DE1962267B2 - Photometric analyzer - Google Patents

Description

fen, with which the analysis of a large number of separate samples continuously and simultaneously can be carried out and in which the steps of volumetric measurement, liquid transfer, the solution mixing, the reaction, the photometric measurement and the data reduction can be performed with a single system.

This object is achieved by means of a photometric analysis device specified at the beginning a motor-driven rotor having a plurality of circularly arranged sample analysis chambers having said sample analysis chambers between a light source and the light examining Devices pass through during the rotation of the rotor, a plurality of circular in axial alignment with the sample analysis chambers arranged loading chambers, the radial are arranged between the sample analysis chambers and the center of rotation of the rotor and a radially extending passage connecting the radially outermost portion of each loading chamber with the corresponding one Sample analysis cuvette connects, taking samples and reagents from said loading chambers said passages to said sample analysis cuvettes as the rotor rotates as a result of the centrifugal force, as well as by means of receiving the output signal of the said light investigation devices 2ur continuous and simultaneous display of the Presence of a common substance in each sample analysis chamber.

According to a particular embodiment, the rotor can be adjacent to the sample analysis chambers have transparent walls.

Further refinements of the invention are the subject of the subclaims.

An exemplary embodiment of the invention will be explained in more detail with reference to the following description using the drawings.

In the drawings shows

F i g. 1 is a schematic representation of a photometric system in accordance with the present invention;

F i g. FIG. 2 is an exploded perspective view of a section of the rotor included in a The system of Figure 1 is used;

F i g. 3 an oscillogram, which with the system according to FIG. 1 was obtained using a 660 ηψ filter and distilled water was used in all cuvettes;

F i g. 4 is an oscillogram which is obtained with the system according to FIG. 1 was obtained, with a 660 ΐημ filter and a uniform solution containing water, ox protein serum and bromophenol blue was used, which was added to cuvettes No. 2 to No. 15 was introduced during the revolution;

FIG. 5 shows an oscillogram which is obtained with the system according to FIG. 1 was obtained, with a 550 ηΐμ filter and a number of increased gauges were used in cuvettes # 3 to # 12;

F i g. 6 an evaluation of the in FIG. 5 contained data;

7 shows an evaluation of the recordings the protein concentration obtained using the system of FIG carrying out the experiment described in Example II.

Fig. 1 shows a roof-shaped rotor assembly which is shown in the exploded view of Fig. 2 in enlarged detail. It consists of a steel rotor body 2 with flanged bolts, a glass rings 3 and 4, a cuvette ring provided with protectors made of polytetrafluoroethylene, retaining rings 6 and 7 made of polytetrafluoroethylene and a flange ring 8. The rings 3, 4, S, 6 and 7 are between the rotor body 2 and the flange ring 8 together

xo quantity pressed to a plurality of radially aligned To form cuvettes 9 in the cuvette ring S provided with protectors. At a distance from each other arranged bores 10, which are axially aligned with the cuvettes 9, are in the rotor body 2, the

Retaining rings 6 and 7 and the flange ring 8 provided to form axially extending passages, which allow the light beam to penetrate through the cuvette. A centrally arranged removable transmission disk 11 is with small

ao radial projections 12 provided, which at a distance are arranged in front of one another on the circumference in order to engage in the cuvettes in the ring 5. It is furthermore a handle 3 is provided for removing the transmission disk 11 from the rotor assembly

to facilitate. The transmission disk 11 is provided with a series of chambers 14 corresponding to each Cell 9 is provided for holding sample liquids and reagents when the motor is not running. the Chambers 14 consist of a plurality of ge

.30 inclined cylindrical bores interconnected at their upper ends and at their lower ends are separated from one another by partitions 15. The partitions 15 prevent the Mixing the sample and the reagent liquids,

when the rotor is at a standstill, while the liquids can flow into the cuvette 9 when the rotor running around. A passage 16 extends from the radially outermost bore of each chamber 14 to the Edge of the corresponding projection 12 to a

To allow passage of the liquid from each chamber to the corresponding cuvette when the The rotor is rotating. A drive motor 17 carries the rotor assembly and displaces it in rotation.

There are a light source and projection devices are provided to a light beam of constant Intensity to throw the rotor assembly 1 crossing at a point that corresponds to the radial positions of the cuvettes 9 and the holes 10 arranged with a gap. The ray of light will oriented so that it penetrates through each bore 10 and each cuvette 9 traverses the beam will. The photometric light source consists of an incandescent lamp 18 with a reflective one

Mirror 19, which is arranged below the rotor assembly 1 and aligned to the light beam to reflect upwards essentially perpendicular to the plane of rotation.

An electronic light detector device 20 is provided and aligned above the rotor assembly 1, to pick up the light penetrating through the cuvette during rotation. the Light detector device 20 electronically provides an output which is proportional to the intensity of the

the light source 18 is light passing through the cuvettes. The light detector 20 consists of a photomultiplier tube, which is arranged immediately above the cuvette circle, around the entire

to receive light emerging upwards through the axially aligned openings.

