Analytical Methods · Lot Data First

The methods we run on every release.

Every commercial lot ships with an RP-HPLC chromatogram, ESI-MS identity confirmation, LC-MS/MS sequence verification, Karl Fischer water content, and the relevant stability time-course. The document is structured to read like a paper's methods section, because that is who reads it.

Lyochem lot release bench — lyophilised research-peptide vials photographed under the analytical-lab inspection lamp; every vial label corresponds to a lot-specific RP-HPLC chromatogram and ESI-MS identity confirmation.

Lot Report · Excerpt

TB-500 · Lot LC-26-B0312

Released

RP-HPLC purity: 98.94 %
ESI-MS [M+H]⁺: 4961.4 Da
MS/MS sequence: Confirmed
Water (KF): 5.2 %
Counter-ion: Acetate
Stability (12 mo): ≥ 97 % retained

The lot report is the document you reference in your supplementary materials and your supplier-qualification file. Every release carries the same scope.

Illustrative lot report excerpt · TB-500 lot LC-26-B0312 · representative scope released with every shipment

Quality you can verify

Lot data first. Sales second.

Every Lyochem lot is released with its own analytical packet — chromatogram, mass spectrum, sequence confirmation, water content, stability. Methods are aligned with USP <1225>, EP, and ICH Q2 — the document a paper reviewer or CRO QA team can read directly off the page.

≥98.5%

HPLC purity floor

MS/MS

Sequence verified

12-mo

-20 °C stability

RP-HPLC + ESI-MS on every release, integrated purity reported at the main retention time, mass within ± 0.5 Da of theoretical.

LC-MS/MS sequence on request — b- and y-ion ladder assignment against the theoretical sequence; full-ladder coverage reported on the data packet.

−20 °C stability programme through 12 months; accelerated 40 °C add-ons on request for first-time supplier qualification.

View a sample data packet (PDF)Inside the analytical programme

Core release tests · On every lot

What every Lyochem release is tested for.

The core release panel runs on every commercial lot before shipment. Method provenance for each row tracks USP <621> (HPLC), USP <1225> (analytical validation), ICH Q2 (analytical method validation), and EP equivalents. The lot report carries the numerical value, the method line, and the release decision against the documented target.

Test	Method	Target	Why we run it
RP-HPLC purity	C18 gradient, 214 nm + 280 nm; USP <621> + ICH Q2 aligned	≥ 98.5 %	Resolves the synthesised peptide from synthesis truncations and late-eluting impurities. The purity number on the lot report is the integrated area % of the main peak; the chromatogram itself is attached to the lot packet.
ESI-MS identity	ESI-MS, monoisotopic [M+H]⁺ vs. theoretical	Confirmed (±0.5 Da)	Confirms the synthesised molecule has the expected molecular weight. Mass alone is necessary but not sufficient (see MS/MS row).
Sequence verification	LC-MS/MS, full b/y-ion ladder for peptides > 5 aa	Sequence confirmed	Directly demonstrates the synthesised sequence matches the labelled sequence. Rules out same-mass truncation-plus-elongation by-products that ESI-MS alone cannot distinguish.
Water content (KF)	Coulometric Karl Fischer	≤ 8.0 %	Lyophilisation-cycle control. Out-of-band water accelerates degradation kinetics during −20 °C storage and is the most common cause of unexpected potency drift at re-test.
Counter-ion	Ion chromatography	Acetate (default) or TFA (specified)	Acetate is the default salt form because residual TFA can suppress observed potency in cell-based bioassays. TFA salt is available when explicitly requested.
Residual solvents	GC headspace	ICH Q3C limits	Catches residuals from the SPPS cleavage, lyophilisation, and purification steps. Relevant for any peptide active going into a downstream bioassay.

Stability programme

What 12 months of stability data look like.

Lyochem maintains a rolling stability programme on the lyophilised lots in the catalogue. The −20 °C 12-month time-course is included with the lot report as standard; accelerated (40 °C / 75 %RH) and solution-stability add-ons ship on request and are recommended for buyers whose workflow holds the active outside cold chain or pre-prepares working stocks.

Test	Method	Target	Why we run it
Lyophilised stability (−20 °C)	Time-course RP-HPLC + ESI-MS through 12 months	≥ 95 % retention at 12 mo	Establishes the practical shelf-life of the released lot under standard cold-chain conditions. The decay envelope is documented; lots that drift outside it before re-test are held.
Accelerated stability (40 °C / 75 %RH)	Time-course RP-HPLC through 1 / 3 / 6 months	Documented decay rate	Predicts the practical shelf-life for buyers whose workflow holds the active outside cold chain (e.g. lab benches, cross-border shipping). On request.
Solution stability	Time-course at reconstitution + 24 / 48 / 72 h in BAC water / PBS	Documented	For buyers who pre-prepare working stocks. Most peptide degradation in research workflows comes from solution-phase handling, not the lyophilised storage.

In vivo add-ons

Endotoxin + microbial limits, on request.

LAL bacterial endotoxin, USP <61>/<62> microbial limits, and granular bioburden testing are not part of the default in vitro release scope — they add 3–5 business days of QC turnaround and are not needed for the bench- scale workflows most academic and CRO buyers run. For in vivo research, however, they are usually a precondition rather than a nice-to-have.

Specify in vivo intent at the inquiry stage and the lot is flagged for the additional release testing; the CoA then carries chemical-purity, sequence-verification, and biological-safety data on a single document, which simplifies IACUC and IRB record-keeping.

Test	Method	Target	Why we run it
Bacterial endotoxin (LAL)	Gel-clot or kinetic chromogenic per USP <85>	< 0.25 EU/mg (or as specified)	Required for any in vivo research workflow — endotoxin contamination directly activates NF-κB reporters, macrophage panels, and many neutrophil-priming assays, confounding the test article's own readout.
Microbial limits	USP <61>/<62>	Compliant	Bioburden control for in vivo or cell-based assay workflows where microbial contamination would confound the readout independently of endotoxin.
Bioburden	Membrane filtration + plate count	Documented CFU/mg	Granular bioburden data for IACUC / IRB record-keeping when the protocol commits to a numerical bioburden ceiling rather than a USP-pass/fail.

Lot report contents

What lands on your bench.

Every commercial release ships with the lot report below. On request we attach the raw analytics — RP-HPLC chromatogram with peak integration, ESI mass spectrum, MS/MS sequence ladder, stability time-series — as the QA work-packet your supplier-qualification dossier or supplementary-materials file needs.

RP-HPLC chromatogram

Peak-integration trace with retention times and integrated areas; the document you cite in supplementary materials.

ESI-MS identity

[M+H]⁺ vs. theoretical with the residual mass error in Da. Confirms molecular weight to ±0.5 Da.

LC-MS/MS sequence ladder

Full b/y-ion ladder for peptides above 5 amino acids. Directly demonstrates synthesised sequence.

Water content (KF)

Coulometric Karl Fischer titration. Lyophilisation-cycle control.

Counter-ion

Acetate by default; TFA on request. Affects solubility, stability, and downstream bioassay readouts.

Stability time-course

−20 °C 12-month lyophilised; accelerated 40 °C and solution-phase on request.

Request a data packet.

First reply under 12 hours from a synthesis chemist or analytical contact — not a templated auto-response. Lot-specific HPLC chromatogram, ESI-MS identity, MS/MS sequence (on request), KF water, and stability data returned with the quote.