Description:Title of the Invention
“A Diagnostic Test Kit for Calcium (Arsenazo III)”
Field of the Invention
The present disclosure relates generally to field of medical devices. More specifically the present invention relates to diagnostic test kit works on photometric method to determine Calcium (Arsenazo III) from blood serum / plasma in vitro for laboratories and medical professionals.
Background of Invention
A calcium blood test measures the amount of calcium in your blood. If there is too much or too little calcium in the blood, it may be a sign of a wide range of medical conditions, such as bone disease, thyroid disease, parathyroid disorders, kidney disease, and other conditions. Calcium is the most abundant mineral element in the body. About 98% of the 1200 g of calcium in the adult is in the form of hydroxyapatite in the skeleton. Hydroxyapatite is a lattice-like crystal composed of calcium, phosphorus, and hydroxide. The remaining calcium is in the extracellular fluid (50%) and in various tissues, especially skeletal muscle. Calcium is maintained within a fairly narrow range from 8.5 to 10.5 mg/dl (4.3 to 5.3 mEq/L or 2.2 to 2.7 mmol/L). Normal values and reference ranges may vary among laboratories as much as 0.5 mg/dl.
Calcium is one of the most important minerals in your body. About 1% of the calcium in your body is in your blood. The rest is stored in your bones and teeth. Having the right amount of calcium in your blood is necessary for your nerves, muscles, and heart to work properly. It also helps blood vessels move blood throughout your body and helps release hormones that affect many body functions. There are two types of calcium blood tests that measure different forms of blood calcium:
An ionized calcium test measures only the "free calcium" in your blood that isn't attached to proteins. An ionized calcium test is more difficult to do, so it's usually ordered if the results of a total calcium test aren't normal. You may also have this test if you have a condition that affects your body's ability to balance the amounts of ionized and bound calcium in your blood, or if you are seriously ill or having surgery. Symptoms of high calcium levels may include: Digestive symptoms, such as:Constipation, Nausea and vomiting, Abdominal (belly) pain, Loss of appetite, Increased thirst, Urinating (peeing) more than usual, Kidney stones, Bone, joint, and muscle symptoms, such as weakness, aches, and pains, Fatigue. A change in mental health, such as depression or confusion Symptoms of low calcium levels may include: Dry skin, coarse hair, and nails that easily break (after a long period of low levels), Muscle cramps, spasms, or stiffness, Tingling in the lips, tongue, fingers, and feet, Irregular heartbeat (arrhythmia), Seizures, if calcium levels are extremely low. The measurement of serum calcium is fraught with possible errors. Several means of contamination might lead to false elevations of serum calcium concentration. Falsely low levels are less common, so if several measurements are obtained, the lowest is usually the most accurate. The precision of the SMAC analysis, an automated colorimetric technique, is usually equal or superior to that of manual analysis. Nevertheless, falsely high or low values may be obtained in patients with liver or renal failure or in patients with lipemic or hemolyzed specimens. Venous occlusion of the arm during venipuncture may increase the total concentration of serum calcium by up to 0.3 mmol/L. This results from an increase in plasma protein concentration caused by hemodynamic changes. Another source of error is posture. If the patient stands up from a supine position, there may be an increase of 0.05 to 0.20 mmol/L in serum calcium. Still another possible source of error is hemolysis. Some methods of measuring calcium are affected by high levels of hemoglobin, and red cells may take up calcium after prolonged contact. If an error is suspected and the measurement is to be redone, the blood should be drawn following an overnight fast because the daily intake of calcium may contribute to the serum calcium concentration as much as 0.15 mmol/L.
The present disclosure relates generally to field of medical devices. More specifically the present invention relates to diagnostic test kit works on photometric method to estimate Calcium (Arsenazo III) from serum / plasma, in vitro for laboratories and medical professionals.
Object of the Invention
The primary object of the invention is to develop a diagnostic test kit to estimate Calcium (Arsenazo III) from blood serum / plasma useful effectively.
The secondary objective of the invention is that the diagnostic test kit for Calcium (Arsenazo III) utilizes photometric method herein the kit configured with suitable reagents like Arsenazo, Imidazole and Calcium as standard, taken in effective concentration of blood serum /plasma.
Another objective of the invention is that the prepared blood serum/plasma samples with reagents estimated for absorbance in photometer at desired wavelength and room temperature.
Further objective of the invention is that Calcium (Arsenazo III) is determined through calculation by the absorbance of sample and standard.
Summary of the Invention

