KR20060113058A - Automatic analysis of volatile basic nitrogen using automated flow injection analysis system - Google Patents

Automatic analysis of volatile basic nitrogen using automated flow injection analysis system Download PDF

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KR20060113058A
KR20060113058A KR1020050035897A KR20050035897A KR20060113058A KR 20060113058 A KR20060113058 A KR 20060113058A KR 1020050035897 A KR1020050035897 A KR 1020050035897A KR 20050035897 A KR20050035897 A KR 20050035897A KR 20060113058 A KR20060113058 A KR 20060113058A
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analysis
basic nitrogen
flow injection
automated flow
volatile basic
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    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
    • G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
    • G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
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    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N35/10—Devices for transferring samples or any liquids to, in, or from, the analysis apparatus, e.g. suction devices, injection devices
    • G01N35/1095—Devices for transferring samples or any liquids to, in, or from, the analysis apparatus, e.g. suction devices, injection devices for supplying the samples to flow-through analysers
    • G01N35/1097—Devices for transferring samples or any liquids to, in, or from, the analysis apparatus, e.g. suction devices, injection devices for supplying the samples to flow-through analysers characterised by the valves
    • G—PHYSICS
    • G11—INFORMATION STORAGE
    • G11C—STATIC STORES
    • G11C7/00—Arrangements for writing information into, or reading information out from, a digital store
    • G11C7/16—Storage of analogue signals in digital stores using an arrangement comprising analogue/digital [A/D] converters, digital memories and digital/analogue [D/A] converters 

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Abstract

An automatic analysis of a volatile basic nitrogen using an automated flow injection analysis system is provided to simplify a structure of the automated flow injection analysis system by not using the other reagents except for an alkali solution in a case of using an ammonia electrode. A method of an automatic analysis for a volatile basic nitrogen is performed by an automated flow injection analysis system which consists of a tubing pump, a solution selective valve, an injection valve, an injection valve actuator, an air solenoid valve, a nitrogen pressure receptacle, a mixed coil, a flow cell, an ammonia electrode, an ammonia meter, a connection device, and a personal computer.

Description

자동화 흐름주입분석장치를 이용한 휘발성염기질소의 자동분석방법{Automatic analysis of volatile basic nitrogen using automated flow injection analysis system}  Automatic analysis of volatile basic nitrogen using automated flow injection analysis system

제1도는 자동화 흐름주입분석장치의 구성도1 is a block diagram of an automated flow injection analysis device

제2도는 플로우셀의 단면도2 is a cross-sectional view of the flow cell

제3도는 휘발성염기질소 자동분석 프로그램의 흐름도3 is a flow chart of the volatile basic nitrogen automatic analysis program

제4도는 돈육시료의 휘발성염기질소 분석에서 미량확산법의 결과와 자동분석법의 결과를 비교한 표 4 is a table comparing the results of microdiffusion and the results of automatic analysis in the analysis of volatile basic nitrogen of pork samples

제5도는 우육시료의 휘발성염기질소 분석에서 미량확산법의 결과와 자동분석법의 결과를 비교한 표5 is a table comparing the results of microdiffusion and the results of automated analysis in the analysis of volatile basic nitrogen of beef samples

본 발명은 자동화 흐름주입분석장치를 이용하여 휘발성염기질소를 자동분석하는 방법에 관한 것이다.The present invention relates to a method for automatically analyzing volatile nitrogen using an automated flow injection analysis device.

식육 또는 어패류와 같은 단백질함유식품의 부패도를 측정하기 위한 한가지 방법으로 휘발성 염기질소를 측정하며, 휘발성 염기질소 측정방법으로써 현재 식품공전에 규정되어 있는 종래의 방법은 미량확산법이다. 그러나, 미량확산법은 적정 법으로서 시험조작이 복잡하고 번거로우며 확산조작과 적정조작에 시간이 많이 소요된다. One method for measuring the degree of corruption of protein-containing foods such as meat and fish and shellfish is to measure volatile basic nitrogen, and the conventional method currently defined in the Food Code as a method of measuring volatile basic nitrogen is microdiffusion. However, the microdiffusion method is a titration method, and the test operation is complicated and cumbersome, and the diffusion and titration operation takes a long time.

