prepare 0.
1 M phosphate buffer having
pH 7.4 ALSO scale down the calculations
ChatGPT said:
To prepare a 0.1 M phosphate buffer at pH 7.4, we typically use a mixture of:
NaH₂PO₄ (sodium dihydrogen phosphate, the acid form)
Na₂HPO₄ (disodium hydrogen phosphate, the base form)
We’ll use the Henderson-Hasselbalch equation to determine the ratio of the two components:
📘 Henderson-Hasselbalch Equation:
pH=pKa+log([Base][Acid])\text{pH} = \text{p}K_a + \log\left(\frac{[\text{Base}]}{[\
text{Acid}]}\right)pH=pKa+log([Acid][Base])
For the phosphate buffer system:
pKa=7.2\text{p}K_a = 7.2pKa=7.2 (for the H₂PO₄⁻ / HPO₄²⁻ system)
Desired pH = 7.4
🔹 Step 1: Calculate Base/Acid Ratio
7.4=7.2+log([Na2HPO4][NaH2PO4])7.4 = 7.2 + \log\left(\frac{[\text{Na}_2\text{HPO}_4]}{[\
text{NaH}_2\text{PO}_4]}\right)7.4=7.2+log([NaH2PO4][Na2HPO4]) log([Base][Acid])=0.2\
log\left(\frac{[\text{Base}]}{[\text{Acid}]}\right) = 0.2log([Acid][Base])=0.2 [Base]
[Acid]=100.2≈1.58\frac{[\text{Base}]}{[\text{Acid}]} = 10^{0.2} \approx 1.58[Acid][Base]
=100.2≈1.58
🔹 Step 2: Let Total Molarity = 0.1 M
Let:
Acid (NaH₂PO₄) = x
Base (Na₂HPO₄) = 1.58x
x+1.58x=0.1⇒2.58x=0.1⇒x=0.12.58≈0.03876 Mx + 1.58x = 0.1 \Rightarrow 2.58x = 0.1 \
Rightarrow x = \frac{0.1}{2.58} \approx 0.03876 \, \
text{M}x+1.58x=0.1⇒2.58x=0.1⇒x=2.580.1≈0.03876M
So:
NaH₂PO₄ = 0.03876 M
Na₂HPO₄ = 0.06124 M
🔹 Step 3: Convert to Grams for 1 Liter
NaH₂PO₄·2H₂O (MW = 156.01 g/mol)
→ 0.03876×156.01≈6.05 g0.03876 \times 156.01 \approx 6.05 \, \
text{g}0.03876×156.01≈6.05g
Na₂HPO₄·2H₂O (MW = 177.99 g/mol)
→ 0.06124×177.99≈10.91 g0.06124 \times 177.99 \approx 10.91 \, \
text{g}0.06124×177.99≈10.91g
✅ For 1 L of 0.1 M Phosphate Buffer at pH 7.4:
6.05 g of NaH₂PO₄·2H₂O
10.91 g of Na₂HPO₄·2H₂O
Dissolve in water and make up to 1 L
🔻 Scale-Down Example: For 100 mL (0.1 L)
Just divide the quantities by 10:
NaH₂PO₄·2H₂O = 0.605 g
Na₂HPO₄·2H₂O = 1.091 g
Dissolve in ~80 mL water, adjust pH if needed, then make up to 100 mL.
Want this in a table format or printable
protocol?
4o
You said:
Want this in a table format or printable protocol? yes
ChatGPT said:
Great! Here's the table format and a printable protocol for preparing 0.1 M phosphate buffer
at pH 7.4, including both full-scale (1 L) and scaled-down (100 mL) versions.
📋 Phosphate Buffer Preparation Table
Molarity Molecular Weight Amount for 1 Amount for 100
Component
(M) (g/mol) L (g) mL (g)
NaH₂PO₄·2H₂O (acid
0.03876 156.01 6.05 0.605
form)
Na₂HPO₄·2H₂O (base
0.06124 177.99 10.91 1.091
form)
🧪 Printable Protocol
Title: Preparation of 0.1 M Phosphate Buffer (pH 7.4)
Buffer System:
NaH₂PO₄·2H₂O (acid) + Na₂HPO₄·2H₂O (base)
pKa = 7.2 (H₂PO₄⁻ / HPO₄²⁻ system)
Target pH: 7.4
Total Buffer Concentration: 0.1 M
📌 For 1 L of Buffer:
1. Weigh:
o 6.05 g of NaH₂PO₄·2H₂O
o 10.91 g of Na₂HPO₄·2H₂O
2. Dissolve both salts in ~800 mL of distilled water.
3. Check pH with a calibrated pH meter.
o Adjust slightly with NaOH or HCl if necessary (usually not needed).
