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How do I compute the +5 charge state m/z for a peptide of 3751.0 Da?

Asked 24 Nov 2025Modified 6 months agoViewed 4.5k times
1

Concretely: +5 · 3751.0 Da.

I want the working, not the result — I need to be able to redo it with different numbers.

I care about the precision as well as the value — I want to know how many figures are real.

How many significant figures are actually justified here?

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KS
askedk_szabo27k2724 Nov 2025
Is the comparison against a supplier certificate or against a second independent result? – lane_transit 8 months ago
Voting to keep this open — it is more specific than it first looks. – h_villanueva 7 months ago
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2 Answers

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m/z = 751.21 at 5+. Electrospray charges a peptide by adding protons, so the observed ion is the neutral mass plus 5 protons, all divided by the charge: (3751 + 5 × 1.00728) ÷ 5 = 3756.036 ÷ 5 = 751.21. The proton term is the one people drop, and because it is z protons over z charges it shifts m/z by 1.007 at every charge state — small, and far larger than the mass accuracy of the instrument. The neighbouring charge state sits at 626.17, and seeing the two of them where they belong is better identity evidence than either one alone. Use the average mass against an average-mass calculation and the monoisotopic mass against a monoisotopic one; mixing them costs you a couple of daltons on a peptide this size.

Stated carefully, the single most important fact about mass spectrometry for peptides is that it measures only the molecular weight and tells you almost nothing about whether the peak is actually your target.

The charge state distribution depends on the solution pH, the structure of the peptide and the source conditions, so the same peptide can look different under different conditions.

The m/z accuracy achievable depends on the mass analyser type — quadrupole gives low accuracy, time-of-flight gives moderate accuracy, and Orbitrap gives high accuracy.

I would not trust a mass result without a good baseline and a blank injection check.

A correct mass is necessary for identity but not sufficient — you also need the chromatography to confirm it.

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VR
answeredv_ramaswamy68k5731 Jan 2026
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Mass spectrometry is an identity technique, not a purity technique, and conflating the two is a common source of false confidence.

The baseline noise on a mass spectrum sets the limit of detection, and a weak signal close to the noise is not reliable evidence for the presence of a species.

A mass shift of minus one hundred and twenty-eight usually means a missing Gln or Lys residue from a synthesis deletion sequence.

Worth noting that source contamination is common and silent, so a result that looks too good to be true often is.

The practical summary: use mass spectrometry for identity, not for purity.

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NT
answeredn_takahashi29k3812 Feb 2026

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