CN104140995A - Ceftriaxone residue detection method - Google Patents

Abstract

A ceftriaxone residue detection method is characterized by comprising the steps that (1) a panel is prepared; (2) a standard solution is prepared, wherein the standard solution is a stock solution and standard diluents; (3) a sample is obtained to be subjected to surface monitoring and air monitoring; (4) a test is carried out. The method is used for detecting ceftriaxone residues on the surface and in the air, the ceftriaxone residues can be detected in time according to the standard, an abnormal residue concentration can be eliminated in time, and air cleanness of a production environment is guaranteed.

Description

A kind of Ceftriaxone method for detecting residue

Technical field

The present invention relates to a kind of residuals content to detection method, be specifically related to a kind of Ceftriaxone method for detecting residue.

Background technology

In order to check that validity and the confirmation of cleaning do not have Ceftriaxone crossed contamination to occur.The present invention has set up Ceftriaxone output zone and other and has had the detection method of Ceftriaxone in the region of potential cross contamination risk.

Summary of the invention

The object of the invention is to, a kind of Ceftriaxone method for detecting residue is provided, to overcome the existing above-mentioned shortcoming and defect of prior art.

The technical problem that will solve required for the present invention, can be achieved through the following technical solutions:

A Ceftriaxone method for detecting residue, is characterized in that, comprising:

(1) preparation is dull and stereotyped;

(2) preparation standard solution, wherein said standardized solution is storing solution and standard diluent;

(3) surface monitoring and air monitering are carried out in sampling;

(4) test

(4.1) typical curve

(4.2) sample adds and cultivates

(4.3) calculate

$b=\frac{\mathrm{\Σ}{x}_i{y}_i-\stackrel{\‾}{x}\mathrm{\Σ}{y}_i}{\mathrm{\Σ}{x}_i^2-\stackrel{\‾}{x}\mathrm{\Σ}{x}_i},a=\stackrel{\‾}{y}-b\stackrel{\‾}{x}$

Y＝bX+a

Accurately measure each antibacterial circle diameter to 0.1 millimeter; With above formula, obtain typical curve regression equation, Y is antibacterial circle diameter, and X is the lg value of Ceftriaxone concentration, by sample antibacterial circle diameter substitution equation, calculates result, and the surperficial sample result of gained should be μ g/m ², air sample is μ g/m ³.

In the present invention, Ceftriaxone method for detecting residue also comprises: (4.4) Acceptable criterion and test frequency

(4.4.1) air monitering

(4.4.2) surface monitoring

(4.4.3) sampling point position distributes;

(4.5) abnormality processing: in accordance with SOP-CB-0008.

Wherein, in step (1), described flat board is that AM1 is dull and stereotyped; AM1 flat board should face with newly joining; Prepare 100 milliliters of No. 1 microbiotic substratum, 121 ℃ of sterilizings 20 minutes, substratum is cooled to 48 ± 2 ℃; Add 2 milliliters of intestinal bacteria inoculation bacterium liquid to 100 milliliter substratum, jog mixes, and avoids producing foam; Substratum is poured into bioassay culture dish, guarantee that the planar surface making is smooth, avoid bubble, be cooled to curing.

Wherein, in step (2), described storing solution: accurately take 40 milligrams of Ceftriaxones in 100 milliliters of volumetric flasks, use pure water to dissolve and constant volume, this solution must face with newly joining;

Described standard diluent: storing solution is made to each dilution standard diluent.

Wherein, in step (3), described surface monitoring: by scraps of paper wiping one particular area of tweezers tweezer 6 mm dias, with 10 * 10cm ²the region irised out of stainless steel framework; The scraps of paper after sampling are positioned on AM1 flat board;

Described air monitering: the filter paper that is 12.7 millimeters by a slice diameter is positioned in suction filter; Regulate vacuum pump rate of air sucked in required to 10 liters of about per minutes, each sampling point extracts 100 litres of air; Taking-up is adopted the excellent scraps of paper and is positioned on AM1 flat board;

Wherein, in step (4.1), the scraps of paper are positioned over to culture medium flat plate, drip standard diluent, each standard diluent drips 2 scraps of paper; With 6 millimeters of scraps of paper, every adds respectively 20 microlitre standard diluents 1～5; With 12.7 millimeters of scraps of paper, every adds respectively 20 microlitre standard diluents 1～4, then adds 80 microlitre pure water to every filter paper.

