the increased reconstitution speeds which are obtained with plexiform stabilized multi parameter control reagents. Presumably, this may be a result of the three-dimensional structure imposed by the plexiform stabilizing means which not only assists water in resurrounding the constituents, but also increases the speed with which proteins unfold. the sugar probably also provides an environment which starkly hinders motion, particularly that of the larger molecules, and thus reduces degradation otherwise resulting therefrom The above, however represents OARED

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theoretical conjecture by the inventor and is not to be construed as a limitation hereof.

Table l presents data depicting lyophilization losses, obtained by comparing a wet value against the value obtained of a reconstituted, lyophilized material. The control material did not have lactose added thereto while the QCS quality control system) did have the plexiform stabilizing means, lactose, added thereto The normal and abnormal columns represent materials having the normal range of constituents and elevated ranges of contituQnts respectively. As may be readily apparent, the loophole-ration losses incurred with the QCS product are significantly less than those incurred in the control. It should be noted, however, that the Asp of 60% is within the accuracy limits of testing since at the normal levels, this only represents a reading difference of l unit.
Thus, the 60~ could just as easily have been 0%.

Tables 2 and 3 present data concerning interference studies for normal and abnormal multi parameter reagents The QCS product had the plexiform stabilization means, lactose, added and the control product did not. The wet column presents values obtained prior to lyophilization while the dry column presents values obtained following reconstitution of a lyophilized material.

Tables 4 and 5 depict accelerated stability data for a lyophilized normal and for a lyophilized abnormal QCS
product respectively. The control column presents the mean of measurements made at 0 time while the test sample columns represent the mean of samples measured following 26 days of storage at 37C. This period represents the equivalent of approximately 3 years when stored at +5C.
Typically, variations around 100% may be attributable to the error ranges normally associated with readings of the particular constituent.
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Similarly, Tables 6 and 7 represent reconstitution stability recovery data for normal and abnormal lactose stabilized multi parameter control reagents.

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Lyophilization Losses Constituent Control QCS
Normal Abnormal Normal Abnormal cup 80% 13% 60% 0 Alp 32~ 35% 0 5 Amy I 3% 0 0 GOT 6% I 0 1 GUT 18~ 17~ 0 4 OK 47% 23~ 0 1 LDH 4% 4% 0 0 Lip 0 18% 0 0 I
CQ, I 64% 28~ 35 OARED

Lo WET DRY
QCS QCS Control Acid Pros 1.0 1.0 0.4~1) 0.2(1) Ask Pros 74 115 96 78 Aimless 50 55 52 51 ~ilirubin 045 ~46 0.5 0.5 BUN 12.7 13.5 14 12.4 Cay 9.1 9.2 9.6 9.2 Cholesterol 161 134 Chloride 102 107 111 106 Albumin 4.05 4022 4,3 4.1 Creatinine 2.1 0.9 2.3 0.9 glucose 90 92 93 86 Iron 102 108 110 112 Lopez 24 20 17 17 My 2.1 2.5 2.3 2.5 p 3.4 3.2 3.6 3.
Sal 9.6 9.6 10.0 9.8 Triglycerides 102 86 TO 6.3 6.0 6.6 5.9 Us I 4.3 5.4 4.4 No 134 137 143 135 R 4.2 4.3 4.6 4.2 H 0.9 0.9 0.94 0.9 To - - 1.66 1.16 TAO - - 1.06 1.10 Courteously - - 9.12 9.4 Dioxin _ _ 3283 03i364 Phenylalanine - - 1.9 1.8 1 - citric acid stabilizer tablet added in addition.

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Abnormal Interference Stud its WET DRY
US Control QCS Control Acid Pros 11.0 11.2 11.0 8.6 Ask Pros 260 265 211 171 10 Aimless 392 386 400 376 Bilirubin 6.5 6.6 6.7 6.5 Cay 12.5 12.7 13.2 12.5 Cholesterol 123 108 124 93 Chloride 116 116 120 116 Albumin 3.1 3.1 3.2 3.1 Creatinine 9.6 8.5 10~4 8.3 Glucose 310 312 324 306 HOD

Iron 237 243 259 244 I.ipase 69 74 68 61 My 4.5 4.9 4.7 5.0 P 7.5 Ed 8.2 7.4 Sal 26 26 27.1 26 30 Triglycerides 70 54 82 63 TO 4.9 4.4 5.1 4.3 US 9.5 8.3 10 8.3 No 147 148 160 150 K 7~15 7.10 702 6.8 35 H 2.4 2.4 2.46 2.5 Co, 17 17 11 6.2 T" Z06 180 T 3.95 3.3 TAO 1.92 1.98 40 Courteously 56.4 47.2 Dioxin 2.61 2.2 Phenylalanine 7.1 6.6 OARED

