CN111289640A - Liquid chromatography detection method for improving detection efficiency by advancing sample introduction - Google Patents

Liquid chromatography detection method for improving detection efficiency by advancing sample introduction Download PDF

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CN111289640A
CN111289640A CN202010124201.8A CN202010124201A CN111289640A CN 111289640 A CN111289640 A CN 111289640A CN 202010124201 A CN202010124201 A CN 202010124201A CN 111289640 A CN111289640 A CN 111289640A
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sample
time
peak
samples
chromatographic
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CN111289640B (en
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伍玉碧
周日尤
李瑛�
曹媛
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Nanjing Kaitong Grain Biochemistry R And D Co ltd
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Nanjing Kaitong Grain Biochemistry R And D Co ltd
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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/04—Preparation or injection of sample to be analysed
    • G01N30/16—Injection
    • G01N30/22—Injection in high pressure liquid systems

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Abstract

A liquid chromatogram detection method for improving detection efficiency by advancing sample introduction is characterized in that a high performance liquid chromatograph is used for detecting a series of similar samples, the retention time of all substances to be detected in the samples is longer, the peak emergence time range of all the substances to be detected in the samples is smaller than the blank time of the samples, in order to effectively utilize the blank time of the samples, the spectral peak time of the previous sample is arranged in the blank time of the next sample, and the advance sample introduction is adopted to improve the detection speed and the utilization rate of the high performance liquid chromatograph.