Wherein, in step (4.2), for surface monitoring, on each sample filter disc, add 20 μ l purified water; For air monitoring, on each sample filter disc, add 100 μ l purified water; At 30～35 ℃, cultivate 20～24 hours; Standard and sample should be on same flat boards.

Beneficial effect of the present invention:

The present invention has set up the residual detection method of Ceftriaxone in surface and air, and the residual of Ceftriaxone can be detected in time according to this standard, and abnormal residual concentration is removed in time, guarantees the air purity of production environment.

Accompanying drawing explanation

Fig. 1 is the sampling point position distribution plan of ground floor.

Fig. 2 is the sampling point position distribution plan on roof.In Fig. 2, the upper left corner direction of arrow is south, and TA3 represents HVAC system exhaust outlet.

Embodiment

Below in conjunction with specific embodiment, the present invention is done to progressive explanation.Should be understood that following examples are only for the present invention is described but not for limiting scope of the present invention.

Embodiment 1

A Ceftriaxone method for detecting residue, comprises the following steps:

1. detect the preparation with AM1 flat board

According to 100 milliliters of No. 1 microbiotic substratum of compound method preparation of supplier, 121 ℃ of sterilizings 20 minutes, substratum is cooled to 48 ± 2 ℃.Add 2 milliliters of intestinal bacteria inoculation bacterium liquid to 100 milliliter substratum, jog mixes, and avoids producing foam.Substratum is poured into bioassay culture dish, guarantee that the planar surface making is smooth, avoid bubble, be cooled to curing.AM1 flat board should face with newly joining.

2. the preparation of standardized solution

Storing solution: accurately take 40 milligrams of Ceftriaxones in 100 milliliters of volumetric flasks, use pure water to dissolve and constant volume.This solution must face with newly joining.

Standard diluent: press the method for table 1, storing solution is made to each dilution standard diluent.

The preparation of table 1 standard diluent

3. sampling

Surface monitoring: use scraps of paper wiping one particular area of tweezers tweezer 6 mm dias (with 10 * 10cm ²the region irised out of stainless steel framework).The scraps of paper after sampling are positioned on AM1 flat board.

Air monitering: the filter paper that is 12.7 millimeters by a slice diameter is positioned in suction filter.Regulate vacuum pump rate of air sucked in required to 10 liters of about per minutes, each sampling point extracts 100 litres of air.Taking-up is adopted the excellent scraps of paper and is positioned on AM1 flat board.

4. test

4.1 typical curve

The scraps of paper are positioned over to culture medium flat plate, drip standard diluent, each standard diluent drips 2 scraps of paper.With 6 millimeters of scraps of paper, every adds respectively 20 microlitre standard diluents 1～5.With 12.7 millimeters of scraps of paper, every adds respectively 20 microlitre standard diluents 1～4, then adds 80 microlitre pure water to every filter paper.

4.2 samples add and cultivate

For surface monitoring, on each sample filter disc, add 20 μ l purified water.For air monitoring, on each sample filter disc, add 100 μ l purified water.At 30～35 ℃, cultivate 20～24 hours.Standard and sample should be on same flat boards.

4.3 calculate

$b=\frac{\mathrm{\Σ}{x}_i{y}_i-\stackrel{\‾}{x}\mathrm{\Σ}{y}_i}{\mathrm{\Σ}{x}_i^2-\stackrel{\‾}{x}\mathrm{\Σ}{x}_i},a=\stackrel{\‾}{y}-b\stackrel{\‾}{x}$

Y＝bX+a

Accurately measure each antibacterial circle diameter to 0.1 millimeter.With above formula, obtain typical curve regression equation, Y is antibacterial circle diameter, and X is the lg value of Ceftriaxone concentration.By sample antibacterial circle diameter substitution equation, calculate result.The surperficial sample result of gained should be μ g/m ², air sample is μ g/m ³.

4.4 Acceptable criterions and test frequency

4.4.1. air monitering/Air monitoring, table 2 is for sampling point is to distributing.

Table 2 sampling point is to distributing

4.4.2 surface monitoring, table 3 sampling point is to distributing

Table 3 sampling point is to distributing

4.4.3 sampling point position distributes.

Fig. 1 is the sampling point position distribution plan of ground floor, the sampling point position distribution plan that Fig. 2 is roof, and sampling point position distribution situation, as depicted in figs. 1 and 2.

4.5 abnormality processing: in accordance with SOP-CB-0008.