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TABLE _ ACCELERATED STABILIT~-VATA- HEFT
Lot Number: ARROWHEAD (NORMAL) Test samples stored at:
37 C. for 26 days Constituent -25C Control Test Samples % Recovery x xxT/xC x 100 Total Protein 6.52 6~56100.6%
gel Albumin 4.51 4.4398%
gel Bilirubin (Direct) 0.10 0.10 100 mg/dl Bilirubin (Total) 0.54 0.55 102%
mg/dl Calcium aye 906 100 mg/dl Chloride 105 105100%
meekly COY 16.5 1~.689%
meekly Creatinine 2.4 2.4100%
mg/dl Glucose 96 80 94%
mg/dl Magnesium 2.2 2~1 95.5%
mg/dl Phosphorus 3.47 3.50101 mg/dl Potassium 4~81 4~6697%
meekly Sodium 143.6 143.7100%
meekly Urea Nitrogen 14.5 14.499%
mg/dl Uric Acid 5.6 5.6100%
mg/dl Acid Phosphates 0.70 0.65 93%
ISLE
Awoke. Phosphates 96 85 88,5 ISLE
Aimless 51 49 I
ISLE
Creative Cannes 204 139 93%
ISLE

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TALL (Continued) ACCELERATED-STABILITY-DATA SHEET
__ __ Lot Number: ARROWHEAD (NORMAL Test samples stored at:
37 C. for 26 days Constituent -25C Control Test Samples % Recovery x xxT/xC x 100 Go 36 35 97%
ISLE
Alpha HOD 247 250 101%
ISLE
LDH 146 137 94%
ISLE
Lopez 12 11 92 ISLE
AT So 50 94 ISLE

ISLE
Cholesterol 165 100 mg/dl Triglycerides 103 99 mg/dl Total Lipids 896 101 To mg/dl 8.38 8.5 101 TO ng~ml 1.44 1.69 117 To Uptake 38.0 38,4 101%
Courteously 10.50 10.37 99%
ug/dl Dioxin 0~99 0.92 93%
ng/ml Iron 113 115 102%
ug/dl TIC 350 397 113%
ug/dl Lithium 0.99 0.98 - 99%
meekly Phenylalanine mg/dl Salicylate 9.9 9.8 99%
mg/dl Protein Electron 6.60 S.45 98%
pharisees Albumin alpha 1 0.39 0.39 100%
Alpha 2 1.33 1.3~ 105~
Beta 1.52 1039 91%
Gamma 1.56 1.79 115%
A/G Russia 1.302 95%

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~20-ACCELERATED STABILITY DATA SHEET
Lot Number: 99A0216RD-l(ABNORMAL) Test samples stored at:
37C for 26 days Constituent -25C Control jest Samples % recovery x xxT/xC x 100 Total Protein 5.04 5~08101%
9/~1 Albumin 3.31 3.30 99.7 gel Bullhorn 0.31 0.36 116 (Direct) mg/dl Bilirubin (Total) 6.80 6.77 99.6 mg/dl Calcium 13~1 13.1100%
mg/dl Chloride 114 115 101%
meekly CO, 11.2 9~6 86 meekly Creatinine 10.4 10.5101%
mg/dl Glucose 324 315 97%
mg/dl Magnesium 4.7 4.6 98 mg/dl Phosphorus 8~02 8.04 100 mg/dl Potassium 7.34 7.26 99%
meekly Sodium 159.4 159.8100%
mEqJL
Urea Nitrogen 53.1 53.7 101 mg/dl Eric Acid 9.9 go 9 mg/dl Acid Phosphates 11~20 11.13 99%
ISLE
Ask. Phosphates 237 224 94.7%
ISLE
Aimless 412 406 98.5%
ISLE
Creative Cannes 751 731 97%
ISLE

ISLE
Alpha HOD 542 554 102 ISLE
1 - Control represents measurements at 0 time.