The remaining electronic components, which are shown schematically in Fig. 1, consist of a proportional tachometer 21 which outputs a voltage signal proportional to the rotor speed a run-up signal generator 22 supplies which feeds a signal to a pulse sampler 23. A rotation detector 24 synchronizes the ramp signal frequency with the rotor speed. The pulse sampler 23, which is determined by the frequency of the run-up signal can be synchronized by the generator 22, speaks proportionally to the in the photodetector device 20 resulting impulses and sorts the impulses as they originated. A pulse peak indicator 25 continuously and simultaneously shows the passage of light through the liquid contents in each cuvette.

In operation, the samples and reagents are first transferred to the chambers 14 when the rotor 1 is at a standstill introduced and then moved centrifugally into the corresponding cuvettes 9 as the rotor rotates. Since the transfer to the cuvette 9 when accelerating the motor for a relatively short period of time happens, all reactions in the cuvettes begin essentially simultaneously and can continuously observed on an oscillating scope or other reading devices 25. In the Providing three cavities in each chamber 14 in the transmission disk 11 can at standstill the rotor can accommodate a sample and two reagents without mixing and rotate As a result of the centrifugal force of the rotor, these will flow off into the corresponding cuvette and mixed. The connections to the cuvettes are made through small passages through the projections 12, as in FIG Figs. 1 and 2 is shown. The transmission disk 11 takes transmission tubes or small, commercially available microliter pipettes. Such devices allow a single reaction or a multiple addition of reactions or reactions in which the reaction time between two additions expires. In reactions in which precipitates are generated, the suspended solids move out of the optical path by centrifugal force, whereby the absorbent can be clearly measured.

In the described rotor, the radial alignment of the cuvettes causes a difference in the tangential speed, which is present between the radially innermost and the radially outermost end of each cuvette. A quick acceleration and deceleration of the rotor during the transfer of the liquid into the cuvette causes a circular Flow of the liquid and increases the mixture. Such a mixture is very desirable because it increases the reaction between the sample and the reagent and gives more consistent results. In practice, the rotor is accelerated quickly in order to transfer the liquid into the cuvettes, quickly delayed to make mixing easier and then back to the desired speed for that Examination accelerated.

example 1

To determine whether reproducible curves with calibration solutions when using the previously described Device could be obtained, a solution containing 1.5 g of crystalline ox protein serum (BSA) and 15 mgm bromophenol blue (BPB) in 100 ml of water diluted with distilled water to a series of solutions containing 10% increment per stock solution. F i g. 3 is setting 5 represents an oscillogram, which shows a typical example using a 660 πιμ filter and distilled water was observed in all cuvettes. The oscillogram was from an oscilloscope obtained with a pulse peak reader 25 described with reference to FIG was provided. The oscillogram of FIG. 4 was made by introducing a solution containing water and BSA-BPB solution in a volume ratio of 1: 1, in cuvettes No. 2 to 15 during the Get rotation. The differences in peak heights, albeit slight, coincided with them direct measurements observed agree. The oscillogram of FIG. 5 was obtained through a complete Series of increased standard measures in cuvettes No. 3 to 12, with a doubling of the Solution that was used in cuvette 12 and also in cuvette 14. The four remaining cuvettes contained only distilled water. The measurements were from photographic enlargements

»5 of the images observed in the oscilloscope obtained and all peaks were in 1 /% T by dividing the first void by each subsequent reading converted. The logarithm of l / T is the extinction, which after subtracting the blank was multiplied by the cuvette factor to get the absorbent substances for one centimeter To get path length. The data thus obtained from the oscillogram of Fig. 5 became evaluated in Fig. 6.

Example 1 2

Another experiment was carried out to show that the system could be used for the following in the Cuvette running reactions can be used. The biuret reaction for protein is a single one One-reagent analysis. which is of general interest and suitable for the efficiency of the transmission disk, the mixture and the efficiency of the system, the absorbing substances to measure and calculate at an early stage in the course of the reactions. The Weichselbaum Biuret Reagent can with protein solutions in the range of different ratios from 0 to 50% of the reagent in the dei Final mixing to be carried out, wöbe: identical solutions are used to unite To obtain calibration curve.

An experiment was performed with 200 microliters of reagent and duplicate protein solutions of 200 microliters, the solutions containing 0.2, 0.4, 0.6, 0.8 and 1% protein. These solvers were placed in the appropriate chambers in the middle disk and when the rotor started up it transfer the cuvettes. Thirty seconds later an oscillogram was made in the same way as wi < obtained in the experiment of Example 1 and di <

Results were evaluated in Fig. 7, wöbe Water was used as a reference measure.

The experimental embodiment of the device which ver was used, allows 15 reactions at the same time; perform and the substances of the samples within very short periods of time after the reac have started to observe and measure. Line greater number of reactions can be thruc '

Use of a larger rotor with a correspondingly larger number of cuvettes and a smaller one Number can be carried out by simply using part of the available cuvettes.