The present disclosure relates a diagnostic kit specifically to determine Calcium (Arsenazo III) concentration in the blood serum/plasma wherein the desired size kit is developed, essentially consist of reagents for reaction with blood serum/plasma samples, detected on photometer at desired wavelength to measure the absorbance and finally calculated to determine Calcium (Arsenazo III) concentration.
A diagnostic test kit for Calcium (Arsenazo III) comprising a test kit characterized in that 50 ml size kit of one compartment cell reagents preferably consist of Arsenazo, Imidazole and Calcium with Calcium as standard estimate Calcium (Arsenazo III) in vitro from blood serum / plasma sample effectively, wherein reagent cell composed of 120 micromol/l of Arsenazo, 100 mmol/l of Imidazole taken with blood serum/ plasma sample from 0.010 ml to 0.015 ml into two clean test tube for test sample and blank sample and Calcium 8 mg/dl sample as standard in third test tube; pipette out the reagent from the kit separately; transferring said reagent into the test tubes respectively at room temperature (37oC) and , uniformly mixed with test sample, blank sample and standard; incubating the reaction mixture for 2 minutes; determining the absorbance of said test sample blank sample and standard Calcium (Arsenazo III) sample on photometer at a wavelength of 650 nm for 30-90 seconds; and finally calculating the absorbance to estimate Calcium (Arsenazo III) from the test serum / plasma sample effectively.

One of the preferred embodiments of the present invention is the unique design of the kit prefilled with the preferred reagent like Arsenazo, Imidazole and Calcium as standard reacts with serum/ plasma Calcium evaluated through the absorbance on photometer.

One of the preferred embodiments of the present invention is that to determine Calcium from the varied concentration more preferably making the test sample, blank sample and Calcium sample as standard to infer the clinical findings effectively.

One of the preferred embodiments of Calcium is estimated in vitro through all in one reagent kit.

Brief Description of Drawings

The following thorough explanation of the various aspects of the invention, taken in conjunction with the corresponding drawing that represents various aspects and other features of the disclosure invention.

Figure 1: Flow chart of method of estimation of Calcium (Arsenazo III)