본 발명에서는 위에 기술한 문제점을 해결하여 휘발성 염기질소 분석을 더 빠르고 더 간편하게 수행할 목적으로 암모니아 전극을 검출기로 채용한 자동화 흐름주입분석 시스템을 구성하여 자동분석하고자 하였다. 흐름주입분석법은 빠르고, 분석 데이타의 신뢰성이 크며 자동화하기가 쉬운 특징이 있다. 암모니아 전극은 암모니아 뿐만 아니라 휘발성 염기질소에 해당하는 휘발성 아민류에도 반응한다. 암모니아와 휘발성 아민류에만 선택적으로 반응하는 반면 시료에 함유된 다른 물질들의 영향은 전혀 받지 않으므로 휘발성 염기질소의 측정에 적합하다. 또한 암모니아 전극을 사용하면 알칼리 용액외의 다른 시약을 사용할 필요가 없기 때문에 흐름주입분석 시스템의 구성이 단순해진다. In the present invention, to solve the above-mentioned problems, to automatically and volatile basic nitrogen analysis to configure an automated flow injection analysis system employing an ammonia electrode as a detector for the purpose of performing faster. Flow injection analysis is fast, reliable in analysis data and easy to automate. The ammonia electrode reacts with volatile amines corresponding to volatile basic nitrogen as well as ammonia. It selectively reacts only with ammonia and volatile amines, but is not affected by other substances in the sample, making it suitable for the determination of volatile basic nitrogen. The use of ammonia electrodes also simplifies the construction of the flow injection analysis system since there is no need to use reagents other than alkaline solutions.

본 발명에서 자동분석을 위하여 제작하여 사용한 자동화 흐름주입분석 시스템의 구성은 도 1 과 같다. 그림에서와 같이 이송용액 운반용 튜빙펌프(Eyela, SMP-23S)(101), 용액선택밸브(Cole-Parmer, H-01367-81)(102), 시료 또는 표준용액 운반용 튜빙펌프(Watson Marlow, 101 U/R)(103), 주입밸브(Rheodyne, 5020)(104), 주입밸브 액튜에이터(Rheodyne, 5701)(105), 에어솔레노이드밸브(Rheodyne, 7163-033)(106), 질소압력통(107), 혼합코일(108), 플로우셀(109), 암모니아전극(Orion, 95-12)(110), 암모니아미터(Suntex, SP-701)(111), 접속장치(Advantech, PCL-812PG)(112) 및 개인용 컴퓨터(113)로 구성하였다. 이 중 플로우셀(109)은 도 2 와 같이 두개의 아크릴판을 가공하여 위(201)와 아래(202)로 붙여서 사용하였으며, 암모니아 전극막 밑부분의 부피는 약 40㎕였다. 테플론 튜빙의 내경은 0.8mm였으며, 주입밸브 루프의 용량은 약 50㎕였다. 이송용액은 5.0M 수산화나트륨을 사용하였으며, 유량은 0.8㎖/min였다. 자동분석 프로그램은 베이직으로 작성하였으며, 프로그램의 흐름도는 도 3 과 같다. 그림에서와 같이 두가지 표준용액 중 더 낮은 농도의 용액인 제1표준용액(0.1mM 또는 1.0mM)의 전압피크치(P1)(301), 더 높은 농도의 용액인 제2표준용액(1.0mM 또는 10.0mM)의 전압피크치(P2)(302) 및 시료의 전압피크치(PS)(303)를 순서대로 검출한 다음 이들을 환산식에 대입하여 시료의 농도를 산출(304)하였다. 환산식은 시료농도(mM) = 10^((PS - P1)/(P2 - P1)) 였다. The configuration of the automated flow injection analysis system manufactured and used for automatic analysis in the present invention is as shown in FIG. 1. As shown in the figure, a transporting tubing pump (Eyela, SMP-23S) 101, a fluid selection valve (Cole-Parmer, H-01367-81) (102), a tubing pump for transporting a sample or a standard solution (Watson Marlow, 101) U / R) (103), injection valve (Rheodyne, 5020) (104), injection valve actuator (Rheodyne, 5701) (105), air solenoid valve (Rheodyne, 7163-033) (106), nitrogen pressure cylinder (107) ), Mixing coil 108, flow cell 109, ammonia electrode (Orion, 95-12) (110), ammonia meter (Suntex, SP-701) (111), connecting device (Advantech, PCL-812PG) ( 112 and a personal computer 113. Of these, the flow cell 109 was used to process two acrylic plates as shown in Figure 2 and attached to the top (201) and bottom 202, the volume of the bottom of the ammonia electrode film was about 40㎛. The internal diameter of the Teflon tubing was 0.8 mm and the volume of the injection valve loop was about 50 μl. 5.0 M sodium hydroxide was used, and the flow rate was 0.8 ml / min. The automatic analysis program was written in BASIC, and the flowchart of the program is shown in FIG. 3. As shown in the figure, the voltage peak value (P1) 301 of the first standard solution (0.1 mM or 1.0 mM), which is the lower concentration of the two standard solutions, and the second standard solution (1.0 mM or 10.0, the higher concentration). mM peak voltage (P2) 302 and the sample peak voltage PS (303) were detected in this order, and these were substituted into the conversion equation to calculate the concentration of the sample (304). The conversion equation was sample concentration (mM) = 10 ^ ((PS-P1) / (P2-P1)) .