In contrast to the subsequent analysis devices no dragging between the samples and the oscilloscope trace observed between each Pmbcn reduction decreased to 0%. With the provision of one or more pure water samples in each row, sample readings showed 0 and 100% transfer during each revolution established. At a rotation speed of 1200 rpm, 20 revolutions per see, whereby 20 series of measurements can be carried out. If an exposure time of one second is selected, an average of 20 readings can be obtained as a result. The time between the peaks is sufficient to digitize the peak heights in the computer averaging to undertake.

If small volumes of liquid are initially added to the rotor, the rotor will become complete stopped and the sample reagent disk is reinstalled. That way you can have reactions which are dependent on consecutive additions. The centrifugal properties of the rotor can, if desired, also be used to deposit special substances and to ensure that the

ίο solutions while the substances are measured, are not cloudy.

The rotor assembly 1 can be made with more or fewer cuvettes than shown or made of different materials, such as B. transparent plastics are produced. The centrally located transmission disc can also be used with more or fewer chambers to accommodate the samples and the reagent liquids provided, whereby the chambers mentioned may differ from the particular form shown.

ao nen.

For this purpose 2 sheets of drawings

309546023

Claims (7)

Patent claims: 1. Photometric analysis device for determining the substance present in a sample, in which a light beam penetrates the sample and is measured by light-analyzing devices which emit an output signal that is proportional to the intensity of the examined Is light, and with which the existence of a common jubility in a plurality of samples can be determined simultaneously and continuously, characterized by a motor-driven rotor that has a Has a plurality of circularly arranged sample analysis chambers between a Light source and the light-examining devices during the rotation of the rotor traverse a plurality of circular shapes in axial alignment with the sample analysis chambers arranged loading chambers radially between the sample analysis chambers and the The center of rotation of the rotor are arranged, and a radially extending passage, the radially outermost part of each loading chamber with the corresponding sample analysis cuvette connects, the samples and reagents from the said lay chambers the said Passages to the named sample analysis cuvettes when the rotor rotates as a result of centrifugal force traverse, as well as by devices for receiving the output signal from the said Lichiiuntersuchungseinrichtung for continuous and simultaneous indication of the presence of a common substance in each sample analysis chamber. 2. Photometric analysis device according to claim 1, characterized in that said Sample analysis chambers made up of a plurality of radially oriented, elongated There are bores or cavities, which are arranged in a circular row above the center of rotation the said rotor assembly are provided. 3. Photometric analysis device according to claim 2, characterized in that the sample analysis chambers are made by interposing a slotted ring between two layers of transparent material. 4. Photometric analysis device according to claim 3, characterized in that the slotted Ring made of polytetrafluoroethylene and the layers mentioned made of a transparent one Material made of glass. 5. Photometric analyzer according to claim 1, characterized in that said Rotor, adjacent to the sample analysis chambers, has transparent walls. 6. Photometric analyzer according to claim 1, characterized in that said Means for examining the light consist of a photomultiplier tube. 7. Photometric analyzer according to claim 1, characterized in that said Devices for the continuous and simultaneous display of the presence of a common substance in each sample analysis chamber consist of an oscilloscope. The invention relates to a photometric analysis device for determining the amount present in a sample Substance in which a light beam penetrates the sample and through light-testing devices is measured, which emit an output signal that is proportional to the intensity of the examined Is light, and with which the presence of a common substance in a plurality of samples determined simultaneously and continuously ίο can be. The term "photometric" used below is not intended to be construed in a restricted sense rather, it should be used generally for the expressions "colorimetric", "fluorimetric" and is "spectrometric" apply. Consistent with it should also the term "photometer" is used in a broad sense for devices that sometimes known as the "colorimeter," "fluorometer," and "spectrometer". The still The term "light", which is also used, encompasses radiant energy in the visible and invisible spectrum as well as radiant energy on special wavelengths is limited. Thus, the invention is intended to encompass systems which employ various types of Use as radiation to get the measurement you want to execute. The need for a photometric system which is capable of a large number of separate Simultaneous sampling existed in clinical and analytical laboratories for a long time. Qualitative and quantitative measurements of metabolic products, hormones, vitamins, Enzymes, minerals, body waste products, bile constituents and stomach contents are daily used in large numbers in such laboratories for diagnosis of diseases as well as for research purposes carried out. A system that could make measurements of this kind quickly, accurately, and cheaply would also bring about great labor and cost savings, combined with improved results. The most previously known instruments of this type were more suitable for analyzes in order than to perform at the same time. The successive analyzes not only limit the number of Analyzes, but in the case of analyzing very small samples, the results usually become unreliable. Another disadvantage of the prior art separate sample analyzers is the requirement that the samples had to be prepared in long steps and in separate apparatus. Such arrangements also limit the number of analyzes since they consume even more time and are costly. Another shortcoming of the prior art photometric instruments is the fact that Quantities of samples, enzymes, or other expensive reagents greater than desirable are required are. This disadvantage affects in some cases continuously working flow monitoring systems so that they become unusable, when a small number of samples are to be analyzed. Another disadvantage is the undesirability the individual treatment of many small, discrete volumes of samples and reagents and the mixture of these at time intervals. The present invention is therefore based on the object of creating a photometric system