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Biological sample processing method, biological sample ferrous ion or total ferric ion determination method and detection kit

A technology of biological samples and ferrous ions, which is applied in the preparation of test samples, sampling, measuring devices, etc., can solve the problems of difficult release and inability to use ferric ions to measure, so as to save manpower and material resources, and the method is simple and accurate. sexual effect

Active Publication Date: 2021-09-28
JILIN UNIV
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  • Abstract
  • Description
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Problems solved by technology

[0005] In the traditional biological sample processing method, the method of releasing the cell content is usually to lyse the cells. At present, there is a general-purpose cell lysate in the market that reacts with tripyridyl triazine to form a milky white precipitate, which cannot be used for the determination of iron ions.
Using the osmotic pressure gradient, the biological sample is placed in double distilled water, and the cells are swelled, but most of the iron ions are combined with proteins, DNA or RNA, and it is difficult to release. Reaction of Tripyridyl Triazine to Form Complexes

Method used

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  • Biological sample processing method, biological sample ferrous ion or total ferric ion determination method and detection kit
  • Biological sample processing method, biological sample ferrous ion or total ferric ion determination method and detection kit
  • Biological sample processing method, biological sample ferrous ion or total ferric ion determination method and detection kit

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0130] Handling of biological samples:

[0131] Step 11, the ovarian cancer Skov3 cell line, cultured for 24 hours to obtain 5 × 10 6 cells, the cultured cells were collected and centrifuged, and the supernatant was discarded to obtain cell pellets.

[0132] Step 12, freezing the collected cell mass in liquid nitrogen overnight and grinding it into fine powder.

[0133] Step 13. Place the biological sample powder obtained in Step 12 in 9% HCL, and ultrasonically shake the sample for 20 minutes after overnight at 4°C. The shaking conditions are 25°C, 40kHz, high frequency.

[0134]Step 14. Place the mixed solution shaken in step 13 in a high-speed centrifuge at 12,000 g at 4°C for 10 minutes, discard the precipitate, and keep the supernatant.

[0135] Determination of the optimum wavelength of the improved TPTZ spectrophotometry:

[0136] Take 1ng / μL ferrous chloride standard stock solution, after being processed by the above-mentioned assay scheme, place it in a microplate ...

Embodiment 2

[0163] Handling of biological samples:

[0164] Step 11, Skov3 cell line, after culturing for 24 hours, 5×10 6 cells, the cultured cells were collected and centrifuged, and the supernatant was discarded to obtain cell pellets.

[0165] Step 12, freezing the collected cell mass in liquid nitrogen overnight and grinding it into fine powder.

[0166] Step 13. Place the biological sample powder obtained in Step 12 in 9% HCL, and ultrasonically shake the sample for 20 minutes after overnight at 4°C. The shaking conditions are 25°C, 40kHz, high frequency.

[0167] Step 14. Place the mixed solution shaken in step 13 in a high-speed centrifuge at 12,000 g at 4°C for 10 minutes, discard the precipitate, and keep the supernatant.

[0168] Determination of the optimum wavelength of the improved TPTZ spectrophotometry:

[0169] Take 1ng / μL ferrous chloride standard stock solution, after being processed by the above-mentioned assay scheme, place it in a microplate reader for full-wavele...

Embodiment 3

[0198] Handling of biological samples:

[0199] Step 11, culture the ovarian cancer Skov3 cell line for 24 hours to obtain 5×10 6 cells, the cultured cells were collected and centrifuged, and the supernatant was discarded to obtain cell pellets.

[0200] Step 12. Place the cell mass in 10% HCL for direct ultrasonic vibration for 1 hour. The vibration condition is 25° C., 50 kHz, high frequency.

[0201] Step 13. Place the mixed solution shaken in step 12 at 10,000 g in a high-speed centrifuge at 0° C., centrifuge for 10 minutes, discard the precipitate, and keep the supernatant.

[0202] Determination of the optimum wavelength of the improved TPTZ spectrophotometry:

[0203] Take 1ng / μL ferrous chloride standard stock solution, after being processed by the above-mentioned assay scheme, place it in a microplate reader for full-wavelength scanning. Double distilled water was used as blank control. The result is as Figure 6 , it can be seen that the optimal detection wavele...

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Abstract

The application provides a biological sample processing method, a biological sample ferrous ion or total iron ion determination method and a detection kit. In the above-mentioned biological sample processing method, the acidic solution can fully swell and crack the cells through the principle of low osmosis, and at the same time can fully release the iron ions in the cells under the action of acid, and do not have other reactions with the chromogenic agent, and do not affect the Measurement of iron ions. The ferrous ion determination method of the above-mentioned biological sample and the total iron ion determination method of the biological sample, by increasing the concentration of the reducing agent, the iron ion is fully reduced, and by increasing the lye, the tripyridyl triazine and the ferrous ion in the solution Iron ions generate stable blue-purple complexes, which can be used for the detection of ferrous ions, which is simple and accurate. The above-mentioned ferrous ion and total iron ion detection kit can be used for the determination of ferrous ion and total iron ion in biological samples.

Description

technical field [0001] The invention relates to the technical field of biological sample detection, in particular to a biological sample processing method, a biological sample ferrous ion determination method, a biological sample total iron ion determination method, a biological sample ferrous ion detection kit and a biological sample total Iron ion detection kit. Background technique [0002] There are many trace metal elements involved in different physiological metabolic activities in organisms. Iron is a mineral that plays a vital role in many biological processes such as DNA synthesis, electron transfer and oxygen transport. Iron is a transition element that can form a range of oxidation states, the most common being ferrous iron (Fe 2+ or ferrous) and ferric (Fe 3+ ). Iron-containing proteins are involved in many reactions, often utilizing transient changes in the oxidation state of iron for chemical reactions. Iron deficiency and excess may cause serious human dy...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/31G01N1/28G01N1/38
CPCG01N1/28G01N1/286G01N1/38G01N21/31G01N2001/2866G01N2001/386
Inventor 周昊函徐巍田佳怡梁琳李乐梁婷
Owner JILIN UNIV
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