이하 자동적정방법을 돈육의 휘발성염기질소 분석에 적용한 내용을 설명한다. 시판 돈육을 실온에 적당한 시간 방치함으로써 부패도가 서로 다른 14가지의 시료를 만들고 각각의 시료에 대하여 미량확산법과 자동화 흐름주입분석법을 동시에 적용하여 비교하였다. 분석에 사용한 시료의 전처리 방법은 다음과 같다. 시료 10.0g을 달아 비이커에 넣고 증류수 50㎖를 첨가하여 잘 저어 섞어 30분간 침출하고 여과한다. 여과액을 5% 황산으로 약산성으로 중화시킨 후 증류수를 넣어 100.0㎖로 만든다. 이렇게 전처리한 시료에 대하여 미량확산법과 자동화 흐름주입분석법으로 분석하였다. 자동화 흐름주입분석법은 위에서 이미 기술하였으므로 이하 미량확산법의 과정을 기술한다. 먼저 확산기(Sibata)를 약간 기울여 놓고 외실의 아래쪽에 전처리한 시료 1.00㎖를 넣은 다음 내실 한쪽에 0.01N 황산 1.00㎖를 넣는다. 확산기 덮개의 갈아 맞추는 부분에 글리세린 소량을 고루 바른 다음 탄산칼륨 포화용액 1.00㎖를 외실의 다른쪽에 넣고 즉시 덮개를 덮어 클립으로 고정하고 확산기 를 전후좌우로 기울이면서 조용히 회전하여 외실의 시료와 탄산칼륨 포화용액을 잘 섞어 섭씨 25도에서 1시간 정치한다. 확산조작을 마친 후 덮개를 열고 내실의 황산용액에 브런스윅 지시약 한방울을 넣고 마이크로뷰렛을 사용하여 0.01N 수산화나트륨 용액으로 적정하여 적정소요량(A㎖)을 구한다. 따로 시료 대신 증류수를 써서 같은 방법으로 공시험을 하여 적정소요량(B㎖)을 구한 다음 환산식에 대입하여 시료농도를 구하였다. 환산식은 시료농도(mM)=10.0×(B-A)×F (F - 0.01N 수산화나트륨 용액의 역가) 였다. 돈육시료에 대한 결과를 도 4 에 표로써 나타내었다. 각각의 결과치는 3회 분석하여 산출한 평균치이다. 표에 나타난 미량확산법의 14개 평균치들과 자동화 흐름주입분석법의 14개 평균치들에 대하여 상관분석과 카이자승검정을 수행하였다. 그 결과 상관계수가 0.993으로서 두가지 방법의 평균치들 사이에 상관성이 아주 높은 것으로 나타났다. 카이자승치는 0.41로서 99.5%의 유의수준에서조차도 두가지 방법사이에 유의차가 인정되지 않았으며, 서로 잘 일치하는 것으로 나타났다. The following describes the application of the automatic titration method to the analysis of volatile nitrogen in pork. By placing the commercial pork at room temperature for a suitable time, 14 samples with different decay rates were prepared, and microdiffusion method and automated flow injection analysis were simultaneously applied to each sample. The pretreatment method of the sample used for analysis is as follows. 10.0 g of the sample is weighed into a beaker, 50 ml of distilled water is added, the mixture is stirred well, leached for 30 minutes and filtered. The filtrate is neutralized slightly with 5% sulfuric acid, and distilled water is added to make 100.0 ml. The pretreated samples were analyzed by microdiffusion and automated flow injection analysis. Automated flow injection analysis has already been described above, so the process of microdiffusion is described below. First, the diffuser (Sibata) is tilted slightly, and 1.00 ml of the pretreated sample is put in the lower part of the outer chamber, and then 1.00 ml of 0.01 N sulfuric acid is put in one of the inner chambers. Apply a small amount of glycerin evenly to the changing part of the diffuser cover, and then add 1.00 ml of saturated potassium carbonate solution to the other side of the outer chamber, and immediately cover the cover and fix it with a clip. Mix the solution well and let stand for 1 hour at 25 degrees Celsius. After the diffusion operation, open the cover, add a drop of Brunswick indicator to the sulfuric acid solution in the chamber, and titrate with 0.01 N sodium hydroxide solution using a microburette to obtain the required amount (Aml). Separately, distilled water was used instead of the sample, and a blank test was carried out in the same manner to obtain an appropriate amount (Bml), and then substituted into a conversion equation to obtain a sample concentration. The conversion formula was sample concentration (mM) = 10.0 x (BA) x F (titer of F-0.01 N sodium hydroxide solution) . The results for the pork sample are shown in Table 4. Each result is the average of three analyzes. Correlation analysis and chi-square test were performed on 14 average values of microdiffusion method and 14 average values of automated flow injection method. The correlation coefficient was 0.993, indicating a very high correlation between the averages of the two methods. The chi-chai score was 0.41, and even at the 99.5% significance level, no significant difference was found between the two methods.