Detailed Description of the Invention

The following description is of exemplary embodiments only and is not intended to limit the scope, applicability or configuration of the invention in any way. Rather, the following description provides a convenient illustration through explanation and figures for implementing exemplary embodiments of the invention. Various changes to the described embodiments may be made in the function and arrangement of the ingredients described without departing from the scope of the invention.
The use of “including”, “comprising” or “having” variations thereof herein is meant to encompass the items listed thereafter and equivalents thereof as well as additional items. The terms “a” and “an” herein do not denote a limitation of quantity, but rather denote the presence of at least one of the referenced items. Further, the use of terms “first”, “second”, and “third”, and the like, herein do not denote any order, quantity, or importance, but rather are used to distinguish one element from another.
The terms used in the invention
mmol: milimole
mmol/l: millimole per liter
md/dl: milligram/deciliter
A calcium blood test measures the amount of calcium in your blood. If there is too much or too little calcium in the blood, it may be a sign of a wide range of medical conditions, such as bone disease, thyroid disease, parathyroid disorders, kidney disease, and other conditions. Calcium is the most abundant mineral element in the body. About 98% of the 1200 g of calcium in the adult is in the form of hydroxyapatite in the skeleton. Hydroxyapatite is a lattice-like crystal composed of calcium, phosphorus, and hydroxide. The remaining calcium is in the extracellular fluid (50%) and in various tissues, especially skeletal muscle. Calcium is maintained within a fairly narrow range from 8.5 to 10.5 mg/dl (4.3 to 5.3 mEq/L or 2.2 to 2.7 mmol/L). Normal values and reference ranges may vary among laboratories as much as 0.5 mg/dl.
A diagnostic test kit for Calcium (Arsenazo III) comprising a test kit characterized in that 50 ml size kit of one compartment cell reagents preferably consist of Arsenazo, Imidazole and Calcium with Calcium as standard estimate Calcium (Arsenazo III) in vitro from blood serum / plasma sample effectively, wherein reagent cell composed of 120 micromol/l of Arsenazo, 100 mmol/l of Imidazole taken with blood serum/ plasma sample from 0.010 ml to 0.015 ml into two clean test tube for test sample and blank sample and Calcium 8 mg/dl sample as standard in third test tube; pipette out the reagent from the kit separately; transferring said reagent into the test tubes respectively at room temperature (37oC) and , uniformly mixed with test sample, blank sample and standard; incubating the reaction mixture for 2 minutes; determining the absorbance of said test sample blank sample and standard Calcium (Arsenazo III) sample on photometer at a wavelength of 650 nm for 30-90 seconds; and finally calculating the absorbance to estimate Calcium (Arsenazo III) from the test serum / plasma sample effectively.
The present disclosure relates a diagnostic kit specifically to determine Calcium (Arsenazo III) concentration in the blood serum/plasma wherein the desired size kit is developed, essentially consists of reagents for reaction with blood serum / plasma samples, detected on photometer at desired wavelength to measure the absorbance and finally calculated the Calcium (Arsenazo III) concentration.
Preferred embodiments of Diagnostic Test Kit For Calcium (Arsenazo III)
i) Diagnostic Kit: The Calcium (Arsenazo III) kit comprises of 50 ml size wherein reagent cell prefilled for the determination of Calcium (Arsenazo III), herein the key reagent like Arsenazo, Imidazole and Calcium as standard reacts with serum/ plasma Calcium (Arsenazo III) used in different concentration, estimates serum / plasma Calcium (Arsenazo III) through the absorbance using photometer of desired wavelength.
ii) Reagents: Blood serum /plasma, Arsenazo, Imidazole and Calcium.
iii) Apparatus / Instrument: Test tubes, Photometer, Incubator etc.
Table no. 01: Preferred embodiments and parameters of the kit with concentrations

Particulars Name of ingredient with concentration
Serum/ plasma sample 0.010 ml to 0.015 ml
Reagent 120 micromol/l of Arsenazo and 100 mmol/l of Imidazole
Standard Calcium 8 mg/dl
Shelf Life 24 months
Kit Size 50 ml
Wavelength 650 nm
Temperature 37oC
Time of Estimation 30 to 90 seconds

Table no. 02: Method to operate Calcium (Arsenazo III)

Particulars Blank Standard Sample
Reagent 1.0 ml 1.0 ml 1.0 ml
Standard -- 0.010 ml --
Serum/plasma test sample -- -- 0.010 to 0.015 ml

General Method to operate the device

Taking the 0.010 to 0.015 ml blood serum/ plasma sample as test sample, reagent as blank sample and Calcium (Arsenazo III) as standard separately in clean test tubes;
pipette out the reagent 120 micromol/l of Arsenazo and 100 mmol/l of Imidazole from the kit separately; transferring said reagent into the test tubes respectively at room temperature (37oC); incubating the reaction mixture for 02 minutes; determining the absorbance of said test sample, blank sample and 8 mg/dl Calcium sample as standard, on photometer at a wavelength of 650 nm for 30-90 seconds; and finally calculating the absorbance to estimate Calcium (Arsenazo III) from the test serum / plasma sample effectively.