4. Make up the volume to 1 L with distilled water.
📌 For 100 mL of Buffer (scaled down):
1. Weigh:
o 0.605 g of NaH₂PO₄·2H₂O
o 1.091 g of Na₂HPO₄·2H₂O
2. Dissolve in ~80 mL of distilled water.
3. Check and adjust pH if needed.
4. Make up to 100 mL with distilled water.
Would you like this as a downloadable PDF or Word document?
4o
prepare 0.1 M potassium phosphate buffer
having pH 7.4 ALSO scale down the calculations
ChatGPT said:
Sure! Here's how to prepare a 0.1 M potassium phosphate buffer at pH 7.4, using KH₂PO₄
(monobasic potassium phosphate, acid) and K₂HPO₄ (dibasic potassium phosphate, base). This
is very similar to the sodium phosphate buffer but with potassium salts instead.
⚗️Potassium Phosphate Buffer (pH 7.4, 0.1 M)
📘 Henderson-Hasselbalch Equation:
pH=pKa+log([Base][Acid])\text{pH} = \text{p}K_a + \log\left(\frac{[\text{Base}]}{[\
text{Acid}]}\right)pH=pKa+log([Acid][Base])
pKa = 7.2 (for H₂PO₄⁻ / HPO₄²⁻)
Desired pH = 7.4
🔹 Step 1: Calculate Base/Acid Ratio
7.4=7.2+log([K2HPO4][KH2PO4])⇒BaseAcid=100.2≈1.587.4 = 7.2 + \log\left(\frac{[\
text{K}_2\text{HPO}_4]}{[\text{KH}_2\text{PO}_4]}\right) \Rightarrow \frac{\text{Base}}{\
text{Acid}} = 10^{0.2} \approx 1.587.4=7.2+log([KH2PO4][K2HPO4])⇒AcidBase
=100.2≈1.58
🔹 Step 2: Molar Proportions
Let:
Acid (KH₂PO₄) = x
Base (K₂HPO₄) = 1.58x
Total Molarity:
x+1.58x=0.1⇒x=0.12.58≈0.03876 Mx + 1.58x = 0.1 \Rightarrow x = \frac{0.1}{2.58} \approx
0.03876 \, \text{M}x+1.58x=0.1⇒x=2.580.1≈0.03876M
So:
KH₂PO₄ = 0.03876 M
K₂HPO₄ = 0.06124 M
🔹 Step 3: Convert to Grams (for 1 L)
Salt Molar Mass (g/mol) Molarity (M) Grams for 1 L
KH₂PO 136.09 0.03876 ≈ 5.27 g
Salt Molar Mass (g/mol) Molarity (M) Grams for 1 L
₄
K₂HPO
174.18 0.06124 ≈ 10.66 g
₄
✅ 📋 Potassium Phosphate Buffer Table
Mol. Wt. Amount for 1 L Amount for 100 mL
Component Molarity (M)
(g/mol) (g) (g)
KH₂PO₄ (acid
0.03876 136.09 5.27 0.527
form)
K₂HPO₄ (base
0.06124 174.18 10.66 1.066
form)
🧪 Printable Protocol
Title: Preparation of 0.1 M Potassium Phosphate Buffer (pH 7.4)
📌 For 1 L of Buffer:
1. Weigh:
o 5.27 g KH₂PO₄
o 10.66 g K₂HPO₄
2. Dissolve in ~800 mL distilled water.
3. Check pH and adjust if necessary with KOH or HCl.
4. Make up to 1 L with distilled water.
📌 For 100 mL of Buffer:
1. Weigh:
o 0.527 g KH₂PO₄
o 1.066 g K₂HPO₄
2. Dissolve in ~80 mL water.
3. Check pH, adjust if needed.
4. Make up to 100 mL with distilled water.
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