Validation Data

Validation Data

1. Yield and Purity of Spin Columns

Sample	Column	Conc.,ng/μL	m,μg	A260/A280	A260/A230
Customer A Buffer	NP30-A	93	9.3	1.8	2
		95	9.5	1.9	2.1
		90	9	1.8	2
Customer A Buffer	Qiagen Miniprep column	83	8.3	1.8	2
		88	8.8	1.8	2.1
		90	9	1.8	2
QIAprep Spin Miniprep Kit Qiagen, Cat. No 27104	NP30-A	100	10	1.8	2
		106	10.6	1.9	2
		104	10.4	1.9	2.1
QIAprep Spin Miniprep Kit Qiagen, Cat. No 27104	Qiagen Miniprep column	102	10.2	1.9	2
		101	10.1	1.8	2
		102	10.2	1.9	2

Results: The performance of NP30-A is equivalent to Qiagen spin columns with both customer and Qiagen buffer.

2. Stability of Spin Columns

Sample	Column	Conc.,ng/μL	m,μg	A260/A280	A260/A230
NP30-A: 1 year of storage	Customer A Kit	141	14.1	1.9	2
		147	14.7	1.8	2.1
		151	15.1	1.9	2.2
NP30-A: New lot.	Customer A Kit	142	14.2	1.8	2.2
		159	15.9	1.8	1.8
		149	14.9	1.8	2.3
NP30-A: 1 year of storage	QIAprep Spin Miniprep Kit Qiagen, Cat. No 27104	168	16.8	1.8	2.0
		178	17.8	1.9	2.1
		160	16.0	1.8	2.0
NP30-A: New lot.	QIAprep Spin Miniprep Kit Qiagen, Cat. No 27104	162	16.2	1.8	2.1
		169	16.9	1.8	2.1
		169	16.9	1.9	2.0

Results: NP30-A show good performance after one year storage.

3. Purity Testing of Silica Membrane

Membrane	Extractant	m/z	Ion	Compound	Source
Biocomma Silica Membrane	Pure Water	304	M+Na+	oleamide	Slip agent in polyethylene films
		97	HSO4-
		97	H2PO4-
	Methanol	360	M+Na+	erucamide	Slip agent in polyethylene films
Filters from Well-known brand	Pure Water	304	M+Na+	oleamide	Slip agent in polyethylene films
		97	HSO4-
		97	H2PO4-
	Methanol	360	M+Na+	erucamide	Slip agent in polyethylene films
		425	M+Na+	Unidentified contaminant
		701	M+Na+		from water and solvent filters

Remarks：
1. The table only records ions that match the “ESI Common Background Ions” reference table.
2. The results have deducted the ions present in the blank, but if a particular m/z ion significantly exceeds the blank, it is recorded.
Conclusion：
Proceed ultrasonic extraction in both water and methanol, mass spectrometry results showed the presence of precipitates in both Biocomma membrane and well-known brand filter. Compound identification was carried out through high-resolution mass spectrometry by full scanning and matching with the “ESI Common Background Ions” reference table. The primary precipitates were HSO4-, H2PO4-, and Na+, and these presence is attributed to the material being glass fiber in the membranes. No other substances were found to precipitate from either of the two membranes, confirming their purity.