The absorbance is calculated using below formula:
Concentration = Absorption of sample / Absorption of standard x 8 mg/dl
(Table No. 1, 2 and Figure No. 1)
Example 1
Taking the 0.010 ml blood serum/ plasma sample as test sample, reagent as blank sample and Calcium (Arsenazo III) as standard separately in clean test tubes;
pipette out the reagent 120 micromol/l of Arsenazo and 100 mmol/l of Imidazole from the kit separately; transferring said reagent into the test tubes respectively at room temperature (37oC); incubating the reaction mixture for 02 minutes; determining the absorbance of said test sample, blank sample and 8 mg/dl Calcium sample as standard, on photometer at a wavelength of 650 nm for 30-90 seconds; and finally calculating the absorbance to estimate Calcium (Arsenazo III) from the test serum / plasma sample effectively.

The absorbance is calculated using below formula:
Concentration = Absorption of sample / Absorption of standard x 8 mg/dl
Example 2
Taking the 0.015 ml blood serum/ plasma sample as test sample, reagent as blank sample and Calcium (Arsenazo III) as standard separately in clean test tubes;
pipette out the reagent 120 micromol/l of Arsenazo and 100 mmol/l of Imidazole from the kit separately; transferring said reagent into the test tubes respectively at room temperature (37oC); incubating the reaction mixture for 02 minutes; determining the absorbance of said test sample, blank sample and 8 mg/dl Calcium sample as standard, on photometer at a wavelength of 650 nm for 30-90 seconds; and finally calculating the absorbance to estimate Calcium (Arsenazo III) from the test serum / plasma sample effectively.

The absorbance is calculated using below formula:
Concentration = Absorption of sample / Absorption of standard x 8 mg/dl
Scope of the Invention

A diagnostic test kit for Calcium (Arsenazo III) principally design to develop a simple method to analyse the Calcium (Arsenazo III) level using photometric method having size of 50 ml size kit of reagent preferably consist of effective concentration of 120 micromol/l of Arsenazo and 100 mmol/l of Imidazole, taken with blood serum/ plasma sample from 0.010 ml to 0.015 ml as test sample and blank sample and Calcium 8 mg/dl sample as standard respectively determines Calcium (Arsenazo III) in vitro from blood serum / plasma sample effectively, could be potential diagnostic and economical method, compare to expensive conventional methods in future.

It is to be understood that the present invention is not limited to the embodiments described above, it should be clear that various modifications and alterations can be made along with various features of one embodiment included in other embodiments, within the scope of the present invention.
, Claims:We claim,
1- A diagnostic test kit for Calcium (Arsenazo III) comprising a test kit characterized in that 50 ml size kit of reagent preferably consist of effective concentration of 120 micromol/l of Arsenazo and 100 mmol/l of Imidazole estimates Calcium (Arsenazo III) in vitro from blood serum / plasma sample taken from 0.010 to 0.015 ml as test sample and 8 mg/dl of Calcium as standard, using photometer effectively.
2- The diagnostic test kit for Calcium (Arsenazo III) as claimed in claim 1, operates through in vitro diagnostic method to determine Calcium (Arsenazo III) comprises the following steps:
taking the 0.010 to 0.015 ml blood serum/ plasma sample as test sample, reagent as blank sample and Calcium (Arsenazo III) as standard separately in clean test tubes;
pipette out the reagent Pipes, Lipoprotein Lipase, Glycerol Kinase and Peroxidase from the kit separately;
transferring said reagent into the test tubes respectively at room temperature (37oC);
incubating the reaction mixture for 10 minutes;
determining the absorbance of said test sample, blank sample and standard Calcium (Arsenazo III) sample on photometer at a wavelength of 510 nm for 30-90 seconds; and
calculating the absorbance to estimate Calcium (Arsenazo III) from the test serum / plasma sample effectively.