Estimation of free and total acidity in a gastric juice sample

Presented by Dr. Nada Soliman

• Biological fluid: gastric juice
 pH  Types of acid

Content

• Quantitative method: titration
 Principle of titration  Type of titration used in free and total acidity estimation  Titrant and indicators used  End points (X1 and X2)  procedure and calculation of free and total acidity.

• Fractional test meal
 procedure  Normal curve  Abnormal curves: 3

Gastric juice
 the most acidic secretion of the body fluid, with a pH= 1.0 (in adult)  Types of acid in gastric juice: 1. HCL: secreted by parietal cells of the gastric mucosa Free HCL Combined HCL (combined with protein) 2. Organic acids: lactic, acetic acid……  Free HCL+ combined HCL+ organic acids: responsible for total acidity  Free HCL: responsible for free acidity of the gastric juice  When the free HCL is neutralized the pH of gastric juice become 3

• Alkaline tide:  Means the presence of an alkaline urine that often follows the ingestion of a meal due to the presence of excess bicarbonate in the blood during HCL secretion

Biological substance of unknown concentration and of known volume (sample)

To determine its conc

A standard solution: titrant

1. 2. 3. 4. Knowing the volume of the titrant needed to react completely with the analyte, helps to determine its concentration The titrant is added drop by drop until the reaction is complete

Another substance Known concentration React with the analyte Volume: measured during the experiment

Definition of titrant: the solution of known concentration added during titration

According to type of the reaction, we have 3 types of titration: 1. Acid-base: HCL with NaOH 2. Precipitation: silver nitrate with barium chloride 3. Oxidation-reduction: uric acid with K permenganate

How we know that the reaction is complete? By observing physical change. the end point Could be:  changes in color  Appearance or disappearance of ppt  Change in electrical properties Definition of end point: Point in titration where physical change manifests itself

For color changes, we use an Indicator: Supplementary chemical compound that changes its color with changes in the medium at the end point

Equipments
• • • • Burettes holders. Pipettes beakers

 Types of titration: acid-base titration  Titrant: 0.1 N NaOH  End point (X1 &X2): color change  Indicators used: • Topfer’s:  Pink color pH <3  Yellow pH>3  Salmon pink pH=3 (X1) • Phenolphthalein:  Colorless at pH <8  Pink at pH= or >8 (X2)

procedure
1. Fill a burette with 0.1 NNaOH solution 2. Pipette 5 ml of the gastric juice into a beaker (using a 5 ml pipette). 3. Add to the contents of the beaker one drop of phenolphthalein indicator colorless 4. Add 2-3 drops of topfer’s indicator pink 5. Titrate against 0.1 NNaOH until the color changes from pink to salmon pink (orange yellow).

6. Stop the titration and take the reading of the burette which represents the end point (= X1) at which all free HCl, present in the 5 ml gastric juice, is neutralized by 0.1N NaOH. Now the pH is 3 7. continue titration until the color changes to pink. Stop the titration and take the reading of the burette from the start which represents the end point(= X2) at which all the total acidity, present in the 5 ml gastric juice, is neutralized by 0.1N NaOH. Now the pH is 8.

Topfer’s
5ml GJ + phenolphthalein
colorless

NaOH pink

X1 X1=number of ml of 0.1 NNaOH required to neutralize the free HCl in 5 ml GJ X2=number of ml of 0.1 NNaOH required to neutralize total acids in 5 ml GJ
Salmon pink

NaOH
X2

pink

Yellowish gradually

• From the previous procedure, X1 and X2 were determined in only 5 ml of the gastric juice sample. • Free acidity is expressed as the number of milliliters of 0.1 N-NaOH required to neutralize the free HCl in 100 ml of gastric juice. • Total acidity is expressed as the number of milliliters of 0.1 N-NaOH required to neutralize the total acids present in 100 ml of gastric juice. • So:  Free acidity = X1 x 100/5 0.1 N-NaOH %  Total acidity =X2 x 100/5 0.1 N-NaOH %

Calculation of free and total acidity

Fractional Test Meal
1. The patient comes fasting and then a stomach tube (Ryle tube) is introduced in the mouth. 2. a fasting sample is aspirated and titrated for determination of free and total acidity. 3. Then the patient takes a standard meal of 1-2 pieces of toast + 2 cups of tea (with sugar but no milk). 4. Samples of 10 ml are then aspirated every 15 or 30 minutes for about 2.5 hours. 5. After all samples are titrated, the amount of free and total acidity can be plotted in a curve against time.

The curve of normal people shows the

following characters:
After 120 minutes, acidity starts to decline gradually. the fasting level is reached at about 150 minutes.

The fasting acidity is neutralized by 20-30 ml 0.1 NNaOH / 100 ml GJ.

15-30 minutes after the meal, the acidity falls to a lower level.

Then the acidity rises again and reaches its maximal value after about 90 minutes.

• AT 15-30 minutes, This decline is due to:
1. Dilution of gastric juice with saliva and tea. 2. Neutralization of gastric juice by NaHCO3 of saliva, protein of the meal and mucin inside the stomach.

• At 90 minutes, the acidity rises again and reaches its maximal value:
o due to stimulation of the gastric glands by the meal.

• at the end of about 150 minutes, This decline is due to:
1. Gastric secretion gets diminished. 2. Gastric contents are evacuated to the duodenum. 3. Gastric contents are neutralized by duodenal regurgitation.

Abnormal curves:
• Hyperacidity (hyperchlorhydria): Acidity is more than normal e.g. in cases of duodenal ulcers and gall bladder diseases. • Hypoacidity (hypochlorhydria): Acidity is below normal e.g. in gastric carcinoma. • Anacidity (achlorhydria): There is complete absence of hydrochloric acid in this condition. The curve obtained is flat. Achlorhydria is compatible with pernicious anemia.

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