TABLE-5 (Continued) ACCELERATED SYBIL DATA-SHEET
Lot Number: 99A0216RD~l(ABNORMAL) Test samples stored at:
37C for 26 days Constituent -25C Control Test Sample % Recovery x xxT/xC x 100 LDH 482 470 97.5 ISLE
Lopez 68 64 94 ISLE
AT 424 394 93%
ISLE
AT 116 103 89%
ISLE
Cholesterol 122 121 99%
mg/dl Triglycerides 78 80 103%
mg/dl Total Lipids 888 851 96%
To 20.34 20.52101 mg/dl To 3.78 3.91103 ng/ml To Uptake 71~0 68.196%
Courteously 56.02 54.g098%
ug/dl Dioxin 2.78 2.80101%
ng/ml Iron 252 253 100 ug/dl ugjdl Lithium 2.60 2.60100%
mEqJL
Salicylate 27.1 27.0 99.6%
mg/dl OARED

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It is to be understood that all of the above discussion is equally applicable to chemistry standard reagents which differ from chemistry controls in that levels or values of constituents in the former is known while in the latter, the constituents may be present anywhere within a given range of values or levels. It is Jo be further understood thaw stability as used herein shall mean later measured activity which is at least 90% of the original measured activity. It is also applicable to other types of clinically useful control reagents such as lipid and cholesterol controls, isoenzyme controls, and coagulation controls, and the like.

As may be readily appreciated by those skilled in the various alterations and substitutions of the alone-mentioned constituents may be made without deviation from the spirit and scope of the instant invention.

oared

Claims (59)

1. In the Claims:
   
   l. A serum based, multi-parameter control reagent comprising:
   
   a) substantially normal blood having all normally present proteins, analytes and enzymes but with substantially all red blood cells, white blood cells and platelets removed; and b) plexiform stabilizing means selected from the group consisting of reducing monosaccharide sugars and reducing disaccharide sugars present in the range of about 2%-8% final volume percent whereby activity of substantially all proteins, analytes and enzymes in solution is maintained for at least 10 days.
2. 2. The reagent of Claim 1 wherein the plexiform stabilizing means is selected from the group consisting of maltose, mannitol, cellobiose, and lactose.
3. 3. The reagent as provided in Claim 1 wherein the plexiform stabilizing means is lactose.
4. 4. The reagent as provided in Claim 3 wherein the lactose is present in a final concentration of about 6%.
5. 5. A stabilized multi-parameter control reagent comprising an aqueous solution of protein and nonprotein analytes and plexiform stabilizing means selected from the group consisting of maltose, mannitol, cellobiose and lactose, said plexiform stabilizing means present in a concentration sufficient to obtain substantially increased stability over that without such plexiform stabilizing means and wherein said concentration is further optimized to preserve maximum enzyme activity.
6. 6. The reagent as provided in Claim 4 further comprising an antimicrobial.
7. 7. The reagent as provided in Claim 6 wherein said antimicrobial agent is chloremphenicol.
8. 8. The reagent of Claim 5 further comprising an antimicrobial agent for substantially inhibiting microbial growth.
9. 9. The reagent as provided in Claim 8 wherein said antimicrobial agent is chloremphenicol.
10. 10. The reagent as provided in Claim 1 in lyophilized form.
11. 11. The reagent of Claim 3 in lyophilized form.
12. 12. The reagent of Claim 4 in lyophilized form.
13. 13. The reagent of Claim 5 in lyophilized form.
14. 14. The reagent of Claim 6 in lyophilized form.
15. 15. The reagent of Claim 9 in lyophilized form.
16. 16. The reagent as provided in Claim 1 further comprising additional proteins, analytes or enzymes whereby the resultant reagent is useful as a multiparameter, abnormal control reagent.
17. 17. The reagent as provided in Claim 3 further comprising additional proteins, analytes and enzymes whereby the resultant control reagent is useful as a multiparameter, abnormal control reagent.
18. 18. The reagent as provided in Claim 4 further comprising additional proteins, analytes or enzymes whereby the resultant reagent is useful as a multiparameter, abnormal control reagent.
19. 19. The reagent as provided in Claim 5 further comprising additional proteins, analytes or enzymes whereby the resultant reagent is useful as a multiparameter, abnormal control reagent.
20. 20. The reagent as provided in Claim 6 further comprising additional proteins, analytes or enzymes whereby the resultant reagent is useful as a multiparameter, abnormal control reagent.
21. 21. The reagent as provided in Claim 9 further comprising additional proteins, analytes or enzymes whereby the resultant reagent is useful as a multiparameter, abnormal control reagent.
22. 22. The reagent as provided in Claim 11 further comprising additional proteins, analytes or enzymes whereby the resultant reagent is useful as a multiparameter, abnormal control reagent.
23. 23. The reagent as provided in Claim 13 further comprising additional proteins, analytes or enzymes whereby the resultant reagent is useful as a multiparameter, abnormal control reagent.
24. 24. The reagent as provided in Claim 15 further comprising additional proteins, analytes and enzymes whereby the resultant control reagent is useful as a multiparameter, abnormal control reagent.
25. 25. A serum based, multiparameter standard comprising:
    