다음으로는 자동적정방법을 우육의 휘발성염기질소 분석에 적용한 내용을 설명한다. 시판 우육을 실온에 적당한 시간 방치함으로써 부패도가 서로 다른 10가지의 시료를 만들고 각각의 시료에 대하여 미량확산법과 자동화 흐름주입분석법을 동시에 적용하여 비교하였다. 분석방법은 돈육의 경우와 동일하며, 분석 결과를 도 5 에 표로써 나타내었다. 각각의 결과치는 3회 분석하여 산출한 평균치이다. 두가지 방법의 평균치들에 대하여 상관분석과 카이자승검정을 수행한 결과 상관계수가 0.996이고 카이자승치는 0.25로서 돈육보다 더 우수한 결과가 얻어졌다. The following describes the application of the automatic titration method to the analysis of volatile nitrogen in beef. By placing the commercially available beef at room temperature for 10 hours, ten samples with different decay rates were prepared, and microdiffusion and automated flow injection analysis were applied to each sample at the same time. The analysis method is the same as that of pork, and the analysis results are shown as a table in FIG. Each result is the average of three analyzes. Correlation analysis and chi-square test were performed on the mean values of the two methods, and the correlation coefficient was 0.996 and chi-square was 0.25, which is better than pork.

돈육과 우육에 대한 실험결과, 두가지 방법으로부터의 분석치 사이의 상관성 이 대단히 높고 적합도가 아주 크므로 자동화 흐름주입분석법으로 미량확산법을 대체할 수 있다고 판단되었다. Experiments on pork and beef showed that the correlation between the analyzes from the two methods was very high and the fit was very good.

미량확산법의 경우 확산시간만 1시간이므로 총 분석시간은 1.5시간 내지 2시간 이상 거리는데 자동화 흐름주입분석장치로 자동분석할 경우에는 한시료의 분석시간을 4분정도까지 단축시킬 수 있다. 또한, 자동분석이므로 시료전처리 후에는 간편하게 분석할 수 있다. 따라서 분석효율을 크게 증가시킬 수 있다. In the microdiffusion method, the diffusion time is only 1 hour, so the total analysis time is 1.5 hours to 2 hours or more. In the case of automatic analysis using an automated flow injection analysis device, the analysis time of one sample can be reduced to about 4 minutes. In addition, it is easy to analyze after sample preparation because it is an automatic analysis. Therefore, the analysis efficiency can be greatly increased.

Claims (1)

휘발성 염기질소 분석에서 튜빙펌프, 용액선택밸브, 주입밸브, 주입밸브 액튜에이터, 에어솔레노이드밸브, 질소압력통, 혼합코일, 플로우셀, 암모니아전극, 암모니아미터, 접속장치 및 개인용 컴퓨터로 구성한 자동화 흐름주입분석장치로 자동분석하는 방법.Automated flow injection analysis consisting of tubing pump, solution selection valve, injection valve, injection valve actuator, air solenoid valve, nitrogen pressure cylinder, mixing coil, flow cell, ammonia electrode, ammonia meter, connecting device and personal computer in volatile basic nitrogen analysis How to auto analyze by device.
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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102043003A (en) * 2010-12-03 2011-05-04 江南大学 Method for measuring mixed base concentration by combining pH electrode and flow injection analysis (FIA)
CN102374989A (en) * 2010-08-20 2012-03-14 北京吉天仪器有限公司 Automatic analyzer for determining ammonia nitrogen in water and automatic analytic method thereof
RU2554799C2 (en) * 2013-11-06 2015-06-27 Федеральное государственное бюджетное научное учреждение "Северо-Кавказский зональный научно-исследовательский институт садоводства и виноградарства" Method of determining total nitrogen by capillary electrophoresis

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JPH07270379A (en) * 1994-03-31 1995-10-20 Tokuyama Corp Determination of chloride ion
JPH08101162A (en) * 1994-09-30 1996-04-16 Tokuyama Corp Quantitative determination method for acetic acid or acetate

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102374989A (en) * 2010-08-20 2012-03-14 北京吉天仪器有限公司 Automatic analyzer for determining ammonia nitrogen in water and automatic analytic method thereof
CN102043003A (en) * 2010-12-03 2011-05-04 江南大学 Method for measuring mixed base concentration by combining pH electrode and flow injection analysis (FIA)
RU2554799C2 (en) * 2013-11-06 2015-06-27 Федеральное государственное бюджетное научное учреждение "Северо-Кавказский зональный научно-исследовательский институт садоводства и виноградарства" Method of determining total nitrogen by capillary electrophoresis

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