Description

Liquid chromatography detection method for improving detection efficiency by advancing sample introduction
Technical Field
The invention relates to the technical field of analysis and detection. In particular to a high performance liquid chromatography detection method.
Background
High Performance Liquid Chromatography (HPLC) has the advantages of high column efficiency, good selectivity, high sensitivity, high analysis speed, good repeatability and the like, is one of the main means of modern analysis technology, and is widely applied to the scientific fields of chemistry, chemical engineering, medicine, biochemistry, environmental protection, agriculture and the like at present.
When the high performance liquid chromatography is used for detection, the time required from sample introduction to the completion of peak emergence of all substances in a sample is the detection time of the sample, and the detection time comprises blank area time and chromatographic area time of the sample.
The blank time is referred to as a blank time. The blank time is the time corresponding to the time from the sample introduction to the beginning of the first chromatographic peak, and no chromatographic peak is present in the time zone. The blank time is effectively an invalid latency.
The chromatographic time is the region from the beginning of the appearance of the first chromatographic peak to the end of the last chromatographic peak in the sample, and the corresponding time width is called the chromatographic peak time.
In actual operation, a larger blank time indicates a longer blank time to wait for each analytical test. Sometimes the blank time is half or even more of the total time of the analysis spectrogram.
In the sample detection of scientific research and enterprise production, especially in the production field of a factory and the trial run stage of a production line, the rapid response to the field sampling result is generally required so as to adjust parameters, process indexes and the like in time. For the production of some enterprises, the speed of analyzing and detecting the speed directly influences the production cost.
While blank time is an ineffective time in the chromatogram, it is also essential in the HPLC assay. How to fully utilize the necessary invalid time is the aim of the invention.
The invention reasonably utilizes the blank time without changing the operating condition of the high performance liquid chromatography, and arranges the peak time of the spectrum of the former sample in the blank time area of the latter sample, thereby shortening the detection time of the sample and improving the use efficiency of the HPLC.
Disclosure of Invention
In HPLC detection, how to reasonably utilize blank time, the invention provides a liquid chromatogram detection method which advances sample introduction to improve detection speed, so as to reduce total detection time and improve detection efficiency.
For clarity, the symbols associated with the present invention are defined as follows:
ta: blank time of a chromatogram in a conventional sample introduction mode of a sample;
tb: and (3) in a conventional sample injection mode, the time from the starting point of the chromatographic peak of the first substance to the peak area of the chromatographic peak of the last substance is up.
According to the invention, a high performance liquid chromatograph is adopted to detect a series of similar samples, the retention time of all substances to be detected in the samples is longer, and the time of the spectral peak area of all the substances in the samples is less than the blank time of the samples.
The invention is realized by the following steps:
(1) after the high performance liquid chromatograph operates stably, a first sample is fed, data are collected, and data collection is stopped 1-2 min before a first chromatographic peak of the first sample appears;
(2) entering a second sample, collecting data, stopping collecting data and storing spectrogram data after the last chromatographic peak of the previous sample is finished;
(3) repeating step (2) starting from the third sample;
(4) and (3) detection results: judging substances corresponding to the chromatographic peaks according to the characteristics and time sequence of the chromatographic peaks without taking the retention time as a basis; and calculating the content of the corresponding substance by using an area normalization method according to the peak area of each chromatographic peak.
Furthermore, the temperature of the chromatographic column and the column, the mobile phase and the flow rate are not changed.
Further, the peak time range of all substances of the sample is less than the blank time of the sample.
Under a conventional sample injection mode, the blank time of a similar sample is Ta, the time of a spectrum peak area is Tb, and as long as Ta is more than Tb, the sample can adopt a liquid chromatography detection method for improving the detection speed by injecting samples in advance.
The blank time can be reduced by adopting a method of sampling in advance, the retention time corresponding to each chromatographic peak is correspondingly shortened compared with the conventional sampling method, but the appearance sequence of the chromatographic peaks of each substance in a spectrogram is kept unchanged, and the substances corresponding to the chromatographic peaks are judged according to the characteristic and the time sequence of the chromatographic peaks instead of the retention time of the conventional sampling when the spectrogram is analyzed. And calculating the content of the corresponding substance by using an area normalization method according to the peak area of each chromatographic peak.
In the process of factory production and production line test operation, a large number of samples of the same type are required to be repeatedly detected, and the detection result is required to be capable of quickly responding, so that the method is particularly suitable for detection of products in different sections and batches in factory production and detection of products in production line test operation.
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FIG. 1A is a schematic diagram of conventional sample injection and pre-injection chromatograms of a sample. Blank time of Ta1The time of the spectral peak is Tb1，Ta1＞Tb1And detecting the sample by adopting a method of sampling in advance.
FIG. 2 is a schematic diagram of conventional sample introduction and pre-sample introduction chromatograms of a class B sample. Blank time of Ta2The time of the spectral peak is Tb2，Ta2＞Tb2And detecting the sample by adopting a method of sampling in advance.
FIG. 3 is a schematic diagram of a cross-over pre-injection chromatogram of a type A sample and a type B sample. Satisfy Ta1＞Tb1，T a2＞Tb2And detecting the two types of samples by adopting a method of cross sampling in advance.
FIG. 4C sample conventional sample chromatogram and detection result.
FIG. 5C sample pre-injection chromatogram and detection results.
FIG. 6 is a conventional sample chromatogram and detection result of class D sample.
FIG. 7A is a chromatogram and detection result of class D sample injected in advance.
Detailed description of the preferred embodiments
The technical features and effects of the present invention will be further illustrated by the following specific embodiments, but the present invention is not limited thereto.
Example 1
A schematic diagram of conventional sample introduction and pre-sample introduction chromatograms of the A-type sample is shown in FIG. 1.
When the A-type sample is subjected to conventional sample injection detection, the sample 1 is fed, and then the sample 2 is fed after blank time and spectrum peak time.
Detection results of conventional sample injection of A-type samples: and calculating the content of the corresponding substance by using an area normalization method according to the peak area of each chromatographic peak by taking the retention time as a basis.
Blank time of A type sample is Ta1The time of the spectral peak is Tb1，Ta1＞Tb1And detecting the sample by adopting a method of sampling in advance. Feeding a sample 1, collecting data, and stopping collecting data 1-2 min before a first chromatographic peak of a first sample appears; feeding a sample 2, collecting data, stopping collecting data and storing spectrogram data after the last chromatographic peak of the sample 1 is peaked; feeding a sample 3, stopping data acquisition and storing spectrogram data after the last chromatographic peak of the sample 2 is peaked; sample 4 was added. And repeating the operation.
Detection results of advanced sample introduction of A-type samples: judging substances corresponding to the chromatographic peaks according to the characteristics and time sequence of the chromatographic peaks without taking the retention time as a basis; and calculating the content of the corresponding substance by using an area normalization method according to the peak area of each chromatographic peak.
Example 2
A schematic diagram of the conventional sample introduction and the pre-sample introduction chromatograms of the B-type sample is shown in FIG. 2.
When the B-type sample is detected by conventional sample introduction, the sample 1 is fed, and the sample 2 is fed after blank time and spectrum peak time.
Detection results of conventional sample injection of B-type samples: and calculating the content of the corresponding substance by using an area normalization method according to the peak area of each chromatographic peak by taking the retention time as a basis.
Blank time of class B sample is Ta2The time of the spectral peak is Tb2，Ta2＞Tb2And detecting the sample by adopting a method of sampling in advance. Feeding a sample 1, collecting data, and stopping collecting data 1-2 min before a first chromatographic peak of a first sample appears; feeding a sample 2, collecting data, stopping collecting data and storing spectrogram data after the last chromatographic peak of the sample 1 is peaked; feeding a sample 3, stopping data acquisition and storing spectrogram data after the last chromatographic peak of the sample 2 is peaked; sample 4 was added. And repeating the operation.
Detection results of advanced sample introduction of B-type samples: judging substances corresponding to the chromatographic peaks according to the characteristics and time sequence of the chromatographic peaks without taking the retention time as a basis; and calculating the content of the corresponding substance by using an area normalization method according to the peak area of each chromatographic peak.
Example 3
The schematic diagram of the advanced cross-sampling chromatogram of the A-type sample and the B-type sample is shown in FIG. 3.
The chromatographic peak patterns of the A-type sample and the B-type sample are different, but both satisfy Ta1＞Tb1，Ta2＞Tb2And detecting the two types of samples by adopting a cross advanced sample introduction method.
Feeding a sample 1, collecting data, and stopping collecting data 1-2 min before a first chromatographic peak of a first sample appears; feeding a sample 2, collecting data, stopping collecting data and storing spectrogram data after the last chromatographic peak of the sample 1 is peaked; feeding a sample 3, stopping data acquisition and storing spectrogram data after the last chromatographic peak of the sample 2 is peaked; sample 4 was added. And repeating the operation.
The detection results of the A-type sample and the B-type sample are in advance subjected to cross sampling: judging substances corresponding to the chromatographic peaks according to the characteristics and time sequence of the chromatographic peaks without taking the retention time as a basis; and calculating the content of the corresponding substance by using an area normalization method according to the peak area of each chromatographic peak.
Example 4
Detecting by high performance liquid chromatography according to conventional sample introduction, wherein the detection conditions are as follows: special H for Bio-Rad sugar analysis+A type ion chromatography column; a difference detector; the mobile phase is 0.33 per mill of dilute sulfuric acid solution prepared by pure water; the flow rate is 0.4 mL/min; the column temperature is 35 ℃; the amount of the sample was 20. mu.L.
The C-type sample is a mixture which is an organic matter mixed solution of polyhydric alcohol; the class D sample is also a mixture, which is an organic mixed solution of polyols.
FIG. 4 is a chromatogram of a conventional sample injection for a class C sample. For the C-type sample, the starting time of the first chromatographic peak is 16.8min, the ending time of the last chromatographic peak is 31.2min, the blank time Ta1 is 16.8min, and the peak time Tb1 is (31.2-16.8) =14.4 min.
Comparison of blank time Ta for class C samples1And its peak time Tb1Ta1 is more than Tb1, and the detection is carried out by adopting a mode of advanced sample injection. The results of the detection are shown in FIG. 5.
FIG. 6 is a conventional sample chromatogram of class D samples. For the D sample, the initial time of the first chromatographic peak is 16.8min, the end time of the last chromatographic peak is 31.0min, and the blank time Ta is216.8min, time Tb of spectral peak region2Is (31.0-16.8) =14.2 min.
Comparison of blank time Ta for class D samples2And its peak time Tb2Ta2 is more than Tb2, and the detection is carried out by adopting a mode of advanced sample injection. The results of the detection are shown in FIG. 7.
Although the present invention has been described with reference to the preferred embodiments, it should be understood that various changes and modifications can be made therein by those skilled in the art without departing from the spirit and scope of the invention as defined in the appended claims.