The material using in the present embodiment

Test bacterial classification: Escherichia coli ATCC 8739/ intestinal bacteria ATCC 8739, to TSA inclined-plane, cultivates intestinal bacteria ATCC 8739 deposit bacterial suspension inoculations 18～24 hours for 30～35 ℃.With 5 milliliters of aseptic peptone physiological saline, wash lower grower, get 1 milliliter, use 10 times of stepwise dilutions of aseptic peptone physiological saline 2 times, final diluent is as inoculation bacterium liquid.

No. 1 microbiotic substratum (Merck or equivalent).

Calibrating filter paper: Φ 6mm surface sample, Φ 12.7mm air sample.

Bioassay culture dish.

Incubator: 30-35 ℃.

Biohazard Safety Equipment.

The precision of slide calliper rule :≤0.1mm.

Tweezers, suction pipe, microscale sampler, test tube, volumetric flask.

Stainless steel framework.

Suction filtration device and vacuum pump.

Above the specific embodiment of the present invention is illustrated, but the present invention is as limit, only otherwise depart from aim of the present invention, the present invention can also have various variations.

Claims (7)

1. a Ceftriaxone method for detecting residue, is characterized in that, comprising:

(1) preparation is dull and stereotyped;

(2) preparation standard solution, wherein said standardized solution is storing solution and standard diluent;

(3) surface monitoring and air monitering are carried out in sampling;

(4) test

(4.1) typical curve

(4.2) sample adds and cultivates

(4.3) calculate

$b=\frac{\mathrm{\Σ}{x}_i{y}_i-\stackrel{\‾}{x}\mathrm{\Σ}{y}_i}{\mathrm{\Σ}{x}_i^2-\stackrel{\‾}{x}\mathrm{\Σ}{x}_i},a=\stackrel{\‾}{y}-b\stackrel{\‾}{x}$

Y＝bX+a

Accurately measure each antibacterial circle diameter to 0.1 millimeter; With above formula, obtain typical curve regression equation, Y is antibacterial circle diameter, and X is the lg value of Ceftriaxone concentration, by sample antibacterial circle diameter substitution equation, calculates result, and the surperficial sample result of gained should be μ g/m ², air sample is μ g/m ³.

2. method according to claim 1, is characterized in that: also comprise: (4.4) Acceptable criterion and test frequency

(4.4.1) air monitering

(4.4.2) surface monitoring

(4.4.3) sampling point position distributes;

(4.5) abnormality processing: in accordance with SOP-CB-0008.

3. method according to claim 1, is characterized in that: in step (1), described flat board is that AM1 is dull and stereotyped; AM1 flat board should face with newly joining; Prepare 100 milliliters of No. 1 microbiotic substratum, 121 ℃ of sterilizings 20 minutes, substratum is cooled to 48 ± 2 ℃; Add 2 milliliters of intestinal bacteria inoculation bacterium liquid to 100 milliliter substratum, jog mixes, and avoids producing foam; Substratum is poured into bioassay culture dish, guarantee that the planar surface making is smooth, avoid bubble, be cooled to curing.

4. method according to claim 1, is characterized in that: in step (2),

Described storing solution: accurately take 40 milligrams of Ceftriaxones in 100 milliliters of volumetric flasks, use pure water to dissolve and constant volume, this solution must face with newly joining;

Described standard diluent: storing solution is made to each dilution standard diluent.

5. method according to claim 1, is characterized in that: in step (3),

Described surface monitoring: by scraps of paper wiping one particular area of tweezers tweezer 6 mm dias, with 10 * 10cm ²the region irised out of stainless steel framework; The scraps of paper after sampling are positioned on AM1 flat board;

Described air monitering: the filter paper that is 12.7 millimeters by a slice diameter is positioned in suction filter; Regulate vacuum pump rate of air sucked in required to 10 liters of about per minutes, each sampling point extracts 100 litres of air; Taking-up is adopted the excellent scraps of paper and is positioned on AM1 flat board.

6. method according to claim 1, is characterized in that: in step (4.1), the scraps of paper are positioned over to culture medium flat plate, drip standard diluent, each standard diluent drips 2 scraps of paper; With 6 millimeters of scraps of paper, every adds respectively 20 microlitre standard diluents 1～5; With 12.7 millimeters of scraps of paper, every adds respectively 20 microlitre standard diluents 1～4, then adds 80 microlitre pure water to every filter paper.

7. method according to claim 1, is characterized in that: in step (4.2), for surface monitoring, add 20 μ l purified water on each sample filter disc; For air monitoring, on each sample filter disc, add 100 μ l purified water; At 30～35 ℃, cultivate 20～24 hours; Standard and sample should be on same flat boards.