Stop-flow analysis

• US 4,399,225 A
• Filed: 08/26/1981
• Issued: 08/16/1983
• Est. Priority Date: 06/14/1978
• Status: Expired due to Term

• Current Assignee: Bifok AB
• Original Assignee: Bifok AB
• Inventors: Ruzicka, Jaromir, Hansen, Elo H.
• Primary Examiner(s): Wyse, Thomas G.
• Application Number:
  US06/296,256
• Time in Patent Office: 720 Days
• Field of Search: 230/230 R, 435/15, 436/34, 436/51-53
• US Class Current: 436/34
• CPC Class Codes: C12Q 1/54 involving glucose or galactose
• CPC Class Codes: G01N 21/272 for following a reaction, e.g. for determining photometrically a reaction rate (photometric cinetic analysis)
• CPC Class Codes: G01N 35/085 Flow Injection Analysis
• CPC Class Codes: Y10T 436/117497 with a continuously flowing sample or carrier stream
• CPC Class Codes: Y10T 436/118339 with formation of a segmented stream

Abstract

A continuous method of quantitatively determining slowly reacting compounds with the use of a single measuring cell, the sample being injected into a continuously flowing carrier solution which is already provided with, or simultaneously with the sample is provided with reagent to form a sample zone in the form of a reproduceable gradient. Said sample zone is led into a measuring cell whereupon the flow is stopped by means of shunting the carrier flow, by stopping the pump, or by means of a valve. The reaction is allowed to take place in the measuring cell while a magnitude, characteristic for the reaction, is registered. The reaction speed can be detected from the incline of the obtained response curve and the amount of the sought-after compounds is calculated. By means of setting the stop at a certain point in the concentration gradient curve and always selecting said point at the same time distance from the peak of the gradient curve during flow, a total reproduceability of the analysis is ensured.

Claims

1. A method for determining the reaction speed of a sample at selected sample concentrations within a sample mixture slug comprising the steps of:

introducing a sample into a flowing carrier stream to form a sample mixture slug having a forward head portion, a rearward tail portion and a central portion, said sample dispersing within said sample mixture slug to form reproduceable concentration gradients in said head and tail portions in which the concentration of said sample varies;

flowing said carrier stream and sample mixture slug through a measuring cell without removing the head portion or tail portion from said sample mixture slug;

stopping the flow of said carrier stream in response to a selected concentration of sample within one of said concentration gradients as said sample mixture slug passes through said measuring cell; and

measuring the reaction of said sample at said selected concentration for a period of time whereby said reaction speed of said sample at said selected sample concentration may be determined.

2. A method according to claim 1 wherein one or more reagents are introduced into said carrier stream to form a reagent carrier stream having reagent uniformly dispersed therein for reaction with sample.

3. A method according to claim 2 wherein said selected sample concentration is determined by measuring a reaction product of said sample and said reagent.

4. A method according to claim 3 wherein samples are sequentially introduced into said flowing carrier stream.

5. A method according to claim 3 wherein said sample concentrations are selected by varying the time period between introduction of said sample into said carrier stream and stopping said carrier stream flow.

6. A method according to claim 2 wherein said reagent is introduced into said carrier stream after said sample is introduced therein.

7. A method according to claim 2 or 6 where said reagent is introduced noncontinuously into said carrier stream as a discrete reagent slug in such a manner to provide mixing of said sample mixing slug and said regent slug in said carrier stream.

8. A method according to claim 1 where the concentration of sample in said concentration gradients increases inward towards the central portion of said sample mixture slug.

9. A method according to claim 8 where the section of said sample mixture zone measured within said measuring cell is selected from the forward or rearward ends of said concentration gradient and not from said central portion.

10. A method according to claim 1 wherein reaction speeds at different sample concentrations within said sample zone are measured by changing the time period between introduction of said sample into said carrier stream and stopping said carrier stream flow.

11. In a method of analysis where a sample is introduced into a flowing carrier stream to form a sample mixture slug having a forward head portion, a rearward tail portion and a central portion, said sample dispersing to form reproducable concentration gradients in said head and tail portions in which the concentration of said sample varies, said sample undergoing a reaction while in said carrier stream, wherein the improvement comprises measuring the reaction speed of said sample at one or more selected sample concentrations within said concentration gradients by stopping the flow of said carrier stream and measuring the reaction of said sample at said selected sample concentrations for a period of time.

12. A method according to claim 11 in which measureable reaction products are produced by said sample reaction, said reaction product formation being related to said sample concentration, wherein said selected sample concentrations are determined indirectly by measurement of said reaction product formation.

13. A method of determining the change in a changing characteristic of a sample at selected sample concentrations comprising the steps of:

introducing a sample into a flowing carrier stream to form a sample mixture slug having a forward head portion, a rearward tail portion and a central portion, said sample dispersing within said sample mixture slug to form reproduceable concentration gradients in said head and tail portions in which the concentration of said sample varies, said sample having a measureable changing characteristic;

flowing said carrier stream and sample mixture slug through a measuring cell without removing the head or tail portion from said sample mixture slug;

stopping the flow of said carrier stream in response to a selected concentration of sample within one of said concentration gradients as said sample mixture slug passes through said measuring cell; and

measuring the changing characteristic of said sample at said selected concentration for a period of time whereby the change in said characteristic of said sample at said selected concentration may be determined.

14. A method according to claim 13 wherein said changing characteristic is a reaction between said sample and one or more reagents added to said sample.

15. A method according to claim 14 wherein said reaction produces a measurable reaction product, said selected sample concentration being determined by measuring said reaction product.