    a) substantially normal blood having substantially all red blood cells, white blood cells and platelets removed and having proteins, analytes and enzymes present in known amounts; and b) plexiform stabilizing means selected from the group consisting of reducing monosaccharide sugars and reducing disaccharide sugars present in the range of about 2% - 8%
    final volume percent whereby activity of substantially all proteins, analytes and enzymes and solution is maintained for at least 10 days.
26. 26. The reagent of Claim 25 wherein the plexiform stabilizing means is selected from the group consisting of maltose, mannitol, cellobiose, and lactose.
27. 27. The reagent as provided in Claim 25 wherein the plexiform stabilizing means is lactose.
28. 28. The reagent as provided in Claim 27 wherein the lactose is present in a final concentration of about 6%.
29. 29. A stabilized multiparameter standard comprising an aqueous solution of known levels of protein and nonprotein analytes and plexiform stabilizing means selected from the group consisting of maltose, mannitol, cellobiose and lactose, said plexiform stabilizing means present in a concentration sufficient to obtain substantially increased stability as compared to a standard without such plexiform stabilizing means.
30. 30. The reagent as provided in Claim 28 further comprising an antimicrobial agent.
31. 31. The reagent as provided in Claim 30 wherein said antimicrobial agent is chloremphenicol.
32. 32. The reagent of Claim 29 further comprising an antimicrobial agent for substantially inhibiting microbial growth.
33. 33. The reagent as provided in Claim 32 wherein said antimicrobial agent is chloremphenicol.
34. 34. The reagent as provided in Claim 25 in lyophilized form.
35. 35. The reagent of Claim 27 in lyophilized form.
36. 36. The reagent of Claim 28 in lyophilized form.
37. 37. The reagent of Claim 29 in lyophilized form.
38. 38. The reagent of Claim 30 in lyophilized form.
39. 39. The reagent of Claim 31 in lyophilized form.
40. 40. A method for stabilizing a multiparameter serum based control reagent obtained from substantially normal blood having all normally present proteins, analytes and enzymes but with substantially all red blood cells, white blood cells and platelets removed comprising adding plexiform stabilizing means selected from the group consisting of re-ducing monosaccharide sugars and reducing disaccharide sugars in an amount sufficient so that the final volume percent of the plexiform stabilizing means is in the range of about 2%-8%.
41. 41. The method as provided in Claim 40 further comprising adding an antimicrobial agent for substantially inhibiting microbial growth.
42. 42. The method as provided in Claim 41 wherein the antimicrobial agent is chloremphenicol.
43. 43. A method for stabilizing a multiparameter abnormal control reagent formed from substantially whole blood having all normally present proteins, analytes and enzymes but with substantially all red blood cells, white blood cells and platelets removed and having added thereto proteins, analytes or enzymes comprising adding plexiform stabilizing means selected from the group consisting of maltose, mannitol, cellobiose and lactose such that the plexiform stabilizing means is present in a final volume percent in the range of about 2%-8%.
44. 44. The method as provided in Claim 43 further comprising adding an antimicrobial agent for substantially inhibiting the microbial growth.
45. 45. The method as provided in Claim 44 wherein the antimicrobial agent is chloremphenicol.
46. 46. A method for stabilizing a multiparameter standard from substantially normal blood having substantially all red blood cells, white blood cells and platelets removed and having known levels of protein and nonprotein analytes comprising adding plexiform stabilizing means selected from the group consisting of maltose, mannitol, cellobiose and lactose such that the plexiform stabilizing means is present in a final volume percent in the range of about 2%-8%.
47. 47. The method as provided in Claim 46 wherein the plexiform stabilizing means is lactose.
48. 48. The method as provided in Claim 47 further comprising adding an antimicrobial agent for substantially inhibiting microbial growth.
49. 43. The method as provided in Claim 48 wherein the antimicrobial agent is chloremphemicol.
50. 50. The method as provided in Claim 40 further comprising removing substantially all water.
51. 51. The method as provided in Claim 41 further comprising removing substantially all water.
52. 52. The method as provided in Claim 42 further comprising removing substantially all water.
53. 53. The method as provided in Claim 43 further comprising removing substantially all water.
54. 54. The method as provided in Claim 44 further comprising removing substantially all water.
55. 55. The method as provided in Claim 45 further comprising removing substantially all water.
56. 56. The method as provided in Claim 46 further comprising removing substantially all water.
57. 57. The method as provided in Claim 47 further comprising removing substantially all water.
58. 58. The method as provided in Claim 48 further comprising removing substantially all water.
59. 59. The method as provided in Claim 49 further comprising removing substantially all water.