Claims (4)

1. A liquid chromatogram detection method for advancing sample introduction and improving detection efficiency uses a high performance liquid chromatograph to detect a series of similar samples, the retention time of all substances to be detected in the samples is longer, and the peak-out time range of all substances to be detected in the samples is less than the blank time of the samples, in order to effectively utilize the blank time of the samples, the spectrum peak time of the previous sample is arranged in the blank time of the next sample, advance sample introduction is adopted to improve the detection speed and the utilization rate of the high performance liquid chromatograph, and the method is characterized by comprising the following steps:
(1) after the high performance liquid chromatograph operates stably, a first sample is fed, data are collected, and data collection is stopped 1-2 min before a first chromatographic peak of the first sample appears;
(2) entering a second sample, collecting data, stopping collecting data and storing spectrogram data after the last chromatographic peak of the previous sample is finished;
(3) repeating step (2) starting from the third sample;
(4) and (3) detection results: judging substances corresponding to the chromatographic peaks according to the characteristics of the chromatographic peaks and the time sequence; and calculating the content of the corresponding substance by using an area normalization method according to the peak area of each chromatographic peak.
2. The method of claim 1, wherein the column temperature, mobile phase and flow rate are constant.
3. The method of claim 1, wherein the sample has an all material peak time range less than the sample blank time.
4. The method according to claim 1, characterized in that the substance corresponding to each chromatographic peak is judged according to the characteristic and time sequence of the chromatographic peak without taking the retention time of conventional sample injection as the basis.
CN202010124201.8A 2020-02-27 2020-02-27 Liquid chromatography detection method for improving detection efficiency by advancing sample introduction Active CN111289640B (en)

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN115453030A (en) * 2022-09-28 2022-12-09 博莱克科技（武汉）有限公司 Method for improving gas chromatography detection efficiency

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2001153853A (en) * 1999-11-26 2001-06-08 Shimadzu Corp Liquid chromatograph
JP2005127814A (en) * 2003-10-23 2005-05-19 Shimadzu Corp Chromatographic analyzer
JP2005257575A (en) * 2004-03-15 2005-09-22 Shimadzu Corp Chromatograph
JP2008076243A (en) * 2006-09-21 2008-04-03 Hitachi High-Technologies Corp Chromatograph device and analysis method

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2001153853A (en) * 1999-11-26 2001-06-08 Shimadzu Corp Liquid chromatograph
JP2005127814A (en) * 2003-10-23 2005-05-19 Shimadzu Corp Chromatographic analyzer
JP2005257575A (en) * 2004-03-15 2005-09-22 Shimadzu Corp Chromatograph
JP2008076243A (en) * 2006-09-21 2008-04-03 Hitachi High-Technologies Corp Chromatograph device and analysis method

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN115453030A (en) * 2022-09-28 2022-12-09 博莱克科技（武汉）有限公司 Method for improving gas chromatography detection efficiency

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